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Sample records for chloramphenicol

  1. Chloramphenicol Injection

    MedlinePlus

    ... treat certain types of serious infections caused by bacteria when other antibiotics cannot be used. Chloramphenicol injection ... antibiotics. It works by stopping the growth of bacteria..Antibiotics such as chloramphenicol injection will not work ...

  2. Chloramphenicol bioassay.

    PubMed

    Bannatyne, R M; Cheung, R

    1979-07-01

    An accurate plate diffusion bioassay for chloramphenicol is described, in which the fast-replicating Beneckea natriegens and 1.5% salt agar are used. Zones of inhibition were well defined after 3 h, and the limit of sensitivity of the method was around 2 mug/ml. The concurrent presence of gentamicin did not influence the assay. The assay is simple to carry out and duplicate assays can be performed with as little as 100 mug of capillary blood.

  3. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... Because of potential antagonism, chloramphenicol should not be administered simultaneously with penicillin.... Because of potential antagonism, chloramphenicol should not be administered simultaneously with...

  4. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... Because of potential antagonism, chloramphenicol should not be administered simultaneously with penicillin.... Because of potential antagonism, chloramphenicol should not be administered simultaneously with...

  5. Killing of Serratia marcescens biofilms with chloramphenicol.

    PubMed

    Ray, Christopher; Shenoy, Anukul T; Orihuela, Carlos J; González-Juarbe, Norberto

    2017-03-29

    Serratia marcescens is a Gram-negative bacterium with proven resistance to multiple antibiotics and causative of catheter-associated infections. Bacterial colonization of catheters mainly involves the formation of biofilm. The objectives of this study were to explore the susceptibility of S. marcescens biofilms to high doses of common antibiotics and non-antimicrobial agents. Biofilms formed by a clinical isolate of S. marcescens were treated with ceftriaxone, kanamycin, gentamicin, and chloramphenicol at doses corresponding to 10, 100 and 1000 times their planktonic minimum inhibitory concentration. In addition, biofilms were also treated with chemical compounds such as polysorbate-80 and ursolic acid. S. marcescens demonstrated susceptibility to ceftriaxone, kanamycin, gentamicin, and chloramphenicol in its planktonic form, however, only chloramphenicol reduced both biofilm biomass and biofilm viability. Polysorbate-80 and ursolic acid had minimal to no effect on either planktonic and biofilm grown S. marcescens. Our results suggest that supratherapeutic doses of chloramphenicol can be used effectively against established S. marcescens biofilms.

  6. Interaction of Chloramphenicol Tripeptide Analogs with Ribosomes.

    PubMed

    Tereshchenkov, A G; Shishkina, A V; Tashlitsky, V N; Korshunova, G A; Bogdanov, A A; Sumbatyan, N V

    2016-04-01

    Chloramphenicol amine peptide derivatives containing tripeptide fragments of regulatory "stop peptides" - MRL, IRA, IWP - were synthesized. The ability of the compounds to form ribosomal complexes was studied by displacement of the fluorescent erythromycin analog from its complex with E. coli ribosomes. It was found that peptide chloramphenicol analogs are able to bind to bacterial ribosomes. The dissociation constants were 4.3-10 µM, which is 100-fold lower than the corresponding values for chloramphenicol amine-ribosome complex. Interaction of the chloramphenicol peptide analogs with ribosomes was simulated by molecular docking, and the most probable contacts of "stop peptide" motifs with the elements of nascent peptide exit tunnel were identified.

  7. 21 CFR 522.390 - Chloramphenicol injection.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Chloramphenicol injection. 522.390 Section 522.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  8. 21 CFR 522.390 - Chloramphenicol injection.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Chloramphenicol injection. 522.390 Section 522.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  9. 21 CFR 522.390 - Chloramphenicol injection.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chloramphenicol injection. 522.390 Section 522.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  10. 21 CFR 522.390 - Chloramphenicol.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Chloramphenicol. 522.390 Section 522.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  11. 21 CFR 522.390 - Chloramphenicol injection.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Chloramphenicol injection. 522.390 Section 522.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS IMPLANTATION OR INJECTABLE DOSAGE FORM NEW ANIMAL DRUGS §...

  12. Characterization of two metagenome-derived esterases that reactivate chloramphenicol by counteracting chloramphenicol acetyltransferase.

    PubMed

    Tao, Weixin; Lee, Myung Hwan; Yoon, Mi-Young; Kim, Jin-Cheol; Malhotra, Shweta; Wu, Jing; Hwang, Eul Chul; Lee, Seon-Woo

    2011-12-01

    Function-driven metagenomic analysis is a powerful approach to screening for novel biocatalysts. In this study, we investigated lipolytic enzymes selected from an alluvial soil metagenomic library, and identified two novel esterases, EstDL26 and EstDL136. EstDL26 and EstDL136 reactivated chloramphenicol from its acetyl derivates by counteracting the chloramphenicol acetyltransferase (CAT) activity in Escherichia coli. These two enzymes showed only 27% identity in amino acid sequence to each other; however both preferentially hydrolyzed short-chain p-nitrophenyl esters (< or =C5) and showed mesophilic properties. In vitro, EstDL136 catalyzed the deacetylation of 1- and 3- acetyl and 1,3-diacetyl derivates; in contrast, EstDL26 was not capable of the deacetylation at C1, indicating a potential regioselectivity. EstDL26 and EstDL136 were similar to microbial hormone-sensitive lipase (HSL), and since chloramphenicol acetate esterase (CAE) activity was detected from two other soil esterases in the HSL family, this suggests a distribution of CAE among the soil microorganisms. The isolation and characterization of EstDL26 and EstDL136 in this study may be helpful in understanding the diversity of CAE enzymes and their potential role in releasing active chloramphenicol in the producing bacteria.

  13. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    .... Treatment of bacterial pulmonary infections, infections of the urinary tract, enteritis, and infections associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

  14. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... Treatment of bacterial pulmonary infections, infections of the urinary tract, enteritis, and infections associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

  15. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    .... Treatment of bacterial pulmonary infections, infections of the urinary tract, enteritis, and infections associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

  16. Chloramphenicol and expression of multidrug efflux pump in Enterobacter aerogenes.

    PubMed

    Ghisalberti, Didier; Masi, Muriel; Pagès, Jean-Marie; Chevalier, Jacqueline

    2005-03-25

    Chloramphenicol has been reported to act as an inducer of the multidrug resistance in Escherichia coli. A resistant variant able to grow on plates containing 64 microg/ml chloramphenicol was obtained from the Enterobacter aerogenes ATCC 13048-type strain. Chloramphenicol resistance was due to an active efflux of this antibiotic and it was associated with resistance to fluoroquinolones and tetracycline, but not to aminoglycoside or beta-lactam antibiotics. MDR in the chloramphenicol-resistant variant is linked to the overexpression of the major AcrAB-TolC efflux system. This overexpression seems unrelated to the global Mar and the local AcrR regulatory pathways.

  17. 21 CFR 524.390a - Chloramphenicol ophthalmic ointment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... bacterial conjunctivitis caused by pathogens susceptible to chloramphenicol. (3) Limitations. Continue... conjunctivitis, attempts should be made to determine through susceptibility testing, which antibiotics will...

  18. 21 CFR 524.390a - Chloramphenicol ophthalmic ointment.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... bacterial conjunctivitis caused by pathogens susceptible to chloramphenicol. (3) Limitations. Continue... conjunctivitis, attempts should be made to determine through susceptibility testing, which antibiotics will...

  19. Structural basis for chloramphenicol tolerance in Streptomyces venezuelae by chloramphenicol phosphotransferase activity.

    PubMed

    Izard, T

    2001-08-01

    Streptomyces venezuelae synthesizes chloramphenicol (Cm), an inhibitor of ribosomal peptidyl transferase activity, thereby inhibiting bacterial growth. The producer escapes autoinhibition by its own secondary metabolite through phosphorylation of Cm by chloramphenicol phosphotransferase (CPT). In addition to active site binding, CPT binds its product 3-phosphoryl-Cm, in an alternate product binding site. To address the mechanisms of Cm tolerance of the producer, the crystal structures of CPT were determined in complex with either the nonchlorinated Cm (2-N-Ac-Cm) at 3.1 A resolution or the antibiotic's immediate precursor, the p-amino analog p-NH(2)-Cm, at 2.9 A resolution. Surprisingly, p-NH(2)-Cm binds CPT in a novel fashion. Additionally, neither 2-N-Ac-Cm nor p-NH(2)-Cm binds to the secondary product binding site.

  20. Type B Chloramphenicol Acetyltransferases Are Responsible for Chloramphenicol Resistance in Riemerella anatipestifer, China

    PubMed Central

    Huang, Li; Yuan, Hui; Liu, Ma-Feng; Zhao, Xin-Xin; Wang, Ming-Shu; Jia, Ren-Yong; Chen, Shun; Sun, Kun-Feng; Yang, Qiao; Wu, Ying; Chen, Xiao-Yue; Cheng, An-Chun; Zhu, De-Kang

    2017-01-01

    Riemerella anatipestifer causes serositis and septicaemia in domestic ducks, geese, and turkeys. Traditionally, the antibiotics were used to treat this disease. Currently, our understanding of R. anatipestifer susceptibility to chloramphenicol and the underlying resistance mechanism is limited. In this study, the cat gene was identified in 69/192 (36%) R. anatipestifer isolated from different regions in China, including R. anatipestifer CH-2 that has been sequenced in previous study. Sequence analysis suggested that there are two copies of cat gene in this strain. Only both two copies of the cat mutant strain showed a significant decrease in resistance to chloramphenicol, exhibiting 4 μg/ml in the minimum inhibitory concentration for this antibiotic, but not for the single cat gene deletion strains. Functional analysis of the cat gene via expression in Escherichia coli BL21 (DE3) cells and in vitro site-directed mutagenesis indicated that His79 is the main catalytic residue of CAT in R. anatipestifer. These results suggested that chloramphenicol resistance of R. anatipestifer CH-2 is mediated by the cat genes. Finally, homology analysis of types A and B CATs indicate that R. anatipestifer comprises type B3 CATs. PMID:28298905

  1. Type B Chloramphenicol Acetyltransferases Are Responsible for Chloramphenicol Resistance in Riemerella anatipestifer, China.

    PubMed

    Huang, Li; Yuan, Hui; Liu, Ma-Feng; Zhao, Xin-Xin; Wang, Ming-Shu; Jia, Ren-Yong; Chen, Shun; Sun, Kun-Feng; Yang, Qiao; Wu, Ying; Chen, Xiao-Yue; Cheng, An-Chun; Zhu, De-Kang

    2017-01-01

    Riemerella anatipestifer causes serositis and septicaemia in domestic ducks, geese, and turkeys. Traditionally, the antibiotics were used to treat this disease. Currently, our understanding of R. anatipestifer susceptibility to chloramphenicol and the underlying resistance mechanism is limited. In this study, the cat gene was identified in 69/192 (36%) R. anatipestifer isolated from different regions in China, including R. anatipestifer CH-2 that has been sequenced in previous study. Sequence analysis suggested that there are two copies of cat gene in this strain. Only both two copies of the cat mutant strain showed a significant decrease in resistance to chloramphenicol, exhibiting 4 μg/ml in the minimum inhibitory concentration for this antibiotic, but not for the single cat gene deletion strains. Functional analysis of the cat gene via expression in Escherichia coli BL21 (DE3) cells and in vitro site-directed mutagenesis indicated that His79 is the main catalytic residue of CAT in R. anatipestifer. These results suggested that chloramphenicol resistance of R. anatipestifer CH-2 is mediated by the cat genes. Finally, homology analysis of types A and B CATs indicate that R. anatipestifer comprises type B3 CATs.

  2. Effects of concurrent drug therapy and of feeding on plasma chloramphenicol levels after oral administration of chloramphenicol in dogs.

    PubMed

    Watson, A D

    1977-01-01

    On separate occasions, five fasted adult greyhounds were dosed orally with 50 mg chloramphenicol/kg, both alone and in conjunction with other drugs. The same five dogs were later given 50 mg chloramphenicol/kg when fed ad lib. Chloramphenicol levels in plasma were determined at intervals after dosing. The intial plasma chloramphenicol levels were higher when the drug was administered concurrently with calcium lactate tablets (50 mg/kg) or a proprietary enteric mixture containing kaolin, pectin, aluminium hydroxide and belladonna extract. Lower plasma levels resulted when the antibiotic was given with dry extract of belladonna (20 mg/dog) or intravenous chlorpromazine (2 mg/kg). There was no significant effect with coated ferrous gluconate tablets (30 mg/kg). Feeding ad lib increased the initial chloramphenicol levels, but produced lower levels subsequently.

  3. Inhibition of existing denitrification enzyme activity by chloramphenicol

    USGS Publications Warehouse

    Brooks, M.H.; Smith, R.L.; Macalady, D.L.

    1992-01-01

    Chloramphenicol completely inhibited the activity of existing denitrification enzymes in acetylene-block incubations with (i) sediments from a nitrate-contaminated aquifer and (ii) a continuous culture of denitrifying groundwater bacteria. Control flasks with no antibiotic produced significant amounts of nitrous oxide in the same time period. Amendment with chloramphenicol after nitrous oxide production had begun resulted in a significant decrease in the rate of nitrous oxide production. Chloramphenicol also decreased (>50%) the activity of existing denitrification enzymes in pure cultures of Pseudomonas denitrificans that were harvested during log- phase growth and maintained for 2 weeks in a starvation medium lacking electron donor. Short-term time courses of nitrate consumption and nitrous oxide production in the presence of acetylene with P. denitrificans undergoing carbon starvation were performed under optimal conditions designed to mimic denitrification enzyme activity assays used with soils. Time courses were linear for both chloramphenicol and control flasks, and rate estimates for the two treatments were significantly different at the 95% confidence level. Complete or partial inhibition of existing enzyme activity is not consistent with the current understanding of the mode of action of chloramphenicol or current practice, in which the compound is frequently employed to inhibit de novo protein synthesis during the course of microbial activity assays. The results of this study demonstrate that chloramphenicol amendment can inhibit the activity of existing denitrification enzymes and suggest that caution is needed in the design and interpretation of denitrification activity assays in which chloramphenicol is used to prevent new protein synthesis.

  4. Electron Microscopy of Chloramphenicol-treated Escherichia coli

    PubMed Central

    Morgan, Councilman; Rosenkranz, Herbert S.; Carr, Howard S.; Rose, Harry M.

    1967-01-01

    Thin sections of Escherichia coli were examined by electron microscopy at sequential intervals after addition and then removal of chloramphenicol. The first changes, occurring at 1 hr after exposure to the drug, were disappearance of the ribosomes and aggregation of the nuclear material toward the center of the bacteria. At 2 hr, aggregates of abnormal cytoplasmic granules first appeared and subsequently increased in size. By 23 hr, amorphous, electron-dense material had accumulated within, and at the periphery of, the nuclear matrix. With the removal of chloramphenicol, the bacteria became normal in appearance, passing through a series of stages that were sequential but not synchronous. At 145 min after removal of chloramphenicol, bacteria were encountered in the process of abnormal division. The influence of deoxyribonucleic acid and ribonucleic acid synthesis, and of energy metabolism, upon the changes seen electron microscopically in chloramphenicol-treated cells, was investigated by selectively inhibiting these functions with hydroxyurea, azauracil, and sodium azide, respectively. Images PMID:5337775

  5. New insights into chloramphenicol biosynthesis in Streptomyces venezuelae ATCC 10712.

    PubMed

    Fernández-Martínez, Lorena T; Borsetto, Chiara; Gomez-Escribano, Juan Pablo; Bibb, Maureen J; Al-Bassam, Mahmoud M; Chandra, Govind; Bibb, Mervyn J

    2014-12-01

    Comparative genome analysis revealed seven uncharacterized genes, sven0909 to sven0915, adjacent to the previously identified chloramphenicol biosynthetic gene cluster (sven0916-sven0928) of Streptomyces venezuelae strain ATCC 10712 that was absent in a closely related Streptomyces strain that does not produce chloramphenicol. Transcriptional analysis suggested that three of these genes might be involved in chloramphenicol production, a prediction confirmed by the construction of deletion mutants. These three genes encode a cluster-associated transcriptional activator (Sven0913), a phosphopantetheinyl transferase (Sven0914), and a Na(+)/H(+) antiporter (Sven0915). Bioinformatic analysis also revealed the presence of a previously undetected gene, sven0925, embedded within the chloramphenicol biosynthetic gene cluster that appears to encode an acyl carrier protein, bringing the number of new genes likely to be involved in chloramphenicol production to four. Microarray experiments and synteny comparisons also suggest that sven0929 is part of the biosynthetic gene cluster. This has allowed us to propose an updated and revised version of the chloramphenicol biosynthetic pathway.

  6. Effect of bio-electrochemical system on the fate and proliferation of chloramphenicol resistance genes during the treatment of chloramphenicol wastewater.

    PubMed

    Guo, Ning; Wang, Yunkun; Yan, Lei; Wang, Xinhua; Wang, Mingyu; Xu, Hai; Wang, Shuguang

    2017-03-30

    Bioelectrochemical systems can effectively degrade antibiotics, but there is the need to better understand the fate of antibiotic resistance bacteria and antibiotic resistance genes during the bioelectrochemical degradation of antibiotics. In this study, a BES was developed as a platform to investigate the fate of chloramphenicol resistance bacteria (CRB) and the expression of chloramphenicol resistance genes (CRGs) under different operating conditions during chloramphenicol biodegradation. The results indicated that chloramphenicol was effectively removed and chloramphenicol removal efficiency could be improved under less chloramphenicol concentration and more negative cathode potential. Higher chloramphenicol concentration enhanced the enrichment of CRB and expression of CRGs. Furthermore, the abundances of CRB were enhanced under more negative cathode potential, the expression of CRGs under less negative cathode potential were induced. However, both the enrichment of CRB and expression of CRGs could be moderated under a medium cathode potential. This result could provide the scientific reference for research about the fate of antibiotic resistance genes in bioelectrochemical systems.

  7. 21 CFR 524.390b - Chloramphenicol ophthalmic solution.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... first 2 days. (2) Indications for use. Treatment of bacterial conjunctivitis caused by organisms susceptible to chloramphenicol. Therapy should be continued for 48 hours after the eye appears normal. (3) Limitations. Therapy for cats should not exceed 7 days. As with other antibiotics, prolonged use may result...

  8. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Chloramphenicol palmitate oral suspension. 520.390c Section 520.390c Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN.... Not for use in animals that are raised for food production. Must not be used in meat-, egg-, or...

  9. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chloramphenicol palmitate oral suspension. 520.390c Section 520.390c Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS §...

  10. 21 CFR 520.390 - Chloramphenicol oral dosage forms.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chloramphenicol oral dosage forms. 520.390 Section 520.390 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS §...

  11. Tetracycline and chloramphenicol efficiency against selected biofilm forming bacteria.

    PubMed

    Liaqat, Iram; Sumbal, Fareeha; Sabri, Anjum Nasim

    2009-08-01

    Despite the constantly increasing need for new antimicrobial agents, antibiotic drug discovery and development seem to have greatly decelerated in recent years. Presented with the significant problem of advancing antimicrobial resistance, the global scientific community has attempted to find alternative solutions; one of the most promising ones is the evaluation and use of old antibiotic compounds. A number of old antibiotic compounds, such as aminoglycosides, chloramphenicol, and tetracycline, are re-emerging as valuable alternatives for the treatment of difficult-to-treat infections. This study examined the in vitro potency for biofilm formation of five isolates (Klebsiella sp., Pseudomonas aeruginosa, Achromobacter sp., Klebsiella pneumoniae, and Bacillus pumilis) and the effects of antibiotics on these biofilms. Furthermore the quantitative analysis of planktonic, loosely attached cells, and their susceptibility to antibiotics was also determined. Twitching motility was observed to determine any effect in the biofilm forming capability of the isolates. All the isolates tested were efficient biofilm-forming strains in the polypropylene and borosilicate test tubes. Standard bacterial enumeration technique and CV staining produced equivalent results both in biofilm and planktonic assays. The biofilm formation of all the strains was affected in the presence of tetracycline or chloramphenicol. Highly significant decrease (P < 0.01) in biofilm formation was observed by treatment with chloramphenicol compared to tetracycline. In addition, the two antibiotics also affected adversely the planktonic and loosely attached cells of all isolates. Thus, testing the effects of older antibiotics on biofilms may supply useful information in addition to standard in vitro testing, particularly in diseases where biofilm formation is involved in the pathogenesis.

  12. Liposomal gel with chloramphenicol: characterisation and in vitro release.

    PubMed

    Pavelić, Zeljka; Skalko-Basnet, Natasa; Jalsenjak, Ivan

    2004-12-01

    The aim of our study was to develop a liposomal carrier system for the local treatment of bacterial vaginosis, capable to efficiently deliver entrapped drug during an extended period of time. Chloramphenicol was entrapped in liposomes composed of egg phosphatidylcholine/egg phosphatidylgycerol-sodium (9:1, molar ratio) and prepared by two different methods, the proliposome method and the polyol dilution method. Both liposome preparations were characterised and compared for particle size, polydispersity, entrapment efficiency and tested for in vitro stability in media that simulate human vaginal conditions (buffer pH 4.5 and vaginal fluid simulant). To achieve application viscosity of liposomes and to further improve their stability, liposomes prepared by the proliposome method were incorporated in the bioadhesive gel made of Carbopol 974P NF resin. In vitro release studies of liposomes incorporated in the gel have shown a prolonged release of entrapped chloramphenicol compared to control gel. Even after 24 hours of incubation in the vaginal fluid simulant, more than 40% of the originally entrapped drug was still retained in the gel. Storage stability studies have proven the ability of the Carbopol 974P NF gel to preserve the original size distribution of incorporated liposomes. All the performed experiments confirm the applicability of liposomes as a novel drug carrier system for the local treatment of bacterial vaginosis.

  13. Photocatalytic degradation and drug activity reduction of Chloramphenicol.

    PubMed

    Chatzitakis, A; Berberidou, C; Paspaltsis, I; Kyriakou, G; Sklaviadis, T; Poulios, I

    2008-01-01

    The photocatalytic degradation of Chloramphenicol, an antibiotic drug, has been investigated in aqueous heterogeneous solutions containing n-type oxide semiconductors as photocatalysts. The disappearance of the organic molecule follows approximately a pseudo-first-order kinetics according to the Langmuir-Hinshelwood model. It was observed that, with TiO(2) P-25 as photocatalyst, quantitative degradation of the organic molecule occurs after 4h of illumination. During this time, the dechlorination of the substrate is complete, while the organic nitrogen was recovered in the form of nitrate and ammonium ions. The effect of temperature on the degradation rate of Chloramphenicol shows similar apparent activation energies for both TiO(2) P-25 and ZnO photocatalysts. The initial apparent photonic efficiency (zeta(0)) of the photo-oxidation and the mineralization under various experimental conditions have been calculated, while the Kirby-Bauer disc diffusion method showed a 100% reduction of the drug activity after 90 min of photocatalytic treatment.

  14. 21 CFR 524.390 - Chloramphenicol ophthalmic and topical dosage forms.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chloramphenicol ophthalmic and topical dosage... HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS OPHTHALMIC AND TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.390 Chloramphenicol ophthalmic and topical dosage forms....

  15. [Real-time UV imaging of chloramphenicol intrinsic dissolution characteristics from ophthalmic in situ gel].

    PubMed

    Chen, Jian-Xiu; Guo, Zhen; Li, Hai-Yan; Wu, Li; He, Zhong-Gui; Hu, Rong-Feng; Zhang, Ji-Wen

    2013-07-01

    In this paper, chloramphenicol was selected as a model drug to prepare in situ gels. The intrinsic dissolution rate of chloramphenicol from in situ gel was evaluated using the surface dissolution imaging system. The results indicated that intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel decreased significantly when the poloxamer concentration increased. The addition of the thickener reduced the intrinsic dissolution rate of chloramphenicol thermosensitive gel, wherein carbomer had the most impact. Different dilution ratios of simulated tear fluid greatly affected gel temperature, and had little influence on the intrinsic dissolution rate of chloramphenicol from the thermosensitive in situ gel. The pH of simulated tear fluid had little influence on the intrinsic dissolution rate of chloramphenicol thermosensitive in situ gel. For the pH sensitive in situ gel, the dissolution rates of chloramphenicol in weak acidic and neutral simulated tear fluids were slower than that in weak alkaline simulated tear fluid. In conclusion, the intrinsic dissolution of chloramphenicol from in situ gel was dependent on formulation and physiological factors. With advantages of small volume sample required and rapid detection, the UV imaging method can be an efficient tool for the evaluation of drug release characteristics of ophthalmic in situ gel.

  16. Phosphorylation of chloramphenicol by a recombinant protein Yhr2 from Streptomyces avermitilis MA4680.

    PubMed

    Rajesh, Thangamani; Sung, Changmin; Kim, Hyeonjeong; Song, Eunjung; Park, Hyung-Yeon; Jeon, Jong-Min; Yoo, Dongwon; Kim, Hyun Joong; Kim, Yong Hyun; Choi, Kwon-Young; Song, Kyung-Guen; Yang, Yung-Hun

    2013-06-15

    Although phosphorylation of chloramphenicol has been shown to occur in the chloramphenicol producer, Streptomyces venezuelae, there are no reports on the existence of chloramphenicol phosphorylase in other Streptomyces species. In the present study, we report the modification of chloramphenicol by a recombinant protein, designated as Yhr2 (encoded by SAV_877), from Streptomyces avermitilis MA4680. Recombinant Yhr2 was expressed in Escherichia coli BL21 (DE3) and the cells expressing this recombinant protein were shown to phosphorylate chloramphenicol to a 3'-O-phosphoryl ester derivative, resulting in an inactivated form of the antibiotic. Expression of yhr2 conferred chloramphenicol resistance to E. coli cells up to 25 μg/mL and in an in vitro reaction, adenosine triphosphate (ATP), guanosine triphosphate (GTP), adenosine diphosphate (ADP) and guanosine diphosphate (GDP) were shown to be the phosphate donors for phosphorylation of chloramphenicol. This study highlights that antibiotic resistance conferring genes could be easily expressed and functionalized in other organisms that do not produce the respective antibiotic.

  17. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells

    SciTech Connect

    Gorman, C.M.; Moffat, L.F.; Howard, B.H.

    1982-09-01

    The authors constructed a series of recombinant genomes which directed expression of the enzyme chloramphenicol acetyltransferase (CAT) in mammalian cells. The prototype recombinant in this series, pSV2-cat, consisted of the beta-lactamase gene and origin of replication from pBR322 coupled to a simian virus 40 (SV40) early transcription region into which CAT coding sequences were inserted. Readily measured levels of CAT accumulated within 48 h after the introduction of pSV2-cat DNA into African green monkey kidney CV-1 cells. Because endogenous CAT activity is not present in CV-1 or other mammalian cells, and because rapid, sensitive assays for CAT activity are available, these recombinants provided a uniquely convenient system for monitoring the expression of foreign DNAs in tissue culture cells. To demonstrate the usefulness of this system, we constructed derivatives of pSV2-cat from which part or all of the SV 40 promoter region was removed. Deletion of one copy of the 72-base-pair repeat sequence in the SV40 promoter caused no significant decrease in CAT synthesis in monkey kidney CV-1 cells; however, an additional deletion of 50 base pairs from the second copy of the repeats reduced CAT synthesis to 11% of its level in the wild type. They also constructed a recombinant, pSVO-cat, in which the entire SV40 promoter region was removed and a unique HindIII site was substituted for the insertion of other promoter sequences.

  18. Rapid surface enhanced Raman scattering detection method for chloramphenicol residues

    NASA Astrophysics Data System (ADS)

    Ji, Wei; Yao, Weirong

    2015-06-01

    Chloramphenicol (CAP) is a widely used amide alcohol antibiotics, which has been banned from using in food producing animals in many countries. In this study, surface enhanced Raman scattering (SERS) coupled with gold colloidal nanoparticles was used for the rapid analysis of CAP. Density functional theory (DFT) calculations were conducted with Gaussian 03 at the B3LYP level using the 3-21G(d) and 6-31G(d) basis sets to analyze the assignment of vibrations. Affirmatively, the theoretical Raman spectrum of CAP was in complete agreement with the experimental spectrum. They both exhibited three strong peaks characteristic of CAP at 1104 cm-1, 1344 cm-1, 1596 cm-1, which were used for rapid qualitative analysis of CAP residues in food samples. The use of SERS as a method for the measurements of CAP was explored by comparing use of different solvents, gold colloidal nanoparticles concentration and absorption time. The method of the detection limit was determined as 0.1 μg/mL using optimum conditions. The Raman peak at 1344 cm-1 was used as the index for quantitative analysis of CAP in food samples, with a linear correlation of R2 = 0.9802. Quantitative analysis of CAP residues in foods revealed that the SERS technique with gold colloidal nanoparticles was sensitive and of a good stability and linear correlation, and suited for rapid analysis of CAP residue in a variety of food samples.

  19. Kinetic studies on enzymatic acetylation of chloramphenicol in Streptococcus faecalis.

    PubMed Central

    Nakagawa, Y; Nitahara, Y; Miyamura, S

    1979-01-01

    The kinetics of chloramphenicol (CP) acetylation by CP acetyltransferase from Streptococcus faecalis was studied. CP was shown to be acetylated enzymatically to its 3-O-acetyl derivative (3-AcCP) in the presence of acetyl coenzyme A, after which 3-AcCP was converted nonenzymatically to its 1-O-acetyl isomer, 1-O-acetyl CP (1-AcCP). At equilibrium, the 1-AcCP and 3-AcCP were present in a 1:4 ratio. Subsequently the diacetylated product, 1,3-O-O-diacetyl CP [1,3-(Ac)2CP], was enzymatically produced from 1-AcCP by the same enzyme. Theoretical calculation of rate constants (k1, k2, k3) for each successive reaction is as follows: (Formula: see text). This calculation gave k1 = 0.4 min-1, k2 = 0.002 min-1, and k3 = 0.016 min-1. Experimental results agreed closely with these calculated values. Images PMID:119483

  20. 21 CFR 524.390d - Chloramphenicol-prednisolone ophthalmic ointment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... bacterial conjunctivitis and ocular inflammation caused by organisms susceptible to chloramphenicol. (3) Limitations. Therapy for cats should not exceed 7 days, prolonged use in cats may produce blood dyscrasia. As... institute appropriate therapy. All topical ophthalmic preparations containing corticosteroids, with...

  1. Post-chemiluminescence determination of chloramphenicol based on luminol-potassium periodate system.

    PubMed

    Yang, Xiao Feng; Li, Nian Bing; Luo, Hong Qun

    2012-01-01

    A post-chemiluminescence (PCL) phenomenon was observed when chloramphenicol was injected into a mixture of luminol and potassium periodate after the chemiluminescence (CL) reaction of luminol-potassium periodate had finished. The possible reaction mechanism was proposed based on studies of the CL kinetic characteristics, the CL spectra, the fluorescence spectra and the UV-vis absorption spectra of the related substances. Based on the PCL reaction, a rapid and sensitive method for the determination of chloramphenicol was established. The linear response range was 6.0 × 10(-7) -1.0 × 10(-5) mol/L, with a correlation coefficient of 0.9986. The relative standard deviation (RSD) for 5.0 × 10(-6) mol/L chloramphenicol was 2.3% (n = 11). The detection limit was 1.6 × 10(-7) mol/L. The method has been applied to the determination of chloramphenicol in pharmaceutical samples with satisfactory results.

  2. Effect of Certain Antimetabolites and of Chloramphenicol on Conjugation of E. coli K-12,

    DTIC Science & Technology

    organisms. The authors studied the effect of chloramphenicol, 5-bromouracil (5-BU), and 8-azaguanine (8-AG) on the conjugation of E . coli K-12 on the transfer of genetic markers from the donor to the recipient.

  3. Biomimetic piezoelectric quartz crystal sensor with chloramphenicol-imprinted polymer sensing layer.

    PubMed

    Ebarvia, Benilda S; Ubando, Isaiah E; Sevilla, Fortunato B

    2015-11-01

    The measurement of banned antibiotic like chloramphenicol is significant for customer protection and safety. The presence of residual antibiotics in foods and food products of animal origin could pose as health hazards and affect food quality for global acceptance. In this study, the potential of a chloramphenicol sensor based on molecularly imprinted polymer (MIP) coupled with a piezoelectric quartz crystal was explored. The MIP was prepared by precipitation polymerization at 60 °C. Methacrylic acid was used as monomer, trimethylolpropane trimethacrylate (TRIM) as crosslinker, and chloramphenicol as the template. Template removal on the resulting polymer was done by extraction using methanol-acetic acid. Characterization of the MIP and NIP were conducted by spectroscopic and microscopic methods. These further supported the imprinting and rebinding process of chloramphenicol to the polymer matrix. The chloramphenicol sensor was devised by spin-coating onto one side of the 10 MHz AT-cut quartz crystal the MIP suspension in polyvinylchloride-tetrahydrofuran (6:2:1 w/w/v) solution. Optimization of sensor response was performed by varying the type of cross-linker, amount of MIP sensing layer, curing time, and pH. The sensor exhibited good sensitivity of about 73 Hz/log (conc., µg mL(-1)) and good repeatability (rsd<10%). A linear relationship (r(2)=0.9901) between frequency shift and chloramphenicol concentration in the range of 1×10(-6) up to 1×10(-1) µg/mL was obtained. The sensor response was highly selective to chloramphenicol than with other compounds of similar chemical structures. Acceptable percent recovery was obtained for real sample analysis using the sensor. The proposed sensor could be a promising low cost and highly sensitive approach for residual chloramphenicol quantification in food products.

  4. CraA, a Major Facilitator Superfamily Efflux Pump Associated with Chloramphenicol Resistance in Acinetobacter baumannii▿

    PubMed Central

    Roca, I.; Marti, S.; Espinal, P.; Martínez, P.; Gibert, I.; Vila, J.

    2009-01-01

    Acinetobacter baumannii has been increasingly associated with hospital-acquired infections, and the presence of multidrug resistance strains is of great concern to clinicians. A. baumannii is thought to possess a great deal of intrinsic resistance to several antimicrobial agents, including chloramphenicol, although the mechanisms involved in such resistance are not well understood. In this work, we have identified a major facilitator superfamily efflux pump present in most A. baumannii strains, displaying strong substrate specificity toward chloramphenicol. PMID:19581458

  5. Randomized comparison of aztreonam and chloramphenicol in treatment of typhoid fever.

    PubMed Central

    Gotuzzo, E; Echevarría, J; Carrillo, C; Sánchez, J; Grados, P; Maguiña, C; DuPont, H L

    1994-01-01

    Patients with clinical typhoid fever plus a blood, bone marrow, or bile positive for Salmonella typhi or Salmonella paratyphi were included in an open clinical trial to compare the efficacy of aztreonam (6 g/day [2 g intravenously every 8 h]) given for 10 days with that of chloramphenicol (50 mg/kg of body weight per day [intravenously or orally]) administered for 14 days. A total of 44 patients, 22 in each group, were included in the study, and both groups were comparable in terms of baseline parameters. All patients randomized to receive chloramphenicol completed the 14 days of treatment, while two patients randomized to receive aztreonam developed an intestinal hemorrhage, and a third patient elected to withdraw from the trial. Defervescence occurred more quickly in the subjects receiving chloramphenicol than in those receiving aztreonam (P < 0.05). All patients in the chloramphenicol group were clinically cured during therapy, while four patients (21%) in the group receiving aztreonam were declared clinical treatment failures. None of the 19 patients receiving aztreonam, compared with 7 of 22 (32%) patients receiving chloramphenicol, had a positive blood culture after 24 h of therapy (P < 0.05). Adverse experiences were unusual and mild. In the study, aztreonam was less effective than chloramphenicol with regard to clinical effectiveness and time of defervescence but was more effective in the elimination of the infecting Salmonella organisms from the bloodstream. PMID:8203854

  6. γ-Cyclodextrin capped silver nanoparticles for molecular recognition and enhancement of antibacterial activity of chloramphenicol.

    PubMed

    Gannimani, Ramesh; Ramesh, Muthusamy; Mtambo, Sphamandla; Pillay, Karen; Soliman, Mahmoud E; Govender, Patrick

    2016-04-01

    Computational studies were conducted to identify the favourable formation of the inclusion complex of chloramphenicol with cyclodextrins. The results of molecular docking and molecular dynamics predicted the strongest interaction of chloramphenicol with γ-cyclodextrin. Further, the inclusion complex of chloramphenicol with γ-cyclodextrin was experimentally prepared and a phenomenon of inclusion was verified by using different characterization techniques such as thermogravimetric analysis, differential scanning calorimetry, (1)H nuclear magnetic resonance (NMR) and two dimensional nuclear overhauser effect spectroscopy (NOESY) experiments. From these results it was concluded that γ-cyclodextrins could be an appropriate cyclodextrin polymer which can be used to functionalize chloramphenicol on the surface of silver nanoparticles. In addition, γ-cyclodextrin capped silver nanoparticles were synthesized and characterized using UV-visible spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy dispersive X-ray analysis (EDX), Fourier transform infrared spectroscopy (FTIR) and zeta potential analysis. Molecular recognition of chloramphenicol by these cyclodextrin capped silver nanoparticles was confirmed by surface enhanced raman spectroscopy (SERS) experiments. Synergistic antibacterial effect of chloramphenicol with γ-cyclodextrin capped silver nanoparticles was evaluated against Pseudomonas aeruginosa (ATCC 27853), Enterococcus faecalis (ATCC 5129), Klebsiella pneumoniae (ATCC 700603) and Staphylococcus aureus (ATCC 43300). The results from the antibacterial experiment were favourable thus allowing us to conclude that the approach of modifying organic drug molecules with cyclodextrin capped inorganic silver nanoparticles could help to enhance the antibacterial activity of them.

  7. Chloramphenicol in children: dose, plasma levels and clinical effects.

    PubMed

    Ismail, R; Teh, L K; Choo, E K

    1998-06-01

    Despite concerns about adverse effects, chloramphenicol (CMC) continues to be used in certain situations and, due to its low therapeutic index and variable pharmacokinetics, therapeutic drug monitoring (TDM) is often recommended. At our centre, CMC finds applications in typhoid and meningitis and TDM is routinely performed. Elsewhere in Malaysia, however, CMC is used without TDM. We therefore decided to evaluate our TDM for CMC in relation to its roles in CMC therapy in children, who constitute most of our patients. Our objective was also to develop strategies to improve our TDM for CMC use. Data were collected from 168 children given CMC for various indications and monitored by the TDM service. Plasma CMC was determined by HPLC and used to adjust doses to maintain concentrations within a range of 10-25 micrograms/ml. Outcomes measured included daily temperatures and haematological indices. Daily doses and plasma CMC varied greatly. Doses averaged 40.5 mg/kg for neonates and 75.5 for older children. Average peak concentrations were therapeutic in 60% and trough in 42%. Average duration of fever was 6.3 days and it was unaffected by plasma CMC. Typhoid was eradicated in 97% but nine children with other diagnoses died. Side-effects were confined to mild reversible haematological abnormalities which developed in 11% of children at plasma concentrations which tended to be high. We conclude that CMC remains useful in children with typhoid. Its use for other indications, however, should be reviewed. Routine TDM for CMC is probably not warranted, at least until a clearer role is defined by well designed prospective studies.

  8. A chromosomal chloramphenicol acetyltransferase determinant from a probiotic strain of Bacillus clausii.

    PubMed

    Galopin, Sébastien; Cattoir, Vincent; Leclercq, Roland

    2009-06-01

    The mechanism of resistance to chloramphenicol was studied in four strains of Bacillus clausii included in a probiotic mixture, which is administered to humans for prevention of gastrointestinal side effects due to oral antibiotic therapy. By cloning experiments, a chloramphenicol acetyltransferase (CAT) gene, cat(Bcl), coding for a putative 228-amino acid CAT protein was identified in B. clausii SIN. The deduced amino acid sequence displayed from 31% to 85% identity with 56 CAT proteins from other Gram-positive bacterial strains. The cat(Bcl) gene was also detected by PCR in the three other B. clausii strains resistant to chloramphenicol, whereas it was absent in the three control strains susceptible to chloramphenicol. Pulse-field gel electrophoresis of total DNA digested by I-CeuI followed by hybridization with a cat-specific probe as well as unsuccessful repeated attempts of in vitro transfer of chloramphenicol resistance to various recipient cells indicated that cat(Bcl) was chromosomally located in all four resistant B. clausii strains.

  9. Observation of residues in tissues of chickens exposed to low dietary concentrations of chloramphenicol.

    PubMed

    Rejtharová, Martina; Rejthar, Libor; Čačková, Katarína; Bureš, Jiří; Vernerová, Eva; Hera, Alfred

    2017-04-01

    To investigate potential residues in tissues arising from naturally occurring low levels of chloramphenicol in plant material, feeding studies were conducted with chickens. A common chicken feed was prepared containing 0, 10, 50 and 200 μg kg(-1) chloramphenicol and levels were confirmed by LC-MS/MS. Four separate groups of broiler chickens, eight animals in each group, were fed all their 35-day life with this contaminated feed. They were allowed ad libitum access to this feed and fresh water. After slaughtering the chickens, the residues in muscle and liver tissues were determined using GC/MS-NCI method. No residues were detected in tissues of animals from groups fed with feed containing 0, 10 or 50 µg kg(-1). Low chloramphenicol residual concentrations were observed in a few of the muscle samples obtained from the group of chickens fed with feed containing chloramphenicol in added concentration 200 µg kg(-1). No residues were detected in the remaining samples of this group. These results indicate that when residues of chloramphenicol are detected it is in all probability through illegal use.

  10. Chloramphenicol – A Potent Armament Against Multi-Drug Resistant (MDR) Gram Negative Bacilli?

    PubMed Central

    2016-01-01

    Introduction Multidrug-resistant gram-negative bacteria cause infections which are hard to treat and cause high morbidity and mortality. Due to limited therapeutic options there is a renewed interest upon older antimicrobials which had fallen into disuse as a result of toxic side effects. One such antibiotic is chloramphenicol which was sidelined due to reports linking its use with the development of aplastic anaemia. Aim A study was conducted to evaluate the susceptibility of chloramphenicol in light of the emerging problem of multi-drug resistant gram negative bacteria (MDR GNB). Materials and Methods A total of 483 MDR GNB of the 650 consecutive Gram Negative Bacteria isolated from various clinical samples of patients admitted at a tertiary care hospital in Jaipur between January-June 2014 were screened for chloramphenicol susceptibility by the disc diffusion method as per CLSI guidelines. Results The MDR GNB isolates were obtained from 217 (45%) urine, 163 (34%) from respiratory samples, 52(11%) from pus, 42 (9%) from blood and 9 (2%) from body fluids. A 68% of the MDR GNB isolates were found to be sensitive to chloramphenicol. Conclusion Clinicians should always check for the local susceptibility of Gram-negative bacteria to chloramphenicol. This antibiotic has a potential to play a role in the therapeutic management of infections due to MDR GNB pathogens. PMID:27042458

  11. Persistence of chloramphenicol residues in chicken muscle tissue after a therapeutic dose administration.

    PubMed

    Rejtharová, Martina; Rejthar, Libor; Bureš, Jiří; Vernerová, Eva; Hera, Alfred

    2017-04-01

    One-day-old chickens were individually orally treated with chloramphenicol at a dose of 100 mg per kg of body weight per day for three consecutive days. After the final treatment, the groups of six birds were sacrificed in seven-day intervals up to 42 days. The muscle tissue collected from the breasts and legs of each bird was individually examined for the presence of chloramphenicol residues using a GC/MS-NCI analytical method, which was validated according to Commission Decision 2002/657/EC. The decision limit (CCα) obtained for the method was 0.05 ng g(-1). The results showed a rapid decrease of chloramphenicol concentration in the muscle tissue after termination of the treatment, but also showed a relatively long persistence of low residue concentrations. Levels of chloramphenicol in muscle tissue averaged 64 ng g(-1) seven days after the final treatment and fell to 0.21 ng g(-1) after 35 days. All animals tested on the 35th day after the final treatment showed detectable chloramphenicol concentrations above the decision limit of the method used. No residues were detected in the animal tissues 42 days after the end of the treatment.

  12. A comparative study of epicillin and chloramphenicol in the treatment of enteric fever.

    PubMed

    Hassau, A; Sippel, J; Farid, Z; Tadros, S S; Brian, M

    1977-01-01

    One hundred patients with acute enteric fever were randomly assigned to treatment with either chloramphenicol 50 mg/kg body-weight or epicillin 1 g six hourly. Eighty-one patients had a positive blood culture for typhoid or paratyphoid bacilli and nineteen had a positive stool culture with a significant Widal titre. All fifty patients in the group treated with chloramphenicol responded, however there was one relapse with bacteraemia. In the group treated with epicillin, six from the total of fifty patients were considred treatment failures. Treatment was considred as a failure if the patient was febrile after ten days treatment or if there was a deterioration despite antibiotic therapy.

  13. Gatifloxacin versus chloramphenicol for uncomplicated enteric fever: an open-label, randomised, controlled trial

    PubMed Central

    Arjyal, Amit; Basnyat, Buddha; Koirala, Samir; Karkey, Abhilasha; Dongol, Sabina; Agrawaal, Krishna Kumar; Shakya, Nikki; Shrestha, Kabina; Sharma, Manish; Lama, Sanju; Shrestha, Kasturi; Khatri, Nely Shrestha; Shrestha, Umesh; Campbell, James I; Baker, Stephen; Farrar, Jeremy; Wolbers, Marcel; Dolecek, Christiane

    2011-01-01

    Summary Background We aimed to investigate whether gatifloxacin, a new generation and affordable fluoroquinolone, is better than chloramphenicol for the treatment of uncomplicated enteric fever in children and adults. Methods We did an open-label randomised superiority trial at Patan Hospital, Kathmandu, Nepal, to investigate whether gatifloxacin is more effective than chloramphenicol for treating uncomplicated enteric fever. Children and adults clinically diagnosed with enteric fever received either gatifloxacin (10 mg/kg) once a day for 7 days, or chloramphenicol (75 mg/kg per day) in four divided doses for 14 days. Patients were randomly allocated treatment (1:1) in blocks of 50, without stratification. Allocations were placed in sealed envelopes opened by the study physician once a patient was enrolled into the trial. Masking was not possible because of the different formulations and ways of giving the two drugs. The primary outcome measure was treatment failure, which consisted of at least one of the following: persistent fever at day 10, need for rescue treatment, microbiological failure, relapse until day 31, and enteric-fever-related complications. The primary outcome was assessed in all patients randomly allocated treatment and reported separately for culture-positive patients and for all patients. Secondary outcome measures were fever clearance time, late relapse, and faecal carriage. The trial is registered on controlled-trials.com, number ISRCTN 53258327. Findings 844 patients with a median age of 16 (IQR 9–22) years were enrolled in the trial and randomly allocated a treatment. 352 patients had blood-culture-confirmed enteric fever: 175 were treated with chloramphenicol and 177 with gatifloxacin. 14 patients had treatment failure in the chloramphenicol group, compared with 12 in the gatifloxacin group (hazard ratio [HR] of time to failure 0·86, 95% CI 0·40–1·86, p=0·70). The median time to fever clearance was 3·95 days (95% CI 3·68–4·68

  14. Electrochemical analysis of chloramphenicol using boron-doped diamond electrode applied to a flow-injection system.

    PubMed

    Chuanuwatanakul, Suchada; Chailapakul, Orawon; Motomizu, Shoji

    2008-04-01

    The electrochemical properties of chloramphenicol at a boron-doped diamond thin-film (BDD) electrode were studied using cyclic voltammetry. The highest current response of chloramphenicol was obtained with phosphate buffer, pH 6 (0.1 M) in 1% ethanol. The relationship between the concentration of chloramphenicol and the current response was linear over the range of 0.1-10 mM (R2=0.9990). The amount of chloramphenicol was analyzed by flow-injection analysis. A thin-layer flow cell equipped with a BDD electrode was used as an amperometric detector, and experiments were carried out at -0.7 V (vs. Ag/AgCl). The linear relationship between the current response and the concentration of chloramphenicol in the range of 0.1-50 microM (R2=0.9948) and the limit of detection of 0.03 microM (S/N=3) were obtained. This method has been successfully applied to the determination of chloramphenicol in sterile eye drops and milk sample by the standard addition method. The average recoveries of chloramphenicol in eye drops were 98.0%, and the average recoveries of chloramphenicol from spiked milk were 93.9-103%.

  15. 75 FR 55810 - Withdrawal of Approval of New Animal Drug Applications; Chloramphenicol, Lincomycin, Pyrantel...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-14

    ... HUMAN SERVICES Food and Drug Administration Withdrawal of Approval of New Animal Drug Applications; Chloramphenicol, Lincomycin, Pyrantel Tartrate, and Tylosin Phosphate and Sulfamethazine AGENCY: Food and Drug Administration, HHS. ACTION: Notice. SUMMARY: The Food and Drug Administration (FDA) is withdrawing approval...

  16. DIFFERENTIATION AND FUNCTIONAL EXPRESSION OF POTENTIAL ANTIBODY-PRODUCING CELLS IN THE PRESENCE OF CHLORAMPHENICOL

    PubMed Central

    Schoenberg, Melvin D.; Moore, Richard D.; Weisberger, Austin S.

    1967-01-01

    Rabbits were immunized with diphtheria toxoid combined with complete Freund's adjuvant. Half of the animals were started on intramuscular injections of chloramphenicol 24 hr before the injection of the antigens. There was a general depression of protein synthesis in the immune system in the presence of chloramphenicol, but a greater effect on the synthesis of antibody than on the synthesis of proteins necessary for reproduction and maturation. In contrast to the finding of antibody in cells of the spleen and in the circulation of the control animals, those animals receiving chloramphenicol did not have measurable circulating antibody, and their spleens contained only a few cells with intracytoplasmic antibody late in the course of the experiment. Cytologically there was maturation of potential antibody-producing cells in the red pulp and nonfollicular white pulp of the spleen while the animals were receiving chloramphenicol. These cells developed more slowly, and were fewer and smaller than those of the control animals. They had numerous small, electron-opaque particles in their cytoplasm early in development. Ribosomes were synthesized, though fewer in number. The endoplasmic reticulum formed more slowly. PMID:10976231

  17. Analysis of conjugation of chloramphenicol and hemoglobin by fluorescence, circular dichroism and molecular modeling

    NASA Astrophysics Data System (ADS)

    Ding, Fei; Liu, Wei; Sun, Ye; Yang, Xin-Ling; Sun, Ying; Zhang, Li

    2012-01-01

    Chloramphenicol is a low cost, broad spectrum, highly active antibiotic, and widely used in the treatment of serious infections, including typhoid fever and other life-threatening infections of the central nervous system and respiratory tract. The purpose of the present study was to examine the conjugation of chloramphenicol with hemoglobin (Hb) and compared with albumin at molecular level, utilizing fluorescence, UV/vis absorption, circular dichroism (CD) as well as molecular modeling. Fluorescence data indicate that drug bind Hb generate quenching via static mechanism, this corroborates UV/vis absorption measurements that the ground state complex formation with an affinity of 10 4 M -1, and the driving forces in the Hb-drug complex are hydrophilic interactions and hydrogen bonds, as derived from computational model. The accurate binding site of drug has been identified from the analysis of fluorescence and molecular modeling, α1β2 interface of Hb was assigned to possess high-affinity for drug, which located at the β-37 Trp nearby. The structural investigation of the complexed Hb by synchronous fluorescence, UV/vis absorption, and CD observations revealed some degree of Hb structure unfolding upon complexation. Based on molecular modeling, we can draw the conclusion that the binding affinity of drug with albumin is superior, compared with Hb. These phenomena can provide salient information on the absorption, distribution, pharmacology, and toxicity of chloramphenicol and other drugs which have analogous configuration with chloramphenicol.

  18. 21 CFR 524.390d - Chloramphenicol-prednisolone ophthalmic ointment.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS OPHTHALMIC AND TOPICAL DOSAGE FORM NEW ANIMAL DRUGS § 524.390d Chloramphenicol-prednisolone ophthalmic ointment. (a) Specifications. Each gram... with other antibiotics, prolonged use may result in overgrowth of nonsusceptible organisms....

  19. Adverse events associated with chloramphenicol use in dogs: a retrospective study (2007-2013).

    PubMed

    Short, J; Zabel, S; Cook, C; Schmeitzel, L

    2014-11-29

    Chloramphenicol is a broad spectrum antibiotic that has been increasingly utilised since the emergence of methicillin-resistant staphylococcal infections. Due to toxicities in humans, use of the drug has been limited. In dogs, gastrointestinal signs are common adverse events described, and bone marrow suppression is possible. The aim of this study was to evaluate the adverse events associated with chloramphenicol in dogs seen by one specialty practice from January 2007 through June 2013. The database was searched for all dogs prescribed chloramphenicol during the time period. Dosage, length of treatment, age and body weight of the dogs were recorded as well as any adverse events that occurred during treatment. A total of 105 cases were evaluated. Thirty-nine dogs experienced at least one adverse event while on the medication. The most commonly noted were gastrointestinal signs and hindlimb weakness. The mean body weight for dogs with hindlimb weakness was 35.3 kg, which was significant. Resolution was documented in 54 per cent of cases when the drug was discontinued. Methicillin-resistant Staphylococcus pseudintermedius on bacterial culture was listed as the reason for chloramphenicol use in 76 per cent of the cases. Based on this information, further prospective studies are recommended to evaluate the reproducibility of this report.

  20. 77 FR 41412 - Determination That CHLOROMYCETIN (Chloramphenicol) Capsules, 250 Milligrams, Were Withdrawn From...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-13

    .... With the approval of additional therapies with less severe adverse drug effects, FDA has determined... HUMAN SERVICES Food and Drug Administration Determination That CHLOROMYCETIN (Chloramphenicol) Capsules, 250 Milligrams, Were Withdrawn From Sale for Reasons of Safety or Effectiveness AGENCY: Food and...

  1. Involvement of the Efflux Pumps in Chloramphenicol Selected Strains of Burkholderia thailandensis: Proteomic and Mechanistic Evidence

    PubMed Central

    Biot, Fabrice V.; Valade, Eric; Garnotel, Eric; Chevalier, Jacqueline; Villard, Claude; Thibault, François M.; Vidal, Dominique R.; Pagès, Jean-Marie

    2011-01-01

    Burkholderia is a bacterial genus comprising several pathogenic species, including two species highly pathogenic for humans, B. pseudomallei and B. mallei. B. thailandensis is a weakly pathogenic species closely related to both B. pseudomallei and B. mallei. It is used as a study model. These bacteria are able to exhibit multiple resistance mechanisms towards various families of antibiotics. By sequentially plating B. thailandensis wild type strains on chloramphenicol we obtained several resistant variants. This chloramphenicol-induced resistance was associated with resistance against structurally unrelated antibiotics including quinolones and tetracyclines. We functionally and proteomically demonstrate that this multidrug resistance phenotype, identified in chloramphenicol-resistant variants, is associated with the overexpression of two different efflux pumps. These efflux pumps are able to expel antibiotics from several families, including chloramphenicol, quinolones, tetracyclines, trimethoprim and some β-lactams, and present a partial susceptibility to efflux pump inhibitors. It is thus possible that Burkholderia species can develop such adaptive resistance mechanisms in response to antibiotic pressure resulting in emergence of multidrug resistant strains. Antibiotics known to easily induce overexpression of these efflux pumps should be used with discernment in the treatment of Burkholderia infections. PMID:21347382

  2. Estimation of the chloramphenicol and cycloheximide inhibition of protein synthesis in brain cholinergic synaptosomes.

    PubMed

    Corbaton, V; Muiño, M T; Fernández-Silva, P; López-Pérez, M J; Montoya, J

    1991-03-15

    Cholinergic synaptosomes have been prepared from sheep brain cortex by means of an immunoaffinity method using a specific anti-(Chol I) antiserum. The [14C]leucine incorporation into proteins of this preparation shows a low cycloheximide and a high chloramphenicol sensitivity. This fact suggests that the mitochondrial protein synthesis system is the only one present in this fraction.

  3. Structural, electronic, thermodynamical and charge transfer properties of Chloramphenicol Palmitate using vibrational spectroscopy and DFT calculations

    NASA Astrophysics Data System (ADS)

    Mishra, Rashmi; Srivastava, Anubha; Sharma, Anamika; Tandon, Poonam; Baraldi, Cecilia; Gamberini, Maria Christina

    2013-01-01

    The global problem of advancing bacterial resistance to newer drugs has led to renewed interest in the use of Chloramphenicol Palmitate (C27H42Cl2N2O6) [Palmitic acid alpha ester with D-threo-(-),2-dichloro-N-(beta-hydroxy-alpha-(hydroxymethyl)-p-nitrophenethyl)acetamide also known as Detereopal]. The characterization of the three polymorphic forms of Chloramphenicol Palmitate (CPP) was done spectroscopically by employing FT-IR and FT-Raman techniques. The equilibrium geometry, various bonding features, and harmonic wavenumbers have been investigated for most stable form A with the help of DFT calculations and a good correlation was found between experimental data and theoretical values. Electronic properties have been analyzed employing TD-DFT for both gaseous and solvent phase. The theoretical calculation of thermodynamical properties along with NBO analysis has also been performed to have a deep insight into the molecule for further applications.

  4. Determination of chloramphenicol residues in shrimps by liquid chromatography-mass spectrometry.

    PubMed

    Ramos, M; Muñoz, P; Aranda, A; Rodriguez, I; Diaz, R; Blanca, J

    2003-07-05

    A liquid chromatographic method with mass spectrometric detection and identification (LC-MS) is presented for the determination of chloramphenicol (CAP) in shrimp tissues. Homogenized shrimp samples were extracted with phosphate buffer (pH 7.0). Clean-up was carried out on a C(18) SPE cartridge. Chloramphenicol was determined by LC-MS-ESI in negative mode. The column used was a Symmetry Shield with a mixture of acetonitrile-water (25:75) as mobile phase. Shrimp samples were fortified at CAP levels between 0.2 and 50 ng g(-1) with 5D-CAP as internal standard. At these levels, accuracies lay between 101 and 110% and between-day reproducibilities were lower than 7.1%, expressed as the variation coefficient of the mean. Limit of decision (CCalpha) was 0.02 ng g(-1). Limit of quantitation (LOQ) was 0.2 ng g(-1).

  5. Dissipation of carbendazim and chloramphenicol alone and in combination and their effects on soil fungal:bacterial ratios and soil enzyme activities.

    PubMed

    Yan, Hu; Wang, Dandan; Dong, Bin; Tang, Feifan; Wang, Baichuan; Fang, Hua; Yu, Yunlong

    2011-07-01

    The dissipation of carbendazim and chloramphenicol alone and in combination and their effects on soil fungal:bacterial ratios and soil enzyme activities were investigated. The results revealed that carbendazim dissipation was little affected by chloramphenicol, whereas chloramphenicol dissipation was found to be retarded significantly by the presence of carbendazim. The inhibitory effect of carbendazim on the fungal:bacterial ratios was increased by the presence of chloramphenicol, and the inhibitory effect of chloramphenicol on neutral phosphatase was increased by the presence of carbendazim. Carbendazim increased soil catalase and urease activities, but this increase was partially diminished by the presence of chloramphenicol. Little interaction was observed between carbendazim and chloramphenicol with regard to their influence on soil invertase. The results obtained in this study suggest that combinations of fungicides and antibiotics may alter the compounds' individual behaviors in soil and their effects on soil enzymes.

  6. The effect of ethidium bromide and chloramphenicol on mitochondrial biogenesis in primary human fibroblasts

    SciTech Connect

    Kao, Li-Pin; Ovchinnikov, Dmitry; Wolvetang, Ernst

    2012-05-15

    The expression of mitochondrial components is controlled by an intricate interplay between nuclear transcription factors and retrograde signaling from mitochondria. The role of mitochondrial DNA (mtDNA) and mtDNA-encoded proteins in mitochondrial biogenesis is, however, poorly understood and thus far has mainly been studied in transformed cell lines. We treated primary human fibroblasts with ethidium bromide (EtBr) or chloramphenicol for six weeks to inhibit mtDNA replication or mitochondrial protein synthesis, respectively, and investigated how the cells recovered from these insults two weeks after removal of the drugs. Although cellular growth and mitochondrial gene expression were severely impaired after both inhibitor treatments we observed marked differences in mitochondrial structure, membrane potential, glycolysis, gene expression, and redox status between fibroblasts treated with EtBr and chloramphenicol. Following removal of the drugs we further detected clear differences in expression of both mtDNA-encoded genes and nuclear transcription factors that control mitochondrial biogenesis, suggesting that the cells possess different compensatory mechanisms to recover from drug-induced mitochondrial dysfunction. Our data reveal new aspects of the interplay between mitochondrial retrograde signaling and the expression of nuclear regulators of mitochondrial biogenesis, a process with direct relevance to mitochondrial diseases and chloramphenicol toxicity in humans. -- Highlights: ► Cells respond to certain environmental toxins by increasing mitochondrial biogenesis. ► We investigated the effect of Chloramphenicol and EtBr in primary human fibroblasts. ► Inhibiting mitochondrial protein synthesis or DNA replication elicit different effects. ► We provide novel insights into the cellular responses toxins and antibiotics.

  7. Two Distinct Major Facilitator Superfamily Drug Efflux Pumps Mediate Chloramphenicol Resistance in Streptomyces coelicolor▿

    PubMed Central

    Vecchione, James J.; Alexander, Blair; Sello, Jason K.

    2009-01-01

    Chloramphenicol, florfenicol, and thiamphenicol are used as antibacterial drugs in clinical and veterinary medicine. Two efflux pumps of the major facilitator superfamily encoded by the cmlR1 and cmlR2 genes mediate resistance to these antibiotics in Streptomyces coelicolor, a close relative of Mycobacterium tuberculosis. The transcription of both genes was observed by reverse transcription-PCR. Disruption of cmlR1 decreased the chloramphenicol MIC 1.6-fold, while disruption of cmlR2 lowered the MIC 16-fold. The chloramphenicol MIC of wild-type S. coelicolor decreased fourfold and eightfold in the presence of reserpine and Phe-Arg-β-naphthylamide, respectively. These compounds are known to potentiate the activity of some antibacterial drugs via efflux pump inhibition. While reserpine is known to potentiate drug activity against gram-positive bacteria, this is the first time that Phe-Arg-β-naphthylamide has been shown to potentiate drug activity against a gram-positive bacterium. PMID:19687245

  8. In vitro comparison of ampicillin-chloramphenicol and ampicillin-cefotaxime against 284 Haemophilus isolates.

    PubMed Central

    Lapointe, J R; Lavallée, C; Michaud, A; Chicoine, L; Joncas, J H

    1986-01-01

    Since November 1982 at the Sainte-Justine Hospital in Montreal, ampicillin and cefotaxime were used in association as initial treatment (greater than or equal to 48 h) for childhood bacterial meningitis. In this report is described the in vitro interaction of the new regimen in comparison with that of the previous ampicillin-chloramphenicol combination against 284 Haemophilus isolates. Among the 156 ampicillin-susceptible, beta-lactamase-negative isolates, synergy was detected in 13 with ampicillin-cefotaxime, and antagonism was detected in only 1; in contrast, synergy was found in only 2 strains with ampicillin-chloramphenicol, and antagonism was found in 15. These differences were statistically significant (P less than 0.01). Such significant differences were not observed among the 128 ampicillin-resistant, beta-lactamase-positive Haemophilus isolates. The synergy of ampicillin-cefotaxime did not contribute to a decrease of the MIC of cefotaxime for 90% of isolates tested, whereas the antagonism of ampicillin-chloramphenicol did not contribute to increase the MIC of ampicillin for 90% of isolates tested. PMID:3486630

  9. Susceptibility of Haemophilus influenzae to chloramphenicol and eight beta-lactam antibiotics.

    PubMed Central

    Thirumoorthi, M C; Kobos, D M; Dajani, A S

    1981-01-01

    We examined the minimal inhibitory concentrations and minimal bactericidal concentrations of chloramphenicol, ampicillin, ticarcillin, cefamandole, cefazolin, cefoxitin, cefotaxime, ceforanide, and moxalactam for 100 isolates of Haemophilus influenzae, 25 of which produced beta-lactamase. Susceptibility was not influenced by the capsular characteristic of the organism. The mean minimal inhibitory concentrations of cefamandole, ticarcillin, and ampicillin for beta-lactamase-producing strains were 3-, 120-, and 400-fold higher than their respective mean minimal inhibitory concentrations for beta-lactamase-negative strains. No such difference was noted for the other antibiotics. We performed time-kill curve studies, using chloramphenicol, ampicillin, cefamandole, cefotaxime, and moxalactam with two concentrations of the antimicrobial agents (4 or 20 times the minimal inhibitory concentrations) and two inoculum sizes (10(4) or 10(6) colony-forming units per ml). The inoculum size had no appreciable effect on the rate of killing of beta-lactamase-negative strains. The rates at which beta-lactamase-producing strains were killed by chloramphenicol, cefotaxime, and moxalactam was not influenced by the inoculum size. Whereas cefamandole in high concentrations was able to kill at 10(6) colony-forming units/ml of inoculum, it had only a temporary inhibiting effect at low drug concentrations. Methicillin and the beta-lactamase inhibitor CP-45,899 were able to neutralize the inactivation of cefamandole by a large inoculum of beta-lactamase-producing H. influenzae. PMID:6974541

  10. Simultaneous Chloramphenicol and Florfenicol Determination by A Validated DLLME-HPLC-UV Method in Pasteurized Milk

    PubMed Central

    Karami-Osboo, Rouhollah; Miri, Ramin; Javidnia, Katayoun; Kobarfard, Farzad

    2016-01-01

    The antibiotic residues in milk are a well-known serious problem and pose several health hazards to consumers. We have described a simple, rapid, and inexpensive DLLME-HPLC/UV technique for the extraction of chloramphenicol and florfenicol residues in milk samples. Under the optimum conditions, linearity of the method was observed over the range 0.02-0.85 µg/L with correlation coefficients > 0.999. The proposed method has been found to have a good limit of detection (signal to noise ratio = 3) for chloramphenicol (12.5 µg/Kg) and florfenicol (12.2 µg/Kg), and precision with relative standard deviation values under 15% (RSD, n = 3). Good recoveries (69.1–79.4%) were obtained for the extraction of the target analytes in milk samples. This simple and economic method has been applied for analyses of 15 real milk samples. Among all samples only one of them was contaminated to florfenicol; 62.4 µg/Kg and contamination to chloramphenicol was not detected. PMID:27980571

  11. Liquid chromatographic determination of multiple sulfonamides, nitrofurans, and chloramphenicol residues in pasteurized milk.

    PubMed

    Perez, Norma; Gutierrez, Rey; Noa, Mario; Diaz, Gilberto; Luna, Hector; Escobar, Irma; Munive, Zenaida

    2002-01-01

    A rapid and selective liquid chromatographic method was developed to detect 6 sulfonamides, 3 nitrofurans, and chloramphenicol residues in pasteurized milk. The 10 drugs were extracted with chloroform-acetone and the organic phase was evaporated; the residues were dissolved in an aqueous sodium acetate buffer solution 0.02M (pH = 4.8), and the fat was removed by washing with hexane. The aqueous layer was collected, filtered, and injected. The 6 sulfonamides and chloramphenicol were detected at 275 nm ultraviolet (UV) using a gradient system starting with sodium acetate buffer solution-acetonitrile (95 + 5) and finishing with sodium acetate buffer solution-acetonitrile (80 + 20). Nitrofurans were detected at 375 nm (UV) isocratically with sodium acetate buffer solution-acetonitrile (80 + 20). For 50 ppb fortified milk, the average recoveries were (sulfathiazole) 65.52%; (sulfamerazine) 75.36%; (sulfamethazine) 93.94%; (sulfachlorpyridazine) 75.94%; (sulfamethoxazole) 85.18%; (sulfamonomethoxine) 83.45%; (chloramphenicol) 104.17%; (nitrofurazone) 91.81%; (furazolidone) 100.76%; and (furaltadone) 72.38%. Method detection limits ranged from 4 ppb (nitrofurazone) to 16 ppb (sulfamethazine). Some matrix interferences (3-7 ppb) were observed only with sulfonamides.

  12. Simultaneous Chloramphenicol and Florfenicol Determination by A Validated DLLME-HPLC-UV Method in Pasteurized Milk.

    PubMed

    Karami-Osboo, Rouhollah; Miri, Ramin; Javidnia, Katayoun; Kobarfard, Farzad

    2016-01-01

    The antibiotic residues in milk are a well-known serious problem and pose several health hazards to consumers. We have described a simple, rapid, and inexpensive DLLME-HPLC/UV technique for the extraction of chloramphenicol and florfenicol residues in milk samples. Under the optimum conditions, linearity of the method was observed over the range 0.02-0.85 µg/L with correlation coefficients > 0.999. The proposed method has been found to have a good limit of detection (signal to noise ratio = 3) for chloramphenicol (12.5 µg/Kg) and florfenicol (12.2 µg/Kg), and precision with relative standard deviation values under 15% (RSD, n = 3). Good recoveries (69.1-79.4%) were obtained for the extraction of the target analytes in milk samples. This simple and economic method has been applied for analyses of 15 real milk samples. Among all samples only one of them was contaminated to florfenicol; 62.4 µg/Kg and contamination to chloramphenicol was not detected.

  13. Matrix molecularly imprinted mesoporous sol-gel sorbent for efficient solid-phase extraction of chloramphenicol from milk.

    PubMed

    Samanidou, Victoria; Kehagia, Maria; Kabir, Abuzar; Furton, Kenneth G

    2016-03-31

    Highly selective and efficient chloramphenicol imprinted sol-gel silica based inorganic polymeric sorbent (sol-gel MIP) was synthesized via matrix imprinting approach for the extraction of chloramphenicol in milk. Chloramphenicol was used as the template molecule, 3-aminopropyltriethoxysilane (3-APTES) and triethoxyphenylsilane (TEPS) as the functional precursors, tetramethyl orthosilicate (TMOS) as the cross-linker, isopropanol as the solvent/porogen, and HCl as the sol-gel catalyst. Non-imprinted sol-gel polymer (sol-gel NIP) was synthesized under identical conditions in absence of template molecules for comparison purpose. Both synthesized materials were characterized by Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FT-IR) and nitrogen adsorption porosimetry, which unambiguously confirmed their significant structural and morphological differences. The synthesized MIP and NIP materials were evaluated as sorbents for molecularly imprinted solid phase extraction (MISPE) of chloramphenicol in milk. The effect of critical extraction parameters (flow rate, elution solvent, sample and eluent volume, selectivity coefficient, retention capacity) was studied in terms of retention and desorption of chloramphenicol. Competition and cross reactivity tests have proved that sol-gel MIP sorbent possesses significantly higher specific retention and enrichment capacity for chloramphenicol compared to its non-imprinted analogue. The maximum imprinting factor (IF) was found as 9.7, whereas the highest adsorption capacity of chloramphenicol by sol-gel MIP was 23 mg/g. The sol-gel MIP was found to be adequately selective towards chloramphenicol to provide the necessary minimum required performance limit (MRPL) of 0.3 μg/kg set forth by European Commission after analysis by LC-MS even without requiring time consuming solvent evaporation and sample reconstitution step, often considered as an integral part in solid phase extraction work-flow. Intra and

  14. Novel mechanism of resistance to oxazolidinones, macrolides, and chloramphenicol in ribosomal protein L4 of the pneumococcus.

    PubMed

    Wolter, Nicole; Smith, Anthony M; Farrell, David J; Schaffner, William; Moore, Matthew; Whitney, Cynthia G; Jorgensen, James H; Klugman, Keith P

    2005-08-01

    Two clinical Streptococcus pneumoniae isolates, identified as resistant to macrolides and chloramphenicol and nonsusceptible to linezolid, were found to contain 6-bp deletions in the gene encoding riboprotein L4. The gene transformed susceptible strain R6 so that it exhibited such resistance, with the transformants also showing a fitness cost. We demonstrate a novel bacterial mechanism of resistance to chloramphenicol and nonsusceptibility to linezolid.

  15. Degradation of Oxytetracycline, Streptomycin, Sulphathiazole and Chloramphenicol Residues in Different Types of Honey.

    PubMed

    Gačić, Milica; Bilandžić, Nina; Šipušić, Đurđica Ivanec; Petrović, Marinko; Kos, Blaženka; Vahčić, Nada; Šušković, Jagoda

    2015-06-01

    Some of the most frequently used antibiotics in apiculture for the treatment of bacterial brood diseases are oxytetracycline, chloramphenicol, sulphathiazole and streptomycin. Therefore, the aim of this research is to determine the degradation kinetics of the residua these antibiotics leave behind in different types of honey stored in dark at 25 °C. The examined honey samples (N=74) originating from the Croatian market had satisfactory physicochemical properties and pollen characteristics. Quantitative analyses of antibiotic residues were done using HPLC and ELISA methods. No antibiotic residues were found in any of the tested acacia, chestnut, floral, meadow and honeydew honey samples (N=74). In the next step, each of the tested honey samples was supplemented with one of the following antibiotics (in mg/kg): oxytetracycline 10, chloramphenicol 200, sulphathiazole 200 or streptomycin 0.5. Relatively high mass fractions of antibiotics have been added to the honey because the stability of each antibiotic is highly dependent on the spiked antibiotic mass fraction and also on its chemical structure. During a 6-month storage in the dark at 25 °C, the dynamics of degradation of the studied antibiotics was proven to differ dependent on the type of honey into which the antibiotic was added. The half-life of oxytetracycline in the acacia, floral, meadow and honeydew honey stored in the dark at 25 °C was 15, 16, 17 and 19 days, respectively, while in the chestnut honey the decomposition failed to be seen even after 60 days of storage. In all examined honey samples, the half-life of chloramphenicol and sulphathiazole was proven to be longer than 6 months. The fastest decomposition was seen of oxytetracycline added into the acacia honey, followed by streptomycin and sulphathiazole added into the same, while the longest decomposition delay was seen when chloramphenicol was added into the floral honey. According to the results obtained using a linear model of degradation

  16. Degradation of Oxytetracycline, Streptomycin, Sulphathiazole and Chloramphenicol Residues in Different Types of Honey

    PubMed Central

    Gačić, Milica; Bilandžić, Nina; Šipušić, Đurđica Ivanec; Petrović, Marinko; Vahčić, Nada; Šušković, Jagoda

    2015-01-01

    Summary Some of the most frequently used antibiotics in apiculture for the treatment of bacterial brood diseases are oxytetracycline, chloramphenicol, sulphathiazole and streptomycin. Therefore, the aim of this research is to determine the degradation kinetics of the residua these antibiotics leave behind in different types of honey stored in dark at 25 °C. The examined honey samples (N=74) originating from the Croatian market had satisfactory physicochemical properties and pollen characteristics. Quantitative analyses of antibiotic residues were done using HPLC and ELISA methods. No antibiotic residues were found in any of the tested acacia, chestnut, floral, meadow and honeydew honey samples (N=74). In the next step, each of the tested honey samples was supplemented with one of the following antibiotics (in mg/kg): oxytetracycline 10, chloramphenicol 200, sulphathiazole 200 or streptomycin 0.5. Relatively high mass fractions of antibiotics have been added to the honey because the stability of each antibiotic is highly dependent on the spiked antibiotic mass fraction and also on its chemical structure. During a 6-month storage in the dark at 25 °C, the dynamics of degradation of the studied antibiotics was proven to differ dependent on the type of honey into which the antibiotic was added. The half-life of oxytetracycline in the acacia, floral, meadow and honeydew honey stored in the dark at 25 °C was 15, 16, 17 and 19 days, respectively, while in the chestnut honey the decomposition failed to be seen even after 60 days of storage. In all examined honey samples, the half-life of chloramphenicol and sulphathiazole was proven to be longer than 6 months. The fastest decomposition was seen of oxytetracycline added into the acacia honey, followed by streptomycin and sulphathiazole added into the same, while the longest decomposition delay was seen when chloramphenicol was added into the floral honey. According to the results obtained using a linear model of

  17. Chromatographic evaluation of polymers imprinted with analogs of chloramphenicol and application to selective solid-phase extraction.

    PubMed

    Schirmer, Christina; Meisel, Hans

    2009-08-01

    To obtain a highly selective material for the antibiotic chloramphenicol, which has several harmful side effects in humans, different molecularly imprinted polymers (MIPs) were prepared. In order to avoid a major traditional drawback associated with MIPs of residual template bleeding, molecules that are structurally related to chloramphenicol were used as templates for polymer synthesis. Chromatographic evaluation indicated that the employed template imparted a significant influence on the recognition properties of the corresponding polymer. A strong retention of chloramphenicol under nonpolar elution conditions (k = 68.03, IF = 17.72) and under aqueous elution conditions (k = 92.44, IF = 1.35) was achieved. After chromatographic evaluation, the MIP was utilized as the recognition sorbent in a solid-phase extraction to determine chloramphenicol using either an organic or aqueous washing solvent. Recoveries of nearly 100% from the chloramphenicol standard solution and nearly 90% from honey samples spiked with chloramphenicol were attained. Furthermore, the applicability of the MIP for sample cleanup was demonstrated.

  18. Development of LC-MS/MS methodology for the detection/determination and confirmation of chloramphenicol, chloramphenicol 3-O-β-d-glucuronide, florfenicol, florfenicol amine and thiamphenicol residues in bovine, equine and porcine liver.

    PubMed

    Fedeniuk, Rick W; Mizuno, Massey; Neiser, Connie; O'Byrne, Collin

    2015-06-01

    A method for the detection and confirmation of organic solvent extractable residues of the neutral, acidic, and basic analytes of the amphenicol class veterinary drugs and selected metabolites was developed and validated. Using a modified QuEChERS extraction with SPE cleanup and LC-MS/MS analysis, limits of detection and confirmation for the different analytes in bovine, equine, and porcine liver ranged from 0.1ng/g for chloramphenicol to 1ng/g for florfenicol amine. Tissue homogenization with an ammonium formate/EDTA solution and subsequent analyte partitioning against 7:3 acetonitrile:isopropanol solution and mixed-mode strong-cation exchange solid-phase extraction cartridge cleanup allowed for the extraction of all compounds from tissues with mean recoveries ranging from 50% (chloramphenicol 3-O-β-d-glucuronide) to 90% (thiamphenicol). Matrix effects ranged from greater than 85% suppression for florfenicol amine to 70% matrix enhancement for chloramphenicol 3-O-β-d-glucuronide. Quantitation and confirmation were accomplished using commercially available penta-deuterated chloramphenicol as internal standard and multiple reaction monitoring (MRM) of two or three transitions per target analyte. Method accuracy was greater than 15% for all compounds except the glucuronide metabolite. Intra-lab method repeatability estimates ranged from 73% RSD for chloramphenicol 3-O-β-d-glucuronide to 14% RSD for chloramphenicol. Only chloramphenicol 3-O-β-d-glucuronide and florfenicol amine at the low end of their calibration ranges (0.25 and 1ng/g, respectively) did not meet AOAC recommended HorRatr guidelines for intra-lab repeatabilities. Preliminary tests show that the method's extraction protocol can be used to recover analytes of the β-agonists, corticosteroids, fluoroquinolones, sulfonamides, and tetracycline drug classes from the same matrices. Requirements for use in national chemical monitoring programs as a detection/confirmatory (florfenicol amine and

  19. Expression of a streptomycete leaderless mRNA encoding chloramphenicol acetyltransferase in Escherichia coli.

    PubMed Central

    Wu, C J; Janssen, G R

    1997-01-01

    The chloramphenicol acetyltransferase (cat) gene from Streptomyces acrimycini encodes a leaderless mRNA. Expression of the cat coding sequence as a leaderless mRNA from a modified lac promoter resulted in chloramphenicol resistance in Escherichia coli. Transcript mapping with nuclease S1 confirmed that the 5' end of the cat message initiated at the A of the AUG translational start codon. Site-directed mutagenesis of the lac promoter or the cat start codon abolished chloramphenicol resistance, indicating that E. coli initiated translation at the 5' terminal AUG of the cat leaderless mRNA. Addition of 5'-AUGC-3' to the 5' end of the cat mRNA resulted in translation occurring also from the reading frame defined by the added AUG triplet, suggesting that a 5'-terminal start codon is an important recognition feature for initiation and establishing reading frame during translation of leaderless mRNA. Addition of an untranslated leader and Shine-Dalgarno sequence to the cat coding sequence increased cat expression in a cat:lacZ fusion; however, the level of expression was significantly lower than when a fragment of the bacteriophage lambda cI gene, also encoding a leaderless mRNA, was fused to lacZ. These results indicate that in the absence of an untranslated leader and Shine-Dalgarno sequence, the streptomycete cat mRNA is translated by E. coli; however, the cat translation signals, or other features of the cat mRNA, provide for only a low level of expression in E. coli. PMID:9352935

  20. Regulatory regions that control expression of two chloramphenicol-inducible cat genes cloned in Bacillus subtilis.

    PubMed

    Duvall, E J; Williams, D M; Mongkolsuk, S; Lovett, P S

    1984-06-01

    Plasmid pPL603 is a promoter cloning vector for Bacillus subtilis and consists of a 1.1-kilobase fragment of Bacillus pumilus DNA inserted between the EcoRI and BamHI sites of pUB110. The gene cat-86, specifying chloramphenicol-inducible chloramphenicol acetyltransferase, is located on the 1.1-kilobase cloned DNA. When pPL603 is present in B. subtilis, cat-86 is unexpressed during vegetative growth but expressed during sporulation. The regulation of cat-86 in pPL603 is due to sequences within two restriction fragments, designated P1 and R1, that precede the main coding portion of the gene. The P1 fragment promotes transcription of cat-86 only during sporulation, whereas the adjacent R1 fragment lacks promoter function but contains sequences essential to chloramphenicol inducibility. A second B. pumilus gene, cat-66, was cloned in B. subtilis and is expressed throughout the vegetative growth and sporulation cycle. The cat-66 coding region is preceded by two adjacent restriction fragments designated as P2 and R2. P1 and P2 are identical in size and share 95% conservation of base sequence. R1 and R2 are also identical in size and share 91% conservation of base sequence. Fragment substitution experiments demonstrate that R2 can functionally replace R1. The substitution of P2 for P1 promotes cat-86 expression throughout vegetative growth and sporulation. Analysis of a derivative of pPL603 in which P2 has replaced P1 demonstrates that P2 promotes transcription of cat-86 during vegetative growth and that P2 contains the start site for transcription of cat-86. Thus, P1 and P2 differ strikingly in vegetative promoter function, yet they differ by single-base substitutions at only 11 positions of 203.

  1. Stability of sulfonamides, nitrofurans, and chloramphenicol residues in preserved raw milk samples measured by liquid chromatography.

    PubMed

    Noa, Mario; Perez, Norima; Gutierrez, Rey; Escobar, Irma; Diaz, Gilberto; Vega, Salvador; Prado, Guadalupe; Urban, Georgina

    2002-01-01

    A stability study was made of 10 antimicrobials: 6 sulfonamides, 3 nitrofurans, and chloramphenicol residues in raw milk samples preserved with 0.1 % potassium dichromate (K2Cr2O7) and 0.05% mercuric bichloride (HgCl2) during cold storage for 7 days. Preserved milk samples fortified with 50 ppb of each antimicrobial were analyzed by liquid chromatography (modified AOAC Method 993.32). Drugs were extracted with chloroform-acetone after solvent evaporation residues were dissolved with aqueous sodium acetate buffer solution (0.02M, pH 4.8), and fat was removed with hexane. Sulfonamides and chloramphenicol were detected at 275 nm (UV) by using a gradient system of sodium acetate buffer solution-acetonitrile starting at 95 + 5 (v/v) and finishing at 80 + 20 (v/v). Nitrofurans were detected at 375 nm (UV) isocratically with sodium acetate buffer solution-acetonitrile (80 + 20, v/v). Residues stability was measured through recovery data. Sulfamethoxazole, sulfachloropyridazine, nitrofurazone, furazolidone, and furaltadone residues remained stable in the presence of either preservative for 7 days. Sulfamethazine and chloramphenicol were not affected by K2Cr2O7, but had significant losses (p <0.05) when HgCl2 was used: 26.2 and 13.4%, respectively. Average recoveries of sulfamonomethoxine, sulfamerazine, and sulfathiazole significantly decreased by Day 7, with losses of 17.1, 17.2, and 23.2% for K2Cr2O7, and 23.3, 20.7, and 48.0% for HgCl2, respectively. During 5 days of cold storage all antimicrobials tested, except sulfathiazole, remained stable in milk samples preserved with 0.1 % K2Cr2O7 or 0.05% HgCl2.

  2. Rapid determination of chloramphenicol residues in aquaculture tissues by immunochromatographic assay.

    PubMed

    Li, Ke; Liu, Liqiang; Xu, ChuanLai; Chu, XiaoGang

    2007-11-01

    An immunochromatographic assay was developed to detect chloramphenicol (CAP) residues in aquaculture tissues. The limit of detection (LOD) was 10 ng g(-1) for detecting CAP spiked in the aquaculture tissues. The results were confirmed by liquid chromatography tandem mass spectrometry (LC/MS/MS) and indicated that there was a good agreement between the two methods. The linear regression equation was y = 1.19x + 0.539 with R(2) = 0.978. The assay time for test was less than 5 min and the method is suitable for rapid testing on-site.

  3. High resolution studies of the Afa/Dr adhesin DraE and its interaction with chloramphenicol.

    PubMed

    Pettigrew, David; Anderson, Kirstine L; Billington, Jason; Cota, Ernesto; Simpson, Peter; Urvil, Petri; Rabuzin, Filip; Roversi, Pietro; Nowicki, Bogdan; du Merle, Laurence; Le Bouguénec, Chantal; Matthews, Stephen; Lea, Susan M

    2004-11-05

    Pathogenic Escherichia coli expressing Afa/Dr adhesins are able to cause both urinary tract and diarrheal infections. The Afa/Dr adhesins confer adherence to epithelial cells via interactions with the human complement regulating protein, decay accelerating factor (DAF or CD55). Two of the Afa/Dr adhesions, AfaE-III and DraE, differ from each other by only three residues but are reported to have several different properties. One such difference is disruption of the interaction between DraE and CD55 by chloramphenicol, whereas binding of AfaE-III to CD55 is unaffected. Here we present a crystal structure of a strand-swapped trimer of wild type DraE. We also present a crystal structure of this trimer in complex with chloramphenicol, as well as NMR data supporting the binding position of chloramphenicol within the crystal. The crystal structure reveals the precise atomic basis for the sensitivity of DraE-CD55 binding to chloramphenicol and demonstrates that in contrast to other chloramphenicol-protein complexes, drug binding is mediated via recognition of the chlorine "tail" rather than via intercalation of the benzene rings into a hydrophobic pocket.

  4. 4-alkoxy and 4-thioalkoxyquinoline derivatives as chemosensitizers for the chloramphenicol-resistant clinical Enterobacter aerogenes 27 strain.

    PubMed

    Gallo, Sandrine; Chevalier, Jacqueline; Mahamoud, Abdallah; Eyraud, Annie; Pagès, Jean-Marie; Barbe, Jacques

    2003-09-01

    Enterobacter aerogenes is a Gram-negative bacteria frequently responsible for nosocomial respiratory tract infections. Strains resistant to chloramphenicol are frequently isolated. Alkoxy and thio-alkoxyquinolines have a potential to act as chemosensitizers that would render multi-drug-resistant (MDR) bacterial infections susceptible to antibiotics to which they were originally resistant. Several new quinoline derivatives have been prepared, characterized and studied for their ability to increase chloramphenicol sensitivity of E. aerogenes 27, a clinical strain that exhibits the MDR phenotype. Drugs investigated were either quinoline ethers or quinoline thio-ethers. Thio-ethers are much more efficient in increasing chloramphenicol sensitivity than other corresponding ethers. In particular, 4-piperidinoethylthio-quinoline increases the strain sensitivity to chloramphenicol by about 20 times at 2 mM concentration. Similarly, sensitivity to quinolone antibiotics dramatically increases. Because these quinoline derivatives act as inhibitors of the drug efflux pump responsible for bacterial resistance to chloramphenicol, they may serve as adjunct to conventional therapy of E. aerogenes infections.

  5. Identification of the chloramphenicol-hydrolyzing enzyme of guinea pig liver as one of the nonspecific carboxylesterases.

    PubMed

    Kuhn, D; Heymann, E

    1982-03-01

    Guinea pig liver has the highest chloramphenicol-hydrolyzing capacity among the livers of various mammals. The enzyme responsible for the hydrolysis of the amide-bond in chloramphenicol is one of the isoenzymes of the microsomal nonspecific carboxylesterases. This isoenzyme is related to the well-known acetanilide-hydrolyzing carboxylesterases/amidases of pig and rat liver. The guinea pig liver enzyme is purified 24-fold starting with microsomes. The purified enzyme is essentially free from other proteins except other carboxylesterase isoenzymes with similar properties. The chloramphenicol-hydrolyzing esterase has an apparent molecular weight of about 180,000, a subunit weight of 60,000 and a pH optimum at 8.5. It also hydrolyzes methyl butyrate and acetanilide and it is completely inhibited by diethyl-4-nitrophenyl phosphate. Two assay procedures for the enzymatic chloramphenicol hydrolysis are described: a thin-layer chromatographic assay using radioactive chloramphenicol and a colorimetric assay utilizing the reaction of the liberated amine with trinitrobenzenesulfonic acid.

  6. Physico-chemical factors affect chloramphenicol efflux and EmhABC efflux pump expression in Pseudomonas fluorescens cLP6a.

    PubMed

    Adebusuyi, Abigail; Foght, Julia

    2013-01-01

    Protein synthesis inhibitors such as chloramphenicol and tetracycline may be inducers of efflux pumps such as MexY in Pseudomonas aeruginosa, complicating their use for the treatment of bacterial infections. We previously determined that chloramphenicol, a substrate of the EmhABC efflux pump in Pseudomonas fluorescens cLP6a, did not induce emhABC expression. In this study, we determined the effect of physico-chemical factors on chloramphenicol efflux by EmhABC, and the expression of emhABC. Efflux assays measuring accumulation of (14)C-chloramphenicol in cell pellets showed that chloramphenicol efflux is dependent on growth temperature, pH and concentration of Mg(2+). These physico-chemical factors modulated the efflux of chloramphenicol by 26 to >50%. All conditions tested that decreased the efflux of chloramphenicol unexpectedly induced transcription of emhABC efflux genes. EmhABC activity also effectively suppressed the deleterious effect of chloramphenicol on the cell membrane of strain cLP6a, which may explain why chloramphenicol is not an inducer of emhABC. Our results suggest that the detrimental effect of an antibiotic on cell membrane integrity and fatty acid composition may be the signal that induces emhABC expression, and that inducers of other bacterial efflux pumps may include environmental factors rather than their substrates per se.

  7. Determination of nitrofuran and chloramphenicol residues by high resolution mass spectrometry versus tandem quadrupole mass spectrometry.

    PubMed

    Kaufmann, A; Butcher, P; Maden, K; Walker, S; Widmer, M

    2015-03-03

    An ultra-high performance liquid chromatography based method, coupled to high resolution mass spectrometry (UHPLC-HRMS), was developed to permit the detection and quantification of various nitrofuran and chloramphenicol residues in a number of animal based food products. This method is based on the hydrolysis of covalently bound metabolites and derivatization with 2-nitrobenzaldehyde. Clean-up is achieved by a liquid/liquid and a reversed phase/solid phase extraction. Not only are the four conventional nitrofurans (nitrofurantoin, furazolidone, nitrofurazone and furaltadone) detected, but also nifursol, nitrovin and nifuroxazide. Furthermore, an underivatizable nitrofuran (nifurpirinol) and another banned drug (chloramphenicol) can be quantified as well. The compounds are detected in the form of their precursor ions, [M+H](+) and [M-H](-), respectively. The mass resolving power of 70,000 FWHM, and the applied mass window ensure sufficient selectivity and sensitivity. Confirmation is obtained by monitoring the HRMS resolved product ions which were derived from the unit-mass resolved precursor ions. The multiplexing capability of the utilized Orbitrap instrument provides not only highly selective, but also sensitive confirmatory signals. This method has been validated according to the CD 2002/657/EC for the following matrices: muscle, liver, kidney, fish, honey, eggs and milk.

  8. Saccharothrix sp. PAL54, a new chloramphenicol-producing strain isolated from a Saharan soil.

    PubMed

    Aouiche, Adel; Sabaou, Nasserdine; Meklat, Atika; Zitouni, Abdelghani; Bijani, Christian; Mathieu, Florence; Lebrihi, Ahmed

    2012-03-01

    An actinomycete strain designated PAL54, producing an antibacterial substance, was isolated from a Saharan soil in Ghardaïa, Algeria. Morphological and chemical studies indicated that this strain belonged to the genus Saccharothrix. Analysis of the 16S rDNA sequence showed a similarity level ranging between 96.9 and 99.2% within Saccharothrix species, with S. longispora DSM 43749(T), the most closely related. DNA-DNA hybridization confirmed that strain PAL54 belonged to Saccharothrix longispora. It showed very strong activity against pathogenic Gram-positive and Gram-negative bacteria responsible for nosocomial infections and resistant to multiple antibiotics. Strain PAL54 secreted the antibiotic optimally during mid-stationary and decline phases of growth. One antibacterial compound was isolated from the culture broth and purified by HPLC. The active compound was elucidated by uv-visible and NMR spectroscopy and by mass spectrometry. The results showed that this compound was a D: (-)-threo chloramphenicol. This is the first report of chloramphenicol production by a Saccharothrix species.

  9. Pyrite-enhanced degradation of chloramphenicol by low concentrations of H2O2.

    PubMed

    Wu, Deli; Liu, Yanxia; Zhang, Zhiyong; Ma, Luming; Zhang, Yalei

    2015-01-01

    A pyrite-catalyzed reaction was used to degrade chloramphenicol. Chloramphenicol could be almost 100% removed within 60 minutes when 1 mM H2O2 and 0.1 g/L pyrite were added at an initial pH=3. During oxidation, intermediates such as nitrobenzaldehyde and dichloroacetamide were identified by gas chromatography/mass spectrometry (GC/MS). The •OH was identified by electron spin-resonance spectroscopy. Pyrite was digested to determine elements by ICP (inductive coupled plasma emission spectrometer). To understand the reaction mechanism and the role of natural pyrite in these processes, techniques including scanning electron microscopy and energy dispersive spectrometry were employed to characterize the solid sample. The results explain that pyrite acts as a 'bond' between Fe3+ and H2O2, and this pathway continues to form •OH and inhibit the quenching reaction. Therefore, pyrite-catalyzed reactions would proceed even in low concentrations of H2O2.

  10. Pharmacokinetics of amikacin and chloramphenicol in the aqueous humor of rabbits.

    PubMed Central

    Mayers, M; Rush, D; Madu, A; Motyl, M; Miller, M H

    1991-01-01

    Composite data describing ocular pharmacokinetics are unreliable because of intersubject variation. To address this problem, an animal model was developed in which multiple aqueous samples from single subjects were obtained. Following direct anterior chamber or intravenous administration of amikacin or chloramphenicol, pharmacokinetic analysis of drug concentrations in the serum and anterior chamber was performed by using a nonlinear least-squares regression program. The number of anterior chamber paracenteses performed did not alter the beta elimination rates or percent penetration into the anterior chamber. The aqueous humor and peripheral-compartment terminal slopes were identical. These data indicate that complete ocular concentration-time curves can be obtained without altering antibiotic pharmacokinetics. Following direct injection into the anterior chamber, the elimination rates for both antibiotics followed a one-compartment model, whereas those following intravenous administration best fit an open, first-order, two-compartment model. Following intravenous administration, the anterior chamber elimination rate constants for both drugs were equal to that of the serum and significantly longer than that following direct injection. The elimination rates of both drugs following direct injection were similar. Systemic administration resulted in drug levels in aqueous humor that persisted longer than those following direct injection. Chloramphenicol, a lipophilic compound, gave higher mean concentrations in aqueous humor than did amikacin. Our model provides a new approach which rigorously examines ocular pharmacokinetics and provides data which suggest that for selected compounds the parenteral route of administration is preferable. PMID:1952850

  11. Synthesis and Evaluation of Chloramphenicol Homodimers: Molecular Target, Antimicrobial Activity, and Toxicity against Human Cells

    PubMed Central

    Kostopoulou, Ourania N.; Magoulas, George E.; Papadopoulos, Georgios E.; Mouzaki, Athanasia; Dinos, George P.; Papaioannou, Dionissios; Kalpaxis, Dimitrios L.

    2015-01-01

    As fight against antibiotic resistance must be strengthened, improving old drugs that have fallen in reduced clinical use because of toxic side effects and/or frequently reported resistance, like chloramphenicol (CAM), is of special interest. Chloramphenicol (CAM), a prototypical wide-spectrum antibiotic has been shown to obstruct protein synthesis via binding to the bacterial ribosome. In this study we sought to identify features intensifying the bacteriostatic action of CAM. Accordingly, we synthesized a series of CAM-dimers with various linker lengths and functionalities and compared their efficiency in inhibiting peptide-bond formation in an Escherichia coli cell-free system. Several CAM-dimers exhibited higher activity, when compared to CAM. The most potent of them, compound 5, containing two CAM bases conjugated via a dicarboxyl aromatic linker of six successive carbon-bonds, was found to simultaneously bind both the ribosomal catalytic center and the exit-tunnel, thus revealing a second, kinetically cryptic binding site for CAM. Compared to CAM, compound 5 exhibited comparable antibacterial activity against MRSA or wild-type strains of Staphylococcus aureus, Enterococcus faecium and E. coli, but intriguingly superior activity against some CAM-resistant E. coli and Pseudomonas aeruginosa strains. Furthermore, it was almost twice as active in inhibiting the growth of T-leukemic cells, without affecting the viability of normal human lymphocytes. The observed effects were rationalized by footprinting tests, crosslinking analysis, and MD-simulations. PMID:26267355

  12. Chloramphenicol succinate, a competitive substrate and inhibitor of succinate dehydrogenase: possible reason for its toxicity.

    PubMed

    Ambekar, C S; Lee, J S K; Cheung, B M Y; Chan, L C; Liang, R; Kumana, C R

    2004-08-01

    From our previous study [Eur. J. Clin. Pharmacol. 56 (2000) 405] we hypothesized that chloramphenicol succinate (CAPS) may be a competitive substrate for succinate dehydrogenase (SDH). It may be oxidized by SDH to release chloramphenicol (CAP), which may inhibit SDH by feed back mechanism. The present ex-vivo/in vitro study was aimed to investigate this possibility by using human tissues (bone marrow and liver samples) and animal tissues (rat liver and kidney). The effect of different SDH activators and specific inhibitors was studied on CAPS metabolism by SDH. The metabolites and reduction products were detected by using HPLC. In marrow samples, CAPS was slowly oxidized to form CAP. The formation of CAP (oxidation product) was enhanced by FAD and low malonate and inhibited by high malonate and 3-NPA. Similar results were obtained with mitochondria from human and rat tissues. These studies suggest that CAPS could be a competitive oxidative substrate and the metabolite CAP could be an inhibitor at the reduction site. Therefore, SDH could be a target molecule responsible for CAPS induced toxicity.

  13. Chloramphenicol with fluid and electrolyte therapy cures terminally ill green tree frogs (Litoria caerulea) with chytridiomycosis.

    PubMed

    Young, Sam; Speare, Rick; Berger, Lee; Skerratt, Lee F

    2012-06-01

    Terminal changes in frogs infected with the amphibian fungal pathogen Batrachochytrium dendrobatidis (Bd) include epidermal degeneration leading to inhibited epidermal electrolyte transport, systemic electrolyte disturbances, and asystolic cardiac arrest. There are few reports of successful treatment of chytridiomycosis and none that include curing amphibians with severe disease. Three terminally ill green tree frogs (Litoria caerulea) with heavy Bd infections were cured using a combination of continuous shallow immersion in 20 mg/L chloramphenicol solution for 14 days, parenteral isotonic electrolyte fluid therapy for 6 days, and increased ambient temperature to 28 degrees C for 14 days. All terminally ill frogs recovered rapidly to normal activity levels and appetite within 5 days of commencing treatment. In contrast, five untreated terminally ill L. caerulea with heavy Bd infections died within 24-48 hr of becoming moribund. Subclinical infections in 15 experimentally infected L. caerulea were cured within 28 days by continuous shallow immersion in 20 mg/L chloramphenicol solution without adverse effects. This is the first known report of a clinical treatment protocol for curing terminally ill Bd-infected frogs.

  14. Influence of tonicity and chloramphenicol on hyperthermic cytotoxicity and cell permeability under various heating rates.

    PubMed

    Morozov, I I; Petin, V G; Dubovick, B V

    1997-01-01

    The cell lethality and permeability induced in Escherichia coli B/r, Escherichia coli Bs-1 and Zygosaccharomyces bailii cells by high temperature (52 degrees C) after heating at different rates (mean s 0.015, 0.25 and 1.50 degrees C per s) and in media of different tonicity and content (isotonic YEP broth versus 0.01 M phosphate buffer, pH 7.0 containing different concentrations of NaCl) and with versus without chloramphenicol (10 micrograms/ml) have been investigated. Hyperthermic treatment in YEP broth of isotonic 0.01 M phosphate buffer resulted in markedly reduced cytotoxicity with decreasing heat rate. The heating rate effect was larger when the cells were treated in YEP broth. Chloramphenicol, which is known to inhibit expression of heat shock proteins in bacteria, did not affect the viability of cells or the development of thermotolerance in cells heated at different heating rates in isotonic phosphate buffer but prevented the development of an additional degree of thermotolerance in cells heated slowly in YEP broth. In contrast, the differential effect of heating rate on cytotoxicity and cell permeability was not demonstrated when cells were heated in hypertonic solution (1M NaCl in phosphate buffer, pH 7.0). It is proposed that heat destabilization of the osmotic cell homeostasis, which is more profound after rapid heating, plays a major part in heat induced cellular lethality.

  15. Azithromycin, fluoroquinolone and chloramphenicol resistance of non-chlamydia conjunctival bacteria in rural community of Ethiopia

    PubMed Central

    Abera, Bayeh; Kibret, Mulugeta

    2014-01-01

    Aim: To determine profiles of non-chlamydia conjunctival bacteria and their antimicrobial susceptibility from adults who underwent trachomatous trichiasis surgery in rural areas of Ethiopia. Materials and Methods: A cross-sectional study was conducted in rural districts in West Gojjam administrative zone. Conjunctival swabs were collected during surgery and transported using Stuart transport broth (Oxoid, UK). Antibiotic susceptibility of conjunctival isolates was determined using the Kirby-Bauer disc-diffusion method. Results: Non-chlamydia pathogenic bacteria were recovered from conjunctiva of 438 (31%) participants before treatment. The isolated conjunctival bacteria were Staphylococcus aureus, coagulase-negative Staphylococci, Streptococcus group (A, C, F and G), Enterococci, Streptococcus pneumoniae, Moraxella spp., Escherichia coli, Citrobacter spp., Proteus spp., Klebsiella spp., Pseudomonas spp. and Enterobacter spp. Overall, resistance rates of 57.8% to azithromycin and 68.5% to chloramphenicol were found. However, 86-94.4% sensitivity was demonstrated to ciprofloxacin and norfloxacin. Moderate sensitivity rates (61.8-78.4%) were observed to ceftriaxone, tetracycline and cotrimoxazole. Conclusion: Fluoroquinolones that have activity against the majority of bacterial isolates were potent at in vitro. However, unacceptably high levels of resistance to azithromycin and chloramphenicol in rural community indicated a need for further study and antimicrobial resistance surveillance. PMID:23571246

  16. An amperometric chloramphenicol immunosensor based on cadmium sulfide nanoparticles modified-dendrimer bonded conducting polymer.

    PubMed

    Kim, Dong-Min; Rahman, Md Aminur; Do, Minh Hien; Ban, Changill; Shim, Yoon-Bo

    2010-03-15

    An amperometric chloramphenicol (CAP) immunosensor was fabricated by covalently immobilizing anti-chloramphenicol acetyl transferase (anti-CAT) antibody on cadmium sulfide nanoparticles (CdS) modified-dendrimer that was bonded to the conducting polymer (poly 5, 2': 5', 2''-terthiophene-3'-carboxyl acid (poly-TTCA)) layer. The AuNPs, dendrimers, and CdS nanoparticles were deposited onto the polymer layer in order to enhance the sensitivity of the sensor probes. The particle sizes were determined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The immobilization of dendrimers, CdS, and anti-CAT were confirmed using energy disruptive spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and quartz crystal microbalance (QCM) techniques. The detection of CAP was based on the competitive immuno-interaction between the free- and labeled-CAP for active sites of the anti-CAT. Hydrazine was used as the label for CAP, and it electrochemically catalyzed the reduction of H(2)O(2) at -0.35 V vs. Ag/AgCl. Under optimized conditions, the proposed immunosensor exhibited a linear range of CAP detection between 50 pg/mL and 950 pg/mL, and the detection limit was 45 pg/mL. The immunosensor was examined in real meat samples for the analysis of CAP.

  17. Final report on key comparison CCQM-K81: Chloramphenicol in pig muscle

    NASA Astrophysics Data System (ADS)

    Polzer, Joachim; Henrion, Andre; Gowik, Petra

    2013-01-01

    Under the auspices of the Organic Analysis Working Group (OAWG) of the Comité Consultatif pour la Quantité de Matière (CCQM) a key comparison, CCQM-K81 'Chloramphenicol in pig muscle', was coordinated by BVL and PTB in 2009/2010. Six NMIs/DIs participated in this comparison. Chloramphenicol (CAP) is an effective broad-spectrum antibiotic which can in principle be used for the treatment of humans and animals. Its use for the treatment of food-producing animals is, however, forbidden worldwide in many countries due to potential severe side effects. The key comparison was the follow-up to the successful pilot study 'CCQM-P90: chloramphenicol in milk'. With this key comparison it was intended to demonstrate the capability of NMIs/DIs to analyse traces of CAP in food at concentration levels resulting from legal requirements for food control. Additionally, the quality of this kind of analysis with respect to compliance with legal requirements for food control methods and the international comparability of measurements should be evaluated in general. The study was classified as a 'track C' study (studies in emerging areas of global interest). For the study incurred lyophilised pig muscle material containing CAP (at a mass-fraction level around the maximum allowable level for import for a number of countries) has been produced as a candidate reference material by BVL and IRMM (Institute for Reference Materials and Measurements, JRC of the European Commission). Animal treatment and slaughtering of the animal to gain incurred muscle material as well as pre-testing of the CAP content was done at the BVL. The IRMM did further processing of the material and testing of homogeneity and stability. Participants were invited to assign the mass fraction of free CAP in the comparison sample. CCQM-K81 demonstrated successfully the capability of the participating laboratories to assign chloramphenicol values in tissue down to residue levels of around 0.3 ng/g (referring to the

  18. Characterization and Uncertainty Assessment of a Certified Reference Material of Chloramphenicol in Methanol (GBW(E)082557)

    PubMed Central

    Zhou, Jian; Song, Yinqing; Wang, Tongtong

    2016-01-01

    Prior to preparation of CRM candidate of chloramphenicol in methanol with a concentration of 100 mg/L, two independent methods including mass balance (MB) and quantitative nuclear magnetic resonance (qNMR) were employed to precisely measure the mass fraction of pure chloramphenicol materials. The mass fraction was assigned to be 99.8% with uncertainty of 0.3%. Homogeneity testing and stability study of chloramphenicol in methanol were examined by using high performance liquid chromatography. Additionally, the uncertainties originating from the process of CRM development were comprehensively evaluated. The experimental results indicate that the property value of this CRM is homogeneous and stable at 4°C for at least six months. The new CRM (GBW(E)082557) can be applicable to calibration of instrument and assurance of accuracy and comparability of results in routine measurement. PMID:27493666

  19. PIXE investigation of in vitro release of chloramphenicol across polyvinyl alcohol/acrylamide hydrogel

    NASA Astrophysics Data System (ADS)

    Rihawy, M. S.; Alzier, A.; Allaf, A. W.

    2011-09-01

    Hydrogels based on polyvinyl alcohol and different amounts of acrylamide monomer were prepared by thermal cross-linking in the solid state. The hydrogels were investigated for drug delivery system applications. Chloramphenicol was adopted as a model drug to study its release behavior. Particle induced X-ray emission was utilized to study the drug release behavior across the hydrogels and a comparison study with ultraviolet measurements was performed. Fourier Transform Infrared measurements were carried out for molecular characterization. The releasing behavior of the drug exhibits a decrease and a subsequent increase in the release rate, as the acrylamide monomer increases. Characterization of the hydrogels has shown a competitive behavior between crosslinking with AAm acrylamide monomer or oligomerized version, depending on the amount added to prepare the hydrogels.

  20. Impact of Ciprofloxacin and Chloramphenicol on the Lipid Bilayer of Staphylococcus aureus: Changes in Membrane Potential

    PubMed Central

    Páez, Paulina L.; Becerra, María C.; Albesa, Inés

    2013-01-01

    The present study was undertaken to explore the interaction of ciprofloxacin and chloramphenicol with bacterial membranes in a sensitive and in a resistant strains of Staphylococcus aureus by using 1-anilino-8-naphthalene sulfonate (ANS). The binding of this probe to the cell membrane depends on the surface potential, which modulates the binding constant to the membrane. We observed that these antibiotics interacted with the bilayer, thus affecting the electrostatic surface potential. Alterations caused by antibiotics on the surface of the bacteria were accompanied by a reduction in the number of binding sites and an increase in the ANS dissociation constant in the sensitive strain, whereas in the ciprofloxacin-resistant strain no significant changes were detected. The changes seen in the electrostatic surface potential generated in the membrane of S. aureus by the antibiotics provide new aspects concerning their action on the bacterial cell. PMID:23762834

  1. Effect of temperature switchover on the degradation of antibiotic chloramphenicol by biocathode bioelectrochemical system.

    PubMed

    Kong, Deyong; Liang, Bin; Lee, Duu-Jong; Wang, Aijie; Ren, Nanqi

    2014-08-01

    Exposure to chloramphenicol (CAP), a chlorinated nitroaromatic antibiotic, can induce CAP-resistant bacteria/genes in diverse environments. A biocathode bioelectrochemical system (BES) was applied to reduce CAP under switched operational temperatures. When switching from 25 to 10°C, the CAP reduction rate (kCAP) and the maximum amount of the dechlorinated reduced amine product (AMCl, with no antibacterial activity) by the biocathode communities were both markedly decreased. The acetate and ethanol yield from cathodophilic microbial glucose fermentation (with release of electrons) was also reduced. Formation of the product AMCl was enhanced by the biocathode dechloridation reaction compared with that produced from pure electrochemical or microbial dechloridation processes. The electrochemical and morphological analyses of cathode biofilms demonstrated that some cathodophilic microbes could adapt to low temperature and play a key role in CAP degradation. The resilient biocathode BES has a potential for the treatment of CAP-containing wastewater in temperature fluctuating environments.

  2. Acute and chronic toxic effects of chloramphenicol on Scenedesmus obliquus and Chlorella pyrenoidosa.

    PubMed

    Zhang, Wei; Sun, Wenfang; An, Shuai; Xiong, Bang; Lin, Kuangfei; Cui, Xinhong; Guo, Meijin

    2013-08-01

    The acute and chronic toxicological effects of Chloramphenicol (CAP) on Scenedesmus obliquus and Chlorella pyrenoidosa are not well understood. The indoor experiments were carried to observe and analyze the CAP induced changes. Results of the observations have showed that CAP exposure could significantly inhibit the growth of Scenedesmus obliquus in almost all the treated groups, while Chlorella pyrenoidosa exhibited less sensitivity. Chlorophyll-a syntheses of Scenedesmus obliquus were all inhibited by CAP exposure, while Chlorella pyrenoidosa displayed obvious stimulation effect. Catalase (CAT) and Superoxide dismutase (SOD) activities of both algae were promoted in all the treatments. The experimental results indicated that the growth and Chlorophyll-a syntheses of Scenedesmus obliquus were more sensitive in response to CAP exposure than that of Chlorella pyrenoidosa. While for CAT and SOD activities, Chlorella pyrenoidosa showed more susceptible. This research provides a basic understanding of CAP toxicity to aquatic organisms.

  3. A highly sensitive europium nanoparticle-based lateral flow immunoassay for detection of chloramphenicol residue.

    PubMed

    Xia, Xiaohu; Xu, Ye; Ke, Rongqin; Zhang, Heng; Zou, Mingqiang; Yang, Wei; Li, Qingge

    2013-09-01

    A europium nanoparticle-based lateral flow immunoassay for highly sensitive detection of chloramphenicol residue was developed. The detection result could be either qualitatively resolved with naked eye or quantitatively analyzed with the assistance of a digital camera. In the qualitative mode, the limit of detection (LOD) was found to be 0.25 ng/mL. In the quantitative mode, the half-maximal inhibition concentration (IC50) was determined to be 0.45 ng/mL and the LOD can reach an ultralow level of 0.03 ng/mL, which is ~100 times lower than that of the conventional colloidal gold-based lateral flow immunoassay. Potential application of the established method was demonstrated by analyzing representative cow milk samples.

  4. Malachite green and chloramphenicol in aquatic products from regions around Dongting Lake in Hunan, China.

    PubMed

    He, Jiang; Cui, Jingzhen

    2016-01-01

    Aquatic products are important sources of animal proteins in human diet, especially in developing countries. As such, the safety of aquatic products is of primary concern. In this study, a standard method is used to detect malachite green (MG) and chloramphenicol (CAP) and to analyse the contents of these banned chemicals in turtle, mandarin fish and grass carp sampled from the region surrounding Dongting Lake area in Hunan, China. Results showed that 10.6% of the samples were MG-positive, most of them turtles. CAP was found in 8.3% of the samples, mostly in mandarin fish. These data indicated that these banned substances are still used in the surveyed area. Hence, adequate strategies must be implemented by the local government to control these banned substances.

  5. Chloramphenicol Derivatives as Antibacterial and Anticancer Agents: Historic Problems and Current Solutions

    PubMed Central

    Dinos, George P.; Athanassopoulos, Constantinos M.; Missiri, Dionissia A.; Giannopoulou, Panagiota C.; Vlachogiannis, Ioannis A.; Papadopoulos, Georgios E.; Papaioannou, Dionissios; Kalpaxis, Dimitrios L.

    2016-01-01

    Chloramphenicol (CAM) is the D-threo isomer of a small molecule, consisting of a p-nitrobenzene ring connected to a dichloroacetyl tail through a 2-amino-1,3-propanediol moiety. CAM displays a broad-spectrum bacteriostatic activity by specifically inhibiting the bacterial protein synthesis. In certain but important cases, it also exhibits bactericidal activity, namely against the three most common causes of meningitis, Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis. Resistance to CAM has been frequently reported and ascribed to a variety of mechanisms. However, the most important concerns that limit its clinical utility relate to side effects such as neurotoxicity and hematologic disorders. In this review, we present previous and current efforts to synthesize CAM derivatives with improved pharmacological properties. In addition, we highlight potentially broader roles of these derivatives in investigating the plasticity of the ribosomal catalytic center, the main target of CAM. PMID:27271676

  6. An improved electrochemical aptasensor for chloramphenicol detection based on aptamer incorporated gelatine.

    PubMed

    Hamidi-Asl, Ezat; Dardenne, Freddy; Blust, Ronny; De Wael, Karolien

    2015-03-27

    Because of the biocompatible properties of gelatine and the good affinity of aptamers for their targets, the combination of aptamer and gelatine type B is reported as promising for the development of biosensing devices. Here, an aptamer for chloramphenicol (CAP) is mixed with different types of gelatine and dropped on the surface of disposable gold screen printed electrodes. The signal of the CAP reduction is investigated using differential pulse voltammetry. The diagnostic performance of the sensor is described and a detection limit of 1.83 × 10(-10) M is found. The selectivity and the stability of the aptasensor are studied and compared to those of other CAP sensors described in literature.

  7. An electrochemical impedimetric aptasensing platform for sensitive and selective detection of small molecules such as chloramphenicol.

    PubMed

    Pilehvar, Sanaz; Dierckx, Tarryn; Blust, Ronny; Breugelmans, Tom; De Wael, Karolien

    2014-07-07

    We report on the aptadetection of chloramphenicol (CAP) using electrochemical impedance spectroscopy. The detection principle is based on the changes of the interfacial properties of the electrode after the interaction of the ssDNA aptamers with the target molecules. The electrode surface is partially blocked due to the formation of the aptamer-CAP complex, resulting in an increase of the interfacial electron-transfer resistance of the redox probe detected by electrochemical impedance spectroscopy or cyclic voltammetry. We observed that the ratio of polarization resistance had a linear relationship with the concentrations of CAP in the range of 1.76-127 nM, and a detection limit of 1.76 nM was obtained. The covalent binding of CAP-aptamer on the electrode surface combined with the unique properties of aptamers and impedimetric transduction leads to the development of a stable and sensitive electrochemical aptasensor for CAP.

  8. An Improved Electrochemical Aptasensor for Chloramphenicol Detection Based on Aptamer Incorporated Gelatine

    PubMed Central

    Hamidi-Asl, Ezat; Dardenne, Freddy; Blust, Ronny; De Wael, Karolien

    2015-01-01

    Because of the biocompatible properties of gelatine and the good affinity of aptamers for their targets, the combination of aptamer and gelatine type B is reported as promising for the development of biosensing devices. Here, an aptamer for chloramphenicol (CAP) is mixed with different types of gelatine and dropped on the surface of disposable gold screen printed electrodes. The signal of the CAP reduction is investigated using differential pulse voltammetry. The diagnostic performance of the sensor is described and a detection limit of 1.83 × 10−10 M is found. The selectivity and the stability of the aptasensor are studied and compared to those of other CAP sensors described in literature. PMID:25825978

  9. Thermoadaptation-directed evolution of chloramphenicol acetyltransferase in an error-prone thermophile using improved procedures.

    PubMed

    Kobayashi, Jyumpei; Furukawa, Megumi; Ohshiro, Takashi; Suzuki, Hirokazu

    2015-07-01

    Enhancing the thermostability of thermolabile enzymes extends their practical utility. We previously demonstrated that an error-prone thermophile derived from Geobacillus kaustophilus HTA426 can generate mutant genes encoding enzyme variants that are more thermostable than the parent enzyme. Here, we used this approach, termed as thermoadaptation-directed enzyme evolution, to increase the thermostability of the chloramphenicol acetyltransferase (CAT) of Staphylococcus aureus and successfully generated a CAT variant with an A138T replacement (CAT(A138T)). This variant was heterologously produced, and its enzymatic properties were compared with those of the wild type. We found that CAT(A138T) had substantially higher thermostability than CAT but had comparable activities, showing that the A138T replacement enhanced protein thermostability without affecting the catalytic activity. Because variants CAT(A138S) and CAT(A138V), which were generated via in vitro site-directed mutagenesis, were more thermostable than CAT, the thermostability enhancement resulting from the A138T replacement can be attributed to both the presence of a hydroxyl group and the bulk of the threonine side chain. CAT(A138T) conferred chloramphenicol resistance to G. kaustophilus cells at high temperature more efficiently than CAT. Therefore, the gene encoding CAT(A138T) may be useful as a genetic marker in Geobacillus spp. Notably, CAT(A138T) generation was achieved only by implementing improved procedures (plasmid-based mutations on solid media); previous procedures (chromosome-based mutations in liquid media) were unsuccessful. This result suggests that this improved procedure is crucial for successful thermoadaptation-directed evolution in certain cases and increases the opportunities for generating thermostable enzymes.

  10. Evaluation of Amphotericin B and Chloramphenicol as Alternative Drugs for Treatment of Chytridiomycosis and Their Impacts on Innate Skin Defenses

    PubMed Central

    Holden, Whitney M.; Ebert, Alexander R.; Canning, Peter F.

    2014-01-01

    Chytridiomycosis, an amphibian skin disease caused by the emerging fungal pathogen Batrachochytrium dendrobatidis, has been implicated in catastrophic global amphibian declines. The result is an alarming decrease in amphibian diversity that is a great concern for the scientific community. Clinical trials testing potential antifungal drugs are needed to identify alternative treatments for amphibians infected with this pathogen. In this study, we quantified the MICs of chloramphenicol (800 μg/ml), amphotericin B (0.8 to 1.6 μg/ml), and itraconazole (Sporanox) (20 ng/ml) against B. dendrobatidis. Both chloramphenicol and amphotericin B significantly reduced B. dendrobatidis infection in naturally infected southern leopard frogs (Rana [Lithobates] sphenocephala), although neither drug was capable of complete fungal clearance. Long-term exposure of R. sphenocephala to these drugs did not inhibit antimicrobial peptide (AMP) synthesis, indicating that neither drug is detrimental to this important innate skin defense. However, we observed that chloramphenicol, but not amphotericin B or itraconazole, inhibited the growth of multiple R. sphenocephala skin bacterial isolates in vitro at concentrations below the MIC against B. dendrobatidis. These results indicate that treatment with chloramphenicol might dramatically alter the protective natural skin microbiome when used as an antifungal agent. This study represents the first examination of the effects of alternative antifungal drug treatments on amphibian innate skin defenses, a crucial step to validating these treatments for practical applications. PMID:24771024

  11. Investigations into the basis of chloramphenicol and tetracycline resistance in Staphylococcus intermedius isolates from cases of pyoderma in dogs.

    PubMed

    Kim, T J; Na, Y R; Lee, J I

    2005-04-01

    A total of 160 Staphylococcus intermedius isolates were recovered from cases of pyoderma in 2002 and were examined for susceptibility to 13 different antimicrobial agents. Ninety per cent (144) of the isolates were resistant to tetracycline, derivatives of which have been used until recently, and 18% (29) were resistant to chloramphenicol which was banned from use 13 years ago. The presence of genes encoding chloramphenicol acetyltransferase (CAT) and tetracycline resistance (tet); tet(K), (L), (M), and (O) were determined by PCR in the 29 chloramphenicol and tetracycline resistant isolates. Seventeen (59%) isolates contained the cat gene while 12 (41%) isolates did not carry the cat gene, implying there may be other genes for chloramphenicol resistance that were not detected by the primers (primer set 1) used in this study. The tet(M) gene was found in 28 (97%) of the resistant S. intermedius isolates, but none contained the tet(O) gene. All 29 isolates carried one or two tet genes; tet(K), (L), and (M), with four different distribution patterns. New PCR products, a 1.1 kb product using primer set 1 and a 0.2 kb product using primer set 2, were cloned and sequenced. A 904 bp fragment of S. aureus plamid pS194, including sequence from the streptomycin adenyltransferase gene (804 bp), was found inserted into the terminal region of the cat gene (GenBank accession no. AY604739), whilst the sequence of 0.2 kb was previously unpublished.

  12. The Identification of Intrinsic Chloramphenicol and Tetracycline Resistance Genes in Members of the Bacillus cereus Group (sensu lato).

    PubMed

    Glenwright, Helen; Pohl, Susanne; Navarro, Ferran; Miro, Elisenda; Jiménez, Guillermo; Blanch, Anicet R; Harwood, Colin R

    2016-01-01

    Bacillus toyonensis strain BCT-7112(T) (NCIMB 14858(T)) has been widely used as an additive in animal nutrition for more than 30 years without reports of adverse toxigenic effects. However, this strain is resistant to chloramphenicol and tetracycline and it is generally considered inadvisable to introduce into the food chain resistance determinants capable of being transferred to other bacterial strains, thereby adding to the pool of such determinants in the gastro-enteric systems of livestock species. We therefore characterized the resistance phenotypes of this strain and its close relatives to determine whether they were of recent origin, and therefore likely to be transmissible. To this end we identified the genes responsible for chloramphenicol (catQ) and tetracycline (tetM) resistance and confirmed the presence of homologs in other members of the B. toyonensis taxonomic unit. Unexpectedly, closely related strains encoding these genes did not exhibit chloramphenicol and tetracycline resistance phenotypes. To understand the differences in the behaviors, we cloned and expressed the genes, together with their upstream regulatory regions, into Bacillus subtilis. The data showed that the genes encoded functional proteins, but were expressed inefficiently from their native promoters. B. toyonensis is a taxonomic unit member of the Bacillus cereus group (sensu lato). We therefore extended the analysis to determine the extent to which homologous chloramphenicol and tetracycline resistance genes were present in other species within this group. This analysis revealed that homologous genes were present in nearly all representative species within the B. cereus group (sensu lato). The absence of known transposition elements and the observations that they are found at the same genomic locations, indicates that these chloramphenicol and tetracycline resistance genes are of ancient origin and intrinsic to this taxonomic group, rather than recent acquisitions. In this context

  13. The Identification of Intrinsic Chloramphenicol and Tetracycline Resistance Genes in Members of the Bacillus cereus Group (sensu lato)

    PubMed Central

    Glenwright, Helen; Pohl, Susanne; Navarro, Ferran; Miro, Elisenda; Jiménez, Guillermo; Blanch, Anicet R.; Harwood, Colin R.

    2017-01-01

    Bacillus toyonensis strain BCT-7112T (NCIMB 14858T) has been widely used as an additive in animal nutrition for more than 30 years without reports of adverse toxigenic effects. However, this strain is resistant to chloramphenicol and tetracycline and it is generally considered inadvisable to introduce into the food chain resistance determinants capable of being transferred to other bacterial strains, thereby adding to the pool of such determinants in the gastro-enteric systems of livestock species. We therefore characterized the resistance phenotypes of this strain and its close relatives to determine whether they were of recent origin, and therefore likely to be transmissible. To this end we identified the genes responsible for chloramphenicol (catQ) and tetracycline (tetM) resistance and confirmed the presence of homologs in other members of the B. toyonensis taxonomic unit. Unexpectedly, closely related strains encoding these genes did not exhibit chloramphenicol and tetracycline resistance phenotypes. To understand the differences in the behaviors, we cloned and expressed the genes, together with their upstream regulatory regions, into Bacillus subtilis. The data showed that the genes encoded functional proteins, but were expressed inefficiently from their native promoters. B. toyonensis is a taxonomic unit member of the Bacillus cereus group (sensu lato). We therefore extended the analysis to determine the extent to which homologous chloramphenicol and tetracycline resistance genes were present in other species within this group. This analysis revealed that homologous genes were present in nearly all representative species within the B. cereus group (sensu lato). The absence of known transposition elements and the observations that they are found at the same genomic locations, indicates that these chloramphenicol and tetracycline resistance genes are of ancient origin and intrinsic to this taxonomic group, rather than recent acquisitions. In this context we

  14. A gel-based visual immunoassay for non-instrumental detection of chloramphenicol in food samples.

    PubMed

    Yuan, Meng; Sheng, Wei; Zhang, Yan; Wang, Junping; Yang, Yijin; Zhang, Shuguang; Goryacheva, Irina Yu; Wang, Shuo

    2012-11-02

    A gel-based non-instrumental immuno-affinity assay was developed for the rapid screening of chloramphenicol (CAP) in food samples with the limit of detection (LOD) of 1 μg L(-1). The immuno-affinity test column (IATC) consisted of a test layer containing anti-CAP antibody coupled gel, and a control layer with anti-HRP antibody coupled gel. Based on the direct competitive immuno-reaction and the horseradish peroxidase enzymatic reaction, the test results could be evaluated visually. Basically, blue color development represented the negative results, while the absence of color development represented the positive results. In this study, CAP spiked samples of raw milk, pasteurized milk, UHT milk, skimmed milk powder, acacia honey, date honey, fish and shrimp were tested. Little or none sample pretreatment was required for this assay. The whole procedure was completed within 10min. In conclusion, the gel-based immuno-affinity test is a simple, rapid, and promising on-site screening method for CAP residues in food samples, with no instrumental requirement.

  15. The changes in pharmacokinetics and conjugation of chloramphenicol in irradiated rats.

    PubMed

    Stoklasová, A; Krízala, J; Ledvina, M

    1978-11-01

    In the serum and the liver of rats levels of chloramphenicol (CAP) following its i.v. administration (200 mg/kg) in the control groups and in the rats irradiated with whole-body air exposure to 500 R were determined with spectrophotometric methods. The CAP-levels in the serum increased in the group of rats 3 days after irradiation, but only during the 1st hour. At later time intervals the values were lower than in the controls. This decrease at the 60th min is striking even in the groups 6 and 9 days after exposure. Free CAP in the liver of rats irradiated 6 and 9 days before was lower at interval 30 min after CAP-administration, but the group irradiated 9 days before was unaltered. However, 120 min after CAP-administration the values of free CAP decreased at all intervals investigated following the irradiation. The levels of conjugated CAP in the liver of the rats 3 and 6 days after exposure were lower than in controls in both intervals after drug administration; but in rats 9 days after irradiation they increased. Our results indicate that the kinetics of CAP is altered and corresponding changes in its conjugation are effected under the condition of acute radiation syndrome.

  16. Rapid Detection of Chloramphenicol Residues in Aquatic Products Using Colloidal Gold Immunochromatographic Assay

    PubMed Central

    Zhou, Chennan; Zhang, Xueyin; Huang, Xinxin; Guo, Xishan; Cai, Qiang; Zhu, Songming

    2014-01-01

    A colloidal gold immunochromatographic assay (GICA) was developed for rapid detection of chloramphenicol (CAP) residues in aquatic products. A nitrocellulose (NC) membrane was used as the carrier, and the polyclonal CAP antibody was used as the marker protein. The average diameter of as-prepared colloidal gold nanoparticles (AuNPs) was about 20 nm. The optimal pH value of colloidal gold solutions and the amount of the antibody of CAP were 8.0 and 7.2 μg/mL, respectively. The CAP antibody was immobilized onto the conjugate pad after purification. The CAP conjugate and goat anti-rabbit IgG (secondary antibody) were coated onto the NC membrane. Next, the non-specific sites were blocked with 1% bovine serum albumin. The minimum detectable concentration of CAP in standard solution is 0.5 ng/mL, with good reproducibility. For the real samples from crucian carps injected with a single-dose of CAP in the dorsal muscles, the minimum detectable concentration of CAP residues was 0.5 μg/kg. The chromatographic analysis time was less than 10 min, and the strip had a long storage lifetime of more than 90 days at different temperatures. The strips provide a means for rapid detection of CAP residues in aquatic products. PMID:25412221

  17. Highly sensitive electrochemical sensor for chloramphenicol based on MOF derived exfoliated porous carbon.

    PubMed

    Xiao, Lili; Xu, Ruiyu; Yuan, Qunhui; Wang, Fu

    2017-05-15

    Benefit from the advantages in costless, simplicity and efficiency, solvent exfoliation has been widely used in preparation of two-dimensional nanosheets with enhanced performances in electronics, photonics, and catalysis. In this work, solvent exfoliation was first applied to prepare exfoliated porous carbon (EPC) from an isoreticular metal-organic framework-8 (IRMOF-8) derived porous carbon (DPC). The obtained EPC with high surface area (1854m(2)g(-1)) and improved dispersibility was used as electrode modifier for glassy carbon electrode (GCE) in square wave voltammetry (SWV) detection of chloramphenicol (CAP). The sensitivity of EPC modified GCE (EPC/GCE) was greatly improved in compare with that of the DPC modification. The corresponding linear ranges are 1×10(-8)-1×10(-6)molL(-1) and 1×10(-6)-4×10(-6)molL(-1). The detection limit was calculated to be 2.9×10(-9)molL(-1) (at a signal-to-noise ratio of 3, S/N=3). In addition, the proposed sensor was successfully applied in the analysis of CAP in honey and achieved satisfying recovery.

  18. Modification by chloramphenicol of diethyl sulphate-induced male recombination frequency in Drosophila melanogaster.

    PubMed

    Miglani, G S; Kaur, N P

    1995-06-01

    To study the effect of chloramphenicol (CPL, an inhibitor of protein synthesis) on diethyl sulphate (DES, a potent mutagen) induced male recombination frequency, the F1 (+/aristaless dumpy black cinnabar, al dp b cn) larvae of D. melanogaster were given a pre- or post-treatment of CPL with DES during the first or second half of larval life. In order to determine sensitivity of different germ cell stages to the induction and modification of male recombination frequency, five 3-day broods were taken from every F1 male. DES showed toxic effect on egg-to-adult development. DES was found to be a potent recombinogen. Several cases of non-reciprocal male recombination were recorded. The most frequent recombinant phenotype observed was b cn followed by cn and al. Majority of the recombinants appeared in clusters suggesting their pre-meiotic origin. DES produced male recombination at a stage where only primary spermatocytes were present in the larval testes. CPL when given as a pre- or post-treatment with DES revealed highest frequency of male recombination in broods that represented effect of treatment on spermatogonia predominantly. CPL enhanced the overall level of male recombination produced by DES in both pre- and post-treatments. The results suggested the role of protein synthesis in induction of male recombination in D. melanogaster. In addition, the present experiments give a methodology of enhancing the frequency of chemically-induced male recombination.

  19. Occurrence of chloramphenicol in cereal straw in north-western Europe.

    PubMed

    Nordkvist, Erik; Zuidema, Tina; Herbes, Rik G; Berendsen, Bjorn J A

    2016-05-01

    Two surveys are presented of straw analysed for naturally occurring chloramphenicol (CAP), a drug banned for use in food-producing animals. In the first study, CAP was analysed by LC-MS/MS and detected in 37 out of 105 straw samples originating from the Netherlands, France, the UK, Germany and Denmark. The highest level found was 6.3 µg kg(-1), the average 0.6 µg kg(-1) and the median 0.2 µg kg(-1). The second study included a method comparison between ELISA and LC-MS/MS and a survey of CAP in cereal straw sampled at farms in all areas of Sweden. A total of 215 samples were screened by ELISA and a subset of 26 samples was also analysed by LC-MS/MS. Fifty-four of the samples contained more than 1 µg kg(-1) CAP and the highest level found was 32 µg kg(-1) (confirmed by LC-MS/MS). The highest contents of CAP in this study were allocated to the Baltic sea coast in the south-eastern part of Sweden (the county of Skåne and the Baltic Sea isle of Gotland). These results indicate a high incidence of CAP in straw in north-west Europe and have a severe impact on the enforcement of European Union legislation.

  20. Electrocatalytic determination of chloramphenicol based on molybdenum disulfide nanosheets and self-doped polyaniline.

    PubMed

    Yang, Ruirui; Zhao, Jinlong; Chen, Meijing; Yang, Tao; Luo, Shizhong; Jiao, Kui

    2015-01-01

    In this paper, a novel molybdenum disulfide (MoS2) intercalated by self-doped polyaniline (SPAN) via ultrasonic exfoliating method was prepared to show outstanding conductivity and synergistic electrocatalytic activity using chloramphenicol (CAP) as a case. In the ultrasonic process, due to the strong π-π(*) stacking interaction and electrostatic repulsion, the negatively charged SPAN served as an intercalator to result in few-layers MoS2 nanosheets, which were exfoliated from bulk MoS2. This nanocomposite was characterized by scanning electron microscopy, transmission electron microscopy and differential pulse voltammetry. The obtained nanocomposite owns large conjugated structure and rich negative charge, which can improve the adsorption of conjugate structured CAP with the detection range from 0.1 to 1000 μmol L(-1). The results also showed that the electrocatalytic responses were further affected by the mass ratio of SPAN-MoS2 and the ultrasonication time. Our electrocatalytic platform could be further applied in the adsorption and detection of other positively charged biomolecules or aromatic molecules.

  1. Occurrence of chloramphenicol in cereal straw in north-western Europe

    PubMed Central

    Nordkvist, Erik; Zuidema, Tina; Herbes, Rik G.; Berendsen, Bjorn J.A.

    2016-01-01

    ABSTRACT Two surveys are presented of straw analysed for naturally occurring chloramphenicol (CAP), a drug banned for use in food-producing animals. In the first study, CAP was analysed by LC-MS/MS and detected in 37 out of 105 straw samples originating from the Netherlands, France, the UK, Germany and Denmark. The highest level found was 6.3 µg kg−1, the average 0.6 µg kg−1 and the median 0.2 µg kg−1. The second study included a method comparison between ELISA and LC-MS/MS and a survey of CAP in cereal straw sampled at farms in all areas of Sweden. A total of 215 samples were screened by ELISA and a subset of 26 samples was also analysed by LC-MS/MS. Fifty-four of the samples contained more than 1 µg kg−1 CAP and the highest level found was 32 µg kg−1 (confirmed by LC-MS/MS). The highest contents of CAP in this study were allocated to the Baltic sea coast in the south-eastern part of Sweden (the county of Skåne and the Baltic Sea isle of Gotland). These results indicate a high incidence of CAP in straw in north-west Europe and have a severe impact on the enforcement of European Union legislation. PMID:27101357

  2. Direct competitive chemiluminescence immunoassays based on gold-coated magnetic particles for detection of chloramphenicol.

    PubMed

    Liang, Xiaohui; Fang, Xiangyi; Yao, Manwen; Yang, Yucong; Li, Junfeng; Liu, Hongjun; Wang, Linyu

    2016-02-01

    Direct competitive chemiluminescence immunoassays (CLIA) based on gold-coated magnetic nanospheres (Au-MNPs) were developed for rapid analysis of chloramphenicol (CAP). The Au-MNPs were modified with carboxyl groups and amino groups by 11-mercaptoundecanoic acid (MUA) and cysteamine respectively, and then were respectively conjugated with CAP base and CAP succinate via an activating reaction using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). NSP-DMAE-NHS, a new and effective luminescence reagent, was employed to label anti-CAP antibody (mAb) as a tracer in direct CLIA for CAP detection using a 'homemade' luminescent measurement system that was set up with a photomultiplier tube (PMT) and a photon counting unit linked to a computer. The sensitivities and limits of detection (LODs) of the two methods were obtained and compared according to the inhibition curves. The 50% inhibition concentration (IC50 ) values of the two methods were about 0.044 ng/mL and 0.072 ng/mL respectively and LODs were approximately 0.001 ng/mL and 0.006 ng/mL respectively. To our knowledge, they were much more sensitive than any traditional enzyme-linked immunosorbent assay (ELISA) ever reported. Moreover, the new luminescence reagent NSP-DMAE-NHS is much more sensitive and stable than luminol and its derivatives, contributing to the sensitivity enhancement.

  3. Conjugation with polyamines enhances the antibacterial and anticancer activity of chloramphenicol

    PubMed Central

    Kostopoulou, Ourania N.; Kouvela, Ekaterini C.; Magoulas, George E.; Garnelis, Thomas; Panagoulias, Ioannis; Rodi, Maria; Papadopoulos, Georgios; Mouzaki, Athanasia; Dinos, George P.; Papaioannou, Dionissios; Kalpaxis, Dimitrios L.

    2014-01-01

    Chloramphenicol (CAM) is a broad-spectrum antibiotic, limited to occasional only use in developed countries because of its potential toxicity. To explore the influence of polyamines on the uptake and activity of CAM into cells, a series of polyamine–CAM conjugates were synthesized. Both polyamine architecture and the position of CAM-scaffold substitution were crucial in augmenting the antibacterial and anticancer potency of the synthesized conjugates. Compounds 4 and 5, prepared by replacement of dichloro-acetyl group of CAM with succinic acid attached to N4 and N1 positions of N8,N8-dibenzylspermidine, respectively, exhibited higher activity than CAM in inhibiting the puromycin reaction in a bacterial cell-free system. Kinetic and footprinting analysis revealed that whereas the CAM-scaffold preserved its role in competing with the binding of aminoacyl-tRNA 3′-terminus to ribosomal A-site, the polyamine-tail could interfere with the rotatory motion of aminoacyl-tRNA 3′-terminus toward the P-site. Compared to CAM, compounds 4 and 5 exhibited comparable or improved antibacterial activity, particularly against CAM-resistant strains. Compound 4 also possessed enhanced toxicity against human cancer cells, and lower toxicity against healthy human cells. Thus, the designed conjugates proved to be suitable tools in investigating the ribosomal catalytic center plasticity and some of them exhibited greater efficacy than CAM itself. PMID:24939899

  4. Hydrolysis of amphenicol and macrolide antibiotics: Chloramphenicol, florfenicol, spiramycin, and tylosin.

    PubMed

    Mitchell, Shannon M; Ullman, Jeffrey L; Teel, Amy L; Watts, Richard J

    2015-09-01

    Antibiotics that enter the environment can present human and ecological health risks. An understanding of antibiotic hydrolysis rates is important for predicting their environmental persistence as biologically active contaminants. In this study, hydrolysis rates and Arrhenius constants were determined as a function of pH and temperature for two amphenicol (chloramphenicol and florfenicol) and two macrolide (spiramycin and tylosin) antibiotics. Antibiotic hydrolysis rates in pH 4-9 buffer solutions at 25°C, 50°C, and 60°C were quantified, and degradation products were characterized. All of the antibiotics tested remained stable and exhibited no observable hydrolysis under ambient conditions typical of aquatic ecosystems. Acid- and base-catalyzed hydrolysis occurred at elevated temperatures (50-60°C), and hydrolysis rates increased considerably below pH 5 and above pH 8. Hydrolysis rates also increased approximately 1.5- to 2.9-fold for each 10°C increase in temperature. Based on the degradation product masses found, the functional groups that underwent hydrolysis were alkyl fluoride, amide, and cyclic ester (lactone) moieties; some of the resultant degradation products may remain bioactive, but to a lesser extent than the parent compounds. The results of this research demonstrate that amphenicol and macrolide antibiotics persist in aquatic systems under ambient temperature and pH conditions typical of natural waters. Thus, these antibiotics may present a risk in aquatic ecosystems depending on the concentration present.

  5. Application of the chloramphenicol acetyltransferase (CAT) diffusion assay to transgenic plant tissues.

    PubMed

    Peach, C; Velten, J

    1992-02-01

    Chloramphenicol acetyltransferase (CAT) activity was quantified in crude extracts from tobacco callus tissues using a modification of a previously reported diffusion assay. We describe here the alterations necessary in applying this rapid and simple assay procedure to plant materials. Due to the high concentration of nonspecific oxidases present in most plant tissues, some type of protective agent is required to maintain enzyme activity. We have tested beta-mercaptoethanol, cysteine, dithiothreitol, ascorbic acid and polyvinyl pyrrolidone as protective agents within the initial extraction buffer. We also investigated the effect of heat (60 degrees C, 10 min) and 5 mM EDTA on CAT activity. The highest CAT activity was obtained using 5 mM cysteine plus 5 mM EDTA in 40 mM Tris-HCl (pH 7.8) as the initial extraction buffer followed by a heat treatment. Using this buffer, CAT activity was stable on ice for more than two hours. In our hands, total acetyl-coenzyme A concentration within the assay mixture was found to be saturating at 250 microM and the Km determined to be 100 microM. Assays performed using the same crude plant extract indicate that 1) duplicate assays show less than 1.5% variation in activities and 2) CAT activity increases linearly with respect to volume of extract used.

  6. A series of shuttle vectors using chloramphenicol acetyltransferase as a reporter enzyme in yeast.

    PubMed

    Mannhaupt, G; Pilz, U; Feldmann, H

    1988-07-30

    Reports from numerous laboratories have shown that the gene coding for the bacterial enzyme chloramphenicol-3-O-acetyltransferase can be used as a reporter gene (cat) in mammalian and plant systems to analyze gene activity at the transcriptional level when combined with endogenous regulatory signals; the enzyme activity can be quantified by a chromatographic or a photometric assay. To adapt this simple and highly sensitive test for the yeast system, we constructed a series of yeast vectors containing the cat gene together with selectable markers for Escherichia coli and yeast; integrating, autonomously replicating and centromere-carrying plasmids were used. We show that the cat gene lacking the endogenous promoter is expressed at low levels in yeast transformants. To demonstrate functional expression of the cat gene placed under the control of a yeast promoter, we chose the PHO5 regulatory region. We found that cat expression was induced via the PHO5 promoter in a manner as observed for the endogenous PHO5 gene, whereas in the repressed state cat expression remained low. Using these vectors, it should be feasible to analyze other sequences conferring promoter activity or other control functions in yeast.

  7. Aptasensing of chloramphenicol in the presence of its analogues: reaching the maximum residue limit.

    PubMed

    Pilehvar, Sanaz; Mehta, Jaytry; Dardenne, Freddy; Robbens, Johan; Blust, Ronny; De Wael, Karolien

    2012-08-07

    A novel, label-free folding induced aptamer-based electrochemical biosensor for the detection of chloramphenicol (CAP) in the presence of its analogues has been developed. CAP is a broad-spectrum antibiotic that has lost its favor due to its serious adverse toxic effects on human health. Aptamers are artificial nucleic acid ligands (ssDNA or RNA) able to specifically recognize a target such as CAP. In this article, the aptamers are fixed onto a gold electrode surface by a self-assembly approach. In the presence of CAP, the unfolded ssDNA on the electrode surface changes to a hairpin structure, bringing the target molecules close to the surface and triggering electron transfer. Detection limits were determined to be 1.6 × 10(-9) mol L(-1). In addition, thiamphenicol (TAP) and florfenicol (FF), antibiotics with a structure similar to CAP, did not influence the performance of the aptasensor, suggesting a good selectivity of the CAP-aptasensor. Its simplicity and low detection limit (because of the home-selected aptamers) suggest that the electrochemical aptasensor is suitable for practical use in the detection of CAP in milk samples.

  8. Potential toxicity of amphenicol antibiotic: binding of chloramphenicol to human serum albumin.

    PubMed

    Chen, Huilun; Rao, Honghao; He, Pengzhen; Qiao, Yongxiang; Wang, Fei; Liu, Haijun; Cai, Minmin; Yao, Jun

    2014-10-01

    Antibiotics are widely used in daily life but their abuse has posed a potential threat to human health. To evaluate the toxicity of chloramphenicol (CAP) at the protein level, the interaction between CAP and human serum albumin (HSA) was investigated by fluorescence, Ultraviolet-visible (UV-Vis) absorption, Fourier transform infrared (FT-IR) spectroscopy and molecular docking methods. Fluorescence data revealed that the fluorescence quenching of HSA by CAP was the result of the formation of CAP-HSA complex, and the binding constant was determined to be 3.196 × 10(4) L mol(-1) at 310 K. The thermodynamic determination indicated that the interaction was driven by enthalpy change and entropy change together, where the multiple hydrogen bonds (CAP and the residues Arg 222 and His 242 of HSA) and van der Waals forces were the dominant binding force. The site marker competition revealed that CAP bound into sub-domain IIA of HSA. The binding of CAP induced the drastic reduction in α-helix conformation and the significant enhancement in β-sheet conformation of HSA. Molecular docking study further confirmed the binding mode obtained by experimental study. This work provides a new quantitative evaluation method for antibiotics to cause the protein damage.

  9. Distribution and chloramphenicol in the bovine genital tract and pharmacokinetic studies of florfenicol in cattle

    SciTech Connect

    Bretzlaff, K.N.

    1986-01-01

    The objectives were to investigate selected aspects of the distribution of chloramphenicol (CAP) in the bovine genital tract and to conduct preliminary pharmacologic studies with florfenicol (FLO), a fluorinated analogue of thiamphenicol, in cattle. After 8 hours' continuous intravenous (IV) infusion of CAP to 7 postpartum cows, steady state plasma-to-genital tissue ratios of CAP were approximately 3. After intrauterine infusion of 20 mg CAP/kg to 3 postpartum cows, approximately 40% of the dose was absorbed into the bloodstream. Tissue concentrations were high at 8 hour postdosing in tissues lining the uterine lumen but were below desired therapeutic concentrations in the myometrium of 2 of the cows. Eighty cows with retained fetal membranes (RFM) were assigned to receive on the following treatments: (1) removal of membranes only; (2) removal plus CAP; (3) nonremoval; (4) nonremoval plus CAP. CAP treatment consisted of 5 g administered IU twice daily for 3 days. The majority of cows in all groups acquired endometritis, although CAP reduced the prevalence and severity of the disease. A quantitative assay for FLO in plasma was developed and validated on a high performance liquid chromatographic (HPLC) system. The pharmacokinetics of FLO determined after IV administration of 50 mg FLO/kg to 5 cows were best described by a three-compartment model. FLO was approximately 18% bound to plasma proteins as determined by equilibrium dialysis and ultrafiltration. In an in vitro system, 5, 125, or 1000 ug/ml of CAP had no effect on neutrophils from 6 cows.

  10. Chloramphenicol stereoisomers need to be distinguished: consequences observed from a proficiency test.

    PubMed

    Sykes, Mark; Czymai, Tobias; Hektor, Thomas; Sharman, Matthew; Knaggs, Michael

    2017-04-01

    Although the use of chloramphenicol (CAP) as a veterinary drug is banned in the European Union and many other countries, monitoring for CAP residues in food is routine. Positive detections are few, but taken extremely seriously. European Union laboratories analysing for CAP should validate methods according to European Commission Decision 2002/657/EC, must be accredited to ISO 17025, and will generally participate in proficiency testing (PT) schemes, such as those offered by the Food Analysis Performance Assessment Scheme (FAPAS®). The FAPAS PTs aim to cover a wide range of relevant matrices including honey, prawns, fish, milk and kidney. Test materials are prepared either by animal dosing studies or by spiking raw matrix. The most common method reported by FAPAS participants used to screen for CAP residues is LC-MS/MS, but ELISA kits are increasingly being used. A recent PT round highlighted that the result obtained might be correlated with the type of analytical method being employed. Follow-up investigations have demonstrated that some of these variations in data are a function of the different stereoisomeric forms of CAP. This paper discusses the implication of this research on method validation requirements and European Union legislation.

  11. Nucleotide sequence and phylogeny of a chloramphenicol acetyltransferase encoded by the plasmid pSCS7 from Staphylococcus aureus.

    PubMed

    Schwarz, S; Cardoso, M

    1991-08-01

    The nucleotide sequence of the chloramphenicol acetyltransferase gene (cat) and its regulatory region, encoded by the plasmid pSCS7 from Staphylococcus aureus, was determined. The structural cat gene encoded a protein of 209 amino acids, which represented one monomer of the enzyme chloramphenicol acetyltransferase (CAT). Comparisons between the amino acid sequences of the pSCS7-encoded CAT from S. aureus and the previously sequenced CAT variants from S. aureus, Staphylococcus intermedius, Staphylococcus haemolyticus, Bacillus pumilis, Clostridium difficile, Clostridium perfringens, Escherichia coli, Shigella flexneri, and Proteus mirabilis were performed. An alignment of CAT amino acid sequences demonstrated the presence of 34 conserved amino acids among all CAT variants. These conserved residues were considered for their possible roles in the structure and function of CAT. On the basis of the alignment, a phylogenetic tree was constructed. It demonstrated relatively large evolutionary distances between the CAT variants of enteric bacteria, Clostridium, Bacillus, and Staphylococcus species.

  12. Adsorption behavior and mechanism of chloramphenicols, sulfonamides, and non-antibiotic pharmaceuticals on multi-walled carbon nanotubes.

    PubMed

    Zhao, Heng; Liu, Xue; Cao, Zhen; Zhan, Yi; Shi, Xiaodong; Yang, Yi; Zhou, Junliang; Xu, Jiang

    2016-06-05

    The adsorption behavior of different emerging contaminants (3 chloramphenicols, 7 sulfonamides, and 3 non-antibiotic pharmaceuticals) on five types of multi-walled carbon nanotubes (MWCNTs), and the underlying factors were studied. Adsorption equilibriums were reached within 12h for all compounds, and well fitted by the Freundlich isotherm model. The adsorption affinity of pharmaceuticals was positively related to the specific surface area of MWCNTs. The solution pH was an important parameter of pharmaceutical adsorption on MWCNTs, due to its impacts on the chemical speciation of pharmaceuticals and the surface electrical property of MWCNTs. The adsorption of ionizable pharmaceuticals decreased in varying degrees with the increased ionic strength. MWCNT-10 was found to be the strongest adsorbent in this study, and the Freundlich constant (KF) values were 353-2814mmol(1-n)L(n)/kg, 571-618mmol(1-n)L(n)/kg, and 317-1522mmol(1-n)L(n)/kg for sulfonamides, chloramphenicols, and non-antibiotic pharmaceuticals, respectively. The different adsorption affinity of sulfonamides might contribute to the different hydrophobic of heterocyclic substituents, while chloramphenicols adsorption was affected by the charge distribution in aromatic rings via substituent effects.

  13. Chloramphenicol binding to human serum albumin: Determination of binding constants and binding sites by steady-state fluorescence

    NASA Astrophysics Data System (ADS)

    Ding, Fei; Zhao, Guangyu; Chen, Shoucong; Liu, Feng; Sun, Ying; Zhang, Li

    2009-07-01

    The interaction between chloramphenicol and human serum albumin (HSA) was studied by fluorescence, UV/vis, circular dichroism (CD) and three-dimensional fluorescence spectroscopy. Fluorescence data revealed that the fluorescence quenching of HSA by chloramphenicol was the result of the formation of drug-HSA complex, and the effective quenching constants ( Ka) were 2.852 × 10 4, 2.765 × 10 4, 2.638 × 10 4 and 2.542 × 10 4 M -1 at 287, 295, 303 and 311 K, respectively. The thermodynamic parameters, enthalpy change (Δ H) and entropy change (Δ S) for the reaction were calculated to be -3.634 kJ mol -1 and 72.66 J mol -1 K -1 according to van't Hoff equation. The results indicated that the hydrophobic and electrostatic interactions played a major role in the binding of drug to HSA. The distance r between donor and acceptor was obtained to be 3.63 nm according to Förster's theory. Site marker competitive experiments indicated that the binding of drug to HSA primarily took place in subdomain IIA. The alterations of HSA secondary structure in the presence of chloramphenicol were confirmed by the evidences from synchronous fluorescence, CD and three-dimensional fluorescence spectra. In addition, the effect of common ions on the binding constants of drug-HSA complex was also discussed.

  14. Molecular characterizations of chloramphenicol- and oxytetracycline-resistant bacteria and resistance genes in mariculture waters of China.

    PubMed

    Dang, Hongyue; Zhao, Jingyi; Song, Linsheng; Chen, Mingna; Chang, Yaqing

    2009-07-01

    In order to gain an understanding of the diversity and distribution of antimicrobial-resistant bacteria and their resistance genes in maricultural environments, multidrug-resistant bacteria were screened for the rearing waters from a mariculture farm of China. Both abalone Haliotis discushannai and turbot Scophthalmus maximus rearing waters were populated with abundant chloramphenicol-resistant bacteria. These bacteria were also multidrug resistant, with Vibriosplendidus and Vibriotasmaniensis being the most predominant species. The chloramphenicol-resistance gene cat II, cat IV or floR could be detected in most of the multidrug-resistant isolates, and the oxytetracycline-resistance gene tet(B), tet(D), tet(E) or tet(M) could also be detected for most of the isolates. Coexistence of chloramphenicol- and oxytetracycline-resistance genes partially explains the molecular mechanism of multidrug resistance in the studied maricultural environments. Comparative studies with different antimicrobial agents as the starting isolation reagents may help detect a wider diversity of the antimicrobial-resistant bacteria and their resistance genes.

  15. The plasmid-encoded chloramphenicol-resistance protein of Rhodococcus fascians is homologous to the transmembrane tetracycline efflux proteins.

    PubMed

    Desomer, J; Vereecke, D; Crespi, M; Van Montagu, M

    1992-08-01

    The nucleotide sequence of the chloramphenicol-resistance gene (cmr) of Rhodococcus fascians NCPPB 1675 (located on the conjugative plasmid pRF2) allowed the identification of two possible open reading frames (ORFs), of which ORF1 was consistent with the mutational analysis. Biochemical analysis of cmr revealed that it does not encode an antibiotic-modifying enzyme. The amino acid sequence of ORF1 predicted a hydrophobic protein, with 12 putative membrane-spanning domains, homologous to proteins involved in the efflux of tetracycline across the plasma membrane. Expression of the cmr gene was induced by addition of chloramphenicol to the growth media. The promoter of this gene was restricted to 50 bp upstream from a 200 bp 5'-untranslated mRNA region, the latter containing two inverted repeats. At the amino acid level, the cmr gene is 52% identical to a previously identified chloramphenicol-resistance determinant in Streptomyces lividans, indicating a wider dispersion of this type of cmr gene among the actinomycetes.

  16. In vitro susceptibility of 104 clinical isolates of Haemophilus influenzae to moxalactam (LY127935), ampicillin, chloramphenicol, and ticarcillin.

    PubMed Central

    Mason, E O; Kaplan, S L; Anderson, D C; Hinds, D B; Feigin, R D

    1980-01-01

    A total of 104 strains of Haemophilus influenzae isolated from pediatric patients over a 1-year period were tested for susceptibility to moxalactam (LY127935), ampicillin, chloramphenicol, and ticarcillin. Of these strains, 30 produced beta-lactamase. LY127935 inhibited 99% of the strains at a concentration of 0.125 microgram/ml; the remaining strain was inhibited by this antibiotic at 0.25 microgram/ml. beta-Lactamase-producing strains were inhibited by ampicillin at greater than or equal to 2 microgram/ml. beta-Lactamase-negative strains were all inhibited by ampicillin at less than or equal to 1 microgram/ml, except for one nontypable strain which required 2 microgram of ampicillin per ml for inhibition. All strains were susceptible to chloramphenicol at less than or equal to 4 microgram/ml. beta-Lactamase-producing strains were less susceptible (geometric mean = 4.702 microgram/ml) to ticarcillin than were strains which did not produce beta-lactamase (geometric mean = 0.331 microgram/ml). LY127935 susceptibility was not influenced by increasing inoculum size, as ws ampicillin susceptibility. Combinations of LY127935 and chloramphenicol or ampicillin were not antagonistic in vitro. PMID:6448579

  17. Single-disk diffusion testing (Kirby-Bauer) of susceptibility of Proteus mirabilis to chloramphenicol: significance of the intermediate category.

    PubMed

    Furtado, G L; Medeiros, A A

    1980-10-01

    The significance of the intermediate category of the single-disk diffusion test (Kirby-Bauer) of antibiotic susceptibility has never been clearly defined. Thirty-two percent of 756 clinical isolates of Proteus mirabilis were of intermediate susceptibility to chloramphenicol, a higher percentage than for any other species. The breakpoint separating susceptible and intermediate isolates nearly bisected the frequency distribution of zone diameters of P. mirabilis but not that of the other species. The breakpoint separating susceptible and intermediate isolates nearly bisected the frequency distribution of zone diameters of P. mirabillis but not that of the other species tested. By serial broth dilution testing, the minimal inhibitory concentrations (MICs) of chloramphenicol of 50 individual isolates of P. mirabilis were 3.9 to 22.1 micrograms/ml (geometric mean, 8.0), whereas the MICs of susceptible Escherichia coli, Klebsiella, and Enterobacter strains were 2.0 to 3.9 micrograms/ml (geometric mean, 2.9). Seventy percent of isolates of P. mirabilis with MICs of 7.8 to 15.6 micrograms/ml were classified as susceptible by disk testing. We conclude that existing Kirby-Bauer breakpoints do not accurately discriminate P. mirabilis isolates that are marginally susceptible to chloramphenicol. These data underscore the difficulty of applying a single set of breakpoints to all species and suggest that species-specific breakpoints would more accurately predict the MIC equivalent of given zone diameters.

  18. Flow-injection chemiluminescence method for the determination of chloramphenicol based on luminol-sodium periodate order-transform second-chemiluminescence reaction.

    PubMed

    Zhuang, Ya-Feng; Zhu, Sheng-Nan; Wei, Wei; Li, Jie-Li

    2011-01-01

    A new chemiluminescence (CL) reaction was observed when chloramphenicol solution was injected into the mixture after the end of the reaction of alkaline luminol and sodium periodate or sodium periodate was injected into the reaction mixture of chloramphenicol and alkaline luminol. This reaction is described as an order-transform second-chemiluminescence (OTSCL) reaction. The OTSCL method combined with a flow-injection technique was applied to the determination of chloramphenicol. The optimum conditions for the order-transform second-chemiluminescence emission were investigated. A mechanism for OTSCL has been proposed on the basis of the chemiluminescence kinetic characteristics, the UV-visible spectra and the chemiluminescent spectra. Under optimal experimental conditions, the CL response is proportional to the concentration of chloramphenicol over the range 5.0 × 10(-7)-5.0 × 10(-5) mol/L with a correlation coefficient of 0.9969 and a detection limit of 6.0 × 10(-8) mol/L (3σ). The relative standard deviation (RSD) for 11 repeated determinations of 5.0 × 10(-6) mol/L chloramphenicol is 1.7%. The method has been applied to the determination of chloramphenicol in pharmaceutical samples with satisfactory results.

  19. Improvement of chloramphenicol production in Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway.

    PubMed

    Vitayakritsirikul, Vipawan; Jaemsaeng, Ratchaniwan; Lohmaneeratana, Karan; Thanapipatsiri, Anyarat; Daduang, Ratama; Chuawong, Pitak; Thamchaipenet, Arinthip

    2016-03-01

    Streptomyces venezuelae ATCC 10712 produces chloramphenicol in small amounts. To enhance chloramphenicol production, two genes, aroB and aroK, encoding rate-limiting enzymes of the shikimate pathway were overexpressed using the expression vector pIJ86 under the control of the strong constitutive ermE* promoter. The recombinant strains, S. venezuelae/pIJ86-aroB and S. venezuelae/pIJ86-aroK, produced 2.5- and 4.3-fold greater amounts respectively of chloramphenicol than wild type at early stationary phase of growth. High transcriptional levels of aroB and aroK genes were detected at the early exponential growth of both recombinant strains and consistent with the enhanced expression of pabB gene encoding an early enzyme in chloramphenicol biosynthesis. The results suggested that the increment of carbon flux was directed towards intermediates in the shikimate pathway required for the production of chorismic acid, and consequently resulted in the enhancement of chloramphenicol production. This work is the first report of a convenient genetic approach to manipulate primary metabolite genes in S. venezuelae in order to increase chloramphenicol production.

  20. Radiation sensitivity of Salmonella isolates relative to resistance to ampicillin, chloramphenicol or gentamicin

    NASA Astrophysics Data System (ADS)

    Niemira, Brendan A.; Lonczynski, Kelly A.; Sommers, Christopher H.

    2006-09-01

    Antibiotic resistance of inoculated bacteria is a commonly used selective marker. Bacteria resistant to the antibiotic nalidixic acid have been shown to have an increased sensitivity to irradiation. The purpose of this research was to screen a collection of Salmonella isolates for antibiotic resistance and determine the association, if any, of antibiotic resistance with radiation sensitivity. Twenty-four clinical isolates of Salmonella were screened for native resistance to multiple concentrations of ampicillin (Amp), chloramphenicol (Chl), or gentamicin (Gm). Test concentrations were chosen based on established clinical minimum inhibitory concentration (MIC) levels, and isolates were classified as either sensitive or resistant based on their ability to grow at or above the MIC. Salmonella cultures were grown overnight at (37 °C) in antibiotic-amended tryptic soy broth (TSB). Native resistance to Gm was observed with each of the 24 isolates (100%). Eight isolates (33%) were shown to be resistant to Amp, while seven isolates (29%) were shown to be resistant to Chl. In separate experiments, Salmonella cultures were grown overnight (37 °C) in TSB, centrifuged, and the cell pellets were re-suspended in phosphate buffer. The samples were then gamma irradiated at doses up to 1.0 kGy. The D10 values (the ionizing radiation dose required to reduce the viable number of microorganisms by 90%) were determined for the 24 isolates and they ranged from 0.181 to 0.359 kGy. No correlation was found between the D10 value of the isolate and its sensitivity or resistance to each of the three antibiotics. Resistance to Amp or Chl is suggested as appropriate resistance marker for Salmonella test strains to be used in studies of irradiation.

  1. A competitive immunoassay for sensitive detection of small molecules chloramphenicol based on luminol functionalized silver nanoprobe.

    PubMed

    Yu, Xiuxia; He, Yi; Jiang, Jie; Cui, Hua

    2014-02-17

    Chloramphenicol (CHL) as a broad-spectrum antibiotic has a broad action spectrum against Gram-positive and Gram-negative bacteria, as well as anaerobes. The use of CHL is strictly restricted in poultry because of its toxic effect. However, CHL is still illegally used in animal farming because of its accessibility and low cost. Therefore, sensitive methods are highly desired for the determination of CHL in foodstuffs. The immunoassays based on labeling as an important tool have been reported for the detection of CHL residues in food-producing animals. However, most of the labeling procedures require multi-step reactions and purifications and thus they are complicated and time-consuming. Recently, in our previous work, luminol functionalized silver nanoparticles have been successfully synthesized, which exhibits higher CL efficiency than luminol functionalized gold nanoparticles. In this work, the new luminol functionalized silver nanoparticles have been used for the labeling of small molecules CHL for the first time and a competitive chemiluminescent immunoassay has been developed for the detection of CHL. Owing to the amplification of silver nanoparticles, high sensitivity for CHL could be achieved with a low detection limit of 7.6×10(-9) g mL(-1) and a wide linear dynamic range of 1.0×10(-8)-1.0×10(-6) g mL(-1). This method has also been successfully applied to determine CHL in milk and honey samples with a good recoveries (92% and 102%, 99% and 107% respectively), indicating that the method is feasible for the determination of CHL in real milk and honey samples. The labeling procedure is simple, convenient and fast, superior to previously reported labeling procedures. The immunoassay is also simple, fast, sensitive and selective. It is of application potential for the determination of CHL in foodstuffs.

  2. Zero valent iron produces dichloroacetamide from chloramphenicol antibiotics in the absence of chlorine and chloramines.

    PubMed

    Chu, Wenhai; Ding, Shunke; Bond, Tom; Gao, Naiyun; Yin, Daqiang; Xu, Bin; Cao, Zhongqi

    2016-11-01

    Dichloroacetamide (DCAcAm) is an important type of nitrogenous disinfection byproduct. This study is the first to report that DCAcAm can be formed in the absence of chlorinated disinfectants (chlorine and chloramines). This can occur through reduction of three chloramphenicol (CAP) antibiotics by zero valent iron (ZVI). The effects of key experimental parameters, including reaction time, ZVI dose, pH, temperature, water type, and the presence of humic acid (HA) on the formation of DCAcAm were ascertained. The DCAcAm yields from three CAPs all presented the trend of increasing first and then decreasing with time and also increased with increasing ZVI dosage. DCAcAm yields from the ZVI reduction route were higher than those resulting from the chlorination of some previously identified DCAcAm precursors. Acidic conditions favored the formation of DCAcAm by the ZVI route. In addition, lower temperatures increased DCAcAm yields at extended contact times (>12 h). DCAcAm formed from the three CAPs in the presence of HA was lower than in the absence of HA. The formation potential of DCAcAm from the reduction of authentic waters spiked with CAPs by ZVI showed good linear correlations with initial concentrations of the three CAPs. This allows the formation of DCAcAm from the reduction of CAPs by ZVI to be predicted. Given that many wastewater and drinking water distribution networks contain unlined cast iron pipes, reactions between CAPs and ZVI may contribute to the formation of DCAcAm in such systems.

  3. Determination of Aflatoxin M1 and Chloramphenicol in Milk Based on Background Fluorescence Quenching Immunochromatographic Assay

    PubMed Central

    Wu, Xiaoxia; Tian, Xiaofeng; Xu, Lihua; Li, Jiutong

    2017-01-01

    Harsh demanding has been exposed on the concentration of aflatoxin M1 (AFM1) and chloramphenicol (CAP) in milk. In this study, we developed a new method based on background fluorescence quenching immunochromatographic assay (bFQICA) to detect AFM1 and CAP in milk. The detection limit for AFM1 was 0.0009 ng/mL, while that for the CAP was 0.0008 ng/mL. The assay variability was determined with 3 AFM1 standards (i.e., 0.25 ng/mL, 0.5 ng/mL, and 1.0 ng/mL), and the actual detection value was 0.2497, 0.5329, and 1.0941, respectively. For the assay variability of 3 CAP standards (i.e., 0.10 ng/mL, 0.30 ng/mL, and 0.50 ng/mL), the actual detection value was 0.0996, 0.3096, and 0.4905, respectively. The recovery rate of AFM1 was 99.7%–101.7%, while that for CAP was 95.3%–97.6%. For the test stability, AFM1 and CAP showed satisfactory test stability even at month 5. Compared with the sensitivity of liquid chromatography-mass spectrometry (LC-MS) method, no statistical difference was noticed in results of the bFQICA. Our method is convenient for the detection of AFM1 and CAP in milk with a test duration of about 8 minutes. Additionally, an internal WiFi facility is provided in the system allowing for quick connection and storage in the intelligent cell phone. PMID:28367449

  4. Bacteria isolated from sewage influent resistant to ciprofloxacin, chloramphenicol and tetracycline.

    PubMed

    Zwenger, Sam R; Gillock, Eric T

    2009-02-01

    This study assessed the presence of antibiotic-resistant bacteria in sewage influent. Resistance was measured by determining the lowest concentration of antibiotic, in micrograms per milliliter (microg mL(- 1)). To determine the minimum inhibitory concentration (MIC), which is used in diagnostic laboratories, we used the Etest, a plastic strip containing an antibiotic concentration gradient. In total, we sampled five sewage treatment plants of various sizes in Kansas and isolated bacteria resistant to three broad-spectrum antibiotics; ciprofloxacin (1-cyclopropyl-6-fluoro-4-oxo-7-piperazin-1-yl-quinoline-3-carboxylic acid), chloramphenicol 2,2-dichlor-N-[(aR, bR)-b-hydroxy-a-hydroxymethyl-4-nitrophenethyl] acetamide), and tetracycline (2-(amino-hydroxy-ethylidene)-4-dimethylamino-6,10,11,12a-tetrahydroxy-6-methyl-4,4a,5,5a-tetrahydrotetracene-1,3,12-trione). In total, 25 Gram-negative isolates were found to be resistant to at least one of the antibiotics tested. Some isolates were multi-drug resistant, regardless of the amount of influent the sewage treatment plant received. A Pseudomonas isolate from the smallest sewage treatment plant (approximately 2 million gallons treated per day) showed resistance to all three antibiotics, albeit at low levels (10 microg mL(- 1)). The largest number of bacteria (6 species) were isolated from the largest sewage treatment plant (45 million gallons per day). Regardless, the results of this study are in agreement with similar studies, antibiotic resistance can persist long after the antibiotics have been forgotten.

  5. Determination of chloramphenicol residues in commercial chicken eggs in the Federal Capital Territory, Abuja, Nigeria.

    PubMed

    Mbodi, Felix E; Nguku, P; Okolocha, E; Kabir, J

    2014-01-01

    The use of antibiotics in poultry can result in residues in eggs. The joint FAO/WHO committee recommended banning the use of chloramphenicol (CAP) in food animals due to its public health hazards of aplastic anaemia, leukaemia, allergy, antibacterial resistance and carcinogenicity. This paper determines the prevalence of CAP residues in chicken eggs and assesses the usage and awareness of its ban amongst poultry farmers in the Federal Capital Territory (FCT), Abuja, Nigeria. A cross-sectional survey of registered poultry farmers in FCT was conducted using questionnaires to determine CAP administration in poultry and awareness of its ban. Pooled egg samples were collected from each poultry farm surveyed and from randomly sampled government-owned markets in FCT. Source of eggs by state were identified by the marketer at the time of collection. Samples were analysed using an enzyme-linked immunosorbent assay (ELISA) technique for the presence of CAP, and prevalence was determined. Of 288 total pooled samples collected, 257 (89.2%) were from the markets and 31 (10.8%) were from poultry farms. A total of 20 (7%) pooled egg samples tested CAP-positive; market eggs originated from 15 (41%) states of the country. Of the market eggs, 16 (6.2%) pooled samples tested positive. Of eggs from poultry farms, four (12.9%) tested positive. Mean CAP concentrations in the positive samples ranged from 0.49 to 1.17 µg kg(-1) (parts per billion). CAP use amongst poultry farmers in FCT was 75.5%; awareness of the CAP ban was 26.3%. Though 66% of veterinarians were unaware of a CAP ban, they were more likely to be aware than other poultry farmers (odds ratio (OR) = 1.4). Farm managers who use CAP were more likely to be aware of CAP ban than the farm managers not using CAP (OR = 5.5; p = 0.04). Establishing a drug residue surveillance and control program and enforcement of CAP legislation/regulation is needful to educate and prohibit the widespread CAP use amongst Nigerian poultry farmers.

  6. Biosynthesis of the dichloroacetyl component of chloramphenicol in Streptomyces venezuelae ISP5230: genes required for halogenation.

    PubMed

    Piraee, Mahmood; White, Robert L; Vining, Leo C

    2004-01-01

    Five ORFs were detected in a fragment from the Streptomyces venezuelae ISP5230 genomic DNA library by hybridization with a PCR product amplified from primers representing a consensus of known halogenase sequences. Sequencing and functional analyses demonstrated that ORFs 11 and 12 (but not ORFs 13-15) extended the partially characterized gene cluster for chloramphenicol (Cm) biosynthesis in the chromosome. Disruption of ORF11 (cmlK) or ORF12 (cmlS) and conjugal transfer of the insertionally inactivated genes to S. venezuelae gave mutant strains VS1111 and VS1112, each producing a similar series of Cm analogues in which unhalogenated acyl groups replaced the dichloroacetyl substituent of Cm. 1H-NMR established that the principal metabolite in the disrupted strains was the alpha-N-propionyl analogue. The sequence of CmlK implicated the protein in adenylation, and involvement in halogenation was inferred from biosynthesis of analogues by the cmlK-disrupted mutant. A role in generating the dichloroacetyl substituent was supported by partial restoration of Cm biosynthesis when a cloned copy of cmlK was introduced in trans into VS1111. Complementation of the mutant also indicated that inactivation of cmlK rather than a polar effect of the disruption on cmlS expression had interfered with dichloroacetyl biosynthesis. The deduced CmlS sequence resembled sequences of FADH2-dependent halogenases. Conjugal transfer of cmlK or cmlS into S. venezuelae cml-2, a chlorination-deficient strain with a mutation mapped genetically to the Cm biosynthesis gene cluster, did not complement the cml-2 lesion, suggesting that one or more genes in addition to cmlK and cmlS is needed to assemble the dichloroacetyl substituent. Insertional inactivation of ORF13 did not affect Cm production, and the products of ORF14 and ORF15 matched Streptomyces coelicolor A3(2) proteins lacking plausible functions in Cm biosynthesis. Thus cmlS appears to mark the downstream end of the gene cluster.

  7. Salmonella typhi resistant to Chloramphenicol, Ampicillin, and Other Antimicrobial Agents: Strains Isolated During an Extensive Typhoid Fever Epidemic in Mexico

    PubMed Central

    Olarte, Jorge; Galindo, Emma

    1973-01-01

    During 1972 a large epidemic, in excess of 10,000 cases, of typhoid fever occurred in Mexico City, Pachuca, and other communities of Mexico. The main characteristic of the epidemic, in addition to the large number of persons affected, was the prevalence of a strain of Salmonella typhi which was highly resistant to chloramphenicol both in vivo and in vitro, and which belonged to a single phage type, Vi degraded approaching type A. Of 493 strains of S. typhi studied during the outbreak, 452 (91.7%) were resistant to chloramphenicol (CM), tetracycline (TC), streptomycin (SM), and sulfonamides (SU). The epidemic strain owes its resistance to an R factor which is easily transferable to Escherichia coli K-12 and which appears to be stable. In the third month of the outbreak, a strain of S. typhi resistant to CM, TC, SM, SU, ampicillin (AM), and kanamycin (KM) was isolated from a patient with severe typhoid fever. During the following 9 months, six additional strains of S. typhi resistant to AM, CM, TC, SM, and SU were also isolated. Transfer experiments to E. coli K-12 indicate that these strains are infected with two different R factors, one causing CM, TC, SM, and SU resistance and the other causing AM or AM and KM resistance. The frequency of transfer of the resistance in overnight crosses was in the order of 10−4 for CM, TC, SM, and SU and 10−6 for AM or AM, and KM. The appearance of these strains resistant both to chloramphenicol and ampicillin was a cause for concern for the clinicians; fortunately, they have remained an infrequent cause of disease. PMID:4602828

  8. Chloramphenicol Biosynthesis: The Structure of CmlS, a Flavin-Dependent Halogenase Shwing a Covalent Flavin-Aspartate Bond

    SciTech Connect

    Podzelinska, K.; Latimer, R; Bhattacharya, A; Vining, L; Zechel, D; Jia, Z

    2010-01-01

    Chloramphenicol is a halogenated natural product bearing an unusual dichloroacetyl moiety that is critical for its antibiotic activity. The operon for chloramphenicol biosynthesis in Streptomyces venezuelae encodes the chloramphenicol halogenase CmlS, which belongs to the large and diverse family of flavin-dependent halogenases (FDH's). CmlS was previously shown to be essential for the formation of the dichloroacetyl group. Here we report the X-ray crystal structure of CmlS determined at 2.2 {angstrom} resolution, revealing a flavin monooxygenase domain shared by all FDHs, but also a unique 'winged-helix' C-terminal domain that creates a T-shaped tunnel leading to the halogenation active site. Intriguingly, the C-terminal tail of this domain blocks access to the halogenation active site, suggesting a structurally dynamic role during catalysis. The halogenation active site is notably nonpolar and shares nearly identical residues with Chondromyces crocatus tyrosyl halogenase (CndH), including the conserved Lys (K71) that forms the reactive chloramine intermediate. The exception is Y350, which could be used to stabilize enolate formation during substrate halogenation. The strictly conserved residue E44, located near the isoalloxazine ring of the bound flavin adenine dinucleotide (FAD) cofactor, is optimally positioned to function as a remote general acid, through a water-mediated proton relay, which could accelerate the reaction of the chloramine intermediate during substrate halogenation, or the oxidation of chloride by the FAD(C4{alpha})-OOH intermediate. Strikingly, the 8{alpha} carbon of the FAD cofactor is observed to be covalently attached to D277 of CmlS, a residue that is highly conserved in the FDH family. In addition to representing a new type of flavin modification, this has intriguing implications for the mechanism of FDHs. Based on the crystal structure and in analogy to known halogenases, we propose a reaction mechanism for CmlS.

  9. Determination of chloramphenicol residue in fish and shrimp tissues by gas chromatography with a microcell electron capture detector.

    PubMed

    Ding, Shuangyang; Shen, Jianzhong; Zhang, Suxia; Jiang, Haiyang; Sun, Zhiwen

    2005-01-01

    A gas chromatography method with microcell electron capture detection was developed for the determination of chloramphenicol residue in fish and shrimp muscle tissues. The tissue samples were extracted with ethyl acetate, defatted with hexane, and derivatized with Sylon BFT [N,O-bis (trimethylsilyl) trifluoroacetamide-trimethylchlorosilane (99 + 1)]. The limit of detection was 0.04 ng/g and the limit of quantitation 0.1 ng/g. Average recoveries were 70.8-90.8% for fish and 69.9-86.3% for shrimp, respectively. The method was validated for the determination of practical samples.

  10. Chloramphenicol encapsulated in poly-ε-caprolactone–pluronic composite: nanoparticles for treatment of MRSA-infected burn wounds

    PubMed Central

    Kalita, Sanjeeb; Devi, Banasmita; Kandimalla, Raghuram; Sharma, Kaustav Kalyan; Sharma, Arup; Kalita, Kasturi; Kataki, Amal Chandra; Kotoky, Jibon

    2015-01-01

    The emergence of methicillin-resistant Staphylococcus aureus (MRSA) infection has increased precipitously over the past several decades, with far-reaching health care and societal costs. MRSA infections in the context of burn wounds lead to invasive disease that could potentially cause mortality. Chloramphenicol is a well-known broad-spectrum bacteriostatic antibiotic that has been used since 1949, but due to its hydrophobicity, poor penetration in skin, fast degradation, and toxicity, its application has been hindered. Furthermore, it has been demonstrated that old antibiotics such as chloramphenicol remained active against a large number of currently prevalent resistant bacterial isolates due to their low-level use in the past. Recently, the novel nanoparticulate drug-delivery system has been used and reported to be exceptionally useful for topical therapeutics, due to its distinctive physical characteristics such as a high surface-to-volume ratio and minuscule size. It helps to achieve better hydrophilicity, bioavailability, and controlled delivery with enhanced therapeutic index, which has resulted in decreased toxicity levels compared to the crude drug. Here, we report a novel chloramphenicol loaded with poly(ε-caprolactone) (PCL)-pluronic composite nanoparticles (CAM-PCL-P NPs), physicochemical characterizations, and its bioactivity evaluation in a MRSA-infected burn-wound animal model. CAM-PCL-P NPs could encapsulate 98.3% of the drug in the nanoparticles and release 81% of the encapsulated drug over 36 days with a time to 50% drug release of 72 hours (51%). Nanoparticle suspensions maintained the initial properties with respect to size and encapsulation efficiency, even after 6 months of storage at 4°C and 25°C, respectively (P>0.05). Significant reduction in the level of toxicity was observed for CAM-PCL-P NPs compared with that of free drug as confirmed from hemolytic activity against human blood erythrocytes and cytotoxicity assay against an MCF-7

  11. A simple sample preparation for simultaneous determination of chloramphenicol and its succinate esters in food products using high-performance liquid chromatography/high-resolution mass spectrometry.

    PubMed

    Amelin, Vasiliy; Korotkov, Anton

    2017-02-01

    A simple method is described for the determination of chloramphenicol and its succinate esters in food products. Examination of food products using high-performance liquid chromatography/high-resolution mass spectrometry showed the presence not only of chloramphenicol but also of its succinate forms. A scheme is proposed for determining chloramphenicol and its succinate esters (calculated as chloramphenicol) in meat (beef, pork, poultry), milk, liver, kidney, eggs, fish and honey. Analytes are extracted from a 1.0 g sample with 5 ml acetonitrile. It was found that using the method of standard addition and diluting the extract with water leads to the elimination of matrix effects and also eliminates errors associated with peak splitting due to the separate elution of the differing forms of the analyte. Validation results were satisfactory, with recoveries from 85% to 111% (meat, milk, liver, kidney, eggs, fish and honey) and a relative standard deviation (RSD) lower than 13% for spiked levels of 0.3, 1.0 and 5 µg kg(-)(1). The limits of detection and quantification (calculated as chloramphenicol for all forms) were 0.1 and 0.3 µg kg(-)(1), respectively. The RSD of the results of the analysis was < 10%. The duration of the analysis was less than 1 h.

  12. Synthesis, spectroscopic, thermal and anticancer studies of metal-antibiotic chelations: Ca(II), Fe(III), Pd(II) and Au(III) chloramphenicol complexes

    NASA Astrophysics Data System (ADS)

    Al-Khodir, Fatima A. I.; Refat, Moamen S.

    2016-09-01

    Four Ca(II), Fe(III), Pd(II) and Au(III) complexes of chloramphenicol drug have been synthesized and well characterized using elemental analyses, (infrared, electronic, and 1H-NMR) spectra, magnetic susceptibility measurement, and thermal analyses. Infrared spectral data show that the chloramphenicol drug coordinated to Ca(II), Pd(II) and Au(III) metal ions through two hydroxyl groups with 1:1 or 1:2 M ratios, but Fe(III) ions chelated towards chloramphenicol drug via the oxygen and nitrogen atoms of amide group with 1:2 ratio based on presence of keto↔enol form. The X-ray powder diffraction (XRD), scanning electron microscope (SEM) and transmission electron microscopy (TEM) techniques were used to identify the nano-size particles of both iron(III) and gold(III) chloramphenicol complexes. The antimicrobial assessments of the chloramphenicol complexes were scanned and collected the results against of some kind of bacteria and fungi. The cytotoxic activity of the gold(III) complex was tested against the human colon carcinoma (HCT-116) and human hepatocellular carcinoma (HepG-2) tumor cell lines.

  13. Comparison between Source-induced Dissociation and Collision-induced Dissociation of Ampicillin, Chloramphenicol, Ciprofloxacin, and Oxytetracycline via Mass Spectrometry.

    PubMed

    Lee, Seung Ha; Choi, Dal Woong

    2013-06-01

    Mass spectrometry (MS) is a very powerful instrument that can be used to analyze a wide range of materials such as proteins, peptides, DNA, drugs, and polymers. The process typically involves either chemical or electron (impact) ionization of the analyte. The resulting charged species or fragment is subsequently identified by the detector. Usually, single mass uses source-induced dissociation (SID), whereas mass/mass uses collision-induced dissociation (CID) to analyze the chemical fragmentations Each technique has its own advantages and disadvantages. While CID is most effective for the analysis of pure substances, multiple- step MS is a powerful technique to get structural data. Analysis of veterinary drugs ampicillin, chloramphenicol, ciprofloxacin, and oxytetracycline serves to highlight the slight differences between SID and CID. For example, minor differences were observed between ciprofloxacin and oxytetracycline via SID or CID. However, distinct fragmentation patterns were observed for ampicllin depending on the analysis method. Both SID and CID showed similar fragmentation spectra but different signal intensities for chloramphenicol. There are several factors that can influence the fragmentation spectra, such as the collision energy, major precursor ion, electrospray mode (positive or negative), and sample homogeneity. Therefore, one must select a fragmentation method on an empirical and case-by-case basis.

  14. Stability-Indicating HPLC Method for Simultaneous Determination of Chloramphenicol, Dexamethasone Sodium Phosphate and Tetrahydrozoline Hydrochloride in Ophthalmic Solution

    PubMed Central

    AlAani, Hashem; Alnukkary, Yasmin

    2016-01-01

    Purpose: A simple stability-indicating RP-HPLC assay method was developed and validated for quantitative determination of Chloramphenicol, Dexamethasone Sodium Phosphate and Tetrahydrozoline Hydrochloride in ophthalmic solution in the presence of 2-amino-1-(4-nitrophenyl)propane-1,3-diol, a degradation product of Chloramphenicol, and Dexamethasone, a degradation product of Dexamethasone Sodium Phosphate. Methods: Effective chromatographic separation was achieved using C18 column (250 mm, 4.6 mm i.d., 5 μm) with isocratic mobile phase consisting of acetonitrile - phosphate buffer (pH 4.0; 0.05 M) (30:70, v/v) at a flow rate of 1 mL/minute. The column temperature was maintained at 40°C and the detection wavelength was 230 nm. Results: The proposed HPLC procedure was statistically validated according to the ICH guideline, and was proved to be stability-indicating by resolution of the APIs from their forced degradation products. Conclusion: The developed method is suitable for the routine analysis as well as stability studies. PMID:27123429

  15. Green synthesized gold nanoparticles decorated graphene oxide for sensitive determination of chloramphenicol in milk, powdered milk, honey and eye drops.

    PubMed

    Karthik, R; Govindasamy, Mani; Chen, Shen-Ming; Mani, Veerappan; Lou, Bih-Show; Devasenathipathy, Rajkumar; Hou, Yu-Shen; Elangovan, A

    2016-08-01

    A simple and rapid green synthesis using Bischofia javanica Blume leaves as reducing agent was developed for the preparation of gold nanoparticles (AuNPs). AuNPs decorated graphene oxide (AuNPs/GO) was prepared and employed for the sensitive amperometric determination of chloramphenicol. The green biosynthesis requires less than 40s to reduce gold salts to AuNPs. The formations of AuNPs and AuNPs/GO were evaluated by scanning electron and atomic force microscopies, UV-Visible and energy dispersive X-ray spectroscopies, X-ray diffraction studies, and electrochemical methods. AuNPs/GO composite film modified electrode was fabricated and shown excellent electrocatalytic ability towards chloramphenicol. Under optimal conditions, the amperometric sensing platform has delivered wide linear range of 1.5-2.95μM, low detection limit of 0.25μM and high sensitivity of 3.81μAμM(-1)cm(-2). The developed sensor exhibited good repeatability and reproducibility, anti-interference ability and long-term storage stability. Practical feasibility of the sensor has been demonstrated in food samples (milk, powdered milk and honey) and pharmaceutical sample (eye drops). The green synthesized AuNPs/GO composite has great potential for analysis of food samples in food safety measures.

  16. Rapid detection of chloramphenicol in animal products without clean-up using LC-high resolution mass spectrometry.

    PubMed

    Xu, Hong; Zhang, Jun; He, Jia; Mi, Jibo; Liu, Lishuang

    2011-10-01

    A rapid confirmatory and quantitative method using liquid chromatography-high resolution mass spectrometry (LC-HRMS) was developed to determine sub-µg/kg levels of chloramphenicol (CAP) in meat products. The sample plus deuterated chloramphenicol internal standard was homogenised, extracted with ethyl acetate, centrifuged and the supernatant evaporated to dryness. The residue was re-dissolved in methanol/5% ammonium acetate solution (20:80, v/v), defatted with hexane and directly injected into the LC-MS. Chromatographic separation was performed on a C(18) column using methanol/water (60:40, v/v) as the mobile phase. CAP was detected using selected ion monitoring of the high accurate mass of the molecular ion [M-H](-) of CAP using a LTQ-Orbitrap mass spectrometer in negative electrospray ionisation mode (ESI(-)). The limit of quantification of the method was 0.1 µg/kg using isotope internal standard. Recoveries of CAP spiked at levels of 0.1-1.0 µg/kg ranged from 73 to 99%, and the relative standard deviation ranged 3.9-8.1%.

  17. Antimicrobial susceptibility testing of pneumococci: determination of Kirby-Bauer breakpoints for penicillin G, erythromycin, clindamycin, tetracycline, chloramphenicol, and rifampin.

    PubMed

    Jacobs, M R; Mithal, Y; Robins-Browne, R M; Gaspar, M N; Koornhof, H J

    1979-08-01

    Antimicrobial susceptibility testing of pneumococci is now essential to monitor for the presence of resistance to agents such as the penicillins, macrolides, lincomycins, chloramphenicol, and tetracycline. In this study, clinical isolates of a selection of resistant South African strains were tested for antimicrobial susceptibility by minimal inhibitory concentration (MIC) determination and by a modified Kirby-Bauer disk diffusion technique, using Mueller-Hinton medium supplemented with 5% horse blood. Disk diffusion breakpoints were determined for penicillin G, erythromycin, clindamycin, tetracycline, chloramphenicol, and rifampin. Reliable results were obtained on disk diffusion for all these agents except for penicillin G. With 6-mug penicillin G disks, zones of strains with intermediate penicillin susceptibility overlapped those of sensitive and resistant strains. With 5-mug methicillin disks, clearer separation of strains based on susceptibility to penicillin G occurred. Strains with zones of <35 mm around penicillin G disks and <25 mm around methicillin disks should have penicillin G MICs determined to confirm their resistance to penicillin G. In view of the potential for pneumococci to be resistant to the agents used in this study, antimicrobial susceptibility of all clinically significant isolates should be determined.

  18. Enhanced inhibition of bacterial biofilm formation and reduced leukocyte toxicity by chloramphenicol:β-cyclodextrin:N-acetylcysteine complex.

    PubMed

    Aiassa, Virginia; Zoppi, Ariana; Becerra, M Cecilia; Albesa, Inés; Longhi, Marcela R

    2016-11-05

    The purpose of this study was to improve the physicochemical and biological properties of chloramphenicol (CP) by multicomponent complexation with β-cyclodextrin (β-CD) and N-acetylcysteine (NAC). The present work describes the ability of solid multicomponent complex (MC) to decrease biomass and cellular activity of Staphylococcus by crystal violet and XTT assay, and leukocyte toxicity, measuring the increase of reactive oxygen species by chemiluminescence, and using 123-dihydrorhodamine. In addition, MC was prepared by the freeze-drying or physical mixture methods, and then characterized by scanning electron microscopy and powder X-ray diffraction. Nuclear magnetic resonance and phase solubility studies provided information at the molecular level on the structure of the MC and its association binding constants, respectively. The results obtained allowed us to conclude that MC formation is an effective pharmaceutical strategy that can reduce CP toxicity against leukocytes, while enhancing its solubility and antibiofilm activity.

  19. Removal of chloramphenicol by macroporous adsorption resins in honey: a novel approach on reutilization of antibiotics contaminated honey.

    PubMed

    Cheng, Ni; Gao, Hui; Deng, Jianjun; Wang, Bini; Xu, Ruihan; Cao, Wei

    2012-09-01

    The effects of different steps in honey production on chloramphenicol (CAP) levels and CAP removal from honey using macroporous adsorption resins (MARs) were investigated in this study. CAP residues in honey were quantified by enzyme-linked immunosorbent assay after each processing step including preheating, filtration, vacuum concentration and pasteurization. Vacuum concentration contributes the most reduction of CAP level (9.9%). Meanwhile, 5 types of MARs (including LSI-1, LSI-2, LSI-3, LS-803, and LS-903) were used in CAP adsorption. The results showed that LS-803 resin had higher adsorption rate of 86% than other resins in removing CAP from honey, and its optimal adsorption time and temperature were 40 min and 55 °C, respectively. The treated honey could be used as feed additive or biomass energy. Therefore, it would be a novel approach to reutilization of antibiotics contaminated honey.

  20. Simultaneous spectrophotometric-multivariate calibration determination of several components of ophthalmic solutions: phenylephrine, chloramphenicol, antipyrine, methylparaben and thimerosal.

    PubMed

    Collado, M S; Mantovani, V E; Goicoechea, H C; Olivieri, A C

    2000-08-16

    The use of multivariate spectrophotometric calibration for the simultaneous determination of several active components and excipients in ophthalmic solutions is presented. The resolution of five-component mixtures of phenylephrine, chloramphenicol, antipyrine, methylparaben and thimerosal has been accomplished by using partial least-squares (PLS-1) and a variant of the so-called hybrid linear analysis (HLA). Notwithstanding the presence of a large number of components and their high degree of spectral overlap, they have been determined simultaneously with high accuracy and precision, with no interference, rapidly and without resorting to extraction procedures using non aqueous solvents. A simple and fast method for wavelength selection in the calibration step is presented, based on the minimisation of the predicted error sum of squares (PRESS) calculated as a function of a moving spectral window.

  1. Purification and structural characterization of a novel antibacterial peptide from Bellamya bengalensis: activity against ampicillin and chloramphenicol resistant Staphylococcus epidermidis.

    PubMed

    Gauri, Samiran S; Mandal, Santi M; Pati, Bikas R; Dey, Satyahari

    2011-04-01

    Increasing tendency of clinical bacterial strains resistant to conventional antibiotics has being a great challenge to the public's health. Antimicrobial peptides, a new class of antibiotics is known to have the activity against a wide range of bacteria resistant to conventional antibiotics. An antimicrobial peptide of 1676 Da was purified from Bellamya bengalensis, a fresh water snail, using ultrafiltration and reversed phase liquid chromatography. The effect of this peptide on Staphylococcus epidermidis resistant to ampicillin and chloramphenicol was investigated; the MIC and MBC values were 8 μg/ml and 16 μg/ml, respectively. Complete sequence of the peptide was determined by tandem mass spectrometry (MS/MS). Further, peptide net charge, hydrophobicity and molecular modeling were evaluated in silico for better understanding the probable mechanisms of action. The peptide showed the specificity to bacterial membranes. Hence, this reported peptide revealed a promising candidate to contribute in the development of therapeutic agent for Staphylococcal infections.

  2. Stimulation of chloramphenicol acetyltransferase mRNA translation by reovirus capsid polypeptide sigma 3 in cotransfected COS cells.

    PubMed Central

    Giantini, M; Shatkin, A J

    1989-01-01

    The mammalian reovirus S4 gene has been implicated in the serotype-dependent inhibition of host cell protein synthesis during viral replication in mouse L cells. To examine the effect(s) of this gene on transcription or translation or both, a DNA copy of the serotype 3 S4 gene was inserted into a eucaryotic expression vector. Cotransfection of COS cells with plasmids containing S4 and the reporter gene, chloramphenicol acetyltransferase (CAT), resulted in a marked stimulation of CAT expression, predominantly at the level of translation. The significance of these findings is discussed in relation to the double-stranded-RNA-binding activity of the S4 gene product, polypeptide sigma 3. Images PMID:2724407

  3. Chloramphenicol versus ampicillin plus gentamicin for community acquired very severe pneumonia among children aged 2-59 months in low resource settings: multicentre randomised controlled trial (SPEAR study)

    PubMed Central

    2008-01-01

    Objective To evaluate whether five days’ treatment with injectable ampicillin plus gentamicin compared with chloramphenicol reduces treatment failure in children aged 2-59 months with community acquired very severe pneumonia in low resource settings. Design Open label randomised controlled trial. Setting Inpatient wards within tertiary care hospitals in Bangladesh, Ecuador, India, Mexico, Pakistan, Yemen, and Zambia. Participants Children aged 2-59 months with WHO defined very severe pneumonia. Intervention Chloramphenicol versus a combination of ampicillin plus gentamicin. Main outcome measures Primary outcome measure was treatment failure at five days. Secondary outcomes were treatment failure defined similarly among all participants evaluated at 48 hours and at 10 and 21 days. Results More children failed treatment with chloramphenicol at day 5 (16% v 11%; relative risk 1.43, 95% confidence interval 1.03 to 1.97) and also by days 10 and 21. Overall, 112 bacterial isolates were obtained from blood and lung aspirates in 110 children (11.5%), with the most common organisms being Staphylococcus aureus (n=47) and Streptococcus pneumoniae (n=22). In subgroup analysis, bacteraemia with any organism increased the risk of treatment failure at 21 days in the chloramphenicol group (2.09, 1.41 to 3.10) but not in the ampicillin plus gentamicin group (1.12, 0.59 to 2.13). Similarly, isolation of S pneumoniae increased the risk of treatment failure at day 21 (4.06, 2.73 to 6.03) and death (5.80, 2.62 to 12.85) in the chloramphenicol group but not in the ampicillin plus gentamicin group. No difference was found in treatment failure for children with S aureus bacteraemia in the two groups, but the power to detect a difference in this subgroup analysis was low. Independent predictors of treatment failure by multivariate analysis were hypoxaemia (oxygen saturation <90%), receiving chloramphenicol, being female, and poor immunisation status. Conclusion Injectable ampicillin plus

  4. Selective inhibition by chloramphenicol of pregnenolone-16. cap alpha. -carbonitrile-inducible rat liver cytochrome P-450 isozymes

    SciTech Connect

    Graves, P.E.; Kaminsky, L.S.; Halpert, J.

    1986-03-01

    Pregnenolone-16 ..cap alpha..-carbonitrile (PCN) has been shown to induce, in male rats, cytochrome P-450 isozymes responsible for the formation of R-10-hydroxywarfarin and R-dehydrowarfarin. Antibodies to the major PCN-inducible isozyme (PB/PCN-E) inhibit both activities in microsomal preparations. Recently the authors have shown that PCN treatment of female rats also induces the formation of both R-warfarin metabolites. However, in both sexes chloramphenicol (CAP) treatment selectively inhibits only the rate of formation of the R-dehydrowarfarin. A decrease in microsomal P-450 content occurs after in vivo administration of CAP to PCN-treated rats of both sexes. This is in contrast to the lack of effect of CAP on P-450 levels in phenobarbital-treated rats. Covalent binding of /sup 14/C-CAP to microsomal protein in vitro was increased 3 to 4-fold following PCN treatment. Chromatographic evidences suggests the presence of at least two PCN-induced isozymes of similar molecular weights in both male and female rat liver microsomes. These data are consistent with the multiplicity of PCN-inducible P-450 in rat liver.

  5. Contribution of the Antibiotic Chloramphenicol and Its Analogues as Precursors of Dichloroacetamide and Other Disinfection Byproducts in Drinking Water.

    PubMed

    Chu, Wenhai; Krasner, Stuart W; Gao, Naiyun; Templeton, Michael R; Yin, Daqiang

    2016-01-05

    Dichloroacetamide (DCAcAm), a disinfection byproduct, has been detected in drinking water. Previous research showed that amino acids may be DCAcAm precursors. However, other precursors may be present. This study explored the contribution of the antibiotic chloramphenicol (CAP) and two of its analogues (thiamphenicol, TAP; florfenicol, FF) (referred to collectively as CAPs), which occur in wastewater-impacted source waters, to the formation of DCAcAm. Their formation yields were compared to free and combined amino acids, and they were investigated in filtered waters from drinking-water-treatment plants, heavily wastewater-impacted natural waters, and secondary effluents from wastewater treatment plants. CAPs had greater DCAcAm formation potential than two representative amino acid precursors. However, in drinking waters with ng/L levels of CAPs, they will not contribute as much to DCAcAm formation as the μg/L levels of amino acids. Also, the effect of advanced oxidation processes (AOPs) on DCAcAm formation from CAPs in real water samples during subsequent chlorination was evaluated. Preoxidation of CAPs with AOPs reduced the formation of DCAcAm during postchlorination. The results of this study suggest that CAPs should be considered as possible precursors of DCAcAm, especially in heavily wastewater-impacted waters.

  6. Cometabolic degradation of chloramphenicol via a meta-cleavage pathway in a microbial fuel cell and its microbial community.

    PubMed

    Zhang, Qinghua; Zhang, Yanyan; Li, Daping

    2017-04-01

    The performance of a microbial fuel cell (MFC) in terms of degradation of chloramphenicol (CAP) was investigated. Approximately 84% of 50mg/L CAP was degraded within 12h in the MFC. A significant interaction of pH, temperature, and initial CAP concentration was found on removal of CAP, and a maximum degradation rate of 96.53% could theoretically be achieved at 31.48°C, a pH of 7.12, and an initial CAP concentration of 106.37mg/L. Moreover, CAP was further degraded through a ring-cleavage pathway. The antibacterial activity of CAP towards Escherichia coli ATCC 25922 and Shewanella oneidensis MR-1 was largely eliminated by MFC treatment. High-throughput sequencing analysis indicated that Azonexus, Comamonas, Nitrososphaera, Chryseobacterium, Azoarcus, Rhodococcus, and Dysgonomonas were the predominant genera in the MFC anode biofilm. In conclusion, the MFC shows potential for the treatment of antibiotic residue-containing wastewater due to its high rates of CAP removal and energy recovery.

  7. Synergy against extensively drug-resistant Acinetobacter baumannii in vitro by two old antibiotics: colistin and chloramphenicol.

    PubMed

    Wei, Wen-Juan; Yang, Hai-Fei

    2017-03-01

    Combination antimicrobial therapy is an important option in the fight against Gram-negative 'superbugs'. This study systematically investigated the synergistic effect of colistin (CST) and chloramphenicol (CHL) in combination against extensively drug-resistant Acinetobacter baumannii (XDR-AB). The microtitre plate chequerboard assay was used to test synergy against 50 XDR-AB clinical strains. Then, three XDR-AB clinical isolates and the type strain A. baumannii ATCC 19606 were chosen for further synergy studies using time-kill assay, mutant prevention concentration (MPC) assay and real-time population analysis profile (PAP) assay. In the chequerboard assays, synergistic or additive effects [defined as a fractional inhibitory concentration index (FICI) of ≤0.5 and 0.5 < FICI < 1, respectively] were observed in all 50 isolates. In further synergy testing, the results of time-kill assays indicated that CST monotherapy produced rapid bacterial killing followed by rapid re-growth, with the emergence of CST resistance; CHL monotherapy was largely ineffective. The combination CST/CHL, however, showed a synergistic effect and enhanced bacterial killing in the four tested strains. It also significantly delayed re-growth and suppressed the emergence of CST resistance. In the MPC assay, a decrease in MPCs for CST was observed in the two CST-susceptible strains. PAP assay showed that both CST-resistant strains were heteroresistant.

  8. A label-free photoelectrochemical aptasensor based on nitrogen-doped graphene quantum dots for chloramphenicol determination.

    PubMed

    Liu, Yong; Yan, Kai; Okoth, Otieno Kevin; Zhang, Jingdong

    2015-12-15

    A photoelectrochemical (PEC) sensing platform for chloramphenicol (CAP) detection was constructed using nitrogen-doped graphene quantum dots (N-GQDs) as transducer species and label-free aptamer as biological recognition element. N-GQDs, synthesized via a facile one-step hydrothermal method, were explored to achieve highly efficient photon-to-electricity conversion under visible light irradiation. The obtained N-GQDs were characterized by transmission electron microscopy (TEM), which displayed a narrow size distribution with a mean diameter of 2.14 nm. The X-ray photoelectron spectroscopic (XPS) and Fourier transform infrared spectroscopic (FT-IR) analysis confirmed that nitrogen was successfully doped in GQDs. The UV-visible absorption spectra indicated that nitrogen doping obviously enhanced the absorption of GQDs in visible light region. As a result, the PEC activity of GQDs was promoted by nitrogen doping. Additionally, the π-conjugated structure of N-GQDs provided an excellent platform for aptamer immobilization via π-π stacking interaction. Such an aptamer/N-GQDs based sensor showed a linear PEC response to CAP concentration in the range of 10-250 nM with a detection limit (3 S/N) of 3.1 nM. The developed PEC aptasensor exhibited high sensitivity and selectivity, good reproducibility and high stability.

  9. Design of elution strategy for simultaneous detection of chloramphenicol and gentamicin in complex samples using surface plasmon resonance.

    PubMed

    Xia, Yinqiang; Su, Rongxin; Huang, Renliang; Ding, Li; Wang, Libing; Qi, Wei; He, Zhimin

    2017-06-15

    For the analysis of massive samples containing multiple analytes, the enhancement of detection efficiency is crucial. In this study, a facile method was developed for sequential detection of chloramphenicol (CAP) and gentamicin (GEN) in complex samples, e.g. milk, using a surface plasmon resonance (SPR)-based biosensor. Based on the immune inhibition format, two conjugates of antigen and bovine serum albumin (BSA)-denoted as CAP-BSA and GEN-BSA-were grafted on the same channel of the SPR sensor chip. Two standard curves for CAP and GEN were separately obtained by first mixing a single antibody with different concentrations of the relevant antigen. Moreover, different regeneration solutions were screened for sequential analysis. An alkaline solution was found to completely remove the antibody against GEN (AbGEN) from the chip, but it exhibited limited ability to dissociate the antibody against CAP (AbCAP). Therefore, alkaline solution and Gly-HCl solutions are successively applied to elute AbGEN and AbCAP, respectively. By gradual elutions, CAP and GEN concentrations were simultaneously calculated with limit of detection values of 5.28ng/mL and 2.26ng/mL, respectively. Furthermore, the spiking milk samples with CAP and GEN validated the assay with recoveries of 77.6-101.1%. Therefore, this method is expected to improve the detection efficiency of SPR biosensors.

  10. A sensitive fluorescent nanosensor for chloramphenicol based on molecularly imprinted polymer-capped CdTe quantum dots.

    PubMed

    Amjadi, Mohammad; Jalili, Roghayeh; Manzoori, Jamshid L

    2016-05-01

    A novel fluorescent nanosensor using molecularly imprinted silica nanospheres embedded CdTe quantum dots (CdTe@SiO2 @MIP) was developed for detection and quantification of chloramphenicol (CAP). The imprinted sensor was prepared by synthesis of molecularly imprinting polymer (MIP) on the hydrophilic CdTe quantum dots via reverse microemulsion method using small amounts of solvents. The resulting CdTe@SiO2 @MIP nanoparticles were characterized by fluorescence, UV-vis absorption and FT-IR spectroscopy and transmission electron microscopy. They preserved 48% of fluorescence quantum yield of the parent quantum dots. CAP remarkably quenched the fluorescence of prepared CdTe@SiO2 @MIP, probably via electron transfer mechanism. Under the optimal conditions, the relative fluorescence intensity of CdTe@SiO2 @MIP decreased with increasing CAP by a Stern-Volmer type equation in the concentration range of 40-500 µg L(-1). The corresponding detection limit was 5.0 µg L(-1). The intra-day and inter-day values for the precision of the proposed method were all <4%. The developed sensor had a good selectivity and was applied to determine CAP in spiked human and bovine serum and milk samples with satisfactory results.

  11. Stimulation of Ribonucleic Acid Synthesis by Chloramphenicol in a rel+ Aminoacyl-Transfer Ribonucleic Acid Synthetase Mutant of Escherichia coli

    PubMed Central

    Yegian, Charles D.; Vanderslice, Rebecca W.

    1971-01-01

    Escherichia coli strain 9D3 possesses a highly temperature-sensitive valyl-transfer ribonucleic acid (tRNA) synthetase (EC 6.1.1.9). Since 9D3 is a rel+ strain, it cannot carry out net RNA synthesis at high temperature. A 100-μg amount of chloramphenicol (CAP) per ml added in the absence of valine cannot stimulate RNA synthesis. Either 300 μg of CAP or 100 μg of CAP plus 50 μg of valine per ml, however, promotes nearly maximal RNA synthesis. These results can be understood as follows. (i) Valyl-tRNA is required for net RNA synthesis, (ii) the synthetase lesion is incomplete, (iii) the rate of mutant acylation of tRNAval at high temperature is valine-dependent, and (iv) the CAP concentration determines the rate of residual protein synthesis. Data are also presented which demonstrate that the rate of net RNA synthesis can greatly increase long after the addition of CAP, if the amount of valyl-tRNA increases. PMID:4942766

  12. Genetic basis of the association of resistance genes mef(I) (macrolides) and catQ (chloramphenicol) in streptococci.

    PubMed

    Mingoia, Marina; Morici, Eleonora; Brenciani, Andrea; Giovanetti, Eleonora; Varaldo, Pietro E

    2014-01-01

    In streptococci mef(I) and catQ, two relatively uncommon macrolide and chloramphenicol resistance genes, respectively, are typically linked in a genetic module designated IQ module. Though variable, the module consistently encompasses, and is sometimes reduced to, a conserved ∼5.8-kb mef(I)-catQ fragment. The prototype IQ module was described in Streptococcus pneumoniae. IQ-like modules have subsequently been detected in Streptococcus pyogenes and in different species of viridans group streptococci, where mef(E) may be found instead of mef(I). Three genetic elements, one carrying the prototype IQ module from S. pneumoniae and two carrying different, defective IQ modules from S. pyogenes, have recently been characterized. All are integrative and conjugative elements (ICEs) belonging to the Tn5253 family, and have been designated ICESpn529IQ, ICESpy029IQ and ICESpy005IQ, respectively. ICESpy029IQ and ICESpy005IQ were the first Tn5253 family ICEs to be described in S. pyogenes. A wealth of new information has been obtained by comparing their genetic organization, chromosomal integration, and transferability. The origin of the IQ module is unknown. The mechanism by which it spreads in streptococci is discussed.

  13. Degradation and antibiotic activity reduction of chloramphenicol in aqueous solution by UV/H2O2 process.

    PubMed

    Zuorro, Antonio; Fidaleo, Marco; Fidaleo, Marcello; Lavecchia, Roberto

    2014-01-15

    The efficacy of the UV/H2O2 process to degrade the antibiotic chloramphenicol (CHL) was investigated at 20 °C using a low-pressure mercury lamp as UV source. A preliminary analysis of CHL degradation showed that the process followed apparent first-order kinetics and that an optimum H2O2 concentration existed for the degradation rate. The first-order rate constant was used as the response variable and its dependence on initial CHL and H2O2 concentrations, UV light intensity and reaction time was investigated by a central composite design based on the response surface methodology. Analysis of response surface plots revealed a large positive effect of radiation intensity, a negative effect of CHL concentration and that there was a region of H2O2 concentration leading to maximum CHL degradation. CHL solutions submitted to the UV/H2O2 process were characterized by TOC and their activity against Escherichia coli and Staphylococcus aureus was assessed. No residual antibiotic activity was detected, even at CHL concentrations higher than those used in the designed experiments. Overall, the obtained results strongly support the possibility of reducing the risks associated with the release of CHL into the environment, including the spread of antibiotic resistance, by the UV/H2O2 process.

  14. Development of an Immunoassay for Chloramphenicol Based on the Preparation of a Specific Single-Chain Variable Fragment Antibody.

    PubMed

    Du, Xin-jun; Zhou, Xiao-nan; Li, Ping; Sheng, Wei; Ducancel, Frédéric; Wang, Shuo

    2016-04-13

    Specific antibodies are essential for the immune detection of small molecule contaminants. In the present study, the heavy and light variable regions (V(H )and V(L)) of the immunoglobulin genes from a hybridoma secreting a chloramphenicol (CAP)-specific monoclonal antibody (mAb) were cloned and sequenced. In addition, the light and heavy chains obtained from the monoclonal antibody were separated using SDS-PAGE and analyzed using Orbitrap mass spectrometry. The results of DNA sequencing and mass spectrometry analysis were compared, and the V(H) and V(L) chains specific for CAP were determined and used to construct a single-chain variable fragment (scFv). This fragment was recombinantly expressed as a soluble scFv-alkaline phosphatase fusion protein and used to develop a direct competitive ELISA. Compared with the parent mAb, scFv exhibits lower sensitivity but better food matrix resistance. This work highlights the application of engineered antibodies for CAP detection.

  15. [Modification of cell damage, caused by variable rate heating, by means of changes in osmotic pressure of the medium or using chloramphenicol].

    PubMed

    Morozov, I I; Petin, V G

    1998-01-01

    We investigated the influence of media with different osmotic pressure and of chloramphenicol on cytotoxic effects of heating with different rate of Escherichia coli B/r and Escherichia coli Bs-1 bacteria, and Zygosaccharomyces bailii yeast cells. It was shown that the hypotonic media appreciably increased cytotoxic action of heating with different rate, and, on the contrary, the hypertonic media inhibited induction of these effects. The inhibitor of protein synthesis chloramphenicol was established not to affect the bacterial thermoresistance in the process of different heating rate. On the basis of analysis of the obtained and literature data, it is proposed that a reason of cell injury dependence of the heating rate may be the availability of dissimilar levels of osmotic homeostasis destabilization in these cells when heated with different rates.

  16. Transcription factor CecR (YbiH) regulates a set of genes affecting the sensitivity of Escherichia coli against cefoperazone and chloramphenicol.

    PubMed

    Yamanaka, Yuki; Shimada, Tomohiro; Yamamoto, Kaneyoshi; Ishihama, Akira

    2016-07-01

    Genomic SELEX (systematic evolution of ligands by exponential enrichment) screening was performed for identification of the binding site of YbiH, an as yet uncharacterized TetR-family transcription factor, on the Escherichia coli genome. YbiH was found to be a unique single-target regulator that binds in vitro within the intergenic spacer located between the divergently transcribed ybiH-ybhGFSR and rhlE operons. YbhG is an inner membrane protein and YbhFSR forms a membrane-associated ATP-binding cassette (ABC) transporter while RhlE is a ribosome-associated RNA helicase. Gel shift assay and DNase footprinting analyses indicated one clear binding site of YbiH, including a complete palindromic sequence of AATTAGTT-AACTAATT. An in vivo reporter assay indicated repression of the ybiH operon and activation of the rhlE operon by YbiH. After phenotype microarray screening, YbiH was indicated to confer resistance to chloramphenicol and cefazoline (a first-generation cephalosporin). A systematic survey of the participation of each of the predicted YbiH-regulated genes in the antibiotic sensitivity indicated involvement of the YbhFSR ABC-type transporter in the sensitivity to cefoperazone (a third-generation cephalosporin) and of the membrane protein YbhG in the control of sensitivity to chloramphenicol. Taken together with the growth test in the presence of these two antibiotics and in vitro transcription assay, it was concluded that the hitherto uncharacterized YbiH regulates transcription of both the bidirectional transcription units, the ybiH-ybhGFSR operon and the rhlE gene, which altogether are involved in the control of sensitivity to cefoperazone and chloramphenicol. We thus propose to rename YbiH as CecR (regulator of cefoperazone and chloramphenicol sensitivity).

  17. Photocatalytic degradation of chloramphenicol in an aqueous suspension of silver-doped TiO2 nanoparticles.

    PubMed

    Shokri, Mohammad; Jodat, Akbar; Modirshahla, Nasser; Behnajady, Mohammad A

    2013-01-01

    In this work, silver-doped TiO2 (Ag/TiO2) nanoparticles were synthesized using a photodeposition technique. The prepared Ag/TiO2 nanoparticles were characterized using TEM, SEM, XRD, and EDX techniques. The characterization of Ag/TiO2 nanoparticles using SEM and EDX techniques revealed the dispersion of Ag metal on the surface of TiO2. The photocatalytic activity of Ag/TiO2 and bare TiO2 in the presence of ultraviolet irradiation was investigated in the removal of chloramphenicol (CAP) as an antibiotic. CAP is a broad-spectrum antibiotic exhibiting activity against both Gram-positive and Gram-negative bacteria, as well as other groups of microorganisms. However, it is, in certain susceptible individuals, associated with serious toxic effects in humans including bone marrow depression, particularly severe in the form of fatal aplastic anaemia. The effects of the operational factors, such as doping content of Ag, photocatalyst dosage and calcination temperature were evaluated in the catalytic activity of Ag/TiO2. The results showed that the photocatalytic efficiency of TiO2 nanoparticles for the degradation of CAP, can be significantly improved by deposition an optimum amount of Ag nanoparticles (0.96 wt%) in the calcination temperature 300 degrees C. It was found that 900 mg/L of Ag/TiO2 is the optimum dosage in the removal of CAP with 20 mg/L initial concentration. The highest removal efficiency of CAP (-100%) at the optimum conditions was observed in 20 min. A mineralization study under optimum conditions showed about 88% reduction in total organic carbon after 120 min of irradiation time.

  18. Successful co-encapsulation of benzoyl peroxide and chloramphenicol in liposomes by a novel manufacturing method - dual asymmetric centrifugation.

    PubMed

    Ingebrigtsen, Sveinung G; Škalko-Basnet, Nataša; de Albuquerque Cavalcanti Jacobsen, Cristiane; Holsæter, Ann Mari

    2017-01-15

    Encapsulation of more than one active pharmaceutical ingredient into nanocarriers such as liposomes is an attractive approach to achieve a synergic drug effect and less complicated dosing schedules in multi-drug treatment regimes. Liposomal drug delivery in acne treatment may improve drug efficiency by targeted delivery to pilosebaceous units, reduce adverse effects and improve patient compliance. We therefore aimed to co-encapsulate benzoyl peroxide (BPO) and chloramphenicol (CAM) into liposomes using the novel liposome processing method - dual asymmetric centrifugation (DAC). Liposomes were formed from soybean lecithin, propylene glycol and distilled water (2:1:2w/v/v ratio), forming a viscous liposome dispersion. Liposomes containing both drugs (BPO-CAM-Lip), single drug (BPO-Lip and CAM-Lip), and empty liposomes were prepared. Drug entrapment of BPO and CAM was determined by a newly developed HPLC method for simultaneous detection and quantification of both drugs. Encapsulation of around 50% for BPO and 60% for CAM respectively was obtained in both single-drug encapsulated formulations (BPO-Lip and CAM-Lip) and co-encapsulated formulations (BPO-CAM-Lip). Liposome sizes were comparable for all liposome formulations, ranging from 130 to 150nm mean diameter, with a polydispersity index <0.2 for all formulations. CAM exhibited a sustained release from all liposomal formulations, whereas BPO appeared retained within the liposomes. BPO retention could be attributed to its poor solubility. However, HaCaT cell toxicity was found dependent on BPO released from the liposomes. In the higher concentration range (4%v/v), liposomal formulations were less cytotoxic than the corresponding drug solutions used as reference. We have demonstrated that DAC is a fast, easy, suitable method for encapsulation of more than one drug within the same liposomes.

  19. The pdx genetic marker adjacent to the chloramphenicol biosynthesis gene cluster in Streptomyces venezuelae ISP5230: functional characterization.

    PubMed

    Magarvey, N; He, J; Aidoo, K A; Vining, L C

    2001-08-01

    The pdx-4 mutation in Streptomyces venezuelae ISP5230 confers a growth requirement for pyridoxal (pdx) and is a marker for the genetically mapped cluster of genes associated with chloramphenicol biosynthesis. A gene regulating salvage synthesis of vitamin B6 cofactors in S. venezuelae was cloned by transforming a pdx-4 mutant host with the plasmid vector pDQ101 carrying a library of wild-type genomic DNA fragments, and by selecting for complementation of the host's pdx requirement. However, the corresponding replicative plasmid could not be isolated. Southern hybridizations and transduction analysis indicated that the complementing plasmid had integrated into the chromosome; after excision by a second crossover, the plasmid failed to propagate. To avoid loss of the recombinant vector, a pdx-dependent Streptomyces lividans mutant, KAA1, with a phenotype matching that of S. venezuelae pdx-4, was isolated for use as the cloning host. Introduction of pIJ702 carrying an S. venezuelae genomic library into S. lividans KAA1, and selection of prototrophic transformants, led to the isolation of a stable recombinant vector containing a 2.5 kb S. venezuelae DNA fragment that complemented requirements for pdx in both S. venezuelae and S. lividans mutants. Sequence analysis of the cloned DNA located an intact ORF with a deduced amino acid sequence that, in its central and C-terminal regions resembled type-I aminotransferases. The N-terminal region of the cloned DNA fragment aligned closely with distinctive helix-turn-helix motifs found near the N termini of GntR family transcriptional regulators. The overall deduced amino acid sequence of the cloned DNA showed 73% end-to-end identity to a putative GntR-type regulator cloned in cosmid 6D7 from the Streptomyces coelicolor A3(2) genome. This location is close to that of pdxA, the first pdx marker in S. coelicolor A3(2) identified and mapped genetically in Sir David Hopwood's laboratory. The S. venezuelae gene and S. coelicolor pdx

  20. Chloramphenicol and tetracycline decrease motility and increase invasion and attachment gene expression in specific isolates of multidrug-resistant Salmonella enterica serovar Typhimurium

    PubMed Central

    Brunelle, Brian W.; Bearson, Bradley L.; Bearson, Shawn M. D.

    2015-01-01

    Salmonella enterica serovar Typhimurium is one of the most common serovars isolated from humans and livestock, and over 35% of these isolates are resistant to three or more antibiotics. Multidrug-resistant (MDR) Salmonella is a public health concern as it is associated with increased morbidity in patients compared to antibiotic sensitive strains, though it is unknown how the antibiotic resistant isolates lead to a more severe infection. Cellular invasion is temporally regulated in Salmonella and normally occurs during late-log and stationary growth. However, our previous work determined that a 30 min exposure to a sub-inhibitory concentration of tetracycline can induce the full invasion phenotype during early-log growth in certain MDR S. Typhimurium isolates. The current study examined whether sub-inhibitory concentrations of other antibiotics could also induce the invasiveness in the same set of isolates. Ampicillin and streptomycin had no effect on invasion, but certain concentrations of chloramphenicol were found to induce invasion in a subset of isolates. Two of the isolates induced by chloramphenicol were also inducible by tetracycline. RNA-seq analyses demonstrated that chloramphenicol and tetracycline both down-regulated motility gene expression, while up-regulating genes associated with attachment, invasion, and intracellular survival. Eleven fimbrial operons were up-regulated, which is notable as only three fimbrial operons were thought to be inducible in culture; six of these up-regulated operons have been reported to play a role in Salmonella persistence in mice. Overall, these data show that the normal progression of the genetic pathways that regulate invasion can be expedited to occur within 30 min due to antibiotic exposure. This altered invasion process due to antibiotics may play a role in the increased intensity and duration of infection observed in patients with MDR Salmonella. PMID:25688233

  1. A high-throughput method for determining chloramphenicol residues in poultry, egg, shrimp, fish, swine and bovine using LC-ESI-MS/MS.

    PubMed

    Rocha Siqueira, Silvia Regina; Luiz Donato, José; de Nucci, Gilberto; Reyes, Felix Guillermo Reyes

    2009-12-01

    A LC-ESI-MS/MS method for determining chloramphenicol residues in fish, shrimp, poultry, eggs, bovine and swine was developed and validated. The samples were extracted with a phosphate extraction solution followed by liquid-liquid extraction with ethyl acetate. The LC was performed on a C18 column at room temperature. For all the matrices, the analytical curves showed r values and recoveries higher than 0.99 and 50%, respectively. The accuracy values lay between 85 and 120% and the precision was lower than 20%. The LOQ was 0.1 ng/g. The method was employed to analyze samples collected in Brazil.

  2. Colorimetric determination of 1-(4'-nitrophenyl)-2-aminopropane-1,3-diol with 2,4,6-trinitrobenzenesulfonic acid in the presence of chloramphenicol.

    PubMed

    Pietta, P G; Agnellini, D; Pace, M

    1979-12-01

    A colorimetric method based on the interaction between the chloramphenicol degradation product 1-(4'-nitrophenyl)-2-aminopropane-1,3-diol and the 2,4,6-trinitrobenzenesulfonic acid reagent was developed. Analytical solutions were reacted with the reagent at pH 9.1 for 20 min at room temperature, and the resulting color was measured at 340 nm. A linear relationship between absorbance and concentration occurred within the 5--25-micrograms/ml range under the conditions studied. Replicate analyses were in good agreement. An average recovery of 99.4 +/- 0.4% was obtained for the synthetic mixtures.

  3. Simultaneous determination of nitroimidazoles, benzimidazoles, and chloramphenicol components in bovine milk by ultra-high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Wang, Yuanyuan; Li, Xiaowei; Zhang, Zhiwen; Ding, Shuangyang; Jiang, Haiyang; Li, Jiancheng; Shen, Jianzhong; Xia, Xi

    2016-02-01

    A sensitive, confirmatory ultra-high performance liquid chromatography-tandem mass spectrometric method was developed and validated to detect 23 veterinary drugs and metabolites (nitroimidazoles, benzimidazoles, and chloramphenicol components) in bovine milk. Compounds of interest were sequentially extracted from milk with acetonitrile and basified acetonitrile using sodium chloride to induce liquid-liquid partition. The extract was purified on a mixed mode solid-phase extraction cartridge. Using rapid polarity switching in electrospray ionization, a single injection was capable of detecting both positively and negatively charged analytes in a 9 min chromatography run time. Recoveries based on matrix-matched calibrations and isotope labeled internal standards for milk ranged from 51.7% to 101.8%. The detection limits and quantitation limits of the analytical method were found to be within the range of 2-20 ng/kg and 5-50 ng/kg, respectively. The recommended method is simple, specific, and reliable for the routine monitoring of nitroimidazoles, benzimidazoles, and chloramphenicol components in bovine milk samples.

  4. Simultaneous determination of chloramphenicol, dexamethasone and naphazoline in ternary and quaternary mixtures by RP-HPLC, derivative and wavelet transforms of UV ratio spectra

    NASA Astrophysics Data System (ADS)

    Hoang, Vu Dang; Hue, Nguyen Thu; Tho, Nguyen Huu; Nguyen, Hue Minh Thi

    2015-03-01

    The application of chemometrics-assisted UV spectrophotometry and RP-HPLC to the simultaneous determination of chloramphenicol, dexamethasone and naphazoline in ternary and quaternary mixtures is presented. The spectrophotometric procedure is based on the first-order derivative and wavelet transforms of ratio spectra using single, double and successive divisors. The ratio spectra were differentiated and smoothed using Savitzky-Golay filter; whereas wavelet transform realized with wavelet functions (i.e. db6, gaus5 and coif3) to obtain highest spectral recoveries. For the RP-HPLC procedure, the separation was achieved on a ZORBAX SB-C18 (150 × 4.6 mm; 5 μm) column at ambient temperature and the total run time was less than 7 min. A mixture of acetonitrile - 25 mM phosphate buffer pH 3 (27:73, v/v) was used as the mobile phase at a flow rate of 1.0 mL/min and the effluent monitored by measuring absorbance at 220 nm. Calibration graphs were established in the range 20-70 mg/L for chloramphenicol, 6-14 mg/L for dexamethasone and 3-8 mg/L for naphazoline (R2 > 0.990). The RP-HPLC and ratio spectra transformed by a combination of derivative-wavelet algorithms proved to be able to successfully determine all analytes in commercial eye drop formulations without sample matrix interference (mean percent recoveries, 97.4-104.3%).

  5. Comparison of different calibration approaches for chloramphenicol quantification in chicken muscle by ultra-high pressure liquid chromatography tandem mass spectrometry.

    PubMed

    Pan, Xiao-Dong; Jiang, Wei; Wu, Ping-Gu

    2015-01-07

    Matrix-dependent signal suppression often occurs in quantitative analysis by ultra-high pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS). In this study, we investigated three calibration methods for compensation of signal suppression on chloramphenicol (CAP) quantification in chicken muscle. The data showed that the spiking recoveries by solvent standard calibration with a stable isotope labelled internal standard (SIL-IS) and matrix-matched standard calibration with a SIL-IS were significantly higher than by external matrix-matched standard calibration (P < 0.05). When the SIL-IS was used, standards prepared in the mobile phase solvent showed no significant difference as those prepared in the matrix (P > 0.05). The limit of detection (LOD) for external matrix matched standard calibration was 0.1 μg kg(-1), and that for SIL-IS calibration (including matrix matched and solvent dissolved standard) was 0.03 μg kg(-1).

  6. In-vitro permeability of the human nail and of a keratin membrane from bovine hooves: penetration of chloramphenicol from lipophilic vehicles and a nail lacquer.

    PubMed

    Mertin, D; Lippold, B C

    1997-03-01

    Lipophilic vehicles and especially nail lacquers are more appropriate for topical application on the nail than aqueous systems because of their better adhesion. This work has, therefore, studied the penetration through the human nail plate of the model compound chloramphenicol from the lipophilic vehicles medium chain triglycerides and n-octanol and from a lacquer based on quaternary poly(methyl methacrylates) (Eudragit RL). The results were compared with data obtained with a keratin membrane from bovine hooves. If the swelling of the nail plate or the hoof membrane is not altered by use of lipophilic vehicles, the maximum flux of the drug is independent of its solubility in the vehicle and is the same as that from a saturated aqueous solution. These vehicles are not able to enter the hydrophilic keratin membrane because of their non-polar character and so cannot change the solubility of the penetrating substance in the barrier. If the concentration of the drug in the nail lacquer is sufficiently high, the maximum flux through both barriers equals that from aqueous vehicles or even exceeds it because of the formation of a supersaturated system. Penetration through the nail plate follows first order kinetics after a lag-time of 400 h. The course of penetration through the hoof membrane is initially membrane-controlled and later becomes a matrix-controlled process because of the membrane's greater permeability. Chloramphenicol is dissolved in the lacquer up to a concentration of 31%. The relative release rates from these solution matrices are independent of the drug concentration but they decrease on changing to a suspension matrix. These results show that drug flux is independent of the character of the vehicle and that penetration of the drug is initially membrane-controlled and changes to being matrix-controlled as the drug content of the lacquer decreases.

  7. Increased formation of halomethanes during chlorination of chloramphenicol in drinking water by UV irradiation, persulfate oxidation, and combined UV/persulfate pre-treatments.

    PubMed

    Wenhai, Chu; Tengfei, Chu; Erdeng, Du; Deng, Yang; Yingqing, Guo; Naiyun, Gao

    2016-02-01

    Ultraviolet/persulfate (UV/PS) has been widely used to generate sulfate radicals for degradation of water organic pollutants in previous studies. However, its impacts on disinfection byproduct formation during post-chlorination of degraded compounds is unclear. The objective of this study was to evaluate the impacts of UV irradiation, PS oxidation, and the combined UV/PS advanced oxidation process (AOP) pre-treatments on halomethane formation during the following chlorination of chloramphenicol (CAP), a model antibiotic commonly found in wastewater-impacted water. Results showed that CAP could be transformed to more trichloromethane (TCM) than monochloromethane (MCM) and dichloromethane (DCM) in the presence of excess chlorine. UV photolysis, PS oxidation and UV/PS AOP all directly decomposed CAP to produce halomethanes (HMs) before post-chlorination. Moreover, UV and UV/PS pre-treatments both enhanced the formation of all the HMs in the subsequent chlorination. PS pre-oxidation decreased the TCM formation during post-chlorination, but increased the yields of MCM, DCM and total HMs. UV pre-irradiation significantly increased the bromide utilization of HMs, whereas UV/PS pre-oxidation decreased the bromine incorporation and utilization of HMs from the chlorination of CAP in a low-bromide water. UV irradiation, PS oxidation, and UV/PS AOP can inactivate pathogens and degrade organic pollutants, but this benefit should be weighed against a potential risk of the increased halomethane formation from degraded organic pollutants with and without post-chlorination.

  8. Quantitative Analysis of Chloramphenicol in Royal Jelly by Column-switching LC-MS/MS Using a Pretreatment Column with a Higher-pressure Capability.

    PubMed

    Kawano, Shin-ichi; Hayakawa, Yoshihiro; Hashi, Yuki; Lin, Jin-Ming

    2015-01-01

    An on-line pretreatment liquid chromatography-tandem mass spectrometry (LC-MS/MS) system was developed for the analysis of chloramphenicol (CAP) in royal jelly. A novel methylcellulose-immobilized restricted access media column with a higher-pressure capability of 60 MPa (MC-ODS HP) was developed for the effective removal of proteins and other compounds in the sample matrix. CAP in a sample solution was extracted in 2 min by the column-switching LC-MS/MS system. The system provides a minimum sample pretreatment along with highly sensitive and reproducible analysis. As a result, the limit of quantitation of CAP was 10 pg/mL (= 0.1 μg/kg royal jelly) and the linear dynamic range was between 10 and 10000 pg/mL (correlation coefficient greater than 0.999). The proposed method meets the requirements of regulations in EU (0.3 μg/kg). The inter-day precision and accuracy of CAP at 100 pg/mL over 3 days were 4.5 and 95.4%, respectively. Compared with the conventional method with a pressure of below 25 MPa, the peak separation in the MRM chromatogram was improved by using smaller particles (1.6 μm) for the analytical ODS column. The LC-MS/MS system with an MC-ODS HP expanded the applicability of the automated pretreatment.

  9. In situ solvothermal growth of metal-organic framework-ionic liquid functionalized graphene nanocomposite for highly efficient enrichment of chloramphenicol and thiamphenicol.

    PubMed

    Wu, Mian; Ai, Youhong; Zeng, Baizhao; Zhao, Faqiong

    2016-01-04

    Here we report a facile in situ solvothermal growth method for immobilization of metal-organic framework-ionic liquid functionalized graphene (MOF-5/ILG) composite on etched stainless steel wire. The X-ray diffraction spectra, scanning electron microscopy and transmission electron microscopy images showed that the metal organic framework possessed good crystal shape and its structure was not disturbed by the introduction of ILG. Moreover, the covalent bond established between the amino group of ILG and the carboxylic group of the metal organic framework improved the mechanical stability and structure uniformity of the microcrystals. The obtained material combined the favorable attributes of both metal-organic framework and ILG, having high surface area (820 m(2)/g) and good adsorption capability. Its adsorption properties were explored by preconcentrating chloramphenicol and thiamphenicol from aqueous solutions prior to gas chromatography-flame ionization detection. The MOF-5/ILG exhibited high enrichment capacity for the analytes as they could interact through π-π and H-bonding interaction. Under the optimum conditions, good linearity (correlation coefficients higher than 0.9981), low limits of detection (14.8-19.5 ng/L), and good precision (relative standard deviations less than 6.0% (n=5)) were achieved. The MOF-5/ILG composite displayed durable property. The method was applied to the determination of two antibiotics in milk, honey, urine and serum samples with acceptable relative recoveries of 82.3-103.2%.

  10. Molybdenum disulfide nanosheets coated multiwalled carbon nanotubes composite for highly sensitive determination of chloramphenicol in food samples milk, honey and powdered milk.

    PubMed

    Govindasamy, Mani; Chen, Shen-Ming; Mani, Veerappan; Devasenathipathy, Rajkumar; Umamaheswari, Rajaji; Joseph Santhanaraj, K; Sathiyan, Anandaraj

    2017-01-01

    We have described a hybrid material that consists of molybdenum disulfide nanosheets (MoS2) coated on functionalized multiwalled carbon nanotubes (f-MWCNTs) for sensitive and selective determination of chloramphenicol (CAP). The MoS2/f-MWCNTs nanocomposite was successfully prepared through a hydrothermal process and its structure was characterized by scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, electrochemical impedance spectroscopy and cyclic voltammetry. The MoS2/f-MWCNTs nanocomposite holds excellent electrochemical properties and it displays excellent electrocatalytic ability to CAP. Under optimized working conditions, the nanocomposite film modified electrode responds linearly to CAP in the concentration range of 0.08-1392μM. The detection limit was obtained as 0.015μM (±0.003). The electrode has high level of selectivity in presence of large excess concentrations of interfering species. In addition, the modified electrode offers satisfactory repeatability, reproducibility and stability. The practical applicability of the electrode was demonstrated in food samples such as, milk, powdered milk and honey samples and the recoveries are agreeable which clearly revealed its practical feasibility in food analysis.

  11. Novel spectrophotometric determination of chloramphenicol and dexamethasone in the presence of non labeled interfering substances using univariate methods and multivariate regression model updating.

    PubMed

    Hegazy, Maha A; Lotfy, Hayam M; Rezk, Mamdouh R; Omran, Yasmin Rostom

    2015-04-05

    Smart and novel spectrophotometric and chemometric methods have been developed and validated for the simultaneous determination of a binary mixture of chloramphenicol (CPL) and dexamethasone sodium phosphate (DSP) in presence of interfering substances without prior separation. The first method depends upon derivative subtraction coupled with constant multiplication. The second one is ratio difference method at optimum wavelengths which were selected after applying derivative transformation method via multiplying by a decoding spectrum in order to cancel the contribution of non labeled interfering substances. The third method relies on partial least squares with regression model updating. They are so simple that they do not require any preliminary separation steps. Accuracy, precision and linearity ranges of these methods were determined. Moreover, specificity was assessed by analyzing synthetic mixtures of both drugs. The proposed methods were successfully applied for analysis of both drugs in their pharmaceutical formulation. The obtained results have been statistically compared to that of an official spectrophotometric method to give a conclusion that there is no significant difference between the proposed methods and the official ones with respect to accuracy and precision.

  12. Novel spectrophotometric determination of chloramphenicol and dexamethasone in the presence of non labeled interfering substances using univariate methods and multivariate regression model updating

    NASA Astrophysics Data System (ADS)

    Hegazy, Maha A.; Lotfy, Hayam M.; Rezk, Mamdouh R.; Omran, Yasmin Rostom

    2015-04-01

    Smart and novel spectrophotometric and chemometric methods have been developed and validated for the simultaneous determination of a binary mixture of chloramphenicol (CPL) and dexamethasone sodium phosphate (DSP) in presence of interfering substances without prior separation. The first method depends upon derivative subtraction coupled with constant multiplication. The second one is ratio difference method at optimum wavelengths which were selected after applying derivative transformation method via multiplying by a decoding spectrum in order to cancel the contribution of non labeled interfering substances. The third method relies on partial least squares with regression model updating. They are so simple that they do not require any preliminary separation steps. Accuracy, precision and linearity ranges of these methods were determined. Moreover, specificity was assessed by analyzing synthetic mixtures of both drugs. The proposed methods were successfully applied for analysis of both drugs in their pharmaceutical formulation. The obtained results have been statistically compared to that of an official spectrophotometric method to give a conclusion that there is no significant difference between the proposed methods and the official ones with respect to accuracy and precision.

  13. Magnetic ionic liquid aqueous two-phase system coupled with high performance liquid chromatography: A rapid approach for determination of chloramphenicol in water environment.

    PubMed

    Yao, Tian; Yao, Shun

    2017-01-20

    A novel organic magnetic ionic liquid based on guanidinium cation was synthesized and characterized. A new method of magnetic ionic liquid aqueous two-phase system (MILATPs) coupled with high-performance liquid chromatography (HPLC) was established to preconcentrate and determine trace amount of chloramphenicol (CAP) in water environment for the first time. In the absence of volatile organic solvents, MILATPs not only has the excellent properties of rapid extraction, but also exhibits a response to an external magnetic field which can be applied to assist phase separation. The phase behavior of MILATPs was investigated and phase equilibrium data were correlated by Merchuk equation. Various influencing factors on CAP recovery were systematically investigated and optimized. Under the optimal conditions, the preconcentration factor was 147.2 with the precision values (RSD%) of 2.42% and 4.45% for intra-day (n=6) and inter-day (n=6), respectively. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.14ngmL(-1) and 0.42ngmL(-1), respectively. Fine linear range of 12.25ngmL(-1)-2200ngmL(-1) was obtained. Finally, the validated method was successfully applied for the analysis of CAP in some environmental waters with the recoveries for the spiked samples in the acceptable range of 94.6%-99.72%. Hopefully, MILATPs is showing great potential to promote new development in the field of extraction, separation and pretreatment of various biochemical samples.

  14. The gene cluster for chloramphenicol biosynthesis in Streptomyces venezuelae ISP5230 includes novel shikimate pathway homologues and a monomodular non-ribosomal peptide synthetase gene.

    PubMed

    He, J; Magarvey, N; Piraee, M; Vining, L C

    2001-10-01

    Regions of the Streptomyces venezuelae ISP5230 chromosome flanking pabAB, an amino-deoxychorismate synthase gene needed for chloramphenicol (Cm) production, were examined for involvement in biosynthesis of the antibiotic. Three of four ORFs in the sequence downstream of pabAB resembled genes involved in the shikimate pathway. BLASTX searches of GenBank showed that the deduced amino acid sequences of ORF3 and ORF4 were similar to proteins encoded by monofunctional genes for chorismate mutase and prephenate dehydrogenase, respectively, while the sequence of the ORF5 product resembled deoxy-arabino-heptulosonate-7-phosphate (DAHP) synthase, the enzyme that initiates the shikimate pathway. A relationship to Cm biosynthesis was indicated by sequence similarities between the ORF6 product and membrane proteins associated with Cm export. BLASTX searches of GenBank for matches with the translated sequence of ORF1 in chromosomal DNA immediately upstream of pabAB did not detect products relevant to Cm biosynthesis. However, the presence of Cm biosynthesis genes in a 7.5 kb segment of the chromosome beyond ORF1 was inferred when conjugal transfer of the DNA into a blocked S. venezuelae mutant restored Cm production. Deletions in the 7.5 kb segment of the wild-type chromosome eliminated Cm production, confirming the presence of Cm biosynthesis genes in this region. Sequencing and analysis located five ORFs, one of which (ORF8) was deduced from BLAST searches of GenBank, and from characteristic motifs detected in alignments of its deduced amino acid sequence, to be a monomodular nonribosomal peptide synthetase. GenBank searches did not identify ORF7, but matched the translated sequences of ORFs 9, 10 and 11 with short-chain ketoreductases, the ATP-binding cassettes of ABC transporters, and coenzyme A ligases, respectively. As has been shown for ORF2, disrupting ORF3, ORF7, ORF8 or ORF9 blocked Cm production.

  15. p-Aminobenzoic acid and chloramphenicol biosynthesis in Streptomyces venezuelae: gene sets for a key enzyme, 4-amino-4-deoxychorismate synthase.

    PubMed

    Chang, Z; Sun, Y; He, J; Vining, L C

    2001-08-01

    Amplification of sequences from Streptomyces venezuelae ISP5230 genomic DNA using PCR with primers based on conserved prokaryotic pabB sequences gave two main products. One matched pabAB, a locus previously identified in S. venezuelae. The second closely resembled the conserved pabB sequence consensus and hybridized with a 3.8 kb NcoI fragment of S. venezuelae ISP5230 genomic DNA. Cloning and sequence analysis of the 3.8 kb fragment detected three ORFs, and their deduced amino acid sequences were used in BLAST searches of the GenBank database. The ORF1 product was similar to PabB in other bacteria and to the PabB domain encoded by S. venezuelae pabAB. The ORF2 product resembled PabA of other bacteria. ORF3 was incomplete; its deduced partial amino acid sequence placed it in the MocR group of GntR-type transcriptional regulators. Introducing vectors containing the 3.8 kb NcoI fragment of S. venezuelae DNA into pabA and pabB mutants of Escherichia coli, or into the Streptomyces lividans pab mutant JG10, enhanced sulfanilamide resistance in the host strains. The increased resistance was attributed to expression of the pair of discrete translationally coupled p-aminobenzoic acid biosynthesis genes (designated pabB/pabA) cloned in the 3.8 kb fragment. These represent a second set of genes encoding 4-amino-4-deoxychorismate synthase in S. venezuelae ISP5230. In contrast to the fused pabAB set previously isolated from this species, they do not participate in chloramphenicol biosynthesis, but like pabAB they can be disrupted without affecting growth on minimal medium. The gene disruption results suggest that S. venezuelae may have a third set of genes encoding PABA synthase.

  16. Rapid intranasal delivery of chloramphenicol acetyltransferase in the active form to different brain regions as a model for enzyme therapy in the CNS

    PubMed Central

    Appu, Abhilash P; Arun, Peethambaran; Krishnan, Jishnu K. S.; Moffett, John R.; Namboodiri, Aryan M. A.

    2015-01-01

    Background The blood brain barrier (BBB) is critical for maintaining central nervous system (CNS) homeostasis by restricting entry of potentially toxic substances. However, the BBB is a major obstacle in the treatment of neurotoxicity and neurological disorders due to the restrictive nature of the barrier to many medications. Intranasal delivery of active enzymes to the brain has therapeutic potential for the treatment of numerous CNS enzyme deficiency disorders and CNS toxicity caused by chemical threat agents. New method The aim of this work is to provide a sensitive model system for analyzing the rapid delivery of active enzymes into various regions of the brain with therapeutic bioavailability. Results We tested intranasal delivery of chloramphenicol acetyltransferase (CAT), a relatively large (75 kD) enzyme, in its active form into different regions of the brain. CAT was delivered intranasally to anaesthetized rats and enzyme activity was measured in different regions using a highly specific High Performance Thin Layer Chromatography (HP-TLC)-radiometry coupled assay. Active enzyme reached all examined areas of the brain within 15 min (the earliest time point tested). In addition, the yield of enzyme activity in the brain was almost doubled in the brains of rats pre-treated with matrix metalloproteinase-9 (MMP-9). Comparison with existing method (s) Intranasal administration of active enzymes in conjunction with MMP-9 to the CNS is both rapid and effective. Conclusion The present results suggest that intranasal enzyme therapy is a promising method for counteracting CNS chemical threat poisoning, as well as for treating CNS enzyme deficiency disorders. PMID:26688469

  17. An aptamer-based effective method for highly sensitive detection of chloramphenicol residues in animal-sourced food using real-time fluorescent quantitative PCR.

    PubMed

    Duan, Ye; Wang, Lihui; Gao, Zhiqiang; Wang, Huishan; Zhang, Hexiao; Li, Hao

    2017-04-01

    Chloramphenicol (CAP) residues can not only harm human health through entering food chain, but also cause the spreading of drug-resistant bacteria, thereby leading to secondary environmental pollution. Therefore, it is in urgent need of establishing an efficient technology to detect CAP residues in animal-sourced food. In this study, a novel sensitive approach for detection of CAP was designed based on a CAP specific aptamer and real-time fluorescent quantitative PCR (qRT-PCR). The CAP specific aptamer was firstly hybridized with a biotin modified complementary probe, and then was immobilized on streptavidin conjugated magnetic beads through biotin. When CAP was added, the aptamer would specifically bind with CAP by forming a hairpin structure and be released from the magnetic beads for CAP detection by qRT-PCR. Factors (i.e., probe strand length, aptamer concentration, NaCl concentration and incubation time) that would influence the determination accuracy of this aptamer-based detection system were optimized. Under the optimized conditions, the present detection system exhibited a high sensitivity toward CAP with a limit of detection of 0.1ng/mL (linear range from 0.1 to 20ng/mL). Moreover, this detection system also showed high selectivity against thiamphenicol (TAP) and florfenicol (FF), which are CAP's structure analogs. Eventually, this detection system was applied for detecting CAP in real spiked milk. The recovery rate of CAP from spiked milk samples ranged from 94.0-102.0%. These results indicated this developed detection system a promising high sensitive and specific method of CAP residues detection in animal-sourced food.

  18. Immunoaffinity column cleanup with LC/MS/MS for the determination of chloramphenicol in honey and prawns: single-laboratory validation.

    PubMed

    Mackie, Jennifer; Marley, Elaine; Donnelly, Carol

    2013-01-01

    A single-laboratory validation was conducted to establish the effectiveness of an immunoaffinity column (IAC) cleanup procedure followed by LC/MS/MS for the determination of chloramphenicol (CAP) in honey and prawns. Honey is dissolved in buffer solution and centrifuged, and an aliquot applied to an IAC. For prawns, a portion of the homogenized sample is shaken with buffer and centrifuged, and an aliquot similarly applied to an IAC. For both matrix extracts, CAP is removed from the IAC with neat methanol, then directly analyzed by electrospray LC/MS/MS in the negative ionization mode using m/z 321 as a precursor ion and m/z 257 and 152 as qualifier and quantifier ions, respectively. Test portions of blank honey and prawns were fortified with CAP to give levels of 0.3, 1.0, and 5.0 microg/kg. Recoveries of CAP on 3 consecutive days ranged from 83-103% for honey and 84-108% for prawns. Based on results for fortified blank matrixes (triplicate at three levels), the RSD for repeatability (RSDr) averaged 8.4% for honey and 4.8% for prawns. The method LOD was 0.05 for prawns and 0.16 microg/kg for honey, both well below the minimum required method performance limit for CAP. The accuracy of the method was demonstrated by participation in proficiency testing, where satisfactory Z-scores were obtained for CAP in incurred samples of both honey and prawns. The method was shown to be applicable to a wide range of other matrixes, including milk, egg, royal jelly, meat, and seafood products.

  19. Isolation and characterization of cyclo-(tryptophanyl-prolyl) and chloramphenicol from Streptomyces sp. SUK 25 with antimethicillin-resistant Staphylococcus aureus activity

    PubMed Central

    Alshaibani, Muhanna M; Jalil, Juriyati; Sidik, Nik M; Edrada-Ebel, Ruangelie; Zin, Noraziah M

    2016-01-01

    Background Zingiber spectabile, commonly known as Beehive Ginger, is used as an ethnobotanical plant in many countries as an appetizer or to treat stomachache, toothache, muscle sprain, and as a cure for swelling, sores and cuts. This is the first report of isolation of Streptomyces strain from the root of this plant. Strain Universiti Kebangsaan 25 (SUK 25) has a very high activity to produce secondary metabolites against methicillin-resistant Staphylococcus aureus (MRSA), which is associated with high morbidity and mortality rates due to acquired multidrug resistance genes and causes medication failure in some clinical cases worldwide. Phylogenetic analysis based on the 16S ribosomal RNA gene sequence exhibited that the most closely related strain was Streptomyces omiyaensis NBRC 13449T (99.0% similarity). Aim This study was conducted to carry out the extraction, identification, and biological evaluation of active metabolites isolated from SUK 25 against three MRSA strains, namely, MRSA ATCC 43300, MRSA ATCC 33591, and MRSA ATCC 49476. Materials and methods The production of secondary metabolites by this strain was optimized through Thronton’s media. Isolation, purification, and identification of the bioactive compounds were carried out using reversed-phase high-performance liquid chromatography, high-resolution mass spectrometry, Fourier transform infrared, and one-dimensional and two-dimensional nuclear magnetic resonance. Results During screening procedure, SUK 25 exhibited good antimicrobial potential against several strains of MRSA. The best biological activity was shown from fraction number VII and its subfractions F2 and F3 with minimum inhibitory concentration values at 16 µg/mL and 8 µg/mL, respectively. These two subfractions were identified as diketopiperazine cyclo-(tryptophanyl-prolyl) and chloramphenicol. Conclusion On the basis of obtained results, SUK 25 isolated from Z. spectabile can be regarded as a new valuable source to produce secondary

  20. Molecularly imprinted matrix solid-phase dispersion for extraction of chloramphenicol in fish tissues coupled with high-performance liquid chromatography determination.

    PubMed

    Guo, Linyuan; Guan, Min; Zhao, Chuande; Zhang, Haixia

    2008-12-01

    The synthesis and evaluation of a molecularly imprinted polymer (MIP) as a selective matrix solid-phase dispersion (MSPD) sorbent, coupled with high-performance liquid chromatography for the efficient determination of chloramphenicol (CAP) in fish tissues are studied. The polymer was prepared using CAP as the template molecule, vinylpyridine as the functional monomer and ethylene glycol dimethacrylate as the cross-linking monomer, and sodium dodecyl sulfate as the surfactant in the presence of water as a solvent by miniemulsion polymerization. The CAP-imprinted polymers and nonimprinted polymers (NIPs) were characterized by Fourier transform IR spectroscopy, scanning electron microscopy, and static adsorption experiments. The CAP-imprinted material prepared showed high adsorption capacity, significant selectivity, and good site accessibility. The maximum static adsorption capacity of the CAP-imprinted and the NIP material for CAP was 78.4 and 59.9 mg g(-1), respectively. The relative selectivity factors of this CAP-imprinted material were larger than 1.9. Several parameters influencing the MSPD process were optimized. Finally, the CAP-imprinted polymers were used as the sorbent in MSPD to determine CAP in three kinds of fishes and resulted in satisfactory recovery in the range 89.8-101.43%. CAP-imprinted polymer as a sorbent in MSPD is better than C18 and attapulgite in terms of both recovery and percent relative standard deviation. The baseline noise was measured from a chromatogram of a blank fish sample which was treated after the MSPD procedure using CAP-imprinted polymer as a sorbent. Signal values of 3 times the noise (signal-to-noise ratio of 3) and 10 times the noise (signal-to-noise ratio of 10) were used to calculate the limit of detection and the limit of quantitation of the calibration curve. The limit of detection for CAP was 1.2 ng g(-1) and the limit of quantitation was 3.9 ng g(-1).

  1. A human parvovirus, adeno-associated virus, as a eucaryotic vector: Transient expression and encapsidation of the procaryotic gene for chloramphenicol acetyltransferase

    SciTech Connect

    Tratschin, J.D.; West, M.H.P.; Sandbank, T.; Carter, B.J.

    1984-10-01

    The authors have used the defective human parvovirus adeno-associated virus (AAV) as a novel eurocaryotic vector (parvector) for the expression of a foreign gene in human cells. The recombinant, pAV2, contains the AAV genome in a pBR322-derived bacterial plasmid. When pAV2 is transfected into human cells together with helper adenovirus particles, the AAV genome is rescued from the recombinant plasmid and replicated to produce infectious AAV particles at high efficiency. To create a vector, we inserted a procaryotic sequence coding for chloramphenicol acetyltransferase (CAT) into derivatives of pAV2 following either of the AAV promoters p/sub 40/ (pAVHiCAT) and p/sub 19/ (pAVBcCAT). When transfected into human 293 cells or HeLa cells, pAVHiCAT expressed CAT activity in the absence of adenovirus. In the presence of adenovirus, this vector produced increased amounts of CAT activity and the recombinant AAV-CAT genome was replicated. In 293 cells, pAVBcCAT expressed a similar amount of CAT activity in the absence or presence of adenovirus and the recombinant AAV-CAT genome was not replicated. In HeLa cells, pAVBcCAT expressed low levels of CAT activity, but this level was elevated by coinfection with adenovirus particles or by cotransfection with a plasmid which expressed the adenovirus early region 1A (E1A) product. The E1A product is a transcriptional activator and is expressed in 293 cells. Thus, expression from two AAV promoters is differentially regulated: expression from p/sub 19/ is increased by E1A, whereas p/sub 40/ yields high levels of constitutive expression in the absence of E1A. Both AAV vectors were packaged into AAV particles by complementation with wild-type AAV and yielded CAT activity when subsequently infected into cells in the presence of adenovirus.

  2. A Study of Electrocatalytic and Photocatalytic Activity of Cerium Molybdate Nanocubes Decorated Graphene Oxide for the Sensing and Degradation of Antibiotic Drug Chloramphenicol.

    PubMed

    Karthik, Raj; Vinoth Kumar, Jeyaraj; Chen, Shen-Ming; Karuppiah, Chelladurai; Cheng, Yi-Hui; Muthuraj, Velluchamy

    2017-02-22

    In this present work, "killing two birds with one stone" strategy was performed for the electrochemical trace level detection and photocatalytic degradation of antibiotic drug chloramphenicol (CAP) using Ce(MoO4)2 nanocubes/graphene oxide (CeM/GO) composite for the first time. The CeM/GO composite was synthesized via simple hydrothermal treatment followed by sonication process. The successful formation of CeM/GO composite was confirmed by several analytical and spectroscopic techniques. The CeM/GO composite modified glassy carbon electrode (GCE) showed excellent electrocatalytic activity toward the reduction of CAP in terms of decrease the potential and increase the cathodic peak current in comparison with different modified and unmodified electrodes. The electrocatalytic reduction of CAP based on the CeM/GO modified GCE exhibited high selectivity, wide linear ranges, lower detection limit, and good sensitivity of 0.012-20 and 26-272 μM, 2 nM ,and 1.8085 μA μM(-1) cm(-2), respectively. Besides, when CeM/GO/GCE was used to analyze the CAP in real samples, such as honey and milk, the satisfactory recovery results were obtained. On the other hand, the CeM/GO composite played excellent catalyst toward the photodegradation of CAP. The obtained results from the UV-vis spectroscopy clearly suggested that CeM/GO composite had high photocatalytic activity compared to pristine Ce(MoO4)2 nanocubes. The degradation efficiency of CeM/GO toward CAP is observed about 99% within 50 min under visible irradiation and it shows a good stability by observing the reusability of the catalyst. The enhanced photocatalytic performance was attributed to the increased migration efficiency of photoinduced electrons and holes.

  3. A homogeneous and "off-on" fluorescence aptamer-based assay for chloramphenicol using vesicle quantum dot-gold colloid composite probes.

    PubMed

    Miao, Yang-Bao; Ren, Hong-Xia; Gan, Ning; Zhou, You; Cao, Yuting; Li, Tianhua; Chen, Yinji

    2016-07-27

    In this work, a novel homogeneous and signal "off-on" aptamer based fluorescence assay was successfully developed to detect chloramphenicol (CAP) residues in food based on the fluorescence resonance energy transfer (FRET). The vesicle nanotracer was prepared through labeling single stranded DNA binding protein (SSB) on limposome-CdSe/ZnS quantum dot (SSB/L-QD) complexes. It was worth mentioning that the signal tracer (SSB/L-QD) with vesicle shape, which was fabricated being encapsulated with a number of quantum dots and SSB. The nanotracer has excellent signal amplification effects. The vesicle composite probe was formed by combining aptamer labeled nano-gold (Au-Apt) and SSB/L-QD. Which based on SSB's specific affinity towards aptamer. This probe can't emit fluoresce which is in "off" state because the signal from SSB/L-QD as donor can be quenched by the Au-aptas acceptor. When CAP was added in the composite probe solution, the aptamer on the Au-Apt can be preferentially bounded with CAP then release from the composite probe, which can turn the "off" signal of SSB/L-QD tracer into "on" state. The assay indicates excellent linear response to CAP from 0.001 nM to 10 nM and detection limit down to 0.3 pM. The vesicle probes with size of 88 nm have strong signal amplification. Because a larger number of QDs can be labeled inside the double phosphorus lipid membrane. Besides, it was employed to detect CAP residues in the milk samples with results being agreed well with those from ELISA, verifying its accuracy and reliability.

  4. Aminoacyl and peptidyl analogs of chloramphenicol as slow-binding inhibitors of ribosomal peptidyltransferase: a new approach for evaluating their potency.

    PubMed

    Michelinaki, M; Mamos, P; Coutsogeorgopoulos, C; Kalpaxis, D L

    1997-01-01

    In a model system derived from Escherichia coli, acetylphenylalanyl-puromycin is produced in a pseudo-first-order reaction between the preformed acetylphenylalanyl/tRNA/poly(U)/ribosome complex (complex C) and excess puromycin. Two aminoacyl analogs [3, Gly-chloramphenicol (CAM): 4, L-Phe-CAM] and two peptidyl analogs (2, L-Phe-Gly-CAM: 5, Gly-Phe-CAM) of CAM (1) were tested as inhibitors in this reaction. Detailed kinetic analysis suggests that these analogs (I) react competitively with complex C and form the complex C*l, which is inactive toward puromycin. C*l is formed via a two-step mechanism in which C*l is the product of a slow conformational change of the initial encounter complex Cl according to the equation C + l reversible Cl reversible C*l. Furthermore, we provide evidence that analog 5 may react further with C*l forming the species C*l2. The values of the apparent association rate constant (K(assoc)) are 1.42 x microM-1 min-1 for 2, 0.55 x microM-1 min-1 for 3, and 0.18 x microM-1 min-1 for 4 and 0.038 x microM-1 min-1 for 5 [corrected]. In the case of analog 5, K(assoc) is a linear function of the inhibitor concentration; when [I] approaches zero, the K(assoc) value is equal to 3.8 x 10(2) M-1 sec-1. Such values allow the classification of CAM analogs as slow-binding inhibitors. According to K(assoc) values, we could surmise that analog 2 is 2.5-fold more potent than 3 and 8-fold more potent than 4. The relative potency of analog 5 is the lowest among the analogs and is dependent on its concentration. The results are compared with previous data and discussed on the basis of a possible retro-inverso relationship between CAM analogs and puromycin.

  5. Impact of persulfate and ultraviolet light activated persulfate pre-oxidation on the formation of trihalomethanes, haloacetonitriles and halonitromethanes from the chlor(am)ination of three antibiotic chloramphenicols.

    PubMed

    Chu, Wenhai; Chu, Tengfei; Bond, Tom; Du, Erdeng; Guo, Yingqing; Gao, Naiyun

    2016-04-15

    Persulfate oxidation processes, with and without activation using ultraviolet light (respectively UV/PS and PS) have the potential to degrade anthropogenic chemicals in water. However, little is known about the impact of PS or UV/PS pre-oxidation on downstream formation of disinfection by-products (DBPs). In this study the three antibiotic chloramphenicols (chloramphenicol and two of its analogues [thiamphenicol and florfenicol], referred to collectively as CAPs), which frequently occur in wastewater-impacted source waters used by drinking water treatment plants, were selected as model antibiotic compounds. The formation of carbonaceous and nitrogenous disinfection by-products, including halomethanes, haloacetonitriles and halonitromethanes, during chlorination and chloramination preceded by PS and UV/PS was investigated. No significant concentrations of haloacetonitriles and halonitromethanes were detected during chlorination. During chloramination chloramphenicol formed a considerable amount of dichloronitromethane (e.g., 3.44 ± 0.33% mol/mol at NH2Cl dose = 1 mM) and trichloronitromethane (e.g., 0.79 ± 0.07% mol/mol at NH2Cl dose = 1 mM), compared with THM and HAN formation. PS pre-oxidation achieved a statistically significant reduction in trichloromethane formation from chlorination, and in HAN and HNM formation from chloramination. Although UV/PS slightly increased dichloroacetonitrile formation during chloramination, it significantly decreased dichloronitromethane and trichloronitromethane formation during chloramination. Overall, the use of PS and UV/PS has the potential to have contrasting impacts on DBP formation in heavily wastewater-impacted waters, depending on the disinfection method. Hence, their application needs to be carefully balanced against the downstream effect on DBP formation.

  6. Development of a Double Nuclear Gene-Targeting Method by Two-Step Transformation Based on a Newly Established Chloramphenicol-Selection System in the Red Alga Cyanidioschyzon merolae

    PubMed Central

    Fujiwara, Takayuki; Ohnuma, Mio; Kuroiwa, Tsuneyoshi; Ohbayashi, Ryudo; Hirooka, Shunsuke; Miyagishima, Shin-Ya

    2017-01-01

    The unicellular red alga Cyanidioschyzon merolae possesses a simple cellular architecture that consists of one mitochondrion, one chloroplast, one peroxisome, one Golgi apparatus, and several lysosomes. The nuclear genome content is also simple, with very little genetic redundancy (16.5 Mbp, 4,775 genes). In addition, molecular genetic tools such as gene targeting and inducible gene expression systems have been recently developed. These cytological features and genetic tractability have facilitated various omics analyses. However, only a single transformation selection marker URA has been made available and thus the application of genetic modification has been limited. Here, we report the development of a nuclear targeting method by using chloramphenicol and the chloramphenicol acetyltransferase (CAT) gene. In addition, we found that at least 200-bp homologous arms are required and 500-bp arms are sufficient for a targeted single-copy insertion of the CAT selection marker into the nuclear genome. By means of a combination of the URA and CAT transformation systems, we succeeded in producing a C. merolae strain that expresses HA-cyclin 1 and FLAG-CDKA from the chromosomal CYC1 and CDKA loci, respectively. These methods of multiple nuclear targeting will facilitate genetic manipulation of C. merolae. PMID:28352279

  7. Determination of chloramphenicol, thiamphenicol and florfenicol in milk and honey using modified QuEChERS extraction coupled with polymeric monolith-based capillary liquid chromatography tandem mass spectrometry.

    PubMed

    Liu, Hsiang-Yu; Lin, Shu-Ling; Fuh, Ming-Ren

    2016-04-01

    A poly(lauryl methacrylate-co-methacrylic acid-co-ethylene glycol dimethacrylate) [LMA-MAA-EDMA] monolithic column was used to simultaneously determine amphenicol antibiotics (chloramphenicol/CAP, thiamphenicol/TAP, and florfenicol/FF) in milk and honey samples by capillary liquid chromatography tandem mass spectrometry (LC-MS/MS). QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was optimized for sample pretreatment. Good linearity (0.1-15 ng g(-1)) and extraction recoveries (95.8-100.2% and 95.6-99.3% for milk and honey samples, respectively; n=3) with minor matrix effect (≦ 5% ion suppression) were obtained. Limits of detection were estimated at 0.02-0.045 ng g(-1). Good intra-day/inter-day precision (0.2-9.1%/0.3-8.7%) and accuracy (90.5-110.0%/93.4-109.3%) were achieved. With more than 200 analyses of real samples, no noticeable carry-over and deterioration of separation efficiency were observed using the monolithic column. The applicability of the developed QuEChERS-capillary LC-MS/MS method was demonstrated by determining the occurrence of CAP, TAP, and FF in various milk and honey samples.

  8. Multi-class method for the determination of nitroimidazoles, nitrofurans, and chloramphenicol in chicken muscle and egg by dispersive-solid phase extraction and ultra-high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Zhang, Zhiwen; Wu, Yuping; Li, Xiaowei; Wang, Yingyu; Li, Hui; Fu, Qin; Shan, Yawen; Liu, Tianhe; Xia, Xi

    2017-02-15

    This study describes the development of a multiresidue method for the efficient identification and quantification of nitroimidazoles, nitrofurans, and chloramphenicol in chicken and egg. After derivatization of nitrofuran metabolites, dispersive-solid phase extraction was used for the extraction of target analytes. An optimization strategy involved the selection of sorbents and extraction solutions for dispersive-solid phase extraction in order to achieve acceptably high recoveries and reduce co-extractives in the final extracts. Analytes were determined by ultra-high performance liquid chromatography-tandem mass spectrometry, in one single injection with a chromatographic run time of 7.5min. Mean recoveries ranged from 86.4% to 116.7% and interday precision was lower than 18%. The limits of quantification were between 0.1 and 0.5μg/kg, which were satisfactory to support surveillance monitoring. Finally, the method was applied to real samples, and metabolite of furazolidone, metronidazole and its metabolite, dimetridazole and its metabolite were detected in both chicken and egg samples.

  9. A novel "dual-potential" electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol.

    PubMed

    Feng, Xiaobin; Gan, Ning; Zhang, Huairong; Yan, Qing; Li, Tianhua; Cao, Yuting; Hu, Futao; Yu, Hongwei; Jiang, Qianli

    2015-12-15

    A novel type of "dual-potential" electrochemiluminescence (ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for simultaneous detection of malachite green (MG) and chloramphenicol (CAP) in one single assay. The SPCE substrate consisted of a common Ag/AgCl reference electrode, carbon counter electrode and two carbon working electrodes (WE1 and WE2). In the system, CdS quantum dots (QDs) were modified on WE1 as cathode ECL emitters and luminol-gold nanoparticles (L-Au NPs) were modified on WE2 as anode ECL emitters. Then the MG aptamer complementary strand (MG cDNA) and CAP aptamer complementary strand (CAP cDNA) were attached on CdS QDs and L-Au NPs, respectively. The cDNA would hybridize with corresponding aptamer that was respectively tagged with cyanine dye (Cy5) (as quenchers of CdS QDs) and chlorogenic acid (CA) (as quenchers of l-Au NPs) using poly(ethylenimine) (PEI) as a bridging agent. PEI could lead to a large number of quenchers on the aptamer, which increased the quenching efficiency. Upon MG and CAP adding, the targets could induce strand release due to the highly affinity of analytes toward aptamers. Meanwhile, it could release the Cy5 and CA, which recovered cathode ECL of CdS QDs and anode ECL of L-Au NPs simultaneously. This "dual-potential" ECL strategy could be used to detect MG and CAP with the linear ranges of 0.1-100 nM and 0.2-150 nM, with detection limits of 0.03 nM and 0.07 nM (at 3sB), respectively. More importantly, this designed method was successfully applied to determine MG and CAP in real fish samples and held great potential in the food analysis.

  10. 21 CFR 524.390b - Chloramphenicol ophthalmic solution.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS OPHTHALMIC AND TOPICAL DOSAGE FORM NEW ANIMAL DRUGS...) Limitations. Therapy for cats should not exceed 7 days. As with other antibiotics, prolonged use may result in... animals. The length of time that residues persist in milk or tissues has not been determined. Federal...

  11. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... every 6 hours. (2) Indications for use. For treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections associated with...

  12. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... every 6 hours. (2) Indications for use. For treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections associated with...

  13. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... weight by mouth every 6 hours. (2) Indications for use—(i) For the treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial......

  14. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... every 6 hours. (2) Indications for use. For treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections associated with...

  15. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... weight by mouth every 6 hours. (2) Indications for use—(i) For the treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial......

  16. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... weight by mouth every 6 hours. (2) Indications for use—(i) For the treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial......

  17. Cellular uptake and covalent binding of nitroso-chloramphenicol

    SciTech Connect

    Murray, T.; Yunis, A.A.

    1981-09-01

    A comparative study of the cellular transport of CAP and its nitroso derivative (NO-CAP) was carried out in Raji cells, a transformed human lymphoblastoid cell line. Both agents were concentrated by the cells by a factor of 3 (cellular/extracellular concentration ratio). The cellular uptake of NO-CAP, like that of CAP, was found to be rapid and temperature-independent. Thus the greater cytotoxicity of NO-CAP is apparently not due to an enhanced uptake of the nitroso derivative relative to CAP. In contrast to the similarity of uptake, NO-CAP becomes covalently bound to both Raji cells and freshly isolated human bone marrow cells to a much higher extent (15-fold). Also, cells previously loaded with CAP or NO-CAP retain three times as much of the nitroso compound during a 24 hr dialysis against a drug-free isotonic solution. The increased binding of NO-CAP to human hematopoietic cells attests to the greater reactivity of the p-substituted aromatic nitroso group and is consistent with the postulate that reduction products of the nitro group of CAP may be responsible for CAP-induced aplastic anemia.

  18. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... every 6 hours. (2) Indications for use. For treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections associated with...

  19. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... dogs—(1) Amount. 25 mg per pound of body weight every 6 hours. (2) Indications for use. For treatment of bacterial pulmonary infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections associated with canine distemper caused by susceptible organisms....

  20. 77 FR 4895 - New Animal Drugs; Chloramphenicol, Diethylcarbamazine Citrate, Hygromycin B, Methoxyflurane...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-01

    ..., Penicillin G, Phenylbutazone, Pyrantel Tartrate, Tylosin Phosphate, and Sulfamethazine AGENCY: Food and Drug...-PEN Walco International, (penicillin G Inc., 15 West benzathine, Putnam, penicillin G Porterville, CA... follows: Sec. 522.1696a Penicillin G benzathine and penicillin G procaine suspension. * * * * *...

  1. Characterisation of a mobilisable plasmid conferring florfenicol and chloramphenicol resistance in Actinobacillus pleuropneumoniae.

    PubMed

    Bossé, Janine T; Li, Yanwen; Atherton, Tom G; Walker, Stephanie; Williamson, Susanna M; Rogers, Jon; Chaudhuri, Roy R; Weinert, Lucy A; Holden, Matthew T G; Maskell, Duncan J; Tucker, Alexander W; Wren, Brendan W; Rycroft, Andrew N; Langford, Paul R

    2015-08-05

    The complete nucleotide sequence of a 7.7kb mobilisable plasmid (pM3446F), isolated from a florfenicol resistant isolate of Actinobacillus pleuropneumoniae, showed extended similarity to plasmids found in other members of the Pasteurellaceae containing the floR gene as well as replication and mobilisation genes. Mobilisation into other Pasteurellaceae species confirmed that this plasmid can be transferred horizontally.

  2. Serum Zinc, Iron, and Copper Concentrations during Typhoid Fever in Man: Effect of Chloramphenicol Therapy

    DTIC Science & Technology

    1974-11-12

    Received Nov. 12I~aepe Dec. 16,1974. Study 1. Nineteen healthy men, inmates of the MS CLINICAL CHEMISTRY , Vol. 21.14o. 4, 1975 - Best Available Copy Maryland...thenap and mserum rm concentrfations. nificant depressions of serum iron and zinc conoen- aM CLINICAL CHEMISTRY , Vol. 21.4o. 4. 10754 *IE EWZX 1~001...evidenc, was in the absence between 30-40% of the zinc bound in serum (16), we CLINICAL CHEMISTRY . Vol. 21. No. 4,1975 531 saw no significant changes in

  3. Beneficial effect of Berberis buxifolia Lam, Ziziphus mistol Griseb and Prosopis alba extracts on oxidative stress induced by chloramphenicol.

    PubMed

    Albrecht, Claudia; Pellarin, Gabriela; Rojas, María José; Albesa, Inés; Eraso, Alberto F

    2010-01-01

    The chemiluminescence of luminol, a measure of oxidative stress, increased immediately as a consequence of reactive oxygen species (ROS) stimulated by this antibiotic. The effect of Ch was dose dependent with maximum stimulus at 8 mg/ml (Vmax); above this concentration the cells began to reduce the production of ROS. The oxidative injury of Ch was counteracted by water extracts of Berberis buxifolia lam, Zizyphus mistol Griseb and Prosopis alba, indigenous fruits from Argentina. The relatively light units (RLU) emitted decreased immediately as a consequence of a protective effect exerted by the extracts of these fruit extracts on blood cells. The three indigenous fruit extracts reduced to a different extent the oxidative injury caused by Ch. B.buxifolia lam exhibited the highest antioxidant capacity followed by Z.mistol Griseb. Water extracts of both fruit extracts were the most effective against the oxidative stress, while P.alba presented better antioxidant capacity in the ethanolic fraction obtained. Hexane extracts showed low protective action on blood cells, with little reduction of area under curve (AUC) of RLU plotted versus time. Leukocytes remained viable in blood samples incubated for 3h with Ch and water extracts of B. buxifolia lam or Z. mistol Griseb (97.1% and 92.5% viability by Trypan blue exclusion, respectively); whereas with Ch only the cells were stressed and viability decreased to 30%. The three fruit extracts protected the viability of leukocytes in parallel with the decrease of ROS. Erythrocytes were not lysed in the presence of Ch.

  4. Chloramphenicol and tetracycline decrease motility and increase invasion and attachment gene expression in specific isolates of multidrug-resistant Salmonella enterica serovar Typhimurium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella enterica serovar Typhimurium (S. Typhimurium) is one of the most common serovars isolated from humans and livestock, and over 35 percent of these isolates are resistant to three or more antibiotics. Multidrug-resistant (MDR) Salmonella is a public health concern as it is associated with i...

  5. Antibacterial application of engineered bacteriophage nanomedicines: antibody-targeted, chloramphenicol prodrug loaded bacteriophages for inhibiting the growth of Staphylococcus aureus bacteria.

    PubMed

    Vaks, Lilach; Benhar, Itai

    2011-01-01

    The increasing development of bacterial resistance to traditional antibiotics has reached alarming levels, thus there is an urgent need to develop new antimicrobial agents. To be effective, these new antimicrobials should possess novel modes of action and/or different cellular targets compared with existing antibiotics. Bacteriophages (phages) have been used for over a century as tools for the treatment of bacterial infections, for nearly half a century as tools in genetic research, for about two decades as tools for the discovery of specific target-binding proteins and peptides, and for almost a decade as tools for vaccine development. We describe a new application in the area of antibacterial nanomedicines where filamentous phages can be formulated as targeted drug-delivery vehicles of nanometric dimensions (phage nanomedicines) and used for therapeutic purposes. This protocol involves both genetic and chemical engineering of these phages. The genetic engineering of the phage coat, which results in the display of a target-specificity-conferring peptide or protein on the phage coat, can be used to design the drug-release mechanism and is not described herein. However, the methods used to chemically conjugate cytotoxic drugs at high density on the phage coat are described. Further, assays to measure the drug load on the surface of the phage and the potency of the system in the inhibition of growth of target cells as well as assessment of the therapeutic potential of the phages in a mouse disease model are discussed.

  6. Comparison of rose bengal-chloramphenicol and modified aureomycin-rose bengal-glucose-peptone agar as media for the enumeration of molds and yeasts in water by membrane filtration techniques.

    PubMed

    Albaum, Silva; Masaphy, Segula

    2009-03-01

    The present study compares the suitability of rose bengalchloramphenicol (RBCh) and modified aureomycin-rose bengal glucose-peptone agar for enumerating fungi recovered from diverse water sources using the membrane filtration method. Both mold and yeast colonies were established more rapidly, with more intensified colony colors and higher RBCh counts.

  7. Antibiotic therapy for bacterial meningitis in children in developing countries.

    PubMed Central

    Kumar, P.; Verma, I. C.

    1993-01-01

    We carried out a study to investigate the effectiveness of chloramphenicol alone as a treatment for bacterial meningitis. A total of 70 consecutive children aged > 3 months with bacterial meningitis, who had been admitted to the paediatric hospital of the All India Institute of Medical Sciences, were randomized to receive chloramphenicol alone or chloramphenicol + penicillin. The two groups were matched with each other. Treatment failure occurred with three (9%) patients in the chloramphenicol-alone group and with four (12.1%) patients in the combination therapy group (P > 0.05). The mean duration of intravenous therapy, the number of intravenous cannulae used per patient, and the incidence of thrombophlebitis were significantly higher for the group that received the combination therapy. Also, the cost of using chloramphenicol + penicillin was four times higher than that of chloramphenicol alone. Hence, chloramphenicol alone was as effective as chloramphenicol + penicillin and much cheaper and more convenient to use. PMID:8490981

  8. UGA can be decoded as tryptophan at low efficiency in Bacillus subtilis.

    PubMed Central

    Lovett, P S; Ambulos, N P; Mulbry, W; Noguchi, N; Rogers, E J

    1991-01-01

    Replacement of cat-86 codon 7 or 144 with the UGA codon permitted the gene to confer chloramphenicol resistance in wild-type Bacillus subtilis. UAA replacements of the same codons resulted in a chloramphenicol-sensitive phenotype in wild-type B. subtilis and a chloramphenicol-resistant phenotype in suppressor-positive strains. N-terminal sequencing showed that UGA at codon 7 was decoded as tryptophan in wild-type cells, at an efficiency of about 6%. Images PMID:1900283

  9. RESTORATION OF NORMAL GLUTAMIC ACID TRANSPORT IN VITAMIN B6-DEFICIENT LACTOBACILLUS PLANTARUM BY ACETATE, AMMONIUM, AND VITAMIN B6,

    DTIC Science & Technology

    GLUTAMIC ACID, * LACTOBACILLUS , VITAMIN B COMPLEX, METABOLIC DISEASES, VITAMIN B COMPLEX, ACETATES, AMMONIUM COMPOUNDS, CHLORAMPHENICOL, DEOXYRIBONUCLEIC ACIDS, AMINO ACIDS, PENICILLINS, CELL WALL, SYNTHESIS, OSMOSIS.

  10. [Experience with vaginal suppositories containing chemotherapeutic agents (author's transl)].

    PubMed

    Török, J; Kószó, E; Altmayer, P; Mezey, G

    1981-01-01

    The authors determined the release of chloramphenicol, oxytetracycline, neomycin and sulphadimidine from different bases (polyoxethene mass, solid fat, 95% solid fat +5% Span 20). The diffusion of chloramphenicol and sulphadimidine was best from hydrophilic bases; that of oxytetracycline and neomycin, from emulsifier-containing lipophil bases. This may be explained by the solubilities of the pharmaca and by interactions between base and active substance.

  11. Isolation and characterization of the grain mold fungi, Cochliobolus and Alternaria spp., from sorghum using semi-selective media and DNA sequence analyses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mold diseases, caused by fungal complexes including Alternaria, Cochliobolus and Fusarium species, limit sorghum grain production. Media were tested by plating Fusarium thapsinum, Alternaria sp. and Curvularia lunata, individually and competitively. Dichloran chloramphenicol rose bengal (DRBC) and m...

  12. Meningitis - meningococcal

    MedlinePlus

    ... is one of the most commonly used antibiotics. Penicillin in high doses is almost always effective. If there is an allergy to penicillin, chloramphenicol may be used. Sometimes, corticosteroids are given ...

  13. Bromocriptine

    MedlinePlus

    ... Bromocriptine is in a class of medications called dopamine receptor agonists. It treats hyperprolactinemia by decreasing the ... ketoconazole (Nizoral); antihistamines; chloramphenicol; dexamethasone (Decadron, Dexpak); other dopamine agonists such as cabergoline (Dostinex), levodopa (Dopar, Larodopa), ...

  14. Mechanisms of antimicrobial resistance and genetic relatedness among enterococci isolated from dogs and cats in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: In this study, mechanisms of antimicrobial resistance and genetic relatedness among resistant enterococci from dogs and cats in the United States were determined. Methods and Results: Enterococci resistant to chloramphenicol, ciprofloxacin, erythromycin, gentamicin, kanamycin, streptomycin,...

  15. FUNDAMENTALS OF THE STUDY OF ANTIBIOTICS,

    DTIC Science & Technology

    ANTIBIOTICS, BIOSYNTHESIS), PHARMACOLOGY, MICROORGANISMS, CLASSIFICATION, PRODUCTION, CULTURE MEDIA, POLYMYXINS, STREPTOMYCINS, TETRACYCLINES ...CHLORAMPHENICOL, OXYTETRACYCLINE , CHLORTETRACYCLINE, PENICILLINS, FUNGI, PLANTS(BOTANY), RESISTANCE(BIOLOGY), PREPARATION, PURIFICATION, AGRICULTURE, CANNING, ECOLOGY, FOOD, MILK, PRESERVATION, USSR

  16. Electrooptical Analysis of Microbial Cell Suspensions forDetermination of Antibiotic Resistance.

    PubMed

    Guliy, Olga I; Bunin, Victor D; Korzhenevich, Vyacheslav I; Volkov, Alexey A; Ignatov, Oleg V

    2016-12-01

    The effects of ampicillin; kanamycin, chloramphenicol, and tetracycline on electrophysical characteristics of cells of sensitive (ampicillin; kanamycin, chloramphenicol) and resistant (ampicillin; kanamycin, chloramphenicol, tetracycline) Escherichia coli strains were studied. Under the action of antibiotics sensitive and resistant E. coli strains acquire different electro-optical properties. Changes in suspension-orientational spectra, that are observed under the action of ampicillin; kanamycin, chloramphenicol, and tetracycline can be used in determination of antibiotic resistance of the studied bacterial strains. In our opinion, the methods of microbial suspension electro-optical analysis can be used in microbiology, mеdicinе, veterinary, and are an effective tool for solving the problems connected with determination of microbial cell antibiotic resistance.

  17. Effect of a Salmonella Group H1 R Factor on Virulence and Response of Infections to Antimicrobial Therapy

    PubMed Central

    Butler, Thomas; Shuster, C. W.; Franco, Amalia

    1979-01-01

    A group H1 R factor encoding resistance to chloramphenicol, streptomycin, sulfonamide, and tetracycline was transferred into Salmonella typhimurium LT-2. The virulence of LT-2 for mice, as assessed by intraperitoneal 50% lethal dose and the number of organisms in the spleen, was not affected by the R factor. On the other hand, the R factor conferred resistance in mouse infections to therapy with chloramphenicol and trimethoprim plus sulfamethoxazole. PMID:380460

  18. [Macrolides in the treatment of children with Mediterranean spotted fever].

    PubMed

    Cascio, Antonio; Colomba, Claudia

    2002-09-01

    Till now there is not a gold standard therapy for Mediterranean spotted fever (MSF) in children. Standard treatment for MSF is the administration of tetracycline or chloramphenicol, however both these drugs can cause significant adverse effects in children (tetracyclines can cause staining of teeth, chloramphenicol severe hematological adverse events such as aplastic anemia, gray baby syndrome and hemolytic anemia in patients with the Mediterranean form of G6PD deficiency). We conducted two randomized clinical trials; the first compared clarithromycin versus chloramphenicol: mean time to defervescence was 36.7 +/- 18.1 h in the clarithromycin group and 47.1+/- 21.9 h in the chloramphenicol group (P= 0.047). The second trial compared clarithromycin versus azithromycin and did not show any statistically significant difference: mean time to defervescence was 46.2 +/- 36.4 h in the clarithromycin group and 39.3 +/- 31.3 h in the azithromycin group (P= 0.34). On the basis of these studies we think that clarithromycin and azithromycin could constitute an acceptable alternative to chloramphenicol and to tetracyclines for the treatment of MSF in children

  19. Synergistic activity of coriander oil and conventional antibiotics against Acinetobacter baumannii.

    PubMed

    Duarte, A; Ferreira, S; Silva, F; Domingues, F C

    2012-02-15

    In this study we investigated the existence of synergistic antibacterial effect between coriander (Coriandrum sativum L.) essential oil and six different antibacterial drugs (cefoperazone, chloramphenicol, ciprofloxacin, gentamicin, tetracycline and piperacillin). The antibacterial activity of coriander oil was assessed using microdilution susceptibility testing and synergistic interaction by checkerboard assays. The association of coriander essential oil with chloramphenicol, ciprofloxacin, gentamicin and tetracycline against Acinetobacter baumannii showed in vitro effectiveness, which is an indicator of a possible synergistic interaction against two reference strains of A. baumannii (LMG 1025 and LMG 1041) (FIC index from 0.047 to 0.375). However, when tested the involvement between coriander essential oil and piperacillin or cefoperazone, the isobolograms and FIC index showed an additive interaction. The in vitro interaction could improve the antimicrobial effectiveness of ciprofloxacin, gentamicin and tetracycline and may contribute to resensitize A. baumannii to the action of chloramphenicol.

  20. Streamlined Synthesis and Assembly of a Hybrid Sensing Architecture with Solid Binding Proteins and Click Chemistry.

    PubMed

    Swift, Brian J F; Shadish, Jared A; DeForest, Cole A; Baneyx, François

    2017-03-13

    Combining bioorthogonal chemistry with the use of proteins engineered with adhesive and morphogenetic solid-binding peptides is a promising route for synthesizing hybrid materials with the economy and efficiency of living systems. Using optical sensing of chloramphenicol as a proof of concept, we show here that a GFP variant engineered with zinc sulfide and silica-binding peptides on opposite sides of its β-barrel supports the fabrication of protein-capped ZnS:Mn nanocrystals that exhibit the combined emission signatures of organic and inorganic fluorophores. Conjugation of a chloramphenicol-specific DNA aptamer to the protein shell through strain-promoted azide-alkyne cycloaddition and spontaneous concentration of the resulting nanostructures onto SiO2 particles mediated by the silica-binding sequence enables visual detection of environmentally and clinically relevant concentrations of chloramphenicol through analyte-mediated inner filtering of sub-330 nm excitation light.

  1. mRNA translation during oocyte maturation plays a key role in development of primordial germ cells in Xenopus embryos.

    PubMed

    Zeynali, Bahman; Dixon, Keith E

    2004-09-01

    It is believed that cytoplasmic localization in the egg is necessary for development of primordial germ cells (PGCs) in Xenopus embryos. In this study, we sought to determine if translation of maternal mRNA during oocyte maturation is involved in the development of PGCs. Donor oocytes were collected from both stimulated (those who receive gonadotropin) and unstimulated females, artificially matured and fertilized using a host transfer technique. Using chloramphenicol (50 microM and 500 microM RNA), RNA translation was inhibited during oocyte maturation. Our results showed that in unstimulated embryos treated with 50 microM chloramphenicol, there was a significant reduction in the number of PGCs reaching genital ridges. In stimulated embryos, however, the number of PGCs was unchanged unless a higher concentration (500 microM) of chloramphenicol was used. From these results it is suggested that maternal mRNA translation during oocyte maturation plays a key role in development of PGCs.

  2. Clinical and Pathological Evaluation of Sulbactam/Ampicillin for Treatment of Experimental Bovine Pneumonic Pasteurellosis

    PubMed Central

    Gifford, Glen A.; Potter, Andrew A.; Babiuk, Lorne A.

    1988-01-01

    An experiment was conducted to evaluate the efficacy of sulbactam/ampicillin for treatment of bovine pneumonic pasteurellosis. Twenty-one Hereford calves were experimentally infected with bovine herpesvirus-1 and an ampicillin-resistant strain of Pasteurella haemolytica, then treated for three days with either sulbactam/ampicillin, chloramphenicol, or a placebo. The treatments were evaluated by comparing clinical illness scores, total sick days, weight changes, mortality rates, and postmortem lung scores between treatment groups. Both antibiotics were highly effective in reducing respiratory disease in the experimentally infected calves. The clinical response to sulbactam/ampicillin treatment was comparable with that of chloramphenicol and was significantly improved compared with the response to the placebo treatment. These findings suggest that the efficacy of sulbactam/ampicillin may be comparable to that of chloramphenicol for treatment of pneumonic pasteurellosis involving ampicillin-resistant strains of P. haemolytica. PMID:17422967

  3. Plasmid coding for transferable drug resistance in bacteria isolated from cultured rainbow trout.

    PubMed Central

    Toranzo, A E; Combarro, P; Lemos, M L; Barja, J L

    1984-01-01

    The occurrence of drug resistance and plasmid-mediated transferability was investigated in 170 strains belonging to eight bacterial groups isolated from cultured rainbow trout. It was found that 87.6% of the strains were resistant to at least one drug, with the highest percentages of resistance being detected for ampicillin (54.7%), sulfadiazine (46.5%), nitrofurantoin (38.2%), and chloramphenicol (37.0%). Six enterobacteria, two Vibrio, and one Aeromonas isolate transferred resistance factors to Escherichia coli K-12. The most common transmissible R factor determined resistance to chloramphenicol and sulfadiazine, demonstrating an association between a specific plasmid and the resistance pattern transferred. The presence of chloramphenicol in fish food was detected by bioassay. In general, transfer frequencies were similar in primary and secondary matings, which indicate the potential water-borne dissemination of these R plasmids. Images PMID:6508296

  4. Antibiotic Resistance Trends in Enteropathogenic Bacteria Isolated in 1985–1987 and 1995–1998 in Barcelona

    PubMed Central

    Prats, Guillermo; Mirelis, Beatriz; Llovet, Teresa; Muñoz, Carmen; Miró, Elisenda; Navarro, Ferran

    2000-01-01

    Trends in resistance to antimicrobial agents used for therapy have been evaluated with 3,797 enteropathogenic bacteria, Campylobacter, Salmonella, Shigella, and Yersinia, between 1985–1987 and 1995–1998. The greater increase in the rate of resistance was observed in Campylobacter jejuni for quinolones (from 1 to 82%) and tetracycline (from 23 to 72%) and in gastroenteric salmonellae for ampicillin (from 8 to 44%), chloramphenicol (from 1.7 to 26%), and trimethoprim-sulfamethoxazole and nalidixic acid (from less than 0.5 to 11%). Multidrug resistance was detected in several Salmonella serotypes. In the 1995–1998 period, 76% of Shigella strains were resistant to trimethoprim-sulfamethoxazole, 43% were resistant to ampicillin, and 39% were resistant to chloramphenicol. Seventy-two percent of Yersinia enterocolitica O3 strains were resistant to streptomycin, 45% were resistant to sulfonamides, 28% were resistant to trimethoprim-sulfamethoxazole, and 20% were resistant to chloramphenicol. PMID:10770742

  5. Direct Introduction of Genes into Rats and Expression of the Genes

    NASA Astrophysics Data System (ADS)

    Benvenisty, Nissim; Reshef, Lea

    1986-12-01

    A method of introducing actively expressed genes into intact mammals is described. DNA precipitated with calcium phosphate has been injected intraperitoneally into newborn rats. The injected genes have been taken up and expressed by the animal tissues. To examine the generality of the method we have injected newborn rats with the chloramphenicol acetyltransferase prokaryotic gene fused with various viral and cellular gene promoters and the gene for hepatitis B surface antigen, and we observed appearance of chloramphenicol acetyltransferase activity and hepatitis B surface antigen in liver and spleen. In addition, administration of genes coding for hormones (insulin or growth hormone) resulted in their expression.

  6. Effects of antibacterials use in aquaculture on biogeochemical processes in marine sediment.

    PubMed

    Ma, Deyi; Hu, Yingying; Wang, Juying; Ye, Sai; Li, Ai

    2006-08-15

    The effects of chloramphenicol on microorganism in marine sediment were studied by spiked experiments in this paper. The results showed that high concentrations of chloramphenicol could inhibit the activities of microorganism in sediment, and that the growth of strains Pseudomons and Acinetobacter in sediment that can degrade organic matters were inhibited apparently. Furthermore, the resistance of heterotrophic bacteria in sediment had developed due to the use of antibacterials. Based on the above results potential environmental effects of antibacterials on microorganism in marine sediment were analyzed.

  7. Silencing of antibiotic resistance in E. coli with engineered phage bearing small regulatory RNAs.

    PubMed

    Libis, Vincent K; Bernheim, Aude G; Basier, Clovis; Jaramillo-Riveri, Sebastián; Deyell, Matthew; Aghoghogbe, Idonnya; Atanaskovic, Iva; Bencherif, Amel Camélia; Benony, Marguerite; Koutsoubelis, Nicolas; Löchner, Anne C; Marinkovic, Zoran S; Zahra, Sarah; Zegman, Yonatan; Lindner, Ariel B; Wintermute, Edwin H

    2014-12-19

    In response to emergent antibiotic resistance, new strategies are needed to enhance the effectiveness of existing antibiotics. Here, we describe a phagemid-delivered, RNA-mediated system capable of directly knocking down antibiotic resistance phenotypes. Small regulatory RNAs (sRNAs) were designed to specifically inhibit translation of chloramphenicol acetyltransferase and kanamycin phosphotransferase. Nonlytic phagemids coding for sRNA expression were able to infect and restore chloramphenicol and kanamycin sensitivity to populations of otherwise resistant E. coli. This modular system could easily be extended to other bacteria with resistance profiles that depend on specific transcripts.

  8. Tryptophanless Death in Bacillus subtilis

    PubMed Central

    Majerfeld, Irene; Barlati, Sergio; Ciferri, Orio

    1970-01-01

    A decline in colony-forming ability is observed in actively growing cultures of a tryptophan arginine auxotroph of Bacillus subtilis after removal of tryptophan (tryptophanless death). This phenomenon can be prevented by simultaneous starvation of the other required amino acid or by chloramphenicol administered in bacteriostatic concentration but not by actinomycin. Addition of tryptophan analogues not only prevents the death but also allows recovery of the cells that have lost the ability to form colonies on solid media. The term tryptophanless death is therefore inappropriate. Chloramphenicol but not actinomycin inhibits the recovery brought about by tryptophan analogues. PMID:4189906

  9. Effect of Haemophilus influenzae infection and moxalactam on platelet function in children.

    PubMed Central

    Kaplan, S L; Courtney, J T; Kenal, K A

    1987-01-01

    In a prospective randomized study, children with Haemophilus influenzae type b meningitis received moxalactam or ampicillin or chloramphenicol. Of 41 children, 6 had prolonged bleeding times (greater than 6 min), and 7 of 9 tested had abnormal platelet aggregation at hospital admission. At the end of therapy, no children in the ampicillin-chloramphenicol group, compared with 5 of 22 moxalactam-treated children (23%) (P = 0.08), had prolonged bleeding times (6.5 to 7.5 min). Our data suggest that H. influenzae meningitis and treatment with moxalactam may each have an effect on platelet function in children. PMID:3579263

  10. [Multi-resistant strains of Salmonella typhi in Spain].

    PubMed

    Usera, M A; Aladueña, A; Jaime, M L; Raya, C; Fuster, C; Planes, A; Bartolomé, R M

    1992-11-01

    Most Salmonella typhi isolated in Spain are susceptible to antibiotics commonly used in its treatment as chloramphenicol, ampicillin and cotrimoxazole. Three multiresistant strains have been isolated from different patients the last two years. Two phage type M1, biotype xylose tetrationate + strains were isolated from blood of two patients in Bembibre (León). One phage type E1a biotype xylose + tetrationate reductase + strain was isolated from blood and faeces of one patient in Barcelona. All strains harboured a 79 Mdal plasmid responsible for multiresistance, chloramphenicol acetyl transferase production and conjugative.

  11. A novel fexA variant from a canine Staphylococcus pseudintermedius isolate that does not confer florfenicol resistance.

    PubMed

    Gómez-Sanz, Elena; Kadlec, Kristina; Feßler, Andrea T; Zarazaga, Myriam; Torres, Carmen; Schwarz, Stefan

    2013-11-01

    Transposon Tn558 integrated in the chromosomal radC gene was detected for the first time in Staphylococus pseudintermedius. It carried a novel fexA variant (fexAv) that confers only chloramphenicol resistance. The exporter FexAv exhibited two amino acid substitutions, Gly33Ala and Ala37Val, both of which seem to be important for substrate recognition. Site-directed mutagenesis that reverted the mutated base pairs to those present in the original fexA gene restored the chloramphenicol-plus-florfenicol resistance phenotype.

  12. A Novel FexA Variant from a Canine Staphylococcus pseudintermedius Isolate That Does Not Confer Florfenicol Resistance

    PubMed Central

    Gómez-Sanz, Elena; Kadlec, Kristina; Feßler, Andrea T.; Zarazaga, Myriam; Schwarz, Stefan

    2013-01-01

    Transposon Tn558 integrated in the chromosomal radC gene was detected for the first time in Staphylococus pseudintermedius. It carried a novel fexA variant (fexAv) that confers only chloramphenicol resistance. The exporter FexAv exhibited two amino acid substitutions, Gly33Ala and Ala37Val, both of which seem to be important for substrate recognition. Site-directed mutagenesis that reverted the mutated base pairs to those present in the original fexA gene restored the chloramphenicol-plus-florfenicol resistance phenotype. PMID:23979755

  13. siRNA Screen Identifies Trafficking Host Factors that Modulate Alphavirus Infection

    DTIC Science & Technology

    2016-05-20

    chloramphenicol acetyltransferase ( CAT ), wild-type Rac1, or variants thereof. Cells were fixed 170 18 h (VEEV) or 24 h (RVFV) after virus inoculation and stained... CAT , wild-type Rac1, or 174 Rac1 K186E. Cells were infected and stained as in (F). 175 176 Rac1 GTPase Function and Rac1:PIP5K1-α Complex...179 established tetracycline-inducible 293 Flp-In T-REx cell lines that express chloramphenicol 180 acetyltransferase ( CAT , used as a control), wild

  14. Ciprofloxacin does not inhibit mitochondrial functions but other antibiotics do.

    PubMed Central

    Riesbeck, K; Bredberg, A; Forsgren, A

    1990-01-01

    At clinical concentrations, ciprofloxacin did not inhibit mitochondrial DNA replication, oxidative phosphorylation, protein synthesis, or mitochondrial mass (transmembrane potential). No difference in supercoiled forms of DNA was observed. The tetracyclines and chloramphenicol inhibited protein synthesis at clinically achievable concentrations, while rifampin, fusidic acid, and clindamycin did not. PMID:2327755

  15. Limited effects of temafloxacin compared with ciprofloxacin on T-lymphocyte function.

    PubMed Central

    Riesbeck, K; Forsgren, A

    1994-01-01

    Temafloxacin increased interleukin-2 production and mRNA levels and enhanced thymidine incorporation in stimulated lymphocyte cultures. Gamma interferon mRNA levels were unaffected. Temafloxacin also stimulated interleukin-2 gene induction, as revealed in a chloramphenicol acetyltransferase reporter gene system. However, temafloxacin exerted significantly weaker effects in these respects than did ciprofloxacin. Images PMID:8031064

  16. Synthesis and biological activity of nifuroxazide and analogs.

    PubMed

    Tavares, L C; Penna, T C; Amaral, A T

    1997-03-01

    Nifuroxazide and thirteen analogs were synthesized from substituted benzoic acids and minimal inhibitory concentrations were determined using the serial dilution tests, in three sequential steps. Nifuroxazide and chloramphenicol were used as reference standards. The tests were performed in TSB against the standard bacterial strain of Staphylococcus aureus ATCC 25923.

  17. Antibiotics

    NASA Astrophysics Data System (ADS)

    Anhalt, John P.

    A 28-year-old man was transferred to our hospital and underwent surgery for resection of an aortic graft infected with Klebsiella pneumoniae. Antimicrobial therapy consisted of amikacin, cefazolin, chloramphenicol, sulfamethoxazole, and trimethoprim. A request for amikacin and sulfamethoxazole assays was received by the laboratory along with information that the patient had received tobramycin until 24 h before the serum was obtained.

  18. Studies on antibacterial activity of Ficus racemosa Linn. leaf extract.

    PubMed

    Mandal, S C; Saha, B P; Pal, M

    2000-06-01

    Extracts of Ficus racemosa Linn. leaves were tested for antibacterial potential against Escherichia coli ATCC 10536, Basillus pumilis ATCC 14884, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa ATCC 25619 and Staphylococcus aureus ATCC 29737. The effects produced by the extracts were significant and were compared with chloramphenicol. The petroleum ether extract was the most effective against the tested organisms.

  19. ANTIBACTERIAL STUDIES ON LEAF EXTRACT OF ELEPHANTOPUS SCABER Linn

    PubMed Central

    Kumar, S. Suresh; Perumal, P.; Suresh, B.

    2004-01-01

    Methanolic extract of Elephantopus scaber Linn was investigated for its antibacterial activity against Staphylococcus aureus (NCIM – 2079), Escherichia coli (NCIM-2067), Bacillus subtilis (NCIM-2063), Pseudomonas aeruginosa (NCIM-2036), Proteus vulgaris (NCIM-2027) at 100 μg/disc by using disc diffusion method. The extract showed significant anti bacterial activity and were compared to chloramphenicol (30 μg/disc). PMID:22557125

  20. Lethal action of quinolones against a temperature-sensitive dnaB replication mutant of Escherichia coli.

    PubMed

    Zhao, Xilin; Malik, Muhammad; Chan, Nymph; Drlica-Wagner, Alex; Wang, Jian-Ying; Li, Xinying; Drlica, Karl

    2006-01-01

    Inhibition of DNA replication in an Escherichia coli dnaB-22 mutant failed to block quinolone-mediated lethality. Inhibition of protein synthesis by chloramphenicol inhibited nalidixic acid lethality and, to a lesser extent, ciprofloxacin lethality in both dnaB-22 and wild-type cells. Thus, major features of quinolone-mediated lethality do not depend on ongoing replication.

  1. Pathogenesis and Management of Gonorrhea

    DTIC Science & Technology

    1987-01-01

    the complaints. The complaints, which range from lower abdomen pain, urgency, frequency and vaginal discharge, are most prominent at the time of...Inheritance of low-level resistance to penicillin, tetracycline, and chloramphenicol in: Neisseria gonorrhoeae. J. Bacteriol. 124:740, (1975). 8

  2. Diagnosis, Treatment, and Prevention of Malaria

    DTIC Science & Technology

    1992-11-01

    Precautions. Candida vaginitis is a troubling and common side effect. Photosensitivity reactions are much less common, and esophageal ulcerations are a rare...crosses into CSF: chloramphenicol or third generation cephalosporin A parenteral antiemetic Bladder catheters Nurses trained to measure vital signs, input...in the evening. Doxycycline use can also be associated with Candida infections, especially vaginitis , nausea, vomiting, and diarrhea. Pyrimethamine

  3. Genome engineering using a synthetic gene circuit in Bacillus subtilis.

    PubMed

    Jeong, Da-Eun; Park, Seung-Hwan; Pan, Jae-Gu; Kim, Eui-Joong; Choi, Soo-Keun

    2015-03-31

    Genome engineering without leaving foreign DNA behind requires an efficient counter-selectable marker system. Here, we developed a genome engineering method in Bacillus subtilis using a synthetic gene circuit as a counter-selectable marker system. The system contained two repressible promoters (B. subtilis xylA (Pxyl) and spac (Pspac)) and two repressor genes (lacI and xylR). Pxyl-lacI was integrated into the B. subtilis genome with a target gene containing a desired mutation. The xylR and Pspac-chloramphenicol resistant genes (cat) were located on a helper plasmid. In the presence of xylose, repression of XylR by xylose induced LacI expression, the LacIs repressed the Pspac promoter and the cells become chloramphenicol sensitive. Thus, to survive in the presence of chloramphenicol, the cell must delete Pxyl-lacI by recombination between the wild-type and mutated target genes. The recombination leads to mutation of the target gene. The remaining helper plasmid was removed easily under the chloramphenicol absent condition. In this study, we showed base insertion, deletion and point mutation of the B. subtilis genome without leaving any foreign DNA behind. Additionally, we successfully deleted a 2-kb gene (amyE) and a 38-kb operon (ppsABCDE). This method will be useful to construct designer Bacillus strains for various industrial applications.

  4. An In Vitro Study on the Effects of Nisin on the Antibacterial Activities of 18 Antibiotics against Enterococcus faecalis

    PubMed Central

    Ling, Junqi; Ma, Jinglei; Huang, Lijia; Zhang, Luodan

    2014-01-01

    Enterococcus faecalis rank among the leading causes of nosocomial infections worldwide and possesses both intrinsic and acquired resistance to a variety of antibiotics. Development of new antibiotics is limited, and pathogens continually generate new antibiotic resistance. Many researchers aim to identify strategies to effectively kill this drug-resistant pathogen. Here, we evaluated the effect of the antimicrobial peptide nisin on the antibacterial activities of 18 antibiotics against E. faecalis. The MIC and MBC results showed that the antibacterial activities of 18 antibiotics against E. faecalis OG1RF, ATCC 29212, and strain E were significantly improved in the presence of 200 U/ml nisin. Statistically significant differences were observed between the results with and without 200 U/ml nisin at the same concentrations of penicillin or chloramphenicol (p<0.05). The checkerboard assay showed that the combination of nisin and penicillin or chloramphenicol had a synergetic effect against the three tested E. faecalis strains. The transmission electron microscope images showed that E. faecalis was not obviously destroyed by penicillin or chloramphenicol alone but was severely disrupted by either antibiotic in combination with nisin. Furthermore, assessing biofilms by a confocal laser scanning microscope showed that penicillin, ciprofloxacin, and chloramphenicol all showed stronger antibiofilm actions in combination with nisin than when these antibiotics were administered alone. Therefore, nisin can significantly improve the antibacterial and antibiofilm activities of many antibiotics, and certain antibiotics in combination with nisin have considerable potential for use as inhibitors of this drug-resistant pathogen. PMID:24586598

  5. Antimicrobial susceptibility to azithromycin among Salmonella enterica isolated from the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Due to emerging resistance to traditional antimicrobial agents such as ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol, azithromycin is increasingly used for the treatment of invasive Salmonella infections. In the present study, 696 isolates of non-Typhi Salmonella collected from human...

  6. Bacteriology of War Wounds at the Time of Injury

    DTIC Science & Technology

    2006-09-01

    Staphylococcus aureus isolates were methi- cillin-resistant S. aureus (MRSA). Conspicuously absent were streptococci (especially Streptococcus ...gangrene between World War I and the Korean War. Implementation of penicillin use after wounding during World War II probably led to the diminution of S...obtained 5 days after surgical therapy and implementation of antimicrobial treatment (typi- cally penicillin with streptomycin sulfate, chloramphenicol so

  7. Geraniol Restores Antibiotic Activities against Multidrug-Resistant Isolates from Gram-Negative Species▿ †

    PubMed Central

    Lorenzi, Vannina; Muselli, Alain; Bernardini, Antoine François; Berti, Liliane; Pagès, Jean-Marie; Amaral, Leonard; Bolla, Jean-Michel

    2009-01-01

    The essential oil of Helichrysum italicum significantly reduces the multidrug resistance of Enterobacter aerogenes, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii. Combinations of the two most active fractions of the essential oil with each other or with phenylalanine arginine β-naphthylamide yield synergistic activity. Geraniol, a component of one fraction, significantly increased the efficacy of β-lactams, quinolones, and chloramphenicol. PMID:19258278

  8. 75 FR 28811 - Office of Biotechnology Activities; Recombinant DNA Research: Proposed Actions Under the NIH...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-24

    ... introduction of drug resistance into a microorganism if the introduction of that drug resistance trait can... resistance trait into an attenuated strain (CO92 lcr-) of Yersinia pestis has been submitted to the NIH... chloramphenicol resistance into these strains, thereby creating lcr- Y. pestis strains that are resistant...

  9. [Side effects of antibiotics].

    PubMed

    Hoigné, R

    1975-03-01

    The clinically severe and newer forms of antibiotic side effects are reviewed. The study covers the following antibiotics: penicillins, cephalosporins, aminoglycosides and polymyxins, tetracyclines, chloramphenicol and thiamphenicol, macrolides and lincomycin, rifamycins and sulfonamides. Special reference is made to (1) hematologic side effects, and (2) general evaluation of drug reactions. The relationship between reaction time and clinical symptoms is of particular practical significance.

  10. Chemotherapy of Bacterial Plasmids

    DTIC Science & Technology

    1979-01-29

    multiresistance to chemotherapeutic drugs, mediated drug resistance are the emergence of strains determined by R-plasmids, causes treatment failures of Haemophilus ... influenzae , resistant to ampicillin [8] of hospital infections, foremost in patients with a or chloramphenicol [9] and of Neisseria gonorrhoeae

  11. Drug-susceptibility of isolates of Brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in Japan in 2009

    PubMed Central

    KAJIWARA, Keita; KOZAWA, Midori; KANAZAWA, Takuya; UETSUKA, Kouji; NAKAJIMA, Hiromi; ADACHI, Yoshikazu

    2015-01-01

    Twenty nine isolates identified as Brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. The isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin G, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with MIC50 values ranging from 0.39 to 3.13. PMID:26596637

  12. The identification of antibacterial compounds for the development of enhanced media for the detection of foodborne fungi.

    PubMed

    Tournas, V H; Kohn, J S; Katsoudas, E J

    2007-08-15

    In an effort to identify a more suitable antibiotic for utilization in mycological media, 12 food borne fungal species from various genera including Alternaria alternata, Aspergillus flavus, A. niger, Eurotium chevalieri, Fusarium moniliforme, Penicillium sp., Rhizopus stolonifer, Saccharomyces cerevisiae, Candida tropicalis, Geotrichum candidum, Rhodotorula glutinis and Kluyveromyces thermotolerans along with 21 chloramphenicol-resistant bacterial isolates from fresh produce and ATCC cultures of Pseudomonas aeruginosa, P. fluorescens, E. coli, Pectobacterium carotovorum, Bacillus cereus and Staphylococcus spp. were tested for their abilities to grow on dichloran rose bengal agar containing various levels of gentamicin, chlortetracycline or chloramphenicol. Results indicated that all fungal isolates except for Rh. glutinis and R. stolonifer grew well on all media tested. Rh. glutinis did not grow on media containing gentamicin whereas R. stolonifer produced very restricted or no growth on these media. All bacterial isolates from fresh produce, P. aeruginosa (ATCC 27853) and P. fluorescens (ATCC BAA-477) grew well at 100, 125 and 150 mg chloramphenicol/liter medium, but they did not grow on media containing chlortetracycline (100, 125, or 150 mg/L) or gentamicin (15, 25, or 35 mg/L). P. aeruginosa (ATCC 10145) grew well on media containing chloramphenicol or gentamicin, but not in the presence of chlortetracycline. P. carotovorum, E. coli, B. cereus and Staphylococcus spp. did not grow on any of the selective media tested.

  13. Fabrication of SERS-active substrates using silver nanofilm-coated porous anodic aluminum oxide for detection of antibiotics.

    PubMed

    Chen, Jing; Feng, Shaolong; Gao, Fang; Grant, Edward; Xu, Jie; Wang, Shuo; Huang, Qian; Lu, Xiaonan

    2015-04-01

    We have developed a silver nanofilm-coated porous anodic aluminum oxide (AAO) as a surface-enhanced Raman scattering (SERS)-active substrate for the detection of trace level of chloramphenicol, a representative antibiotic in food systems. The ordered aluminum template generated during the synthesis of AAO serves as a patterned matrix on which a coated silver film replicates the patterned AAO matrix to form a 2-dimensional ordered nanostructure. We used atomic force microscopy and scanning electron microscopy images to determine the morphology of this nanosubstrate, and characterized its localized surface plasmon resonance by ultraviolet-visible reflection. We gauged the SERS effect of this nanosubstrate by confocal micro-Raman spectroscopy (782-nm laser), finding a satisfactory and consistent performance with enhancement factors of approximately 2 × 10(4) and a limit of detection for chloramphenicol of 7.5 ppb. We applied principal component analysis to determine the limit of quantification for chloramphenicol of 10 ppb. Using electromagnetic field theory, we developed a detailed mathematical model to explain the mechanism of Raman signal enhancement of this nanosubstrate. With simple sample pretreatment and separation steps, this silver nanofilm-coated AAO substrate could detect 50 ppb chloramphenicol in milk, indicating good potential as a reliable SERS-active substrate for rapid detection of chemical contaminants in agricultural and food products.

  14. Identification of a Novel G2073A Mutation in 23S rRNA in Amphenicol-Selected Mutants of Campylobacter jejuni

    PubMed Central

    Naren, Gaowa; Li, Hui; Xia, Xi; Wu, Congming; Shen, Jianzhong; Zhang, Qijing; Wang, Yang

    2014-01-01

    Objectives This study was conducted to examine the development and molecular mechanisms of amphenicol resistance in Campylobacter jejuni by using in vitro selection with chloramphenicol and florfenicol. The impact of the resistance development on growth rates was also determined using in vitro culture. Methods Chloramphenicol and florfenicol were used as selection agents to perform in vitro stepwise selection. Mutants resistant to the selective agents were obtained from the selection process. The mutant strains were compared with the parent strain for changes in MICs and growth rates. The 23S rRNA gene and the L4 and L22 ribosomal protein genes in the mutant strains and the parent strain were amplified and sequenced to identify potential resistance-associated mutations. Results C. jejuni strains that were highly resistant to chloramphenicol and florfenicol were obtained from in vitro selection. A novel G2073A mutation in all three copies of the 23S rRNA gene was identified in all the resistant mutants examined, which showed resistance to both chloramphenicol and florfenicol. In addition, all the mutants selected by chloramphenicol also exhibited the G74D modification in ribosomal protein L4, which was previously shown to confer a low-level erythromycin resistance in Campylobacter species. The mutants selected by florfenicol did not have the G74D mutation in L4. Notably, the amphenicol-resistant mutants also exhibited reduced susceptibility to erythromycin, suggesting that the selection resulted in cross resistance to macrolides. Conclusions This study identifies a novel point mutation (G2073A) in 23S rRNA in amphenicol-selected mutants of C. jejuni. Development of amphenicol resistance in Campylobacter likely incurs a fitness cost as the mutant strains showed slower growth rates in antibiotic-free media. PMID:24728007

  15. Elimination of the amphibian chytrid fungus Batrachochytrium dendrobatidis by Archey's frog Leiopelma archeyi.

    PubMed

    Bishop, Phillip J; Speare, Rick; Poulter, Russell; Butler, Margi; Speare, Benjamin J; Hyatt, Alex; Olsen, Veronica; Haigh, Amanda

    2009-03-09

    Archey's frog Leiopelma archeyi is a critically endangered New Zealand endemic species. The discovery of the emerging infectious disease, chytridiomycosis, in wild populations of this frog raised concern that this disease may drive the species to extinction. Twelve wild-caught Archey's frogs naturally infected with the amphibian chytrid fungus Batrachochytrium dendrobatidis were monitored in captivity by observing clinical signs, measuring weight gain, and performing repeated PCR tests. Eight frogs were treated with topical chloramphenicol, without PCR results being available, for B. dendrobatidis at the day of entry of the frog into the trial. Eleven of the 12 frogs (92%) cleared their infection within 3 mo of capture, even though they were held at 15 degrees C and in high humidity, conditions that are ideal for the survival and propagation of B. dendrobatidis. B. dendrobatidis in the remaining frog tested positive for the fungus was eliminated after treatment with topical chloramphenicol. None of the 8 frogs exposed to chloramphenicol showed any acute adverse reactions. Archey's frog appears to have a low level of susceptibility to the clinical effects of chytridiomycosis. Individual frogs can eliminate B. dendrobatidis and Archey's frog can apparently be treated with topical chloramphenicol with no acute adverse reactions. However, the small number of specimens treated here requires that more extensive testing be done to confirm the safety of chloramphenicol. The significance of the amphibian chytrid fungus for wild populations of Archey's frog needs to be determined by a longitudinal study in an infected wild population to correlate the presence of B. dendrobatidis in individual frogs. Such a study should occur over a period of at least 3 yr with clinical assessment and monitoring of survival, growth and body condition parameters.

  16. Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part II: In vitro antimicrobial activity evaluation of a bacteriophage cocktail and several antibiotics.

    PubMed

    Santos, T M A; Gilbert, R O; Caixeta, L S; Machado, V S; Teixeira, L M; Bicalho, R C

    2010-01-01

    The use of pathogenic-specific antimicrobials, as proposed by bacteriophage therapy, is expected to reduce the incidence of resistance development. Eighty Escherichia coli isolated from uteri of Holstein dairy cows were phenotypically characterized for antimicrobial resistance to ampicillin, ceftiofur, chloramphenicol, florfenicol, spectinomycin, streptomycin, and tetracycline by broth microdilution method. The lytic activity of a bacteriophage cocktail against all isolates was performed by a similar method. Additionally, the effect of different concentrations of antimicrobials and multiplicities of infections (MOI) of the bacteriophage cocktail on E. coli growth curve was measured. Isolates exhibited resistance to ampicillin (33.7%), ceftiofur (1.2%), chloramphenicol (100%), and florfenicol (100%). All strains were resistant to at least 2 of the antimicrobial agents tested; multidrug resistance (>or=3 of 7 antimicrobials tested) was observed in 35% of E. coli isolates. The major multidrug resistance profile was found for ampicillin-chloramphenicol-florfenicol, which was observed in more than 96.4% of the multidrug-resistant isolates. The bacteriophage cocktail preparation showed strong antimicrobial activity against multidrug-resistant E. coli. Multiplicity of infection as low as 10(-4) affected the growth of the E. coli isolates. The ratio of 10 bacteriophage particles per bacterial cell (MOI=10(1)) was efficient in inhibiting at least 50% of all isolates. Higher MOI should be tested in future in vitro studies to establish ratios that completely inhibit bacterial growth during longer periods. All isolates resistant to florfenicol were resistant to chloramphenicol and, because florfenicol was recently introduced into veterinary clinics, this finding suggests that the selection pressure of chloramphenicol, as well as other antimicrobials, may still play a relevant role in the emergence and dissemination of florfenicol resistance in E. coli. The bacteriophage

  17. The In Vitro Antimicrobial Effects of Lavandula angustifolia Essential Oil in Combination with Conventional Antimicrobial Agents

    PubMed Central

    de Rapper, Stephanie; Viljoen, Alvaro

    2016-01-01

    The paper focuses on the in vitro antimicrobial activity of Lavandula angustifolia Mill. (lavender) essential oil in combination with four commercial antimicrobial agents. Stock solutions of chloramphenicol, ciprofloxacin, nystatin, and fusidic acid were tested in combination with L. angustifolia essential oil. The antimicrobial activities of the combinations were investigated against the Gram-positive bacterial strain Staphylococcus aureus (ATCC 6538) and Gram-negative Pseudomonas aeruginosa (ATCC 27858) and Candida albicans (ATCC 10231) was selected to represent the yeasts. The antimicrobial effect was performed using the minimum inhibitory concentration (MIC) microdilution assay. Isobolograms were constructed for varying ratios. The most prominent interaction was noted when L. angustifolia essential oil was combined with chloramphenicol and tested against the pathogen P. aeruginosa (ΣFIC of 0.29). Lavendula angustifolia essential oil was shown in most cases to interact synergistically with conventional antimicrobials when combined in ratios where higher volumes of L. angustifolia essential oil were incorporated into the combination. PMID:27891157

  18. Antibiotics and immunity: effects of antibiotics on mitogen responsiveness of lymphocytes and interleukin-2 production.

    PubMed

    Ibrahim, M S; Maged, Z A; Haron, A; Khalil, R Y; Attallah, A M

    1988-12-01

    The immunomodulating properties of antimicrobial drugs may have important implications in prescriptive practice. This is particularly so for patients whose immune system has been compromised. In this study, tetracycline, cephalothin, rifampicin, polymyxin B and nitrofurantoin reduced mitogen responsiveness of both B and T lymphocytes of mouse spleen cells and human peripheral blood lymphocytes in vitro in a dose-dependent fashion. Ampicillin, chloramphenicol, gentamicin, streptomycin and erythromycin had no effect. In the in vivo study none of the antibiotics affected mouse spleen cell transformation in response to mitogen. The addition of interleukin-2 (IL-2) did not prevent the effect of the antibiotics tested on human lymphocytes in vitro. Cephalothin, chloramphenicol and gentamicin decreased IL-2 production by mouse spleen cells in vitro.

  19. Antibiotics and immunity: effects of antibiotics on natural killer, antibody dependent cell-mediated cytotoxicity and antibody production.

    PubMed

    Ibrahim, M S; Maged, Z A; Haron, A; Khalil, R Y; Attallah, A M

    1987-12-01

    We studied the effects of antibiotics on natural killer (NK), antibody dependent cell-mediated cytotoxicity (ADCC) and immunoglobulin production. When human peripheral blood lymphocytes were incubated overnight with the antibiotic before the assay, nitrofurantoin significantly reduced NK but not ADCC activity. Nitrofurantoin also suppressed both spontaneous and interferon-enhanced NK activities in a dose-dependent fashion. Though it did not affect spontaneous ADCC activity, nitrofurantoin suppressed interferon enhancement of ADCC. Chloramphenicol significantly decreased the number of plaque forming cells in mice. In addition to chloramphenicol, tetracycline, rifampicin, cephalothin, polymyxin B and nitrofurantoin reduced mitogen-induced polycloned immunoglobulin synthesis. Results of this study may have clinical relevance, especially in treating immunocompromised patients.

  20. Minimal inhibitory concentrations of antimicrobial agents against Actinobacillus pleuropneumoniae.

    PubMed Central

    Nadeau, M; Larivière, S; Higgins, R; Martineau, G P

    1988-01-01

    Forty-five isolates of Actinobacillus pleuropneumoniae were tested for susceptibility to 12 antimicrobial agents using a microdilution method for the minimal inhibitory concentration determinations. These results confirmed the high prevalence of A. pleuropneumoniae strains resistant to antibiotics as reported earlier using the disc diffusion method (Kirby-Bauer method). While 36% of the isolates were resistant to the penicillins, 47% were resistant to chloramphenicol and 68% were resistant to tetracycline. Minimal inhibitory concentrations for the resistant isolates were approximately 32 times higher than those for the susceptible isolates to the above antibacterial agents. The isolates were in general weakly susceptible or resistant to spectinomycin, lincomycin, tiamulin and spiramycin whereas most of them were susceptible to gentamicin, trimethoprim and erythromycin. The susceptibility pattern was similar throughout the 1980 to 1984 period. The 14 serotype 5 isolates were more resistant to tetracycline but less resistant to chloramphenicol and the penicillins than the 28 serotype 1 isolates. PMID:3167716

  1. [Hydrogen metabolism in Anabaena variabilis in the dark].

    PubMed

    Gogotov, I N; Kosiak, A V

    Cells and extracts of the cyanobacterium Anabaena variabilis are capable of hydrogen absorption in the dark in the presence of H-acceptors with various redox potentials. Preliminary adaptation of the cells to anaerobic conditions has no effect on the process. A. variabilis can also evolve hydrogen in the dark. Reduced methylviologen (RMV), formiate, pyruvate, and glucose may be substrates for hydrogen evolution by the cells. The extracts evolve hydrogen in the presence of RMV, benzylviologen, azocarmine, or NAD (P) H + ATP. No adaptation of the cells to anaerobic conditions is required for hydrogen evolution from RMV, and chloramphenicol has no effect on the process. The rate of hydrogen evolution is however higher in the cells adapted to anaerobic conditions. Production of hydrogen from pyruvate by the cells adapted and non-adapted to anaerobic conditions was detected only after their incubation with the substrate during 2--3 hours, and chloramphenicol inhibited the process.

  2. Antibiotic resistance of Campylobacter in raw retail chickens and imported chicken portions.

    PubMed Central

    Wilson, I. G.

    2003-01-01

    Campylobacter isolates from raw retail chickens (n = 434) sampled between 1998 and 2000 were tested for resistance to 12 antibiotics. Among 208 campylobacters tested, more than 90% of isolates were susceptible to 4 out of 9 antibiotics (nalidixic acid, erythromycin, chloramphenicol and gentamicin). Most campylobacters were resistant to 3 antibiotics and multiple resistance was found in 4%. Ciprofloxacin resistance was 11%. Campylobacter contamination (28%) in imported chickens (n = 150) was almost half that found in local whole chickens (50%), but the resistance of imported isolates (n = 42) was similar to that of local campylobacters. Resistance in isolates from imported chicken breasts was generally more common, but to only 4 antibiotics. Resistance patterns of chicken isolates were compared to human clinical isolates (n = 494), and a greater similarity was found between the clinical and local isolates than with imported campylobacters. Lower chloramphenicol resistance was found in clinical Campylobacter isolates than in those from chicken sources. PMID:14959786

  3. In vitro interactions of Peucedanum officinale essential oil with antibiotics.

    PubMed

    Miladinović, Dragoljub L; Ilić, Budimir S; Kocić, Branislava D; Miladinović, Ljiljana C; Marković, Marija S

    2015-01-01

    The chemical composition and antibacterial activity of Peucedanum officinale L. (Apiaceae) essential oil were examined, as well as the association between it and antibiotics: tetracycline, streptomycin and chloramphenicol. The interactions of the essential oil with antibiotics were evaluated using the microdilution checkerboard assay. Monoterpene hydrocarbons, with α-phellandrene as the dominant constituent, were the most abundant compound class of the essential oil of P. officinale. The researched essential oil exhibited slight antibacterial activity against the tested bacterial strains in vitro. On the contrary, essential oil of P. officinale possesses a great synergistic potential with chloramphenicol and tetracycline. Their combinations reduced the minimum effective dose of the antibiotic and, consequently, minimised its adverse side effects. In addition, investigated interactions are especially successful against Gram-negative bacteria, the pharmacological treatment of which is very difficult nowadays.

  4. Global surveillance of antibiotic sensitivity of Vibrio cholerae*

    PubMed Central

    O'Grady, F.; Lewis, M. J.; Pearson, N. J.

    1976-01-01

    Strains of Vibrio cholerae—1156 from various parts of the world—were examined by standardized antibiotic sensitivity tests in one centre, to determine the global incidence of antibiotic resistance in this organism and to assess the extent to which differences in methods of sensitivity testing might be responsible for discrepancies in the reported incidence of resistant strains. Of the strains examined, 1127 were fully sensitive to ampicillin, chloramphenicol, tetracycline, furazolidone, and three different sulphonamides, 27 showed stable and reproducible resistance to one or more of these agents, and 2 proved to contain a minority of cells with unstable, presumably plasmid-borne, resistance to chloram-phenicol. Unstable resistance to antibiotics may be common in V. cholerae but rarely recognized, and may account for some of the discrepancies in the reported incidence of resistant strains. PMID:1088100

  5. Liposomes containing drugs for treatment of vaginal infections.

    PubMed

    Pavelić, Z; Skalko-Basnet, N; Jalsenjak, I

    1999-08-01

    To develop a novel vaginal delivery system, able to effectively deliver entrapped drugs during an extended period of time at the site of action, liposomes made of phosphatidylcholine were prepared by two different methods, namely the polyol dilution method and the proliposome method. Liposomes containing three commonly applied drugs in the treatment of vaginal infections: clotrimazole, metronidazole and chloramphenicol were tested for in vitro stability (in buffers at pH 4.5 and 5.9 representing pre- and postmenopausal vaginal pH). In situ stability (in the presence of cow vaginal mucosa) showed that after 6 h incubation (at 37 degrees C), liposomes retained more than 40% of originally entrapped clotrimazole, 28% of entrapped metronidazole or 37% of entrapped chloramphenicol. In vitro and in situ stability studies confirmed the applicability of liposomes as a carrier system for vaginal delivery. Even after 24 h of incubation in the presence of vaginal mucosa liposomes retained sufficient amounts of entrapped drugs.

  6. A study in vitro of the sensitivity to antibiotics of Bacteroides fragilis.

    PubMed

    Ingham, H R; Selkon, J B; Codd, A A; Hale, J H

    1968-07-01

    During a two-year period of observation Bacteroides species were isolated from specimens of pus and vaginal swabs from 115 patients in this hospital. Thirty-five representative strains proved on examination to be Bacteroides fragilis. Minimal inhibitory and minimal bactericidal concentrations of six antibiotics for these strains were determined. All strains were resistant to streptomycin, neomycin, and polymyxin, slightly sensitive to penicillin and ampicillin, and fully sensitive to tetracycline, chloramphenicol, erythromycin, and lincomycin. The minimum bactericidal concentrations of chloramphenicol, erythromycin, and lincomycin were two to four times the minimal inhibitory concentrations. Tetracycline failed to exert any consistent bactericidal effect.The treatment of patients with infections caused by B. fragilis is discussed in the light of the findings in vitro.

  7. Advances on the chromatographic determination of amphenicols in food.

    PubMed

    Guidi, Letícia R; Tette, Patrícia A S; Fernandes, Christian; Silva, Luiza H M; Gloria, Maria Beatriz A

    2017-01-01

    Antibiotics are widely used in veterinary medicine to treat and prevent diseases and their residues can remain in food of animal origin causing adverse effects to human health. Amphenicols (chloramphenicol, thiamphenicol, and florfenicol) may be found in foodstuffs, although the use of chloramphenicol has been prohibited in many countries due to its high toxicity. Since these antibiotics are usually present at trace levels in food, sensitive and selective techniques are required to detect them. This paper reviews analytical methods used since 2002 for the quantitative analysis of amphenicols in food. Sample preparation and separation/detection techniques are described and compared. The advantages and disadvantages of these procedures are discussed. Furthermore, the worldwide legislation and occurrence of these antibiotics in food matrices as well as future trends are also presented.

  8. Direct identification and recognition of yeast species from clinical material by using albicans ID and CHROMagar Candida plates.

    PubMed Central

    Baumgartner, C; Freydiere, A M; Gille, Y

    1996-01-01

    Two chromogenic media, Albicans ID and CHROMagar Candida agar plates, were compared with a reference medium, Sabouraud-chloramphenicol agar, and standard methods for the identification of yeast species. This study involved 951 clinical specimens. The detection rates for the two chromogenic media for polymicrobial specimens were 20% higher than that for the Sabouraud-chloramphenicol agar plates. The rates of identification of Candida albicans for Albicans ID and CHROMagar Candida agar plates were, respectively, 37.0 and 6.0% after 24 h of incubation and 93.6 and 92.2% after 72 h of incubation, with specificities of 99.8 and 100%. Furthermore, CHROMagar Candida plates identified 13 of 14 Candida tropicalis and 9 of 12 Candida krusei strains after 48 h of incubation. PMID:8789038

  9. Low level of cross-resistance between triclosan and antibiotics in Escherichia coli K-12 and E. coli O55 compared to E. coli O157.

    PubMed

    Braoudaki, Maria; Hilton, Anthony Craig

    2004-06-15

    Misuse of biocides has encouraged the emergence of resistance and cross-resistance in certain strains. This study investigated resistance of triclosan-adapted Escherichia coli K-12 and E. coli O55 to antimicrobial agents and compared these to E. coli O157:H7. Cross-resistance in E. coli K-12 and E. coli O55 was observed however to a lesser extent than in E. coli O157:H7. Triclosan-adapted E. coli K-12 demonstrated cross-resistance to chloramphenicol, whereas triclosan-adapted E. coli O55 exhibited resistance to trimethoprim. In comparison, E. coli O157:H7 was resistant to chloramphenicol, tetracycline, amoxicillin, amoxicillin/clavulanic acid, trimethoprim, benzalkonium chloride and chlorohexidine suggesting strain specific rather than general resistance mechanisms.

  10. Correlation Between the Rate of Ribonucleic Acid Synthesis and the Level of Valyl Transfer Ribonucleic Acid in Mutants of Escherichia coli

    PubMed Central

    Kaplan, Sam

    1969-01-01

    By use of a mutant of Escherichia coli with a partially thermolabile transfer ribonucleic acid (tRNA) synthase, it was possible to regulate the rate of RNA synthesis over a 10-fold range. The addition of chloramphenicol to cultures kept at the nonpermissive temperature stimulated RNA synthesis. The longer the culture was kept at the nonpermissive temperature prior to addition of chloramphenicol, the lower was the resulting rate of RNA synthesis. The decrease in the rate of incorporation of labeled uracil into RNA was correlated with the decrease in the level of valyl tRNA. Additional experiments provided evidence which may be interpreted as indicating that valyl tRNA does not, by itself, react with the RNA-forming system. PMID:4891259

  11. Antibiotic resistance monitoring in heterotrophic bacteria from anthropogenic-polluted seawater and the intestines of oyster Crassostrea hongkongensis.

    PubMed

    Wang, Rui Xuan; Wang, AnLi; Wang, Jiang Yong

    2014-11-01

    A total of 1,050 strains of heterotrophic bacteria isolated from farming seawater and the intestines of oyster species Crassostrea hongkongensis were tested for resistance to 10 antibiotics by the Kirby-Bauer diffusion method. The resistant rates of seawater-derived bacteria to chloramphenicol, enrofloxacin, and ciprofloxacin were low (less than 20%), whereas the bacteria obtained from oysters showed low resistance to chloramphenicol and enrofloxacin. Many strains showed high resistant rates (more than 40%) to furazolidone, penicillin G, and rifampin. A total of 285 strains from farming seawater and oysters were resistant to more than three antibiotics. Several strains showed resistance to more than nine antibiotics. Furthermore, the peak resistant rates of the seawater-derived strains to multiple antibiotics overlapped in April, June, September, and November, and those of oyster-derived strains overlapped during April, July, and September. The multi-resistant rate patterns of strains from farming seawater and oyster intestines were similar.

  12. Emergence of tetracycline resistance due to a multiple drug resistance plasmid in Vibrio cholerae O139.

    PubMed

    Yamamoto, T; Nair, G B; Takeda, Y

    1995-04-01

    Of the 173 clinical strains of Vibrio cholerae O139 isolated from India, Bangladesh, and Thailand tested, six strains from India were resistant to tetracycline, ampicillin, chloramphenicol, kanamycin, and gentamicin. These six strains harbored a self-transmissible plasmid that mediated resistance to tetracycline, ampicillin, chloramphenicol, kanamycin, gentamicin, sulfamethoxazole, trimethoprim, and O/129. The multiple drug resistance plasmids were 200 kb in size and belonged to the incompatibility group C. Although a majority of the O139 strains (94.8%) were highly resistant to streptomycin, sulfamethoxazole, trimethoprim, and O/129, the tetracycline-susceptible strains so far tested were plasmid-negative. The data suggest the existence of two distinct multiple antimicrobial agent resistance (MAR) patterns in V. cholerae O139.

  13. Antibiotic sensitivity patterns among Indian strains of avian Pasteurella multocida.

    PubMed

    Shivachandra, S B; Kumar, A A; Biswas, A; Ramakrishnan, M A; Singh, Vijendra P; Srivastava, S K

    2004-11-01

    An investigation was carried out to study the antibiotic sensitivity of avian strains of Pasteurella multocida and to select an effective antimicrobial agent for control of avian pasteurellosis in India. A total of 123 strains of P. multocida recently isolated from different avian species (chicken, duck, turkey, quail, and goose), from different regions of India were subjected to antibiotic sensitivity tests using 20 different antibiotics. Absolute resistance was observed against sulfadiazine. The studies indicated that the strains were most sensitive to chloramphenicol (73.98%), followed by enrofloxacin (71.54%), lincomycin (64.23%), norfloxacin (61.79%) and doxycycline-HCl (56.91%). The majority of the strains were found to exhibit intermediate sensitivity. Chloramphenicol was selected and suggested for treatment. Antibiogram studies also revealed the emergence of multidrug-resistant strains of P. multocida among Indian poultry.

  14. Antimicrobial susceptibility of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from various animals.

    PubMed

    Rubin, Joseph E; Ball, Katherine R; Chirino-Trejo, Manuel

    2011-02-01

    This study characterized the antimicrobial susceptibility of 221 Staphylococcus aureus isolated from various species, and 60 canine Staphylococcus pseudintermedius isolated from 1986 through 2000 at the Western College of Veterinary Medicine (WCVM). Resistance of S. aureus was most common to penicillin (31%) and tetracycline (14%); resistance of S. pseudintermedius to penicillin was present in 8% and to tetracycline in 34% of isolates. Resistance to trimethoprim/sulfamethoxazole was only seen among S. pseudintermedius, and there was no resistance to amoxicillin/clavulanate, ampicillin/sulbactam, cephalothin, amikacin, gentamicin, enrofloxacin, chloramphenicol, or rifampin among any isolate. Inducible clindamycin resistance was found in both S. aureus and S. pseudintermedius, highlighting the need for careful interpretation of culture and susceptibility test results. There were significant differences in the minimum inhibitory concentrations of penicillin, ciprofloxacin, enrofloxacin, clindamycin, erythromycin, chloramphenicol, and tetracycline between avian, bovine, equine, and porcine isolates.

  15. [Rocky Mountain spotted fever in Brazil].

    PubMed

    del Sá DelFiol, Fernando; Junqueira, Fábio Miranda; da Rocha, Maria Carolina Pereira; de Toledo, Maria Inês; Filho, Silvio Barberato

    2010-06-01

    Although the number of confirmed cases of spotted fever has been declining in Brazil since 2005, the mortality rate (20% to 30%) is still high in comparison to other countries. This high mortality rate is closely related to the difficulty in making the diagnosis and starting the correct treatment. Only two groups of antibiotics have proven clinical effectiveness against spotted fever: chloramphenicol and tetracyclines. Until recently, the use of tetracyclines was restricted to adults because of the associated bone and tooth changes in children. Recently, however, the American Academy of Pediatrics and various researchers have recommended the use of doxycycline in children. In more severe cases, chloramphenicol injections are often preferred in Brazil because of the lack of experience with injectable tetracycline. Since early diagnosis and the adequate drug treatment are key to a good prognosis, health care professionals must be better prepared to recognize and treat spotted fever.

  16. Purification and characterization of a 4-hydroxybiphenyl UDP-glucuronosyltransferase from rat liver microsomes

    SciTech Connect

    Styczynski, B.; Green, M.; Coffman, B.; Puig, J.; Tephly, T. )

    1991-03-11

    A phenobarbital-inducible rat liver microsomal UDP-glucuronosyltransferase (4-HBP UDPGT) which catalyzes the glucuronidation of 4-hydroxybiphenyl has been purified to homogeneity. The apparent subunit molecular weight of this protein is 52,500 as determined by SDS-PAGE. 4-HBP UDPGT was shown to react with 4-hydroxybiphenyl, p-nitrophenol and 4-methylumbelliferone, but did not react with morphine, testosteron or chloramphenicol. Upon treatment with Endoglycosidase H, the 4-HBP UDPGT underwent about a 2,000 dalton decrease in subunit molecular weight, suggesting that this protein in N-glycosylated. Western blot analysis has revealed immunorecognition of 4-HBP UDPGT by antibodies raised in rabbit against rat 3{alpha}- and 17{beta}-hydroxysteroid UDPGTs. Additionally, the authors have obtained the N-terminal amino acid sequence of 4-HBP UDPGT. These data provide evidence which suggests that this protein is different from another UDPGT previously shown to react with 4-hydroxybiphenyl, testosterone and chloramphenicol.

  17. Synthesis, Characterization, Electrochemical Studies, and In Vitro Antibacterial Activity of Novel Thiosemicarbazone and Its Cu(II), Ni(II), and Co(II) Complexes

    PubMed Central

    Khan, Salman A.; Asiri, Abdullah M.; Al-Amry, Khalid; Malik, Maqsood Ahmad

    2014-01-01

    Metal complexes were prepared by the reaction of thiosemicarbazone with CuCl2, NiCl2, CoCl2, Cu(OAc)2, Ni(OAc)2, and Co(OAc)2. The thiosemicarbazone coordinates to metal through the thionic sulfur and the azomethine nitrogen. The thiosemicarbazone was obtained by the thiosemicarbazide with 3-acetyl-2,5-dimethylthiophene. The identities of these compounds were elucidated by IR, 1H, 13C-NMR, and GC-MS spectroscopic methods and elemental analyses. The antibacterial activity of these compounds was first tested in vitro by the disc diffusion assay against two Gram-positive and two Gram-negative bacteria, and then the minimum inhibitory concentration (MIC) was determined by using chloramphenicol as reference drug. The results showed that compound 1.1 is better inhibitor of both types of tested bacteria as compared to chloramphenicol. PMID:24523641

  18. A Novel Method for Preparation of Gold NanoBipyramids Using Microwave Irradiation and Its Application in Immunosensors

    NASA Astrophysics Data System (ADS)

    Huynh, Trong Phat; Ngo, Vo Ke Thanh; Nguyen, Dang Giang; Nguyen, Hoang Phuong Uyen; Nghiem, Quoc Dat; Lam, Quang Vinh; Huynh, Thanh Dat

    2016-05-01

    Gold nanobipyramids (NBPs) have attracted attention for producing smart sensing devices as diagnostic tools in biotechnological and medical applications, because they show more advantageous plasmonic properties than comparable gold nanorods. Normally, NBPs were synthesized using seed-mediated growth process at room temperature. In this report, our group describes a method for synthesising of NBPs using microwave irradiation with ascorbic acid reduction and cetyltrimethylammonium bromide + silver nitrate (AgNO3) as capping agents. The advantages of this method are a highly effective approach to fast and uniform NBPs. The product was characterized by ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, and x-ray powder diffraction. As an application in quartz crystal microbalance immunosensors, NBPs is conjugated with the chloramphenicol antibodies for signal amplification to detect chloramphenicol residuals in the QCM system.

  19. Synergistic antibacterial activity of Salvia officinalis and Cichorium intybus extracts and antibiotics.

    PubMed

    Stefanović, Olgica D; Stanojević, Dragana D; Comić, Ljiljana R

    2012-01-01

    Synergistic activity of Salvia officinalis and Cichorium intybus extracts and commonly used antibiotics, amoxicillin and chloramphenicol, were evaluated. Interactions between plant extracts and antibiotics were tested by checkerboard method and interpreted as FIC index. Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 and clinical isolates Staphylococcus aureus, Bacillus subtilis, Enterobacter cloacae, Klebsiella pneumoniae, Escherichia coli and Proteus mirabilis were used. Salvia officinalis showed better synergistic capacity than Cichorium intybus. Synergistic interactions were observed between amoxicillin and acetone or ethyl acetate extract of Salvia officinalis and between chloramphenicol and ethyl acetate extract of Salvia officinalis. In the presence of sub-inhibitory concentration (1/4 MIC to 1/32 MIC) of sage extracts, the MIC values of antibiotics were decreased by 2- to 10-fold. Synergism was observed against all test bacteria, except Escherichia coli. The combinations of acetone and ethyl acetate extract from Cichorium intybus and antibiotics resulted in additive and indifferent effects against tested bacteria.

  20. Construction of two selectable markers for integrative/conjugative plasmids in Flavobacterium columnare

    NASA Astrophysics Data System (ADS)

    Zhang, Jin; Zou, Hong; Wang, Liangfa; Huang, Bei; Li, Nan; Wang, Guitang; Nie, Pin

    2012-03-01

    Flavobacterium columnare, the etiological agent of columnaris disease, is one of the most important and widespread bacterial pathogens of freshwater fish. In this study, we constructed two artificial selectable markers (chloramphenicol and spectinomycin resistance) for gene transfer in F. columnare. These two new artificial selectable markers, which were created by placing the chloramphenicol or spectinomycin resistance gene under the control of the native acs regulatory region of F. columnare, were functional in both F. columnare and Escherichia coli. The integrative/conjugative plasmids constructed by using these markers were introduced into F. columnare G4 via electroporation or conjugation. The integrated plasmid DNA was confirmed by Southern blotting and PCR analysis. These two markers can be employed in future investigations into gene deletion and the pathogenicity of virulence factors in F. columnare.

  1. Quinazoline derivatives are efficient chemosensitizers of antibiotic activity in Enterobacter aerogenes, Klebsiella pneumoniae and Pseudomonas aeruginosa resistant strains.

    PubMed

    Chevalier, Jacqueline; Mahamoud, Abdallah; Baitiche, Milad; Adam, Elissavet; Viveiros, Miguel; Smarandache, Adriana; Militaru, Andra; Pascu, Mihail L; Amaral, Leonard; Pagès, Jean-Marie

    2010-08-01

    Amongst the three series of quinazoline derivatives synthesised and studied in this work, some molecules increase the antibiotic susceptibility of Gram-negative bacteria presenting multidrug-resistant phenotypes. N-alkyl compounds induced an increase in the activity of chloramphenicol, nalidixic acid and sparfloxacin, which are substrates of the AcrAB-TolC and MexAB-OprM efflux pumps in clinical isolates. These molecules are able to increase the intracellular concentration of chloramphenicol in efflux pump-overproducing strains. Their activity depends on the antibiotic structure, suggesting that different sites may be involved for the recognition of substrates by a given efflux pump. Quinazoline molecules exhibiting a nitro functional group are more active, and structure-activity relationship studies may be undertaken to identify the pharmacophoric group involved in the AcrB and MexB affinity sites.

  2. High level multiple antibiotic resistance among fish surface associated bacterial populations in non-aquaculture freshwater environment.

    PubMed

    Ozaktas, Tugba; Taskin, Bilgin; Gozen, Ayse G

    2012-12-01

    Freshwater fish, Alburnus alburnus (bleak), were captured from Lake Mogan, situated in Ankara, during spring. The surface mucus of the fish was collected and associated bacteria were cultured and isolated. By sequencing PCR-amplified 16S RNA encoding genes, the isolates were identified as members of 12 different genera: Acinetobacter, Aeromonas, Bacillus, Brevundimonas, Gordonia, Kocuria, Microbacterium, Mycobacterium, Pseudomonas, Rhodococcus, and Staphylococcus, in addition to one strain that was unidentified. The mucus-dwelling bacterial isolates were tested for resistance against ampicillin, kanamycin, streptomycin and chloramphenicol. About 95% of the isolates were found to be resistant to ampicillin, 93% to chloramphenicol, and 88% to kanamycin and streptomycin. A Microbacterium oxydans and the unidentified environmental isolate were resistant to all four antibiotics tested at very high levels (>1600 μg/ml ampicillin and streptomycin; >1120 μg/ml kanamycin; >960 μg/ml chloramphenicol). Only a Kocuria sp. was sensitive to all four antibiotics at the lowest concentrations tested (3.10 μg/ml ampicillin and streptomycin; 2.15 μg/ml kanamycin; 1.85 μg/ml chloramphenicol). The rest of the isolates showed different resistance levels. Plasmid isolations were carried out to determine if the multiple antibiotic resistance could be attributed to the presence of plasmids. However, no plasmid was detected in any of the isolates. The resistance appeared to be mediated by chromosome-associated functions. This study indicated that multiple antibiotic resistance at moderate to high levels is common among the current phenotypes of the fish mucus-dwelling bacterial populations in this temperate, shallow lake which has not been subjected to any aquaculturing so far but under anthropogenic effect being in a recreational area.

  3. Salmonella enterica Typhimurium fljBA operon stability: implications regarding the origin of Salmonella enterica I 4,[5],12:i:.

    PubMed

    Tomiyama, M P O; Werle, C H; Milanez, G P; Nóbrega, D B; Pereira, J P; Calarga, A P; Flores, F; Brocchi, M

    2015-12-29

    Salmonella enterica subsp enterica serovar 4,5,12:i:- has been responsible for many recent Salmonella outbreaks worldwide. Several studies indicate that this serovar originated from S. enterica subsp enterica serovar Typhimurium, by the loss of the flagellar phase II gene (fljB) and adjacent sequences. However, at least two different clones of S. enterica 4,5,12:i:- exist that differs in the molecular events responsible for fljB deletion. The aim of this study was to test the stability of the fljBA operon responsible for the flagellar phase variation under different growth conditions in order to verify if its deletion is a frequent event that could explain the origin and dissemination of this serovar. In fact, coding sequences for transposons are present near this operon and in some strains, such as S. enterica Typhimurium LT2, the Fels-2 prophage gene is inserted near this operon. The presence of mobile DNA could confer instability to this region. In order to examine this, the cat (chloramphenicol acetyltransferase) gene was inserted adjacent to the fljBA operon so that deletions involving this genomic region could be identified. After growing S. enterica chloramphenicol-resistant strains under different conditions, more than 104 colonies were tested for the loss of chloramphenicol resistance. However, none of the colonies were sensitive to chloramphenicol. These data suggest that the origin of S. enterica serovar 4,5,12:i:- from Typhimurium by fljBA deletion is not a frequent event. The origin and dissemination of 4,5,12:i:- raise several questions about the role of flagellar phase variation in virulence.

  4. Role of OmpA in the Multidrug Resistance Phenotype of Acinetobacter baumannii

    PubMed Central

    Fàbrega, Anna; Roca, Ignasi; Sánchez-Encinales, Viviana; Vila, Jordi; Pachón, Jerónimo

    2014-01-01

    Acinetobacter baumannii has emerged as a nosocomial pathogen with an increased prevalence of multidrug-resistant strains. The role of the outer membrane protein A (OmpA) in antimicrobial resistance remains poorly understood. In this report, disruption of the ompA gene led to decreased MICs of chloramphenicol, aztreonam, and nalidixic acid. We have characterized, for the first time, the contribution of OmpA in the antimicrobial resistance phenotype of A. baumannii. PMID:24379205

  5. Synthesis of novel quinoxalinone derivatives by conventional and microwave methods and assessing their biological activity.

    PubMed

    Nasir, Waqar; Munawar, Munawar Ali; Ahmed, Ejaz; Sharif, Ahsan; Ahmed, Saeed; Ayub, Amjad; Khan, Misbahul Ain; Nasim, Faizul Hassan

    2011-10-01

    In this study, twenty-one arylaminoquinoxalinone derivatives were synthesized and their antibacterial activities against Staphylococci aureus, Pseudomonas aureus, Escherichia coli, Bacillus subtilis, Salmonella typhi, and Shigella pneumoniae were evaluated relative to known antibiotics; augmentin, ampicillin, and chloramphenicol. The insecticidal activities of the prepared compounds were also investigated against Tribolium castaneum using permethrin as a standard insecticide. The derivatives were synthesized using both conventional and microwave techniques. Their structures were confirmed using spectral techniques and elemental analysis.

  6. [The antimicrobial activity of a new wound-healing compound ointment].

    PubMed

    Iakushchenko, V A; Biriukova, S V; Dziuban, N F; Piminov, A F; Volianskiĭ, Iu L; Prokopenko, T S

    1996-01-01

    Drug composition containing polyethyleneoxide PEO-400 and PEO-1500 (8:2), antimicrobe agent-chloramphenical as well as new antiinflammatory mean of plant origin EOL in concentration of 5% has been created. Introduction of EOL to the composition of the drug under elaboration permitted decreasing chloramphenicol concentration 2-4 times as compared to concentrations used in contemporary ointments. This has lowered toxic and allergenic properties of the drug suggested for healing the infected wounds.

  7. Draft Genome Sequence of the First NDM-1-Producing Providencia stuartii Strain Isolated in Portugal.

    PubMed

    Manageiro, Vera; Sampaio, Daniel A; Pereira, Patrícia; Rodrigues, Paulo; Vieira, Luís; Palos, Carlos; Caniça, Manuela

    2015-09-24

    We report here the draft genome sequence of the first NDM-1-producing Providencia stuartii strain isolated in Portugal. Sequence analyses revealed the presence of an incompatibility group A/C2 (IncA/C2) plasmid and of diverse acquired genes conferring resistance to β-lactams, aminoglycosides, tetracycline, macrolides, chloramphenicol, and sulfonamides. This sequence contributes to the evaluation of the spread of NDM-1 producers.

  8. Wound infection by multiresistant Staphylococcus sciuri identified by molecular methods.

    PubMed

    Coimbra, Daniel G; Almeida, Alda G C S; Jùnior, Jorge B O; da Silva, Luiz A F; Pimentel, Beatriz J; Gitaì, Daniel L G; Moreira, Luciana S; Silva-Filho, Eurìpedes A; de Andrade, Tiago G

    2011-10-01

    We describe a case of wound infection by multidrug-resistant Staphylococcus sciuri in a patient admitted to hospital for injuries in Agreste Alagoas, Brazil, identified through broad-spectrum PCR and sequencing of 16S rDNA gene. Due to its high resistance profile, the infection was characterized as methicillin-resistant Staphylococcus presenting sensitive only to vancomycin and chloramphenicol. The injury resulting from trauma associated with infection resulted in amputation of the infected limb.

  9. Tsutsugamushi disease presenting with aortic valve endocarditis: a case report and literature review

    PubMed Central

    Yu, Shujie; Yu, Xianguan; Zhou, Bin; Liu, Dinghui; Wang, Min; Zhang, Hui; Qian, Xiaoxian

    2016-01-01

    Tsutsugamushi disease is a zoonotic disease caused by Orientia tsutsugamushi in which humans are accidental hosts. Infective endocarditis associated with Tsutsugamushi disease has not been previously reported. We are describing a case of Tsutsugamushi disease presenting with aortic valve endocarditis. The clinical data of a 67-year-old female with O. tsutsugamushi-induced aortic valve endocarditis was summarized retrospectively and analyzed with a literature review. Treatment of O. tsutsugamushi-induced aortic valve endocarditis with chloramphenicol is recommended. PMID:28078179

  10. Nodular panniculitis in a female toy poodle.

    PubMed

    Post, K

    1983-05-01

    A four month old female toy Poodle showing multiple nodules 2 to 3 cm in diameter, some of which had ruptured over the neck and trunk area, was diagnosed as having nodular panniculitis. The animal had been treated with a course of chloramphenicol and ampicillin for one month with no response. The diagnosis was based on history, clinical signs, laboratory data and dramatic response to the addition of corticosteroid to the antibiotic therapy.

  11. Development and characterisation of highly antibiotic resistant Bartonella bacilliformis mutants

    PubMed Central

    Gomes, Cláudia; Martínez-Puchol, Sandra; Ruiz-Roldán, Lidia; Pons, Maria J.; del Valle Mendoza, Juana; Ruiz, Joaquim

    2016-01-01

    The objective was to develop and characterise in vitro Bartonella bacilliformis antibiotic resistant mutants. Three B. bacilliformis strains were plated 35 or 40 times with azithromycin, chloramphenicol, ciprofloxacin or rifampicin discs. Resistance-stability was assessed performing 5 serial passages without antibiotic pressure. MICs were determined with/without Phe-Arg-β-Napthylamide and artesunate. Target alterations were screened in the 23S rRNA, rplD, rplV, gyrA, gyrB, parC, parE and rpoB genes. Chloramphenicol and ciprofloxacin resistance were the most difficult and easiest (>37.3 and 10.6 passages) to be selected, respectively. All mutants but one selected with chloramphenicol achieved high resistance levels. All rifampicin, one azithromycin and one ciprofloxacin mutants did not totally revert when cultured without antibiotic pressure. Azithromycin resistance was related to L4 substitutions Gln-66 → Lys or Gly-70 → Arg; L4 deletion Δ62–65 (Lys-Met-Tyr-Lys) or L22 insertion 83::Val-Ser-Glu-Ala-His-Val-Gly-Lys-Ser; in two chloramphenicol-resistant mutants the 23S rRNA mutation G2372A was detected. GyrA Ala-91 → Val and Asp-95 → Gly and GyrB Glu474 → Lys were detected in ciprofloxacin-resistant mutants. RpoB substitutions Gln-527 → Arg, His-540 → Tyr and Ser-545 → Phe plus Ser-588 → Tyr were detected in rifampicin-resistant mutants. In 5 mutants the effect of efflux pumps on resistance was observed. Antibiotic resistance was mainly related to target mutations and overexpression of efflux pumps, which might underlie microbiological failures during treatments. PMID:27667026

  12. ANTIBACTERIAL ACTIVITY OF ROOT EXTRACT OF TRIANTHEMA DECANDRA

    PubMed Central

    Jaswanth, A.; Jagannathan, K.; Heisonrobert, S. Jerry; Loganathan, V.; Manimaran, S.; Ruckmani, K.

    2002-01-01

    Methanolic extract of Trianthema decandra was investigated for its antibacterial activity against staphylococcus aureus (NCIM 2079), Escherichia coli (NCIM 2065), Bacillus subtilis (NCIM 2063), Pseudomonas aeruginosa (NICIM 2036) and Proteus vulgaris (NICIM 2027) at 100 μg/disc using disc diffusion method. The extract showed significant antibacterial activity and were comparable to Chloramphenicol (30/ μg/disc). Our findings confirm the traditional therapeutic claims for this herb. PMID:22557045

  13. Sexually Transmitted Diseases

    DTIC Science & Technology

    1989-12-01

    a. 50% asymptomatic or very mild symptoms b. endocervicitis-- vaginal discharge (10) c. urethritis--painful urination d. pelvic inflammatory disease... chloramphenicol , gentamicin, or lincomycin may be effective 3. sexual partners should be examined and treated if required Prevention (106) 1. use of condoms 2...may be asymptomatic in both women and men 2. may present as vaginal discharge and soreness, burning and itching (111-112) 3. the discharge is often

  14. Determination of asymptomatic carrier rate of beta-haemolytic group B Streptococcus in vaginas of pregnant women in Ibadan, Nigeria.

    PubMed

    Olanisebe, S B; Adetosoye, A I

    1986-04-01

    Bacteriological studies of 500 vaginal swabs from pregnant women in second and third trimester from 4 government hospitals in Ibadan showed that 8 (1.6 per cent) were found to be culture-positive for Group B Streptococcus. Five (1.0 per cent) were found to harbour group D streptococci. All the 8 group B Streptococcus isolates were sensitive to erythromycin, methicillin, penicillin G, ampicillin and chloramphenicol in decreasing order. They were resistant to tetracycline, sulphafurazole and streptomycin.

  15. Melibiose is hydrolyzed exocellularly by an inducible exo-alpha-galactosidase in Azotobacter vinelandii.

    PubMed

    Wong, T Y

    1990-07-01

    Azotobacter vinelandii hydrolyzed melibiose exocellularly, leading to an accumulation of free glucose and galactose in the medium. This enzyme could also be induced by galactose, raffinose, and stachyose. The alpha-galactosidase activity could be detected quantitatively by using p-nitrophenyl-alpha-galactopyranoside as a substrate for intact cells. Chloramphenicol totally inhibited the induction of this enzyme. However, benzyl alcohol inhibited the secretion of this enzyme but did not inhibit the biosynthesis of the enzyme.

  16. Successful treatment of amoebic meningoencephalitis in a Chinese living in Hong Kong.

    PubMed

    Wang, A; Kay, R; Poon, W S; Ng, H K

    1993-09-01

    Primary amoebic meningoencephalitis due to Naegleria fowleri was found in a 38-year-old Chinese man living in Hong Kong who presumably acquired the infection from swimming in a hot spring in neighbouring China. Amoebic cysts were identified in tissue taken from a brain abscess. The patient responded to surgical drainage and a 6-week course of amphotericin B, rifampicin and chloramphenicol. This is one of 6 cases of successful treatment of primary amoebic meningoencephalitis documented in the medical literature.

  17. Salmonellosis in Indonesia: Phage-Type of Salmonella oranienburg Obtained from Hospitalized Patients in Jakarta, Indonesia

    DTIC Science & Technology

    1980-09-15

    difference of two proportions) and greater overall reistance (all antibiotics combined P = 0.-001, even when tetracycline isI excluded; P = 0-022 ((’hi...susceptible to dehydration. Phage type I was sig- nificantly more resistant than phage type II to the individual antibiotics : tetra- cycline, chloramphenicol...kanamycin and neomycin. However, there was no difference in their respective antibiotic resistance patterns as measured by disk and MIC assay. All

  18. Intermediate Susceptibility to Ciprofloxacin among Salmonella enterica Serovar Typhi Isolates in Lima, Peru

    PubMed Central

    Lejon, Veerle; Horna, Gertrudis; Astocondor, Lizeth; Vanhoof, Raymond; Bertrand, Sophie; Jacobs, Jan

    2014-01-01

    Thirty-three Salmonella enterica serovar Typhi blood isolates from Lima, Peru (2008 to 2012), were fully susceptible to trimethoprim-sulfamethoxazole, chloramphenicol, ceftriaxone, and tetracycline; 8/33 (24.2%) showed intermediate susceptibility to ciprofloxacin carrying mutations in the quinolone resistance-determining region of the gyrA gene (Ser83-Phe and Asp87-Asn) and in the gyrB gene (Ser464-Phe). PMID:24371234

  19. Effect of cooking and cold storage on biologically active antibiotic residues in meat.

    PubMed Central

    O'Brien, J. J.; Campbell, N.; Conaghan, T.

    1981-01-01

    An investigation was undertaken to see if cooking or cold storage would destroy or decrease the level of biologically active antibiotic in tissues from animals given therapeutic doses of antibiotic on three occasions prior to slaughter. The effects of cooking and cold storage on the biological activity of the residues of ampicillin, chloramphenicol, oxytetracycline, streptomycin and sulphadimidine were varied; in some instances the effects were minimal, in others nil. PMID:7310129

  20. Gene Disruption by Homologous Recombination in the Xylella fastidiosa Citrus Variegated Chlorosis Strain

    PubMed Central

    Gaurivaud, Patrice; Souza, Leonardo C. A.; Virgílio, Andrea C. D.; Mariano, Anelise G.; Palma, Renê R.; Monteiro, Patrícia B.

    2002-01-01

    Mutagenesis by homologous recombination was evaluated in Xylella fastidiosa by using the bga gene, coding for β-galactosidase, as a model. Integration of replicative plasmids by homologous recombination between the cloned truncated copy of bga and the endogenous gene was produced by one or two crossover events leading to β-galactosidase mutants. A promoterless chloramphenicol acetyltransferase gene was used to monitor the expression of the target gene and to select a cvaB mutant. PMID:12200328

  1. Identification of a TAAT-containing motif required for high level expression of the COL1A1 promoter in differentiated osteoblasts of transgenic mice

    NASA Technical Reports Server (NTRS)

    Dodig, M.; Kronenberg, M. S.; Bedalov, A.; Kream, B. E.; Gronowicz, G.; Clark, S. H.; Mack, K.; Liu, Y. H.; Maxon, R.; Pan, Z. Z.; Upholt, W. B.; Rowe, D. W.; Lichtler, A. C.

    1996-01-01

    Our previous studies have shown that the 49-base pair region of promoter DNA between -1719 and -1670 base pairs is necessary for transcription of the rat COL1A1 gene in transgenic mouse calvariae. In this study, we further define this element to the 13-base pair region between -1683 and -1670. This element contains a TAAT motif that binds homeodomain-containing proteins. Site-directed mutagenesis of this element in the context of a COL1A1-chloramphenicol acetyltransferase construct extending to -3518 base pairs decreased the ratio of reporter gene activity in calvariae to tendon from 3:1 to 1:1, suggesting a preferential effect on activity in calvariae. Moreover, chloramphenicol acetyltransferase-specific immunofluorescence microscopy of transgenic calvariae showed that the mutation preferentially reduced levels of chloramphenicol acetyltransferase protein in differentiated osteoblasts. Gel mobility shift assays demonstrate that differentiated osteoblasts contain a nuclear factor that binds to this site. This binding activity is not present in undifferentiated osteoblasts. We show that Msx2, a homeodomain protein, binds to this motif; however, Northern blot analysis revealed that Msx2 mRNA is present in undifferentiated bone cells but not in fully differentiated osteoblasts. In addition, cotransfection studies in ROS 17/2.8 osteosarcoma cells using an Msx2 expression vector showed that Msx2 inhibits a COL1A1 promoter-chloramphenicol acetyltransferase construct. Our results suggest that high COL1A1 expression in bone is mediated by a protein that is induced during osteoblast differentiation. This protein may contain a homeodomain; however, it is distinct from homeodomain proteins reported previously to be present in bone.

  2. Outcome in three groups of patients with typhoid fever in Indonesia between 1948 and 1990.

    PubMed

    van den Bergh, E T; Gasem, M H; Keuter, M; Dolmans, M V

    1999-03-01

    The outcome in three groups of patients with bacteriologically confirmed typhoid fever caused by Salmonella typhi, treated during three episodes between 1948 and 1990 in Java, Indonesia, was compared by retrospective analysis of hospital records. The study population consisted of three groups of patients. Group I (n = 50) was treated in Batavia (the present Jakarta) from 1948 to 1950, Group II (n = 61) in Yogyakarta from 1952 to 1956, Group III (n = 105) in Semarang from 1989 to 1990. Main outcome measures were days until defervescence, early relapses during hospitalization, duration of hospital stay, complications and mortality. Group I received supportive treatment only, Group II low doses of chloramphenicol (total 12.5 g) and Group III full doses of chloramphenicol (total 27 g); occasionally other antibiotics were used. In Group I, II and III the mean number of days until defervescence was 16, 8 and 6 and the mean number of days in hospital 43, 47 and 15, respectively. Mortality was 26%, 10% and 5% and complications occurred in 38%, 18% and 13%, respectively. Between Group I and Group II the differences in mortality and complications were statistically significant (P < 0.05). Compared to Group I the proportion of early relapses was higher in Group II, but was zero in Group III. There were significantly fewer gastrointestinal complications in Group II than in Group I (P < 0.01) and even fewer in Group III. When no antibiotic against S. typhi was available, typhoid fever had a protracted course, and only 74% of patients survived. Even with low dosages of chloramphenicol, defervescence was earlier and mortality and complications decreased dramatically, but early relapses were frequent. Full doses of chloramphenicol for a sufficient period of time only slightly reduced mortality and complications further, but eliminated early relapses completely.

  3. A Literature Review - Problem Definition Studies on Selected Toxic Chemicals. Volume 6. Occupational Health and Safety and Environmental Aspects of Urea-Formaldehyde Resins

    DTIC Science & Technology

    1978-04-01

    such as penicillin, streptomycin, chloramphenicol, tetracycline, erythromycin, neomycin, polymyxin, chlor- tetracycline, oxytetracycline , novobi’ocin...so=-e nitrogen in the polymers was converted to nitrogen gas (N2 ) by the flaming coabustion. The stability of urea-form.aldehyde resins against...armonia (12). The stability of urea-formaldehyde resins against ultraviolet irradiation was studied by applying a film of the resin on the i•r plate

  4. Risk of Relapse Associated with Doxycycline Therapy for Scrub Typhus

    DTIC Science & Technology

    1981-01-01

    143150-9 202 Rickettsiae ard Rickettsial Diseases I. INTRODUCTION Chloramphenicol (Chloromycetin) or oxytetracycline (Terra- mycin) are the drugs...doses of oxytetracycline in a double blind, randomized trial conducted in a population of scrub typhus patients who were exposed in the Pescadores... oxytetracycline , 500 mg every 6 hrs for 7 days (OXY, 23 patients). Drug and placebo were manufactured in such a way and administered every 6 hrs for 7

  5. Transmission, Control and Treatment of Infectious Diseases of Military Importance in Equatorial Asia

    DTIC Science & Technology

    1989-06-22

    antibiotic shown to cause unacceptable risk as a prophylactic agent. Subsequently tetracycline replaced chloramphenicol. Our study was undertaken to...determine and compare the efficacy of each of two different doxycycline, a long acting tetracycline , regimes for the prevention of scrub typhus in a...The Effect of Tetracycline Therapy on the Host Defense of Mice Infected with Ehrlichia sennetsu Ehrlichia sennetsu is the etiological agent of a well

  6. Multiple Clones within Multidrug-Resistant Salmonella enterica Serotype Typhimurium Phage Type DT104

    PubMed Central

    Markogiannakis, Antonis; Tassios, Panayotis T.; Lambiri, Maria; Ward, Linda R.; Kourea-Kremastinou, Jenny; Legakis, Nicholas J.; Vatopoulos, Alkiviadis C.

    2000-01-01

    Six distinct clones were present among Greek multidrug-resistant Salmonella enterica serotype Typhimurium phage type DT104, since isolates belonging to resistance phenotypes including the ACSSuT (ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline) core could be distinguished with respect to their pulsed-field gel electrophoresis patterns, int1 integron structures, and presence or absence of antibiotic resistance genes ant(3")-Ia, pse-1, and tem-1. PMID:10699039

  7. (Electro)Sensing of Phenicol Antibiotics-A Review.

    PubMed

    Pilehvar, Sanaz; Gielkens, Kristoffer; Trashin, Stanislav A; Dardenne, Freddy; Blust, Ronny; De Wael, Karolien

    2016-10-25

    The presence of residues from frequent antibiotic use in animal feed can cause serious health risks by contaminating products meant for human consumption such as meat and milk. The present paper gives an overview of the electrochemical methods developed for the detection of phenicol antibiotic residues (chloramphenicol, thiamphenicol, and florfenicol) in different kinds of foodstuffs. Electrochemical sensors based on different biomolecules and nanomaterials are described. The detection limit of various developed methods with their advantages and disadvantages will be highlighted.

  8. [Transport of phenylalanine and tyrosine in Brevibacterium linens: specificity and incorporation into proteins].

    PubMed

    Boyaval, P; Moreira, E; Desmazeaud, M J

    1984-04-01

    The specificity of phenylalanine and tyrosine carriers was investigated using actively metabolizing cells of Brevibacterium linens. The cellular protein synthesis of resting cells was very weakly inhibited, even with high concentrations of chloramphenicol or tetracycline. The nonaromatic amino acids were weak inhibitors for these carriers, while fluorinate analogues of phenylalanine and tyrosine were very potent competitive inhibitors. In practice these analogues cannot be used to replace amino acids to evaluate transport without incorporation because they are incorporated in cellular proteins.

  9. Mitochondrial inheritance and disease.

    PubMed

    Fine, P E

    1978-09-23

    Spontaneously occurring variants of the D.N.A. content of mitochondria may be responsible for human disease. Among the prime candidates for such a mitochondrial aetiology are certain drug-induced blood dyscrasias, particularly that due to chloramphenicol. Because mitochondria are generally inherited from the female parent, such disorders should be clustered among matroclinally related individuals. The clinical manifestations of such diseases are a function of the manner in which mitochondria are allocated to somatic cells and tissues during development.

  10. The Biochemistry and Physiology of Bacterial Adhesion to Surfaces

    DTIC Science & Technology

    1984-01-20

    proteins may be similar to the "adhesins" employed by enterotoxigenic E. coli strains to adhere to intestinal epithelium (Vosbeck et al., 1979; 1982...chloramphenicol, tetracycline) reduced the adhesiveness of several E. coli strains to intestine monolayer cells. The DNA synthesis inhibitor nalidxic acid made... strains are not encapsulated, since glycocalyces and capsules make bacteria hydrophilic (Hogt et al. 1982). The nature of the hydrophobic moiety at the

  11. Development and characterisation of highly antibiotic resistant Bartonella bacilliformis mutants.

    PubMed

    Gomes, Cláudia; Martínez-Puchol, Sandra; Ruiz-Roldán, Lidia; Pons, Maria J; Del Valle Mendoza, Juana; Ruiz, Joaquim

    2016-09-26

    The objective was to develop and characterise in vitro Bartonella bacilliformis antibiotic resistant mutants. Three B. bacilliformis strains were plated 35 or 40 times with azithromycin, chloramphenicol, ciprofloxacin or rifampicin discs. Resistance-stability was assessed performing 5 serial passages without antibiotic pressure. MICs were determined with/without Phe-Arg-β-Napthylamide and artesunate. Target alterations were screened in the 23S rRNA, rplD, rplV, gyrA, gyrB, parC, parE and rpoB genes. Chloramphenicol and ciprofloxacin resistance were the most difficult and easiest (>37.3 and 10.6 passages) to be selected, respectively. All mutants but one selected with chloramphenicol achieved high resistance levels. All rifampicin, one azithromycin and one ciprofloxacin mutants did not totally revert when cultured without antibiotic pressure. Azithromycin resistance was related to L4 substitutions Gln-66 → Lys or Gly-70 → Arg; L4 deletion Δ62-65 (Lys-Met-Tyr-Lys) or L22 insertion 83::Val-Ser-Glu-Ala-His-Val-Gly-Lys-Ser; in two chloramphenicol-resistant mutants the 23S rRNA mutation G2372A was detected. GyrA Ala-91 → Val and Asp-95 → Gly and GyrB Glu474 → Lys were detected in ciprofloxacin-resistant mutants. RpoB substitutions Gln-527 → Arg, His-540 → Tyr and Ser-545 → Phe plus Ser-588 → Tyr were detected in rifampicin-resistant mutants. In 5 mutants the effect of efflux pumps on resistance was observed. Antibiotic resistance was mainly related to target mutations and overexpression of efflux pumps, which might underlie microbiological failures during treatments.

  12. [Distribution and changes of antibiotic susceptibility of genus Haemophilus (author's transl)].

    PubMed

    Kosakai, N; Oguri, T

    1976-02-01

    We studied on the distribution and changes of antibiotic susceptibility of H. influenzae, H. parainfluenzae and H. parahaemolyticus isolated from clinical materials, mainly from sputum and pharyngeal swabs. In this study we used 132 strains of H. influenzae, 89 strains of H. parainfluenzae and 43 strains of H. parahaemolyticus isolated during January and June of 1975, and estimated the susceptibility for the following eighteen antibiotics by the agar plate dilution method: ampicillin, amoxicillin, ciclacillin, sulbenicillin, carbenicillin, cephalothin, cefazolin, ceftezole, cephalexin, streptomycin, kanamycin, gentamicin, dibekacin, tetracycline, doxycycline, chloramphenicol, thiamphenicol and colistin. We compared these with previously reported results and observed the changes of antibiotic susceptibility. Ampicillin has the strongest antibiotic activity on three species of Haemophilus and the activity of four cephalosporins was weakest. Among three species H. parahaemolyticus was most susceptible and H. influenzae least susceptible to cephalosporins. Antibiotic activity of cyclacillin was rather weak. Other twelve antibiotics have good activity on Haemophilus. We could not find any ampicillin-resistant strain, but found five (3.8%) streptomycin-resistant, one (0.8%) kanamycin-resistant, eleven (8.3%) tetracycline-resistant, and seven (5.3%) chloramphenicol-resistant strains of H. influenzae. Six years ago we found five (9.6%) streptomycin-resistant and one (1.9%) tetracycline-resistant strains, but no resistant strain to other antibiotics. Tetracycline- and chloramphenicol-resistant strains are supposed to have a tendency to increase. There were very few strains which were resistant to more than two antibiotics among H. influenzae. We found a few strains resistant to tetracycline or chloramphenicol among H. parainfluenzae and H. parahaemolyticus, and one strain of H. parainfluenzae was less susceptible to ampicillin.

  13. The Influence of Primary and Secondary DNA Structure in Deletion and Duplication between Direct Repeats in Escherichia Coli

    PubMed Central

    Trinh, T. Q.; Sinden, R. R.

    1993-01-01

    We describe a system to measure the frequency of both deletions and duplications between direct repeats. Short 17- and 18-bp palindromic and nonpalindromic DNA sequences were cloned into the EcoRI site within the chloramphenicol acetyltransferase gene of plasmids pBR325 and pJT7. This creates an insert between direct repeated EcoRI sites and results in a chloramphenicol-sensitive phenotype. Selection for chloramphenicol resistance was utilized to select chloramphenicol resistant revertants that included those with precise deletion of the insert from plasmid pBR325 and duplication of the insert in plasmid pJT7. The frequency of deletion or duplication varied more than 500-fold depending on the sequence of the short sequence inserted into the EcoRI site. For the nonpalindromic inserts, multiple internal direct repeats and the length of the direct repeats appear to influence the frequency of deletion. Certain palindromic DNA sequences with the potential to form DNA hairpin structures that might stabilize the misalignment of direct repeats had a high frequency of deletion. Other DNA sequences with the potential to form structures that might destabilize misalignment of direct repeats had a very low frequency of deletion. Duplication mutations occurred at the highest frequency when the DNA between the direct repeats contained no direct or inverted repeats. The presence of inverted repeats dramatically reduced the frequency of duplications. The results support the slippage-misalignment model, suggesting that misalignment occurring during DNA replication leads to deletion and duplication mutations. The results also support the idea that the formation of DNA secondary structures during DNA replication can facilitate and direct specific mutagenic events. PMID:8325478

  14. Antimicrobial susceptibility of Escherichia coli isolated from shrimp (Litopenaeus vannamei) and pond environment in northeastern Brazil.

    PubMed

    Vieira, Regine H S Dos F; Carvalho, Edirsana M R; Carvalho, Fatima C T; Silva, Camila M; Sousa, Oscarina V; Rodrigues, Dalia P

    2010-04-01

    This study aimed to test the susceptibility of Escherichia coli strains isolated from the water, bottom sediments and individuals cultivated in shrimp farm ponds, to antibiotics belonging to different families, namely B-Lactams: Imipenem (IPM; 10 micro g), Ampicillin (AMP; 10 micro g), Cephalothin (CEP; 30 micro g), Cefoxitin (FOX; 30 micro g), Ceftriaxone (CRO; 30 micro g); Tetracycline: Tetracycline (TCY; 30 micro g); Aminoglycosides: Gentamicin (GEN; 10 micro g), Amikacin (AMK; 30 micro g); Chloramphenicol: Chloramphenicol (CHO; 30 micro g); Fluoroquinolones: Ciprofloxacin (CIP; 5 micro g); Nitrofurans: Nitrofurantoin (NIT; 300 micro g); Sulfonamides: Trimethoprim-Sulfamethoxazole (SXT; 30 micro g); Quilononas: Nalidixic Acid (NAL; 30 micro g). In the laboratory, the method of dissemination (Test Kirby-Bauer) was performed in order to fulfill the antibiogram tests. The results showed high indices of resistance to Imipenem, Cephalothin and Ampicillin. Chloramphenicol, Nitrofurantoin, Cefoxitin, Ceftiaxone and Ciprofloxacin have displayed the highest index of sensitive strains. The antibiotic resistance index (ARI) and the multiple resistance index (MAR) varied within the ranges of 0.068-0.077 and 0.15-0.39, respectively. More than 90.5% of strains of Escherichia coli showed a variety of resistance profiles to the tested antibiotics. The high indices of resistance may be a consequence of indiscriminate use of antibiotics, but also the transfer of resistance through mobile genetic elements found in shrimp farms.

  15. Physiological Aspects of Modification and Restoration of Chromosomal Synthesis in Bacteria After X Irradiation

    PubMed Central

    Billen, Daniel; Hewitt, Roger

    1965-01-01

    Billen, Daniel (The University of Texas, Houston), and Roger Hewitt. Physiological aspects of modification and restoration of chromosomal synthesis in bacteria after X irradiation. J. Bacteriol. 90:1218–1225. 1965.—A study was made of the effect of amino acid deprivation or chloramphenicol on the character of postirradiation deoxyribonucleic acid (DNA) replication in bacteria with the use of radioisotopes and 5-bromouracil as a density label. CsCl density-gradient studies of DNA showed that postirradiation incubation of amino acid-requiring Escherichia coli in an amino acid-free medium interfered with continued linear chromosomal replication. In the presence of the required amino acids, linear chromosomal replication was shown to resume. Addition of chloramphenicol was found to prevent this resumption. Deletion of the required amino acids or the presence of chloramphenicol in a fully supplemented medium allowed the detection of altered DNA synthesis in bacteria at X-ray doses as low as 500 r. The character of the limited DNA made in the presence of the density label after irradiation is described. The results are interpreted as showing that the synthesis of a protein(s) is required for restoration of linear chromosomal replication in the irradiated cells. PMID:5321477

  16. Breath hydrogen concentration and small intestinal malabsorption in calves.

    PubMed

    Holland, R E; Herdt, T H; Refsal, K R

    1986-09-01

    Breath hydrogen concentrations were measured to assess intestinal carbohydrate malabsorption in preruminating calves. Oral administration of 1.25 g of lactulose (a nonabsorbable carbohydrate)/kg to calves produced breath hydrogen concentrations significantly (P less than 0.001) higher than values determined after calves were fed milk and before the treatment was given. This indicates that, in the calf, fermentation of nonabsorbed carbohydrates results in increased breath hydrogen values. To induce small intestinal malabsorption, chloramphenicol was administered orally at 50 mg/kg, 2 times a day, to 5 calves for 3 days. Before therapy was started, each calf was fitted with a duodenal cannula to facilitate collection of intestinal mucosal biopsy samples during treatment. Chloramphenicol therapy significantly (P less than 0.001) increased breath hydrogen concentrations from those values measured after calves were fed milk alone. Concurrently, chloramphenicol administration significantly decreased intestinal villous length (P less than 0.001) and D-xylose absorption (P less than 0.05), compared with those values before treatment was given. These results demonstrate that decreased intestinal absorptive capacity is associated with an increase in breath hydrogen concentrations and that breath hydrogen may be useful in evaluating malabsorption in calves with naturally occurring enteric disease.

  17. Inactivation and regrowth of antibiotic-resistant bacteria by PAA disinfection in the secondary effluent of a municipal wastewater treatment plant.

    PubMed

    Huang, Jing Jing; Xi, Jing Ying; Hu, Hong Ying; Tang, Fang; Pang, Yu Chen

    2013-10-01

    Inactivation and microbial regrowth of penicillin-, ampicillin-, cefalexin-, tetracycline-, chloramphenicol-, and rifampicin-resistant bacteria were studied to explore risks associated with selection and regrowth of antibiotic-resistant bacteria after PAA disinfection. The results showed that after exposure to 20 mg/L PAA for 10 min, inactivation of ampicillin-resistant bacteria reached 2.3-log, which was significantly higher than that of total heterotrophic bacteria with a decrease of 2.0-log. In contrast, inactivation of tetracycline- resistant bacteria was significantly less efficient, reaching only 1.1-log. Chloramphenicol-and tetracycline-resistant bacteria, as well as total heterotrophic bacteria regrew more than 10 fold compared to those in the untreated wastewater sample with 22 h stilling culture after exposure to 2 or 5 mg/L PAA as for 10 min. Selection and potential regrowth of tetracycline-and chloramphenicol-resistant bacteria are potential risks when utilizing PAA disinfection, which may induce the spread of specific antibiotic-resistant bacteria in reclaimed water.

  18. Studies of the development of congenital anomalies in rats. III. Effects of inhibition of mitochondrial energy systems on embryonic development.

    PubMed

    Mackler, B; Grace, R; Tippit, D F; Lemire, R J; Shepard, T H; Kelley, V C

    1975-12-01

    Pregnant rats were treated with various inhibitors of mitochondrial oxidative energy metabolism and with lowered oxygen tension, and the embryo fetuses examined for the occurrence of congenital malformations and for changes in enzymatic activities. Treatment with all agents tested resulted in the production of skeletal anomalies. Sodium phenobarbital was the most teratogenic of the drugs tested and produced a high incidence of malformations which included cleft palate, tail anomalies, spinal retroflexion, domed head, and facial hypoplasia. Diphenylhydantoin produced a low incidence of syndactyly and oligodactyly. In addition to its effects on fetal growth and development chloramphenicol appeared to interfere with implantation. Tissue preparations from embryos exposed to sodium phenobarbital and chloramphenicol showed markedly lowered levels of DPNH oxidase activity. Cytochrome oxidase activity was also markedly lowered in the preparations from chloramphenicol-exposed embryos. Enzyme activities in preparations from embryos exposed to malonate and diphenylhydantoin appeared unaffected, although the drugs are strong inhibitors of electron transport in vitro; the lack of apparent effect may be due to the fact that both drugs do not bind to the enzyme preparations and were diluted 100- to 200-fold during preparation and assay of the tissue homogenates.

  19. Comparison of antimicrobial resistance in Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes strains isolated from organic and conventional poultry meat.

    PubMed

    Miranda, J M; Vázquez, B I; Fente, C A; Calo-Mata, P; Cepeda, A; Franco, C M

    2008-12-01

    The presence of Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes was determined in 55 samples of organic poultry meat and in 61 samples of conventional poultry meat. A total of 220 E. coli, 192 S. aureus, and 71 L. monocytogenes strains were analyzed by an agar disk diffusion assay for their resistance to ampicillin, cephalothin, chloramphenicol, ciprofloxacin, doxycycline, fosfomycin, gentamicin, nitrofurantoin, streptomycin, and sulfisoxazole (E. coli); chloramphenicol, ciprofloxacin, clindamycin, doxycycline, erythromycin, gentamicin, nitrofurantoin, oxacillin, and sulfisoxazole (S. aureus); and chloramphenicol, doxycycline, erythromycin, gentamicin, sulfisoxazole, and vancomycin (L. monocytogenes). The results indicated a significantly higher (P < 0.0001) prevalence of E. coli but not of S. aureus and L. monocytogenes in organic poultry meat as compared with conventional poultry meat. E. coli isolated from organic poultry meat exhibited lower levels of antimicrobial resistance against 7 of the 10 antimicrobials tested as compared with isolates recovered from conventional meat. In the case of S. aureus and L. monocytogenes isolated from conventional poultry, antimicrobial resistance was significantly higher only for doxycycline as compared with strains isolated from organic poultry. In the case of E. coli, the presence of multiresistant strains was significantly higher (P < 0.0001) in conventional poultry meat as compared with organic poultry meat. Organically farmed poultry samples showed significantly lower development of antimicrobial resistance in intestinal bacteria such as E. coli.

  20. Concurrence of cat and tet genes in multiple antibiotic-resistant bacteria isolated from a sea cucumber and sea urchin mariculture farm in China.

    PubMed

    Dang, Hongyue; Song, Linsheng; Chen, Mingna; Chang, Yaqing

    2006-11-01

    A basic understanding of abundance and diversity of antibiotic-resistant microbes and their genetic determinants is necessary for finding a way to prevent and control the spread of antibiotic resistance. For this purpose, chloramphenicol and multiple antibiotic-resistant bacteria were screened from a mariculture farm in northern China. Both sea cucumber and sea urchin rearing ponds were populated with abundant antibiotic-resistant bacteria, especially marine vibrios. Sixty-five percent chloramphenicol-resistant isolates from sea cucumber harbored a cat gene, either cat IV or cat II, whereas 35% sea urchin isolates harbored a cat gene, actually cat II. The predominant resistance determinant cat IV gene mainly occurred in isolates related to Vibrio tasmaniensis or Pseudoalteromonas atlantica, and the cat II gene mainly occurred in Vibrio splendidus-like isolates. All the cat-positive isolates also harbored one or two of the tet genes, tet(D), tet(B), or tet(A). As no chloramphenicol-related antibiotic was ever used, coselection of the cat genes by other antibiotics, especially oxytetracycline, might be the cause of the high incidence of cat genes in the mariculture farm studied.

  1. [Antimicrobial activity of ornidazole and 6 other antibiotics against anaerobic bacteria].

    PubMed

    Alados, J C; Martínez-Brocal, A; Miranda, C; Rojo, M D; García, V; Domínguez, M C; de la Rosa, M

    1991-04-01

    The antimicrobial susceptibility of 235 anaerobic bacterial strains to ornidazole, metronidazole, chloramphenicol, clindamycin, penicillin, cefoxitin and imipenem has been studied using agar-dilution technique. Ornidazole and metronidazole were active against 88.6% and 86% of gram-positive cocci. Overall, 99.1% of Bacteroides group fragilis, and 91.3% of non-fragilis Bacteroides were also sensitive to both drugs. We did not find any Clostridium perfringens resistant strain. Cefoxitin and penicillin showed good activity against all Clostridium perfringens strains, and also against 97.7% and 92.5% of gram-positive cocci. We found one single imipenem resistant strain among gram-positive bacteria. Bacteroides fragilis also showed sensitivity to penicillin (41.5%), cefoxitin (85.7%) and imipenem (97.1%). Clindamycin was active against Clostridium perfringens (90.9%), gram-positive cocci (86.7%) and imipenem (68.6%). Chloramphenicol showed good activity against Clostridium perfringens (100%), gram-positive cocci (95.5%) and Bacteroides spp. (99.4%). Our results showed an overall good activity of all the seven drugs tested against anaerobic gram-positive microorganisms. Of notice, we found a good activity of chloramphenicol, imipenem, metronidazole and ornidazole against Bacteroides spp.

  2. PubMed Central

    Archimbault, P.; Boutier, C.; Fellous, R.

    1980-01-01

    Serum concentrations and factors affecting the blood bioavailability of chloramphenicol in bovine The authors have compared the serum concentrations and the factors affecting blood bioavailability of chloramphenicol after intramuscular administration of canadian commercial preparations containing 500 mg/mL of antibiotic. The animals (dairy cows and heifers) received each drug (20 mg/kg) in one or two injection sites. The serum samples, analysed by colorimetric or microbiological methods, showed that considerable differences in concentration exist between the two methods. The evolution of biodisponibility factors proved identical in both cases. It appears that therapeutic levels of chloramphenicol are reached only by drug A for four to five hours. The usual dosage (2-10 mg/kg), by intramuscular route, is not sufficient to attain these active concentrations using the other drugs. However, the important variability obtained during the experiment and reflected in the standard deviation values, has not proved that drug A has a better bioavailability based on the criteria of the only microbiological analysis. PMID:7260826

  3. [Changes of resistant phenotype and CRISPR/Cas system of four Shigella strains passaged for 90 times without antibiotics].

    PubMed

    Zhang, B; Hong, L J; Duan, G C; Liang, W J; Yang, H Y; Xi, Y L

    2017-02-10

    Objective: To explore the stability of resistant phenotypes and changes of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) gene system on four Shigella strains in the absence of antibiotics. Methods: Four clinical isolated Shigella strains that resistant to different antibiotics were consecutive passaged for 90 times without antibiotics. Agar dilution method was used to determine the minimum inhibitory concentration of Shigella strains. After sequence analysis with PCR, CRISPR Finder and Clustal X 2.1 were applied to identify the changes of CRISPR loci in the Shigella strains. Results: After the consecutive transfer of 90 generations, sensitivity to certain antibiotics of four Shigella strains with different drug resistant spectrums increased. Mel-sf1998024/zz resistance to ampicillin, cephalexin, cefotaxime, chloramphenicol decreased, mel-s2014026/sx resistance to norfloxacin, trimethoprim decreased, mel-sf2004004/sx drug resistance to ampicillin, cefuroxime, cefotaxime, chloramphenicol, trimethoprim decreased and mel-sf2013004/bj resistance to chloramphenicol decreased. The spacer of which matched gene codes Cas and its upstream repeat in 3'end of CRISPR3 got lost in mel-sf1998024/zz and mel-sf2013004/bj. Conclusions:Shigella strains could reduce or lose their resistance to some antibiotics after consecutive transfers, without the interference of antibiotics. CRISPR3 locus had dynamic spacers in Shigella strains while CRISPR3 locus and cas genes might have been co-evolved.

  4. Antibiotic combinations for controlling colistin-resistant Enterobacter cloacae.

    PubMed

    Lima, Thais Bergamin; Silva, Osmar Nascimento; de Almeida, Keyla Caroline; Ribeiro, Suzana Meira; Motta, Dielle de Oliveira; Maria-Neto, Simone; Lara, Michelle Brizolla; Filho, Carlos Roberto Souza; Ombredane, Alicia Simalie; de Faria Junior, Celio; Parachin, Nadia Skorupa; Magalhães, Beatriz Simas; Franco, Octávio Luiz

    2017-02-01

    Enterobacter cloacae is a Gram-negative bacterium associated with high morbidity and mortality in intensive care patients due to its resistance to multiple antibiotics. Currently, therapy against multi-resistant bacteria consists of using colistin, in spite of its toxic effects at higher concentrations. In this context, colistin-resistant E. cloacae strains were challenged with lower levels of colistin combined with other antibiotics to reduce colistin-associated side effects. Colistin-resistant E. cloacae (ATCC 49141) strains were generated by serial propagation in subinhibitory colistin concentrations. After this, three colistin-resistant and three nonresistant replicates were isolated. The identity of all the strains was confirmed by MALDI-TOF MS, VITEK 2 and MicroScan analysis. Furthermore, cross-resistance to other antibiotics was checked by disk diffusion and automated systems. The synergistic effects of the combined use of colistin and chloramphenicol were observed via the broth microdilution checkerboard method. First, data here reported showed that all strains presented intrinsic resistance to penicillin, cephalosporin (except fourth generation), monobactam, and some associations of penicillin and β-lactamase inhibitors. Moreover, a chloramphenicol and colistin combination was capable of inhibiting the induced colistin-resistant strains as well as two colistin-resistant clinical strains. Furthermore, no cytotoxic effect was observed by using such concentrations. In summary, the data reported here showed for the first time the possible therapeutic use of colistin-chloramphenicol for infections caused by colistin-resistant E. cloacae.

  5. Mediterranean spotted fever: clinical and laboratory characteristics of 415 Sicilian children

    PubMed Central

    Colomba, Claudia; Saporito, Laura; Polara, Valentina Frasca; Rubino, Raffaella; Titone, Lucina

    2006-01-01

    Background Mediterranean spotted fever (MSF) is an acute febrile, zoonotic disease caused by Rickettsia conorii and transmitted to humans by the brown dogtick Rhipicephalus sanguineus. Nearly four hundred cases are reported every year (mainly from June to September) on the Italian island of Sicily. The aim of the study was to analyze the clinical and laboratory characteristics of patients with MSF and the efficacy of the drugs administered. Methods Our study was carried out on 415 children with MSF, during the period January 1997 – December 2004, at the "G. Di Cristina" Children's hospital in Palermo, Sicily, Italy. On admission patients' clinical history, physical and laboratory examination and indirect immunofluorescence antibody test (IFAT) for Rickettsia conorii were performed. Diagnosis was considered confirmed if the patients had an MSF diagnostic score greater than or equal to 25 according to the Raoult's scoring system. All patients were treated with chloramphenicol or with macrolides (clarithromycin or azithromycin). Results Fever, rash and tache noire were present in 386 (93%), 392 (94.5%) and 263 (63.4%) cases respectively. Eighteen (4.6%) children showed atypical exanthema. Chloramphenicol and newer macrolides all appeared to be effective and safe therapies. Conclusion Clinical features of 415 children with MSF were similar to those reported by other authors except for a lower incidence of headache, arthralgia and myalgia and a higher frequency of epato-splenomegaly. Concerning therapy, clarithromycin can be considered a valid alternative therapy to tetracyclines or chloramphenicol especially for children aged < eight years. PMID:16553943

  6. Human Immunodeficiency Virus Tat-Activated Expression of Poliovirus Protein 2A Inhibits mRNA Translation

    NASA Astrophysics Data System (ADS)

    Sun, Xiao-Hong; Baltimore, David

    1989-04-01

    To study the effect of poliovirus protein 2A on cellular RNA translation, the tat control system of human immunodeficiency virus (HIV) was used. Protein 2A was expressed from a plasmid construct (pHIV/2A) incorporating the HIV long terminal repeat. Protein synthesis was measured by using chloramphenicol acetyltransferase as a reporter gene driven by the Rous sarcoma virus long terminal repeat. When HIV/2A was contransfected with the reporter, addition of a tat-producing plasmid caused at least a 50-fold drop in chloramphenicol acetyltransferase synthesis. A HeLa cell line carrying HIV/2A was established. In it, tat expression caused more than a 10-fold drop in chloramphenicol acetyltransferase synthesis from the reporter plasmid. Furthermore, 2A induction by tat caused cleavage of the cellular translation factor P220, a part of eukaryotic translation initiation factor 4F. Thus protein 2A can, by itself, carry out the inhibition of cellular protein synthesis characteristic of a poliovirus infection. Also, the HIV tat activation provides a very effective method to control gene expression in mammalian cells.

  7. Synthesis and Assessment of Antibacterial Activities of Ruthenium(III) Mixed Ligand Complexes Containing 1,10-Phenanthroline and Guanide

    PubMed Central

    Hailemariam, Tizazu

    2016-01-01

    In this work, two complexes of ruthenium(III) ([Ru(phen)2Cl2]Cl·2H2O and [Ru(phen)2(G)Cl]2Cl·H2O) were synthesized from 1,10-phenanthroline alone as well as from both 1,10-phenanthroline and guanide. The synthesis was checked using halide test, conductance measurement, and spectroscopic (ICP-OES, FTIR, and UV/Vis) analysis. Their in vitro antibacterial activities were also investigated on two Gram-positive (Staphylococcus aureus (S. aureus) and methicillin resistant Staphylococcus aureus (MRSA)) and two Gram-negative (Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae)) bacteria. These complexes showed wide-range better activities than the commercially available controls (Chloramphenicol and Ciprofloxacin) against even the most drug resistant K. pneumoniae. [Ru(phen)2(G)Cl]2Cl·H2O inhibited S. aureus, MRSA, E. coli, and K. pneumoniae by 17.5%, 27.4%, 16%, and 52%, respectively, better than Chloramphenicol. It also inhibited these pathogens by 5.9%, 5.1%, 2.3%, and 17.2%, respectively, better than Ciprofloxacin. Similarly, [Ru(Phen)2(Cl)2]Cl·2H2O inhibited these pathogens by 11%, 8.7%, 0.1%, and 31.2%, respectively, better than Chloramphenicol. Therefore, after in vivo cytotoxicity investigations, these compounds can be considered as potential antibiotic drugs. PMID:27833473

  8. Synthesis and evaluation of inhibitors of bacterial drug efflux pumps of the major facilitator superfamily.

    PubMed

    Okandeji, Babajide O; Greenwald, Daniel M; Wroten, Jessica; Sello, Jason K

    2011-12-15

    Inhibitors of drug efflux pumps have great potential as pharmacological agents that restore the drug susceptibility of multidrug resistant bacterial pathogens. Most attention has been focused on the discovery of small molecules that inhibit the resistance nodulation division (RND) family drug efflux pumps in Gram-negative bacteria. The prototypical inhibitor of RND-family efflux pumps in Gram-negative bacteria is MC-207,110 (Phe-Arg-β-naphthylamide), a C-capped dipeptide. Here, we report that C-capped dipeptides inhibit two chloramphenicol-specific efflux pumps in Streptomyces coelicolor, a Gram-positive bacterium that is a relative of the human pathogen Mycobacterium tuberculosis. Diversity-oriented synthesis of a library of structurally related C-capped dipeptides via an Ugi four component reaction and screening of the resulting compounds resulted in the discovery of a compound that is threefold more potent as a suppressor of chloramphenicol resistance in S. coelicolor than MC-207,110. Since chloramphenicol resistance in S. coelicolor is mediated by major facilitator superfamily drug efflux pumps, our findings provide the first evidence that C-capped dipeptides can inhibit drug efflux pumps outside of the RND superfamily.

  9. Antibiotic-dependent induction of Pseudomonas putida DOT-T1E TtgABC efflux pump is mediated by the drug binding repressor TtgR.

    PubMed

    Terán, Wilson; Felipe, Antonia; Segura, Ana; Rojas, Antonia; Ramos, Juan-Luis; Gallegos, María-Trinidad

    2003-10-01

    Pseudomonas putida is well known for its metabolic capabilities, but recently, it has been shown to exhibit resistance to a wide range of antibiotics. In P. putida DOT-T1E, the TtgABC efflux pump, which has a broad substrate specificity, extrudes antibiotics such as ampicillin, carbenicillin, tetracycline, nalidixic acid, and chloramphenicol. We have analyzed the expression of the ttgABC efflux pump operon and its regulatory gene, ttgR, in response to several structurally unrelated antibiotics at the transcriptional level and investigated the role of the TtgR protein in this process. ttgABC and ttgR are expressed in vivo at a moderate basal level, which increases in the presence of hydrophobic antibiotics like chloramphenicol and tetracycline. In vitro experiments show that, in the absence of inducers, TtgR binds to a palindromic operator site which overlaps both ttgABC and ttgR promoters and dissociates from it in the presence of chloramphenicol and tetracycline. These results suggest that the TtgR repressor is able to bind to structurally different antibiotics, which allows induction of TtgABC multidrug efflux pump expression in response to these antimicrobial agents. This is the first case in which the expression of a drug transporter of the resistance-nodulation-division family has been shown to be regulated directly by antibiotics.

  10. Anoxic survival potential of bivalves: (arte)facts.

    PubMed

    de Zwaan, Albertus; Babarro, Jose M F; Monari, Marta; Cattani, Otello

    2002-03-01

    The anoxic survival time of the bivalves Chamelea gallina, Cerastoderma edule and Scapharca inaequivalvis from two different ecosystems and differing anoxia tolerances was studied in static (closed) and flow-through systems. The antibiotics chloramphenicol, penicillin and polymyxin were added, and molybdate (specific inhibitor of the process of sulfate reduction). Survival in (near) anoxic seawater of Chamelea was studied in a static system by comparing untreated seawater with autoclaved seawater and untreated clams with clams incubated in well-aerated seawater, containing the broad-spectrum antibiotic chloramphenicol, prior to the anoxic survival test. With untreated clams and natural seawater (median mortality time 2.4 days) a decrease in pH and exponential accumulation of sulfide and ammonium was observed in the anoxic medium, indicating excessive growth of (sulfate reducing) bacteria. In sterilized seawater LT50 (2.1 days) was not significantly different and again considerable amounts of ammonium and sulfide accumulated. However, pre-treatment of clams with chloramphenicol resulted in an increase of LT50 (11.0 days) by approximately fivefold. Accumulation of ammonium and sulfide was retarded, but was finally even stronger than in the medium containing untreated clams. Median mortality times were 2.5 and 2.4 days for Chamelea and 2.7 and 2.9 days for Cerastoderma for static and flow-through incubations, respectively. Addition of chloramphenicol increased strongly survival time in both systems with corresponding values of 11.0 and 16.3 days for Chamelea, and 6.4 and 6.5 days for Cerastoderma. LT50 of Scapharca in anoxic seawater was 14.4 days. Chloramphenicol and penicillin increased median survival time to 28.5 and 28.7 days, respectively, whereas polymyxin displayed no effect (LT50=13.6 days). Molybdate added to artificial sulfate free seawater blocked biotic sulfide formation, but did not improve survival time (LT50=13.7 days). Overall the results indicate

  11. Salicylate-inducible antibiotic resistance in Pseudomonas cepacia associated with absence of a pore-forming outer membrane protein.

    PubMed Central

    Burns, J L; Clark, D K

    1992-01-01

    The most common mechanism of antibiotic resistance in multiply resistant Pseudomonas cepacia is decreased porin-mediated outer membrane permeability. In some gram-negative organisms this form of antibiotic resistance can be induced by growth in the presence of weak acids, such as salicylates, which suppress porin synthesis. To determine the effects of salicylates on outer membrane permeability of P. cepacia, a susceptible laboratory strain, 249-2, was grown in 10 mM sodium salicylate. Antibiotic susceptibility and uptake, as well as outer membrane protein patterns, were compared between strain 249-2 grown with and without salicylates. The MICs of chloramphenicol, trimethoprim, ciprofloxacin, and ceftazidime were compared between organisms grown in standard and salicylate-containing medium and are as follows: chloramphenicol, 12.5 versus 100 micrograms/ml; trimethoprim, 0.78 versus 3.125 micrograms/ml; ciprofloxacin, 0.4 versus 1.56 micrograms/ml; ceftazidime, 3.125 versus 3.125 micrograms/ml. The permeability of beta-lactam antibiotics was calculated from the rate of hydrolysis of the chromogenic cephalosporin, PADAC. There was no significant difference between strains grown in the presence and absence of salicylate. By using high-pressure liquid chromatography quantitation of loss from culture medium, the effect of 10 mM salicylate on the cellular permeability of chloramphenicol was measured in strain 249-2 by introduction of a plasmid which encodes production of chloramphenicol acetyltransferase. After 1 h of incubation, 18.5% +/- 1.54% versus 70.1% +/- 3.52%, and after 2 h, 4.20% +/- 1.65% versus 41.90% +/- 2.16% remained in supernatants from organisms grown in the absence and presence of 10 mM salicylate, respectively. Outer membrane protein pattern analysis demonstrated the absence of a protein of apparent molecular weight of 40,000 when strain 249-2 was grown in the presence of 10 mM salicylate. To determine whether this protein functioned as a porin

  12. Waterborne typhoid fever in Dade County, Florida. Clinical and therapeutic evaluation of 105 bacteremic patients.

    PubMed

    Hoffman, T A; Ruiz, C J; Counts, G W; Sachs, J M; Nitzkin, J L

    1975-10-01

    An extensive outbreak of waterborne typhoid fever occurred in 1973 at a migrant labor camp in Dade County, Florida. Blood cultures from 105 of the 188 patients with proved or presumptive cases of typhoid fever grew Salmonella typhi. The clinical and laboratory findings in these patients were reviewed. Fever, usually with temperatures above 38.8 degrees C and of the sustained type, was a primary manifestation of disease, although a majority of the patients also complained of headache and gastroenteric symptoms. Hepatic or splenic enlargement was present in 52 per cent and 42 per cent, respectively, whereas rose spots were detected in only 13 per cent. The total leukocyte count was normal in 74 per cent, but serum levels of liver and muscle enzymes were frequently elevated. Gastrointestinal, pulmonary and neurologic complications were infrequent; circulatory failure was not observed. Defervescence in response to antibiotic therapy was variable; however, the median response among 68 patients who received chloramphenicol was two days less than that in 34 patients treated with ampicillin. There was one possible treatment failure with ampicillin. The relapse rate of 10 per cent in chloramphenicol-treated patients was not significantly greater than the 3 per cent rate among those treated with ampicillin. Serologic studies for typhoid fever were of limited diagnostic value since the titer of agglutinins was 1:160 or higher in 49 per cent of the serums obtained before treatment, and a fourfold rise in titer occurred in only 24 per cent of 57 patients studied. The serologic response to chloramphenicol treatment did not differ from that to ampicillin.

  13. Characterization of plasmid-borne afa-3 gene clusters encoding afimbrial adhesins expressed by Escherichia coli strains associated with intestinal or urinary tract infections.

    PubMed

    Le Bouguenec, C; Garcia, M I; Ouin, V; Desperrier, J M; Gounon, P; Labigne, A

    1993-12-01

    The afa gene clusters encode afimbrial adhesins (AFA) that are expressed by uropathogenic and diarrhea-associated Escherichia coli strains and belong to a family of hemagglutinins recognizing the Dr blood group antigen as a receptor. This family so far includes AFA-I and AFA-III as well as the Dr and F1845 adhesins (B. Nowicki, A. Labigne, S. Moseley, R. Hull, S. Hull, and J. Moulds, Infect. Immun. 58:279-281, 1990). Reported in this work is the genetic organization of the afa-3 gene cluster cloned from a uropathogenic E. coli strain (A30) which expressed a subtype of AFA designated AFA-III. The amino acid sequence of AFA-III was deduced from the nucleotide sequence of the afaE3 gene and was found to be highly homologous to that of the Dr adhesin (98.1% identity). A polymerase chain reaction assay was developed to detect the presence of afa-3 gene clusters in E. coli strains. Study of the genetic support of the afa-3 gene clusters in the strains which showed positive amplification revealed that they were always located on large, 100-kb plasmids whether the strains originated from patients with cystitis or with diarrhea. Moreover, the cloned afa-3 gene clusters from A30 and from the diarrhea-associated strain AL845 appeared to be carried by 9-kb plasmid regions which displayed a similar genetic organization. Chloramphenicol was reported to be a potent inhibitor of receptor binding by the Dr adhesin (Nowicki et al., Infect. Immun. 58:279-281, 1990). AFA-III expressed by strains AL845 and AL847 appeared to mediate, like the Dr adhesin, chloramphenicol-sensitive hemagglutination, whereas AFA-III produced by A30 conferred chloramphenicol-resistant adherence. A comparison of the sequences of these four proteins indicated that the amino acid at position 52 of the processed AFA could be part of the receptor-binding domain.

  14. Presence of Multidrug Resistant Enteric Bacteria in Dairy Farm Topsoil

    PubMed Central

    Burgos, J. M.; Ellington, B. A.; Varela, M. F.

    2008-01-01

    In addition to human and veterinary medicine, antibiotics are extensively used in agricultural settings, such as for treatment of infections, growth enhancement and prophylaxis in food animals, leading to selection of drug and multidrug resistant bacteria. In order to help circumvent the problem of bacterial antibiotic resistance, it is first necessary to understand the scope of the problem. However, is it not fully understood how widespread antibiotic resistant bacteria are in agricultural settings. The lack of such surveillance data is especially evident in dairy farm environments, such as soil. It is also unknown to what extent various physiological modulators, such as salycilate, a component of aspirin and known model modulator of multiple antibiotic resistance (mar) genes, influence bacterial multidrug resistance. We isolated and identified enteric soil bacteria from local dairy farms within Roosevelt County, NM, determined the resistance profiles to antibiotics associated with mar, such as chloramphenicol, nalidixic acid, penicillin G and tetracycline. We then purified and characterized plasmid DNA and detected mar phenotypic activity. The minimal inhibitory concentrations (MICs) of antibiotics for the isolates ranged between 6 - >50 μg/mL for chloramphenicol, 2–8 μg/mL for nalidixic acid, 25- >300 μg/mL for penicillin G and 1- > 80 μg/mL for tetracycline. On the other hand, the many of the isolates had significantly enhanced MICs for the same antibiotics in the presence of 5 mM salycilate. Plasmid DNA extracted from 12 randomly chosen isolates ranged in size between 6 and 12.5kb and in several cases conferred resistances to chloramphenicol and penicillin G. It is concluded that enteric bacteria from dairy farm topsoil are multi-drug resistant and harbor antibiotic resistance plasmids. A role for dairy topsoil in zoonosis is suggested, thus implicating this environment as a reservoir for bacterial resistance development against clinically relevant

  15. Rocky Mountain spotted fever.

    PubMed

    Kamper, C A; Chessman, K H; Phelps, S J

    1988-02-01

    The epidemiology, pathogenesis, clinical features, and treatment of Rocky Mountain spotted fever are reviewed. Rocky Mountain spotted fever is a severe infection caused by Rickettsia rickettsii transmitted to man by various species of ticks. High-incidence areas exist in the southeast and south central United States. Only 60-70% of patients with the disease report a history of tick bite or exposure to tick-infested areas. The disease is initially characterized by fever, headache, gastrointestinal complaints, myalgia, and a generalized rash. In several days generalized vasculitis may lead to periorbital edema and nonpitting edema of the face and extremities. Central nervous system involvement is common. Because signs and symptoms associated with the disease are nonspecific, the diagnosis is often delayed or missed. Traditionally diagnostic confirmation relied on serologic testing, but an indirect fluorescent antibody assay will soon be commercially available. Rocky Mountain spotted fever is usually treated with the rickettsiostatic agents chloramphenicol or tetracycline, but few comparative data on these agents in patients with the disease are available. For patients who cannot tolerate oral medications, intravenous chloramphenicol sodium succinate is the preferred treatment; chloramphenicol is also the drug of choice for children less than eight years of age. Otherwise, oral tetracycline hydrochloride is the drug of choice. Antibiotic therapy should be continued for 7-10 days or until the patient is afebrile for two to five days. All cases of Rocky Mountain spotted fever must be reported to the Centers for Disease Control. The best ways to decrease the morbidity and mortality of the disease are to increase awareness of its signs and symptoms and to prevent exposure to ticks.

  16. Flow cytometric method for the assessment of the minimal inhibitory concentrations of antibacterial agents to Mycoplasma agalactiae.

    PubMed

    Assunção, Patrícia; Antunes, Nuno T; Rosales, Ruben S; de la Fe, Christian; Poveda, Carlos; Poveda, José B; Davey, Hazel M

    2006-10-01

    In this study, flow cytometry was evaluated for the determination of the minimal inhibitory concentrations (MIC) of seven antibacterial agents (enrofloxacin, ciprofloxacin, gentamicin, streptomycin, chloramphenicol, oxytetracycline, and tylosin) on Mycoplasma (M.) agalactiae. Flow cytometry was able to detect M. agalactiae inhibition from 6 h postincubation, although it seems that definitive MIC values determined by flow cytometry were only possible at 12-h postincubation. However, the results obtained by the traditional method were only obtained at 24 h, when a visible change in the medium had occurred. At 24 h, both methods gave the same result for six antibacterial agents (enrofloxacin, ciprofloxacin, gentamicin, streptomycin, chloramphenicol, and oxytetracycline); whereas flow cytometry gave slightly higher MIC for tylosin. This was attributed to the fact that the M. agalactiae growth that had occurred in the tubes containing tylosin was not enough to visibly change the color of the medium. Futhermore, flow cytometry detected that inhibitory concentrations of oxytetracycline, chloramphenicol, and tylosin as judged at 24 h were not able to inhibit the M. agalactiae growth after 48 h. MIC values of enrofloxacin and ciprofloxacin were sufficient only to maintain the total counts per milliliter throughout the time matched samples, whereas higher concentrations of theses antibacterial agents reduced the total counts per milliliter over the course of the experiment. The main advantage of the flow cytometric method is that MIC results for M. agalactiae can be obtained in a shorter time than is possible with the traditional method. The method presented makes identification of resistant populations of M. agalactiae possible and, unlike the traditional method, allows the effect of each antibacterial agent to be determined in real-time at the single-cell level.

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

    PubMed

    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.

  18. Antimicrobial resistance of Salmonella spp. isolated from food

    PubMed

    Mąka, Łukasz; Popowska, Magdalena

    This review summarizes current data on resistance among Salmonella spp. isolates of food origin from countries in different regions of the world. The mechanisms of resistance to different groups of antimicrobial compounds are also considered. Among strains resistant to quinolones and/or fluoroquinolones the most prevalent mechanism is amino acid substitutions in quinolone resistance-determining region (QRDR) of genes gyrA, parC but mechanism of growing importance is plasmid-mediated quinolone resistance (PMQR) associated with genes qnrA, qnrB, qnrC, qnrD, qnrS but frequency of their detection is different. Resistance to sulfonamides is mostly associated with genes sul1 and sul2, while resistance to trimethoprim is associated with various variants of dhfr ( dfr) genes. Taking into account Salmonella spp. strains isolated from food, resistance to β-lactams is commonly associated with β-lactamases encoding by blaTEM genes. However strains ESBL and AmpC – positive are also detected. Resistance to aminoglicosides is commonly result of enzymatic inactivation. Three types of aminoglycoside modifying enzyme are: acetyltransferases (AAC), adenyltransferases (ANT) and phosphotransferases (APH). Resistance to tetracyclines among Salmonella spp. isolated from food is most commonly associated with active efflux. Among numerous genetic determinants encoding efflux pumps tetA, tetB, tetC, tetD, tetE and tetG are reported predominatingly. One of the most common mechanisms of resistance against chloramphenicol is its inactivation by chloramphenicol acetyltrasferases (CATs), but resistance to this compound can be also mediated by chloramphenicol efflux pumps encoded by the genes cmlA and floR. It is important to monitor resistance of Salmonella isolated from food, because the globalization of trade, leading to the long-distance

  19. Antimicrobial Resistance Profiles of Salmonella spp. from Agricultural Environments in Fruit Production Systems.

    PubMed

    Gomba, Annancietar; Chidamba, Lizyben; Korsten, Lise

    2016-09-01

    Foodborne disease outbreaks involving fresh produce have increased in recent years. The risk of infection from contaminated food is worsened by the increased prevalence of antibiotic-resistant strains. This study evaluated the prevalence of antibiotic resistance in Salmonella isolates (n = 263) from agricultural production systems through to the final packed product. Salmonella isolates were preliminarily identified by matrix-assisted laser desorption ionization-time-of-flight mass spectroscopy (MALDI-TOF MS) and API 20E and identities confirmed by invA gene polymerase chain reaction. Antimicrobial susceptibility was performed with 15 antimicrobial agents using the Kirby-Bauer disk diffusion test. Of the 263 Salmonella isolates assessed, 59.3% were resistant to one or more antimicrobials. The most frequently detected resistance was against chloramphenicol and kanamycin (46.7%), trimethoprim-sulfamethoxazole (28%), and streptomycin (14%), and the less frequently detected resistance was toward ampicillin (1.14%), amikacin (0.76%), and amoxicillin-clavulanic acid (0.38%). Multiple antimicrobial resistance (MAR) (resistance to ≥3 antibiotics) was found in 48.7% (76/156) isolates. The most common MAR phenotype was to chloramphenicol and trimethoprim/sulfamethoxazole-kanamycin (43.6%). Resistance to chloramphenicol, kanamycin, or trimethoprim/sulfamethoxazole was only observed in MAR phenotypes. All isolates were susceptible to ceftiofur, cefoxitin, ceftriaxone, ciprofloxacin, nalidixic acid, gentamicin, and tetracycline. This study confirms the importance of fresh produce production environments as potential reservoirs and fresh produce as carriers of antibiotic-resistant Salmonella spp. with significant clinical importance. Further studies to evaluate the actual level of health risk from these pathogens should include characterization of the antibiotic resistance determinant genes among the isolates.

  20. Clonal relationships among penicillin-susceptible, multiresistant serotype 6B Streptococcus pneumoniae isolates recovered in Greece and France.

    PubMed

    Syrogiannopoulos, G A; Doit, C; Grivea, I N; Geslin, P; Bingen, E

    2001-01-01

    In January 1996 the emergence of penicillin-susceptible, multiresistant serotype 6B Streptococcus pneumoniae isolates resistant to chloramphenicol, tetracycline, erythromycin, clindamycin and trimethoprim-sulfamethoxazole was observed in young carriers in the city of Patras, located in the southwestern region of Greece. Later, a significant spread of pneumococci with this unusual phenotype was noted in carriers living in various other areas of the country. Using restriction fragment length polymorphism of the ribosomal RNA genes, clonal relationships were found between these Greek strains and serotype 6B penicillin-susceptible, multiresistant pneumococci isolated in France between January 1992 and September 1996. The French and Greek isolates appear to have a common ancestry.

  1. In vitro antibiotic susceptibility of Neisseria gonorrhoeae in Jakarta, Indonesia.

    PubMed

    Lesmana, M; Lebron, C I; Taslim, D; Tjaniadi, P; Subekti, D; Wasfy, M O; Campbell, J R; Oyofo, B A

    2001-01-01

    Antibiotic susceptibilities were determined for 122 Neisseria gonorrheae isolates obtained from 400 sex workers in Jakarta, Indonesia, and susceptibilities to ciprofloxacin, cefuroxime, cefoxitin, cefotaxime, ceftriaxone, chloramphenicol, and spectinomycin were found. All isolates were resistant to tetracycline. A number of the isolates demonstrated decreased susceptibilities to erythromycin (MIC >/= 1.0 microg/ml), thiamphenicol (MIC >/= 1.0 microg/ml), kanamycin (MIC >/= 16.0 microg/ml), penicillin (MIC >/= 2.0 microg/ml), gentamicin (MIC >/= 16.0 microg/ml), and norfloxacin (MIC = 0.5 microg/ml). These data showed that certain antibiotics previously used in the treatment of gonorrhea are no longer effective.

  2. Susceptibility of Listeria monocytogenes isolated from food in Italy to antibiotics.

    PubMed

    Aureli, Paolo; Ferrini, Anna Maria; Mannoni, Veruscka; Hodzic, Snjezana; Wedell-Weergaard, Christina; Oliva, Brunello

    2003-06-25

    The susceptibility of 148 strains of Listeria monocytogenes isolated from food to antibiotics currently used in veterinary and human therapy was determined by standard agar dilution and disk diffusion methods. The antibiotics included amikacin, amoxicillin, cefazolin, chloramphenicol, erythromycin, flumequine, fosfomycin, gentamicin, kanamycin, lincomycin, oxytetracycline, rifampicin, spiramycin, streptomycin, tetracycline, tobramycin and vancomycin. Soussy's breakpoints and MIC(50)-MIC(90) values were used to classify the strains into sensitive, moderately sensitive and resistant groups. This work is part of a wider surveillance program on listeriosis started in Italy in 1995.

  3. In vitro studies of Dermatophilus congolensis antimicrobial susceptibility by determining minimal inhibitory and bacteriocidal concentrations.

    PubMed

    Hermoso de Mendoza, J; Arenas, A; Rey, J; Alonso, J M; Gil, M C; Naranjo, G; Hermoso de Mendoza, M

    1994-01-01

    The Minimal Inhibitory Concentration (MIC) and Minimal Bacteriocidal Concentration (MBC) of 19 antimicrobials on 16 isolates of D. congolensis were determined. The potential field efficacy of the agents was evaluated by comparing the results with serum levels of drug unbound to proteins and the in vitro and in vivo findings of other authors. A modified standard microtechnique was used for serial dilution-antimicrobial sensitivity and found to be easy and reproducible. Erythromycin, spiramycin, penicillin G, ampicillin, chloramphenicol, the streptomycin, amoxicillin, the tetracyclines and novobiocin had high serum concentrations in comparison with their MBCs and were shown to have potential use for the treatment of dermatophilosis.

  4. [Sensitivity of Kingella kingae to antibiotics].

    PubMed

    Prère, M F; Seguy, M; Vezard, Y; Lareng, M B

    1986-06-01

    Kingella kingae is a small Gram negative rod of the Neisseriaceae family, formerly called Moraxella kingae. This microorganism is found occasionally in the oral cavity and is capable of causing infections. We report three cases of septic arthritis in children due to K. kingae. In vitro susceptibility of the recovered strains was tested using determination of MICs in agar. The strains were susceptible to penicillin, ampicillin, ticarcillin, cephalothin, cefotaxime, gentamicin, chloramphenicol, tetracycline, trimethoprim-sulfamethoxazole, and pefloxacin, less susceptible to erythromycin and resistant to lincomycin (MIC 32 mg/l).

  5. Growth of moderately halophilic bacteria isolated from sea water using phenol as the sole carbon source.

    PubMed

    Muñoz, J A; Pérez-Esteban, B; Esteban, M; de la Escalera, S; Gómez, M A; Martínez-Toledo, M V; González-López, J

    2001-01-01

    Moderately halophilic bacteria utilizing phenol as the sole carbon source were isolated by selective enrichment from sea water. The isolate (Gram-negative motile rods) was identified as Deleya venusta. It grew well in the presence of up to 1600 mg/L of phenol and 8% NaCl under aerobic conditions. When the cells were treated with chloramphenicol prior to the addition of phenol they did not utilize added phenol, even after prolonged incubation. Thus, the enzymes necessary for phenol metabolism appeared to be inducible.

  6. Alkaline phosphatase activity of Escherichia coli starved in sterile lake water microcosms.

    PubMed

    Ozkanca, R; Flint, K P

    1996-03-01

    Escherichia coli grown in high or low phosphate medium was inoculated into a lake water starvation medium. The viable count decreased at 37 degrees C but not at the lower temperatures over 70 d. Alkaline phosphatase was monitored using a colorimetric assay with pNPP as the substrate. Derepression of the enzyme occurred in cultures starved for > 30 d in the lake water and within 5 d in lake water microcosms supplemented with carbon and nitrogen sources where there was rarely an increase in viable count. Chloramphenicol prevented the synthesis of alkaline phosphatase suggesting that, even under starvation conditions, de novo synthesis of the enzyme occurs.

  7. Inducible repair system in Haemophilus influenzae unaccompanied by mutation. [uv

    SciTech Connect

    Notani, N.K.; Setlow, J.K.

    1980-07-01

    Weigle reactivation of ultraviolet-irradiated HPlc1 phage was observed after ultraviolet or mitomycin C treatment of Haemophilus influenzae cells. The amount of reactivation was considerably increased when the treated cells were incubated in growth medium before infection. The presence of chloramphenicol during this incubation abolished the reactivation. No mutation of this phage accompanied the reactivation. When cells were treated so as to produce a maximal reactivation of phage, neither reactivation nor mutation of cells was observed. It is concluded that H. influenzae has an inducible repair system that is not accompanied by mutation.

  8. Recombination in recA cells between direct repeats of insertion element IS1.

    PubMed Central

    Braedt, G

    1985-01-01

    The IS1 sequences that flank the Tn9 chloramphenicol acetyltransferase gene as direct repeats recombine after transformation into an Escherichia coli recA strain. The recombination requires the lambda pL promoter on the plasmid. A plasmid that contains mutant IS1 elements does not recombine. These results indicate that this recombination requires an IS1-specific gene product. The recombinational activity of IS1 may resolve transient cointegrates formed during the transposition of IS1. I discuss a possible role for the lambda pL promoter. Images PMID:2985536

  9. Kinetics of genetic recombination in Escheria coli K-12: competition between genetic integration and degradation.

    PubMed

    Ou, J T; Wood, T H

    1973-12-01

    The effect of shifting zygotes from a rich medium into a poor medium (shift-down) on the frequency of recombination has been examined; our results suggest that protein syntheses other than those required for growth may be involved. Experiments with chloramphenicol support this notion, and further suggest that there is a competition between genetic integration and degradation in the recipient cells. By inhibiting protein synthesis one can shift the competition in favor of either integration or segregation, depending on the physiological state of the zygotes. By calculating the time required for segregation as one generation, we conclude that the recombination chromosome is homozygous at the time of integration.

  10. Effect of seven antibiotics on the growth and reproduction of Heliothis subflexa X H. virescens interspecific hydrids and backcross males

    SciTech Connect

    LaChance, L.E.; Karpenko, C.P.

    1981-09-01

    To test the hypothesis that a maternally transmitted cytoplasmic microorganism is involved in male hybrid sterility found in H. subflexa (Guenee) X H. virescens (F.) hybrid and backcross progeny, we reared H. subflexa, hybrids and backcross progeny on larval diets containing high concentrations of tetracycline, penicillin G, chloramphenicol, ampicillin, gentamicin, rifampin, and 5-iodo-2'-deoxyuridine. The insects tolerated relatively high concentrations (from 120 mg/liter to 6 g/liter) of antibiotics in the larval diet and showed virtually no changes in larval or pupal developmental time, adult lifespan, or fertility. Hybrid and backcross males reared on such adulterated diets were as sterile as those reared on conventional diets.

  11. Transferring Gus gene into intact rice cells by low energy ion beam

    NASA Astrophysics Data System (ADS)

    Zengliang, Yu; Jianbo, Yang; Yuejin, Wu; Beijiu, Cheng; Jianjun, He; Yuping, Huo

    1993-06-01

    A new technique of transferring genes by low energy ion beam has been reported in this paper. The Gus and CAT (chloramphenicol acetyltransferase) genes, as "foreign" genetic materials, were introduced into the suspension cells and ripe embryos or rice by implantation of 20-30 keV Ar + at doses ranging from 1 × 10 15 to 4 × 10 15 ions/cm 2. The activities of CAT and Gus were detected in the cells and embryos after several weeks. The results indicate that the transfer was a success.

  12. Empirical Treatment of Shigella Dysentery with Trimethoprim: Five-Day Course Versus Single Dose

    DTIC Science & Technology

    1987-06-18

    been ade- used because it is 4-5 times more expensive than quately evaluated in the treatment of shigellosis. ampicillin, sulfa drugs , chloramphenicol...tablets of a sulfa drug the preceding day and NMC, NCR, work units 3M464758D849.- 622 OLDFIELD ET AL. BH.062 and 3Ml61102BSIO.AA.421. The views 1974. A...because of increasing resistance to commonly ducted a randomized double-blind placebo con- prescribed drugs . Sulfonamides were the drug of trolled

  13. Susceptibilities of Listeria species isolated from food to nine antimicrobial agents.

    PubMed Central

    Franco Abuín, C M; Quinto Fernández, E J; Fente Sampayo, C; Rodríguez Otero, J L; Domínguez Rodríguez, L; Cepeda Sáez, A

    1994-01-01

    The agar dilution method was used to determine the activities of gentamicin, erythromycin, streptomycin, chloramphenicol, ampicillin, sulfamethazine, cephalothin, penicillin G, and tetracycline against 73 strains belonging to the genus Listeria (L. innocua, L. seeligeri, and L. monocytogenes). All strains were isolated from raw milk, cheese, the dairy processing plant, poultry, and the poultry slaughterhouse. Gentamicin, ampicillin, and erythromycin, of which the MICs for 90% of the strains tested for all three species were < or = 5.96 micrograms/ml, were found to be the most active agents studied. Most of the L. innocua strains isolated from poultry and the poultry slaughterhouse were resistant to tetracycline. PMID:7979303

  14. Study of coagulase positive and negative Staphylococci in clinical samples.

    PubMed

    Deepak, S; Samant, S A; Urhekar, A D

    1999-10-01

    A total of 983 patients samples (pus, urine, sputum, throat swabs, blood culture, CSF, vaginal swabs, ear swabs) were studied for a period of one year. The incidence of coagulase positive Staphylococci was highest in pus and the incidence of coagulase negative Staphylococci was highest in conjunctival swabs. Antibiotic Sensitivity pattern of routinely used antibiotics was studied. Coagulase positive Staphylococci was found to be most sensitive to Ciprofloxacin followed by Norfloxacin and Gentamycin. Coagulase negative Staphylococci showed maximum sensitivity to Ciprofloxacin followed by Norfloxacin and Chloramphenicol.

  15. In vitro antimicrobial susceptibility of Actinobacillus actinomycetemcomitans.

    PubMed Central

    Slots, J; Evans, R T; Lobbins, P M; Genco, R J

    1980-01-01

    The agar dilution technique was used for determination of the antibiotic susceptibilities of 57 oral isolates and 2 nonoral isolates of Actinobacillus actinomycetemcomitans. Tetracycline, minocycline, and chloramphenicol inhibited more than 96% of the strains tested at a concentration of less than or equal to 2 micrograms/ml; 89% of the strains were inhibited by 2 micrograms of carbenicillin per ml. The other antimicrobial agents tested were less active. Approximately 10% of the A. actinomycetemcomitans strains were resistant to ampicillin, erythromycin, and penicillin G at concentrations of 32 to 64 micrograms/ml. These data suggest that tetracycline and minocycline may be valuable drugs in the treatment of A. actinomycetemcomitans infections. PMID:6903116

  16. Plasmid-determined resistance to fosfomycin in Serratia marcescens.

    PubMed Central

    Mendoza, C; Garcia, J M; Llaneza, J; Mendez, F J; Hardisson, C; Ortiz, J M

    1980-01-01

    Multiple-antibiotic-resistant strains of Serratia marcescens isolated from hospitalized patients were examined for their ability to transfer antibiotic resistance to Escherichia coli by conjugation. Two different patterns of linked transferable resistance were found among the transconjugants. The first comprised resistance to carbenicillin, streptomycin, and fosfomycin; the second, and more common, pattern included resistance to carbenicillin, streptomycin, kanamycin, gentamicin, tetracycline, chloramphenicol, sulfonamide, and fosfomycin. The two types of transconjugant strains carried a single plasmid of either 57 or 97 megadaltons in size. Both of these plasmids are present in parental S. marcescens strains resistant to fosfomycin. The 57-megadalton plasmid was transformed into E. coli. Images PMID:7004337

  17. Chloroquinolines block antibiotic efflux pumps in antibiotic-resistant Enterobacter aerogenes isolates.

    PubMed

    Ghisalberti, Didier; Mahamoud, Abdallah; Chevalier, Jacqueline; Baitiche, Milad; Martino, Michèle; Pagès, Jean-Marie; Barbe, Jacques

    2006-06-01

    Efflux mechanisms protect bacterial cells by pumping out toxic compounds and actively contribute to bacterial multidrug resistance. Agents inhibiting efflux pumps are of interest for the control of multidrug-resistant bacterial infections. Herein we report the effects of new chloroquinoline derivatives that render resistant Enterobacter aerogenes isolates noticeably more susceptible to structurally unrelated antibiotics. In addition, some of these chloroquinolines increase the intracellular concentration of chloramphenicol. Some of the molecules tested in this work are able to inhibit the main efflux pump (AcrAB-TolC), which is involved in E. aerogenes antibiotic resistance.

  18. Continuous-flow automation of the Lactobacillus casei serum folate assay.

    PubMed Central

    Tennant, G B

    1977-01-01

    A method is described for the continuous-flow automation of the serum folate assay using Lactobacillus casei. The total incubation period is approximately four hours. The growth response of the organism to folate is estimated by measuring the rate of reduction of 2,3,5-triphenyl tetrazolium chloride (TTC). A simple continuous culture apparatus is used to grow the inoculum. Supplementation of the assay medium is necessary to obtain parallel results. A statistical assessment shows a favourable comparison with the whole-serum tube assay using a chloramphenicol resistant strain of L. casei. The method is less sensitive to inhibitory substances than the tube assay. PMID:415069

  19. First detection of oqxAB in Salmonella spp. isolated from food.

    PubMed

    Wong, Marcus Ho Yin; Chen, Sheng

    2013-01-01

    Food-borne salmonellosis is an important public health problem worldwide and the second leading cause of food-borne illnesses in Hong Kong. In this study, the prevalence and antimicrobial resistance of Salmonella in meat products in Hong Kong were determined. Interestingly, a plasmid-mediated quinolone resistance (PMQR) gene combination, oqxAB, which mediates resistance to nalidixic acid, chloramphenicol, and olaquindox, was for the first time detectable on the chromosomes of two Salmonella enterica serovar Derby isolates. Further surveillance of oqxAB in Salmonella will be needed.

  20. Isolation and Antimicrobial Testing of Aeromonas spp., Citrobacter spp., Cronobacter spp., Enterobacter spp., Escherichia spp., Klebsiella spp., and Trabulsiella spp. from the Gallbladder of Pigs.

    PubMed

    Evangelopoulou, Grammato; Filioussis, Georgios; Kritas, Spyridon; Kantere, Maria; Burriel, Angeliki R

    2015-01-01

    The presence of Gram-negative bacteria species, other than Salmonella spp., in the gallbladder of pigs was examined. Isolated Gram-negative bacteria were assigned to species using the Microgen™ GnA+B-ID Systems. Of the 64 isolated strains 43 were identified as Escherichia coli, seven as Enterobacter spp., three each as Klebsiella spp., Citrobacterfreundii, Aeromonas hydrophila and Cronobacter sakazakii and one each as Escherichiafergusonii and Trabulsiella guamensis. Their antibiograms showed very high resistance to ampicillin, amoxicillin, tetracycline, chloramphenicol and sulfamethoxazole/trimethoprim. It was concluded that the pigs' gallbladder is a reservoir of potentially pathogenic Gram-negative bacteria for pork consumers.

  1. Susceptibility of Pittsburgh pneumonia agent (Legionella micdadei) and other newly recognized members of the genus Legionella to nineteen antimicrobial agents.

    PubMed Central

    Pasculle, A W; Dowling, J N; Weyant, R S; Sniffen, J M; Cordes, L G; Gorman, G M; Feeley, J C

    1981-01-01

    The susceptibilities of 11 strains representing the five recognized species of Legionella were determined by agar dilution testing on buffered charcoal-yeast extract agar. All of the legionellae tested were susceptible to rifampin, erythromycin, rosaramycin, chloramphenicol, and the aminoglycosides and were resistant to clindamycin and vancomycin. Susceptibilities to penicillins and cephalosporins were variable. Legionella micdadei, Legionella bozemanii, and Legionella gormanii were susceptible to these agents, but minimal inhibitory concentrations for each species were different. Legionella dumoffii resembled Legionella pneumophila in being resistant to penicillin, cephalothin, and cephamandole and susceptible to moxalactam and cefoxitin. All species except L. micdadei produced beta-lactamase. PMID:7325645

  2. Survey of in vitro susceptibilities of Vibrio cholerae O1 and O139 to antimicrobial agents.

    PubMed Central

    Yamamoto, T; Nair, G B; Albert, M J; Parodi, C C; Takeda, Y

    1995-01-01

    Vibrio cholerae O139 (173 strains) and O1 (221 strains) were tested for their in vitro susceptibilities to 39 antimicrobial agents. Both O139 and O1 strains were highly susceptible to azithromycin, cephems, minocycline, penems, and newer fluoroquinolones. O139 strains (94.8%), O1 Indian El Tor strains (97%), and Bangladeshi El Tor strains (50%) were highly resistant to streptomycin, sulfamethoxazole, and trimethoprim and moderately resistant to chloramphenicol and furazolidone, in sharp contrast to O1 Peruvian El Tor and O1 classical strains. Some Bangladeshi El Tor strains (43.3%) showed tetracycline resistance as well. PMID:7695314

  3. Antibiotics in South Indian coastal sea and farmed prawns (Penaeus monodon).

    PubMed

    Palaniyappan, Venkatesh; Nagalingam, Arun Kumar; Ranganathan, Hari Prasad; Kandhikuppam, Krishnamoorthy Bharathi; Kothandam, Hari Prasath; Vasu, Soumya

    2013-01-01

    Sulphonamides and chloramphenicol antibiotics were analysed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in sea and farmed prawn (Penaeus monodon) samples obtained from the coastal region of southern India during 2011-2012. Average recoveries were 77-99% and precision was between 1% and 8%. The results revealed that in sea prawn samples neither of the two antibiotics was detected, but in farmed samples from coastal Andhra Pradesh some sulphonamides were detected in a concentration range greater than the maximum residual limit as set by Council Directive 2377/90 EC.

  4. Evaluation of a dehydrated test strip for the detection of yeasts.

    PubMed

    Davies, R R; Savage, M A

    1975-09-01

    Use of a dehydrated test strip for the detection of yeasts is compared with traditional culture on Sabouraud's agar containing 50 mug/ml chloramphenicol. While the selective medium of the strip is satisfactory for the isolation of species of Candida, Torulopsis glabrata grows only very slowly. The strip has the advantage of a long storage life without deterioration, but a high cost may preclude general usage. The numbers of yeasts collected by a bacteriological swab disadvantages of the selective medium, and the value of direct microscopy in the examination of vaginal swabs are discussed.

  5. [Complication following the vaginal use of potassium permanganate for an abortion].

    PubMed

    Atanasov, A; Katsulov, A; Pranchev, N; Katranushkova, N

    1981-01-01

    Complications after abortifacient use of potassium permanganate in a 22-year old woman are described. Approximately 1 month prior to admission, the patient placed in the vagina a bag with potassium permanganate for 12 hours. Gynecological examination showed stenosis of the vagina, enlargement of the uterus, and adhesions in the cervix and vagina. The patient underwent surgical separation of the vagina, daily irrigations of the vagina, and postoperative administration of ampilillin. Microbiological analysis of the vaginal discharge showed the presence of bacteroides melaninogenics and Peptostreptococcus sensitive to ampicillin, chloramphenicol, and cephalosporin. These findings indicate the role of chemical injury of the vagina in pathogenesis of the endogenous anaerobic genital infection.

  6. Genetic control of prophage induction in Haemophilus influenzae after exposure to psoralen plus near-uv light

    SciTech Connect

    George, M.; Notani, N.K.

    1980-09-01

    Prophage S2 could be induced by psoralen plus near-uv light (PNUV) from a wild-type strain of Haemophilus influenzae, from uv light-sensitive strains uvr-1 and uvr-2, and PNUV-sensitive strains PSO1 and PSO7, but not from a recombination-deficient strain, rec-1. The levels of prophage induction were comparable in the wild type and an ATP-dependent DNase-deficient strain, KW31, even though the PNUV-induced degradation in the latter strain was considerably lower. Prophage induction could be observed even with chloramphenicol present before, during, and 30 min after PNUV treatment.

  7. Anaerobic oxidation of acetylene by estuarine sediments and enrichment cultures

    USGS Publications Warehouse

    Culbertson, Charles W.; Zehnder, Alexander J. B.; Oremland, Ronald S.

    1981-01-01

    Acetylene disappeared from the gas phase of anaerobically incubated estuarine sediment slurries, and loss was accompanied by increased levels of carbon dioxide. Acetylene loss was inhibited by chloramphenicol, air, and autoclaving. Addition of 14C2H2 to slurries resulted in the formation of 14CO2 and the transient appearance of 14C-soluble intermediates, of which acetate was a major component. Acetylene oxidation stimulated sulfate reduction; however, sulfate reduction was not required for the loss of C2H2 to occur. Enrichment cultures were obtained which grew anaerobically at the expense of C2H2.

  8. Sensitivity to amphotericin B of a Naegleria sp. isolated from a case of primary amoebic meningoencephalitis.

    PubMed

    Carter, R F

    1969-07-01

    An amoeba of the genus Naegleria causing fatal meningoencephalitis in a human subject has been investigated for its sensitivity to several drugs. Penicillin, sulphadiazine, chloramphenicol, oxytetracycline hydrochloride, streptomycin, methotrexate, emetine, quinine, and metronidazole had no effect on the organism in vitro at levels in excess of those likely to be attained therapeutically in the brain. Amphotericin B was highly amoebicidal in vitro and protected mice from infection with the organism. Used in high dosage by the intraventricular as well as the intravenous route, this drug might be successful in the treatment of further cases of primary amoebic meningoencephalitis.

  9. Anaerobic Oxidation of Acetylene by Estuarine Sediments and Enrichment Cultures

    PubMed Central

    Culbertson, Charles W.; Zehnder, Alexander J. B.; Oremland, Ronald S.

    1981-01-01

    Acetylene disappeared from the gas phase of anaerobically incubated estuarine sediment slurries, and loss was accompanied by increased levels of carbon dioxide. Acetylene loss was inhibited by chloramphenicol, air, and autoclaving. Addition of 14C2H2 to slurries resulted in the formation of 14CO2 and the transient appearance of 14C-soluble intermediates, of which acetate was a major component. Acetylene oxidation stimulated sulfate reduction; however, sulfate reduction was not required for the loss of C2H2 to occur. Enrichment cultures were obtained which grew anaerobically at the expense of C2H2. PMID:16345714

  10. Multiresistance of Staphylococcus xylosus and Staphylococcus equorum from Slovak Bryndza cheese.

    PubMed

    Mikulášová, Mária; Valáriková, Jana; Dušinský, Roman; Chovanová, Romana; Belicová, Anna

    2014-05-01

    Staphylococcus xylosus, Staphylococcus equorum, and Staphylococcus epidermidis strains were isolated from Bryndza cheese and identified using PCR method. The antimicrobial susceptibility of these strains was assessed using disc diffusion method and broth microdilution method. The highest percentage of resistance was detected for ampicillin and oxacillin, and in contrary, isolates were susceptible or intermediate resistant to ciprofloxacin and chloramphenicol. Fourteen of the S. xylosus isolates (45%) and eleven of the S. equorum isolates (41%) exhibited multidrug resistance. None of the S. epidermidis isolate was multiresistant. The phenotypic resistance to oxacillin was verified by PCR amplification of the gene mecA.

  11. Optimum management of Citrobacter koseri infection.

    PubMed

    Deveci, Aydin; Coban, Ahmet Yilmaz

    2014-09-01

    Low virulent Citrobacter koseri can cause life threatening infections. Neonates and other immunocompromised patients are particularly susceptible to infection from C. koseri. Any infection due to C. koseri mandates antimicrobial therapy based on the sensitivity of the pathogen microorganism. Various types of antibiotics, including aminoglycosides carbapenems, cephalosporins, chloramphenicol and quinolones, are used for the treatment of C. koseri infections. The rational choice of antimicrobial therapy for Citrobacter infections is a challenge for clinicians because there is a sustained increase in antibacterial resistance. We reviewed antimicrobial agents used for C. koseri infections in this review.

  12. Antibiotic resistance and serotypes of 100 Streptococcus pneumoniae strains isolated in a children's hospital in Barcelona, Spain.

    PubMed Central

    Latorre, C; Juncosa, T; Sanfeliu, I

    1985-01-01

    A total of 100 Streptococcus pneumoniae strains with various penicillin G susceptibilities, isolated in Barcelona, Spain, from different pediatric sources during 1983 and the first 4 months of 1984, were tested for susceptibility to tetracycline, chloramphenicol, erythromycin, clindamycin, vancomycin, and rifampin. The isolates were distributed in nine patterns of antibiotic resistance, and 15 different serotypes were encountered. The high incidence of resistance to multiple antibiotics clearly indicates the need to perform antibiotic susceptibility testing of all pneumococcal isolates with proved pathologic significance to avoid therapeutic failure. PMID:3834840

  13. Development of a plasmid vector for easy selection of strong promoters.

    PubMed

    Piñeiro, S A; Sordelli, D O; Centrón, D

    2000-05-01

    A promoter vector pACPR33 for Escherichia coli based on the promotorless ampicillin-resistance gene from pBR322 has been constructed. The promoter of the ampicillin-resistance gene was deleted and replaced by a suitable multiple cloning site. Molecular cloning of promoters into the polylinker resulted in activation of the ampicillin resistance in E. coli. The plasmid contains a functional origin of DNA replication and a tetracycline resistance gene for E. coli, and a chloramphenicol resistance gene for S. aureus. The vector permitted direct detection of promoter activity, especially strong promoters, by easy iodometric determination of beta-lactamase activity in liquid or solid media.

  14. Revised ciprofloxacin breakpoints for Salmonella Typhi: its implications in India.

    PubMed

    Balaji, V; Sharma, A; Ranjan, P; Kapil, A

    2014-01-01

    The rise of multidrug resistant strains of Salmonella Typhi in the last decade of the previous century led to the use of fluoroquinolones as the drug of choice. However, over the past few years fluoroquinolone resistance has been increasingly reported. In accordance with the revised Clinical and Laboratory Standards Institute (CLSI) breakpoints, only 3% of the isolates were susceptible to ciprofloxacin in comparison to 95% as per the earlier guidelines when 488 isolates collected between 2010 and 2012 were re-interpreted. Interestingly, re-emergence of strains susceptible to chloramphenicol, ampicillin and cotrimoxazole is being seen. Amidst the changing susceptibility profile, azithromycin remains a promising alternative.

  15. Extended Spectrum Beta Lactamase producing Cephalosporin resistant Salmonella Typhi, reported from Rawalpindi, Pakistan.

    PubMed

    Munir, Tehmina; Lodhi, Munir; Ansari, Jawad Khaliq; Andleeb, Saadia; Ahmed, Mushtaq

    2016-08-01

    Typhoid is endemic in many parts of southeast Asia. Due to the resistance of the organism to first line of antibiotics (ampicillin, chloramphenicol, cotrimoxazole) as well as to fluoroquinolones, third generation cephalosporins have been in use for the empiric treatment of typhoid for years. However an increasing incidence of Salmonella Typhi is being reported sporadically from various regions. We report a case of typhoid due to Salmonella Typhi which was non-responsive to treatment with a cephalosporin, was found to be multidrug resistant and resistant to ciprofloxacin and third generation cephalosporin as well. The patient was finally treated successfully with intravenous administration of a carbapenem.

  16. Rickettsial infection of the central nervous system: the role of prompt antimicrobial therapy.

    PubMed

    Shaked, Y

    1991-04-01

    Rickettsial diseases of man, which are prevalent in all the continents (except Antartica) continue to be a major health problem in tropical and temperate parts of the world. Rickettsioses must be considered in every traveller seeking medical attention soon after returning from endemic areas, since the disease can be associated with significant morbidity. When a definite diagnosis has been made by specific serological tests, prompt antimicrobial therapy is indicated. A tetracycline should be regarded as the drug of choice due to its high efficacy, low toxicity, superior in vitro activity and the possibility of relapse which can follow chloramphenicol therapy.

  17. Conjugative transfer of cadmium resistance plasmids in Rhodococcus fascians strains.

    PubMed

    Desomer, J; Dhaese, P; Van Montagu, M

    1988-05-01

    The presence of a 138-kilobase plasmid (pD188) correlated with increased resistance to cadmium in Rhodococcus fascians D188. This plasmid could be transferred by a conjugation-like system in matings between R. fascians strains. Transconjugants expressed the cadmium resistance and could be used as donors in subsequent matings. Four other R. fascians strains (NCPPB 1488, NCPPB 1675, NCPPB 2551, and ATCC 12974) could also be used as donors for cadmium resistance in matings. Strain NCPPB 1675 showed a 100% cotransfer of cadmium and chloramphenicol resistance markers.

  18. Improved electroporation of Rhodococcus equi.

    PubMed

    Sekizaki, T; Tanoue, T; Osaki, M; Shimoji, Y; Tsubaki, S; Takai, S

    1998-02-01

    The condition of an electroporation method was re-evaluated for the introduction of foreign plasmid DNA into Rhodococcus equi. The method is based on an electroporation of the bacteria made competent by culturing in a broth containing glycine and by heat shock at 50 degrees C. Transformation of R. equi could be achieved with a chloramphenicol-resistant shuttle vector originating from Rhodococcus fascians at an efficiency of about 10(4) transformants/microgram DNA. The bacteria were also shown to become competent when they were incubated with a chemical transformation buffer prior to washing with an electroporation buffer.

  19. Matrix effect on the analysis of amphenicols in fish by liquid chromatography-tandem mass spectrometry (LC-MS/MS)

    NASA Astrophysics Data System (ADS)

    Guidi, L. R.; Tette, P. A. S.; Evangelista, W. P.; Fernandes, C.; Glória, M. B. A.

    2015-01-01

    Matrix effect is an important parameter to be investigated during the development and validation of a method for the quantitative determination of contaminants in food. The objective of this study was to evaluate the matrix effect, through statistical tests, in the quantification of amphenicols in fish by HPLC-MS/MS. The study was performed by comparing the standard curves prepared in solvent solutions and in a fish sample previously known to be free of amphenicols. Since matrix effect was observed for the three analytes, calibration curves for quantification of chloramphenicol, thiamphenicol and florfenicol should be constructed using the matrix.

  20. In vitro drug susceptibility pattern of Mycoplasma alligatoris isolated from symptomatic American alligators (Alligator mississippiensis).

    PubMed

    Helmick, Kelly E; Brown, Daniel R; Jacobson, Elliott R; Brown, Mary B

    2002-06-01

    A recently described mycoplasma, Mycoplasma alligatoris, was isolated from dead American alligators (Alligator mississippiensis) that had demonstrated clinical signs of lethargy, anorexia, bilateral ocular discharge, edema. paraparesis, and polyarthritis. The in vitro minimum inhibitory concentration for nine antibacterial agents was determined through serial dilution in broth and plate culture for M. alligatoris isolates. The inhibitory concentration obtained for doxycycline, enrofloxacin, sarafloxacin, oxytetracycline, tilmicosin, and tylosin (< 1 microg/ml) was lower than that of clindamycin (1-8 microg/ml), chloramphenicol (8-16 microg/ml), and erythromycin (32-138 microg/ml).

  1. Vinylpyrrolidone-co-(meth)acrylic acid inserts for ocular drug delivery: synthesis and evaluation.

    PubMed

    Barbu, Eugen; Sarvaiya, Indrajeetsinh; Green, Keith L; Nevell, Thomas G; Tsibouklis, John

    2005-09-15

    Copolymeric hydrogels constituting of vinylpyrrolidone and methacrylic or acrylic acid repeat units have been prepared and investigated for their ability to act as controlled release vehicles in ophthalmic drug delivery. The materials were synthesized by radical-induced polymerization in the presence of N,N'-methylenebisacrylamide crosslinker, and the influences of network composition and drug solubility upon the swelling properties, adhesion behavior, and drug release characteristics were studied. In vitro release experiments showed that some of these materials could be useful vehicles for the delivery of drugs such as pilocarpine or chloramphenicol, while in vivo studies, using the rabbit model, confirmed their high potential for the controlled ocular delivery of pilocarpine hydrochloride.

  2. Transient Transfection and Expression in the Obligate Intracellular Parasite Toxoplasma gondii

    NASA Astrophysics Data System (ADS)

    Soldati, Dominique; Boothroyd, John C.

    1993-04-01

    Toxoplasma gondii is a protozoan pathogen that produces severe disease in humans and animals. This obligate intracellular parasite provides an excellent model for the study of how such pathogens are able to invade, survive, and replicate intracellularly. DNA encoding chloramphenicol acetyltransferase was introduced into T. gondii and transiently expressed with the use of three vectors based on different Toxoplasma genes. The ability to introduce genes and have them efficiently and faithfully expressed is an essential tool for understanding the structure-function relation of genes and their products.

  3. [In vitro antibiotic sensitivity of French strains of Mycoplasma bovis].

    PubMed

    Poumarat, F; Martel, J L

    1989-01-01

    The in vitro activity of 15 antibiotics was tested with 30-90 Mycoplasma bovis representative strains of bovine lung pathology in France. The distribution of minimal inhibitory concentration (MIC) is homogeneous with low values for spectinomycin, lincomycin, tylosin, gentamicin and baytril, intermediate for chloramphenicol and neomycin, high for nalidixic acid, Flumequine and erythromycin. The MIC distribution is heterogeneous with intermediate values for spiramycin and tetracyclines, and high values for streptomycin. For the later antibiotics, the heterogeneity of the susceptibility suggests a mechanism of acquired resistance.

  4. Characterization of a Plasmid-Encoded Type IV Secretion System in Campylobacter jejuni 81-176

    DTIC Science & Technology

    2004-01-01

    jejuni 81-176, pTet, is a conjugative R factor encoding tetracycline resistance (Batchelor et al., 2004), and is likely related to the tetO...chloramphenicol per ml, 25 µg of kanamycin per ml, 20 µg of streptomycin per ml, 20 µg of tetracycline per ml, and 10 µg of trimethoprim per ml. Plasmids...shows the qualitative results from a yeast 2- hybrid screen using Cjp5/VirB11 as both bait and prey. Colonies were subcloned on 54 minimal

  5. Antimicrobial therapy of experimental Legionella micdadei pneumonia in guinea pigs.

    PubMed Central

    Pasculle, A W; Dowling, J N; Frola, F N; McDevitt, D A; Levi, M A

    1985-01-01

    Several antimicrobial agents were evaluated for activity against experimental Legionella micdadei pneumonia in guinea pigs. Erythromycin, rifampin, doxycycline, and sulfamethoxazole-trimethoprim produced significant reductions in mortality. Penicillin, cefazolin, cefoxitin, chloramphenicol, and gentamicin were not efficacious even though, at the doses administered, the peak concentrations of these agents in serum substantially exceeded their MICs for the test strain. It is suggested that the poor performance of the latter group of agents resulted from poor penetration into cells in which L. micdadei was multiplying. PMID:3878688

  6. Three-year prospective study of intestinal pathogens in Madrid, Spain.

    PubMed Central

    Velasco, A C; Mateos, M L; Más, G; Pedraza, A; Díez, M; Gutiérrez, A

    1984-01-01

    During the period July 1980 through June 1983, in a General Hospital in Madrid, the following organisms were detected from 6,970 patients with gastroenteritis: 710 Salmonella spp. 506 Campylobacter jejuni, 379 Shigella spp., 12 Yersinia enterocolitica, 1,466 rotavirus, 134 Giardia lamblia, and 4 Entamoeba histolytica. Chloramphenicol showed good activity against most tested strains of Salmonella spp., Shigella spp., and C. jejuni. The incidence of Salmonella spp. and Shigella spp. was very marked in the hot dry months of the year, rotavirus predominated during the cold months, and no seasonal variations of importance were seen for C. jejuni and G. lamblia. PMID:6092418

  7. Salmonella serovars and antimicrobial resistance in strains isolated from wild animals in captivity in Sinaloa, Mexico.

    PubMed

    Silva-Hidalgo, Gabriela; López-Valenzuela, Martin; Juárez-Barranco, Felipe; Montiel-Vázquez, Edith; Valenzuela-Sánchez, Beatriz

    2014-08-01

    The aim of the present study was to evaluate the frequency of antibiotic resistance in Salmonella spp. strains from wild animals in captivity at the Culiacan Zoo and the Mazatlan Aquarium in Sinaloa, Mexico. We identified 17 different Salmonella enterica serovars at a prevalence of 19.90% (Culiacan Zoo) and 6.25% (Mazatlan Aquarium). Antibiotic sensitivity tests revealed that, of the 83 strains studied, 100% were multidrug resistant (MDR). The drugs against which the greatest resistance was observed were: penicillin, erythromycin, dicloxacillin, ampicillin, cephalothin, and chloramphenicol. We therefore conclude that MDR is common among Salmonella isolates originating from wild animals in captivity in Sinaloa.

  8. Antimicrobial Resistance of Diarrheagenic Escherichia coli Isolated from Children under the Age of 5 Years from Ifakara, Tanzania

    PubMed Central

    Vila, Jordi; Vargas, Martha; Casals, Climent; Urassa, Honorato; Mshinda, Hassan; Schellemberg, David; Gascon, Joaquim

    1999-01-01

    Diarrhea caused by multidrug-resistant bacteria is an important public health problem among children in developing countries. The prevalence and antimicrobial susceptibility of diarrheagenic Escherichia coli in 346 children under 5 years of age in Ifakara, Tanzania, were studied. Thirty-eight percent of the cases of diarrhea were due to multiresistant enterotoxigenic E. coli, enteroaggregative E. coli, or enteropathogenic E. coli. Strains of all three E. coli categories showed high-level resistance to ampicillin, tetracycline, co-trimoxazole, and chloramphenicol but were highly susceptible to quinolones. Guidelines for appropriate use of antibiotics in developing countries need updating. PMID:10582903

  9. Expression of Aequorea green fluorescent protein in plant cells.

    PubMed

    Hu, W; Cheng, C L

    1995-08-07

    The coding region of the green fluorescent protein (GFP) from Aequorea victoria has been fused to the cauliflower mosaic virus 35S promoter and introduced into maize leaf protoplasts. Transient expression of GFP was observed. In addition, the coding region of GFP was fused to an Arabidopsis heat shock promoter and co-transformed with another construct in which GFP has been replaced with chloramphenicol acetyltransferase (CAT). The heat-induced expression of GFP in maize protoplasts parallels that of CAT. While GFP was expressed in both dark-grown and green maize leaf protoplasts, no green fluorescence was observed in similarly transformed Arabidopsis protoplasts.

  10. Selection of multiple-antibiotic-resistant (mar) mutants of Escherichia coli by using the disinfectant pine oil: roles of the mar and acrAB loci.

    PubMed Central

    Moken, M C; McMurry, L M; Levy, S B

    1997-01-01

    Mutants of Escherichia coli selected for resistance to the disinfectant pine oil or to a household product containing pine oil also showed resistance to multiple antibiotics (tetracycline, ampicillin, chloramphenicol, and nalidixic acid) and overexpressed the marA gene. Likewise, antibiotic-selected Mar mutants, which also overexpress marA, were resistant to pine oil. Deletion of the mar or acrAB locus, the latter encoding a multidrug efflux pump positively regulated in part by MarA, increased the susceptibility of wild-type and mutant strains to pine oil. PMID:9420057

  11. Production of superoxide dismutases from Proteus mirabilis and Proteus vulgaris.

    PubMed

    Dayton, T M; Diefenbach, K A; Fuller, M L; Valtos, J; Niederhoffer, E C

    1996-04-01

    Proteus mirabilis and Proteus vulgaris expressed a combination of superoxide dismutase (Sod) activities, which was assigned to FeSod1, FeSod2 and MnSod for P. mirabilis, and FeSod, MnSod and CuZnSod for P. vulgaris. Production of the Sod proteins was dependent on the availability of iron, whether cells were grown under anaerobiosis or aerobiosis and growth phase. Nalidixic acid and chloramphenicol inhibited cell growth and the iron- and dioxygen-dependent production of Sod. These results support the involvement of metal ions and redox status in the production of Proteus Sods.

  12. [Genetic transfer of methycilline resistance in Staphylococus epidermidis and Staphylococcus aureus strains in mixed cultures].

    PubMed

    Młynarczyk, A; Młynarczyk, G; Jeljaszewicz, J

    1999-01-01

    In mixed cultures of staphylococci a transfer of the resistance to methicillin and penicillinase plasmids as well as tetracycline and chloramphenicol plasmids was investigated. It was shown that the resistance to methicillin was transferred in mixed cultures from one strain of S. aureus to another and from S. epidermidis to S. aureus. In both cases transfer of methicillin resistance required, the presence of penicillinase plasmid in recipient or donor strain. In the case of other markers transmission was independent. Moreover it was shown that the transfer of resistance genes in mixed cultures was mediated by bacteriophage of the serologic group A.

  13. Identification and Characterization of MYC Regulatory Elements: Links to Prostate Cancer

    DTIC Science & Technology

    2011-06-01

    mg/mL kanamycin and 20 mg/mL chloramphenicol. Positive colonies were first identified by polymerase chain reaction (PCR) using beta-globin and lacZ...purpose of this proposal is to identify and characterize prostate enhancers within the prostate cancer associated intervals on 8q24 using a combination of...the need to investigate the expression patterns of other nearby genes. Task 1b, 1c and 1d: Use progressively smaller DNA fragments in in vivo mouse

  14. Studies on the antimicrobial activity of Nigella sativa seed (black cumin).

    PubMed

    Hanafy, M S; Hatem, M E

    1991-09-01

    Filter paper discs impregnated with the diethyl ether extract of Nigella sativa seeds (25-400 micrograms extract/disc) caused concentration-dependent inhibition of Gram-positive bacteria represented by Staphylococcus aureus. Gram-negative bacteria represented by Pseudomonas aeruginosa and Escherichia coli (but not Salmonella typhimurium) and a pathogenic yeast Candida albicans. The extract showed antibacterial synergism with streptomycin and gentamicin and showed additive antibacterial action with spectinomycin, erythromycin, tobramycin, doxycycline, chloramphenicol, nalidixic acid, ampicillin, lincomycin and sulphamethoxyzole-trimethoprim combination. The extract successfully eradicated a non-fatal subcutaneous staphylococcal infection in mice when injected at the site of infection.

  15. Clonal relationship among Vibrio cholerae O1 El Tor strains isolated in Somalia.

    PubMed

    Scrascia, Maria; Pugliese, Nicola; Maimone, Francesco; Mohamud, Kadigia A; Grimont, Patrick A D; Materu, Sadiki F; Pazzani, Carlo

    2009-03-01

    One hundred and three Vibrio cholerae O1 strains, selected to represent the cholera outbreaks which occurred in Somalia in 1998-1999, were characterized by random amplified polymorphic DNA patterns, ribotyping, and antimicrobial susceptibility. All strains showed a unique amplified DNA pattern and 2 closely related ribotypes (B5a and B8a), among which B5a was the more frequently identified. Ninety-one strains were resistant to ampicillin, chloramphenicol, spectinomycin, streptomycin, sulfamethoxazole, and trimethoprim, conferred, except for spectinomycin, by a conjugative plasmid IncC. These findings indicated that the group of strains active in Somalia in the late 1990s had a clonal origin.

  16. Outer Membrane Proteins form Specific Patterns in Antibiotic-Resistant Edwardsiella tarda

    PubMed Central

    Peng, Bo; Wang, Chao; Li, Hui; Su, Yu-bin; Ye, Jin-zhou; Yang, Man-jun; Jiang, Ming; Peng, Xuan-xian

    2017-01-01

    Outer membrane proteins of Gram-negative bacteria play key roles in antibiotic resistance. However, it is unknown whether outer membrane proteins that respond to antibiotics behave in a specific manner. The present study specifically investigated the differentially expressed outer membrane proteins of an antibiotic-resistant bacterium, Edwardsiella tarda, a Gram-negative pathogen that can lead to unnecessary mass medication of antimicrobials and consequently resistance development in aquaculture and a spectrum of intestinal and extraintestinal diseases in humans. The comparison of a clinically isolated strain to the laboratory derived kanamycin-, tetracycline-, or chloramphenicol-resistant strains identified their respective outer membrane proteins expression patterns, which are distinct to each other. Similarly, the same approach was utilized to profile the patterns in double antibiotic-resistant bacteria. Surprisingly, one pattern is always dominant over the other as to these three antibiotics; the pattern of chloramphenicol is over tetracycline, which is over kanamycin. This type of pattern was also confirmed in clinically relevant multidrug-resistant bacteria. In addition, the presence of plasmid encoding antibiotic-resistant genes also alters the outer membrane protein profile in a similar manner. Our results demonstrate that bacteria adapt the antibiotic stress through the regulation of outer membrane proteins expression. And more importantly, different outer membrane protein profiles were required to cope with different antibiotics. This type of specific pattern provides the rationale for the development of novel strategy to design outer membrane protein arrays to identify diverse multidrug resistance profiles as biomarkers for clinical medication. PMID:28210241

  17. Molecular characterization of Vibrio cholerae outbreak strains with altered El Tor biotype from southern India

    PubMed Central

    Jain, M.; Kumar, P.; Jiang, S. C.

    2009-01-01

    Forty-four Vibrio cholerae isolates collected over a 7-month period in Chennai, India in 2004 were characterized for gene traits, antimicrobial susceptibility and genomic fingerprints. All 44 isolates were identified as O1 El Tor Ogawa, positive for various toxigenic and pathogenic genes viz. ace, ctxB, hlyA, ompU, ompW, rfbO1, rtx, tcpA, toxR and zot. Nucleotide sequencing revealed the presence of cholera toxin B of classical biotype in all the El Tor isolates, suggesting infection of isolates by classical CTXΦ. Antibiogram analysis showed a broad-spectrum antibiotic resistance that was also confirmed by the presence of resistant genes in the genomes. All isolates contained a class 1 integron and an SXT constin. However, isolates were sensitive to chloramphenicol and tested negative for the chloramphenicol resistant gene suggesting a deletion in SXT constin. Fingerprinting analysis of isolates by ERIC- and Box PCR revealed similar DNA patterns indicating the clonal dissemination of a single predominant V. cholerae O1 strain throughout the 2004 outbreak in Chennai. Electronic supplementary material The online version of this article (doi:10.1007/s11274-009-0171-7) contains supplementary material, which is available to authorized users. PMID:20495624

  18. Metal and antibiotic resistance of bacteria isolated from the Baltic Sea.

    PubMed

    Moskot, Marta; Kotlarska, Ewa; Jakóbkiewicz-Banecka, Joanna; Gabig-Cimińska, Magdalena; Fari, Karolina; Wegrzyn, Grzegorz; Wróbel, Borys

    2012-09-01

    The resistance of 49 strains of bacteria isolated from surface Baltic Sea waters to 11 antibiotics was analyzed and the resistance of selected strains to three metal ions (Ni2+, Mn2+, Zn2+) was tested. Most isolates belonged to Gammaproteobacteria (78%), while Alphaproteobacteria (8%), Actinobacteria (10%), and Bacteroidetes (4%) were less abundant. Even though previous reports suggested relationships between resistance and the presence of plasmids or the ability to produce pigments, no compelling evidence for such relationships was obtained for the strains isolated in this work. In particular, strains resistant to multiple antibiotics did not carry plasmids more frequently than sensitive strains. A relation between resistance and the four aminoglycosides tested (gentamycin, kanamycin, neomycin, and streptomycin), but not to spectinomycin, was demonstrated. This observation is of interest given that spectinomycin is not always classified as an aminoglycoside because it lacks a traditional sugar moiety. Statistical analysis indicated relationships between resistance to some antibiotics (ampicillin and erythromycin, chloramphenicol and erythromycin, chloramphenicol and tetracycline, erythromycin and tetracycline), suggesting the linkage of resistance genes for antibiotics belonging to different classes. The effects of NiSO4, ZnCl2 and MnCl2 on various media suggested that the composition of Marine Broth might result in low concentrations of Mn2+ due to chemical interactions that potentially lead to precipitation.

  19. Antibiotic Resistance Patterns of Gram-Negative Psychrotrophic Bacteria from Bulk Tank Milk.

    PubMed

    Decimo, Marilù; Silvetti, Tiziana; Brasca, Milena

    2016-04-01

    Bacterial resistance to antibiotics is a major global health problem and resistance of Pseudomonadaceae and Enterobacteriaceae is a serious concern. We investigated the prevalence of drug-resistance in a total of 80 psychrotrophic strains from bulk milk belonging to Pseudomonas genus (n. 63) and Enterobacteriaceae group (n. 17). All the strains were tested against 16 antibiotics. Pseudomonas were further investigated for their sensitivity against 12 additional antibiotics. Pseudomonas showed a high susceptibility toward fluoroquinolones, aminoglycosides, and piperacillin and, to a lesser extent, to imipenem, ceftazidime, cefepime. Thirty-five out of 63 Pseudomonas strains were susceptible to meropenem, while among antibiotics for which recommended breakpoints are not yet available, 55% of Pseudomonas strains had no inhibition halo in presence of nitrofurantoin, highlighting a resistance toward this drug. The results obtained in this study indicate a high efficiency of fluoroquinolones, chloramphenicol (94%), and kanamycin (76%) for Enterobacteriaceae while a high prevalence of resistant strains was found to ampicillin (13/17). Serratia marcescens is highly susceptible to fluoroquinolones, chloramphenicol, and kanamycin. Moreover, mupirocin seems to be the new antibiotic with the less efficacy for Enterobacteriaceae, with 41% of strains without halo, pointing out an important resistance. Further knowledge on resistance to known and new antibiotics among Pseudomonas species and Enterobacteriaceae of milk origin was acquired.

  20. Safety assessment of Tetragenococcus halophilus isolates from doenjang, a Korean high-salt-fermented soybean paste.

    PubMed

    Jeong, Do-Won; Heo, Sojeong; Lee, Jong-Hoon

    2017-04-01

    We assessed the safety of 49 Tetragenococcus halophilus strains isolated from doenjang in Korea. Minimum inhibitory concentration assays showed that all strains can be considered as susceptible to ampicillin, erythromycin, penicillin G, tetracycline, and vancomycin, but resistant to ciprofloxacin based on the Enterococcus breakpoint values provided by the European Committee on Antimicrobial Susceptibility testing in 2015. Ciprofloxacin resistance was sufficiently high to consider the potential for acquisition of transmissible determinants. Two strains exhibiting potentially acquired resistance to chloramphenicol and gentamicin, and chloramphenicol alone, were identified. None of the strains exhibited α-hemolytic activity or biofilm formation; two strains exhibited weak β-hemolytic activity. Doenjang isolates produced an average of 3338.6 ppm of tyramine in the laboratory, considerably higher than the levels produced by two reference strains. All of the test strains exhibited similar cadaverine, histamine, and putrescine production patterns. Most T. halophilus strains could grow at a NaCl concentration >18%, exhibited acid production at 15% NaCl, and expressed strain-specific protease and lipase activities. The potential acquisition of transmissible determinants for antibiotic resistance and tyramine production identified in this study necessitate the need for a thorough safety assessment of T. halophilus before it can be considered for use in food fermentation processes.

  1. Serologic test systems development. Progress report, October 1, 1978-September 30, 1979

    SciTech Connect

    Seawright, G.L.; Sanders, W.M.; Hollstein, U.; Butler, J.E.; Mills, K.W.; Despommier, D.D.; Zimmerman, W.J.; Martinez, E.; Hindman, K.R.; Payne, R.J.

    1980-12-01

    Work has continued on the development and automation of enzyme immunoassays (EIA) for detecting diseases and toxic agents in food animals. Further evaluations were made of the Technicon Autoanalyzer II(AAII) for conducting totally automated EIAs. The problems investigated were machine carryover and assay variation. Modifications greatly reduced or eliminated carryover and produced acceptable levels of test variation. The EIA for swine trichinosis was significantly improved by the use of a new, partially purified antigen preparation. The result was improved detection of early seroconversions and reduced probability for false negatives and false positives. The amplified EIA was adapted as a diagnostic test for bovine brucellosis and studies were initiated for differentiating vaccinated and infected animals. Preliminary data indicate that the IgG/sub 1/ response may be diagnostic but further studies are necessary. Development of the EIA for detecting low molecular weight contaminants and residues in food products was also initiated. Compounds studied were the antibiotics chloramphenicol, tetracycline, and gentamicin; the mycotoxin, aflatoxin, and the shale oil toxin, 2-aminofluorene. Results indicate that chloramphenicol nonspecifically binds to antibody and interferes with antibody activity. Thus, the test is not yet satisfactory. Initial attempts to automate the gentamicin test were unsuccessful because of machine carryover but modifications of the AAII have produced encouraging preliminary data. Work is continuing on the development of EIAs for all of the compounds mentioned above. (ERB)

  2. PubMed Central

    Nicolas, P.; Cavallo, J. D.; Fabre, R.; Martet, G.

    1998-01-01

    Studying the susceptibility of 189 Neisseria meningitidis strains to penicillin, amoxicillin, cefotaxime, ceftriaxone, chloramphenicol and rifampicin by determination of minimum inhibitory concentrations (MICs) by agar dilution (reference method), E-test and disc diffusion method on Mueller-Hinton agar at 37 degrees C with 5% CO2 enabled us to standardize the antibiograms. While MIC determination by agar dilution is still the reference method, it is possible to obtain exact or approximate MIC values using the E-test. For laboratories that cannot determine penicillin MICs, it is impossible to detect strains that are relatively resistant to penicillin (RRP strains: 0.1 < or = MIC < or = 1 mg/l) using a 10-U penicillin disc. A 1 microgram-oxacillin disc allows MIC to be determined in most cases when the oxacillin inhibition zone is < or = 10 mm. Such strains must be sent to a reference laboratory for exact MIC determination. Based on our results and literature data on pharmacokinetics, we propose critical concentrations for these various antibiotics as well as critical diameters for chloramphenicol and rifampicin discs. PMID:9803590

  3. Optimal expression condition of recombinant RAP.

    PubMed

    Zhang, Jie; Zhang, Hong; Bi, Hao; Liu, Zhiguo; Guo, Jianli; Qu, Shen

    2007-02-01

    In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.

  4. Response of bacterial isolates from Antarctic shallow sediments towards heavy metals, antibiotics and polychlorinated biphenyls.

    PubMed

    Lo Giudice, Angelina; Casella, Patrizia; Bruni, Vivia; Michaud, Luigi

    2013-03-01

    The response of bacterial isolates from Antarctic sediments to polychlorinated biphenyls (Aroclor 1242 mixture), heavy metal salts (cadmium, copper, mercury and zinc) and antibiotics (ampicillin, chloramphenicol, kanamycin and streptomycin) was investigated. Overall, the ability to growth in the presence of Aroclor 1242 as a sole carbon source was observed for 22 isolates that mainly belonged to Psychrobacter spp. Tolerance to the heavy metals assayed in this study was in the order of Cd > Cu > Zn > Hg and appeared to be strictly related to the metal concentrations, as determined during previous chemical surveys in the same area. With regards to antibiotic assays, the response of the isolates to the tested antibiotics ranged from complete resistance to total susceptibility. In particular, resistances to ampicillin and chloramphenicol were very pronounced in the majority of isolates. Our isolates differently responded to the presence of toxic compounds primarily based on their phylogenetic affiliation and secondarily at strain level. Moreover, the high incidence of resistance either to metal or antibiotics, in addition to the capability to grow on PCBs, confirm that bacteria are able to cope and/or adapt to the occurrence pollutants even in low human-impacted environments.

  5. [Standardization of the Neisseria meningitidis antibiogram. Detection of strains relatively resistant to penicillin].

    PubMed

    Nicolas, P; Cavallo, J D; Fabre, R; Martet, G

    1998-01-01

    Studying the susceptibility of 189 Neisseria meningitidis strains to penicillin, amoxicillin, cefotaxime, ceftriaxone, chloramphenicol and rifampicin by determination of minimum inhibitory concentrations (MICs) by agar dilution (reference method), E-test and disc diffusion method on Mueller-Hinton agar at 37 degrees C with 5% CO2 enabled us to standardize the antibiograms. While MIC determination by agar dilution is still the reference method, it is possible to obtain exact or approximate MIC values using the E-test. For laboratories that cannot determine penicillin MICs, it is impossible to detect strains that are relatively resistant to penicillin (RRP strains: 0.1 < or = MIC < or = 1 mg/l) using a 10-U penicillin disc. A 1 microgram-oxacillin disc allows MIC to be determined in most cases when the oxacillin inhibition zone is < or = 10 mm. Such strains must be sent to a reference laboratory for exact MIC determination. Based on our results and literature data on pharmacokinetics, we propose critical concentrations for these various antibiotics as well as critical diameters for chloramphenicol and rifampicin discs.

  6. Antibiotic resistance and mechanisms implicated in clinical enterococci in a Tunisian hospital.

    PubMed

    Klibi, N; Gharbi, S; Masmoudi, A; Ben Slama, K; Poeta, P; Zarazaga, M; Fendri, C; Boudabous, A; Torres, C

    2006-02-01

    Susceptibility testing for 15 antibiotics was performed in a series of 191 clinical enterococci recovered in a Tunisian Hospital during 2000-2003. Species detected were the following ones (number of isolates): E. faecalis (139), E. faecium (41), E. casseliflavus (5), E. gallinarum (3), E. avium (2) and E. hirae (1). The percentages of antibiotic resistance detected were as follows (E. faecalis/ E. faecium/ other species) : penicillin (0/ 73/ 9%), tetracycline (78/ 44/ 54%), chloramphenicol (52/ 29/ 27%), erythromycin (66/ 100/ 82%), spiramycin (84/ 83/ 64%), pristinamycin (100/ 0/ 73%), trimethoprim-sulfamethoxazole (88/ 78/ 91%), rifampicin (72/ 41/ 0%), vancomycin (0/ 0/ 36%), teicoplanin (0/ 0/ 0%), high-level-resistance for gentamicin (24/ 29/ 45%), streptomycin (34/ 56/ 55%) and kanamycin (41/ 68/ 55%). Increased vancomycin minimum inhibitory concentrations (MICs) were only detected in E. casseliflavus and E. gallinarum isolates (MIC range 8-24 microg/ml). The erm(B), catA, tet(M), aac(6')-aph(2''), aph(3')-IIIa, and ant(6)-Ia genes were detected in 91%, 32%, 86%, 98%, 100%, and 72% of the E. faecium and E. faecalis isolates resistant to erythromycin, chloramphenicol, tetracycline and high-level-resistant to gentamicin, kanamycin and streptomycin, respectively. A total of 20 unrelated pulsed-field-gel-electrophoresis patterns were found in the series of 46 high-level gentamicin-resistant E. faecalis and E. faecium isolates of this study.

  7. [Antibiotic sensitivity of meningococci isolated from patients and carriers].

    PubMed

    Dabernat, H; Delmas, C; Lareng, M B

    1984-06-01

    Ninety-two strains of Neisseria meningitidis (twenty-four recovered from CSF, four from blood, and sixty-four from oropharyngeal swabs) were tested for susceptibility to penicillin, ampicillin, minocycline, chloramphenicol, rifampicin, erythromycin, spiramycin, josamycin, sulphamethoxazole, and trimethoprim. Antibiotic activity (MIC) was determined by agar dilution. Among the 92 cultures, strains were in the following decreasing order: group B, undefined groups, and group C. 75% of strains recovered from carriers and 50% of those recovered from CSF were susceptible to 8 mg/l of sulphamethoxazole. No strain was susceptible to trimethoprim (MIC 8 mg/l). All strains were susceptible to penicillin (range MICs: 0.007-0.25; mean MIC: 0.06), ampicillin (0.015-0.5; 0.12), minocycline (0.03-0.05; 0.25), chloramphenicol (0.12-4; 0.5). and rifampin (0.007-0.25; 0.015). In vitro activity of macrolides was influenced by culture conditions. With 5 to 10% CO2, MICs were 2 to 4 times greater than those culture conditions. With 5 to 10% CO2, MICs were 2 to 4 times greater than those observed without CO2. Josamycin was more active than erythromycin and spiramycin.

  8. [Practical problems raised by antibiotic minimal inhibitory concentration determination in Legionella species (author's transl)].

    PubMed

    Bornstein, N; Roudier, C; Fleurette, J

    1982-06-01

    Because of culture medium constitution and growth conditions of Legionella species, antibiotic Minimal inhibitory Concentration (MIC) determination is difficult and does not fit with the accepted standards. The following strains have been tested : references strains : Staphylococcus aureus Oxford CNCM 53154, S. aureus 209P CNCM 53156, E. coli CNCM A224, E. coli CNCM 7324. L. pneumophila, 9 strains representative of the 6 serogroups. L. micdadei, L. dumoffii. 5 clinical isolates of L. pneumophila serogroup 1 (3 isolated in our Laboratory and 2 in Belgium). MIC of 17 antibiotics have been determined, among them macrolids not tested before. The method used was the Agar Dilution Method, with several media compared. MIC values are different in function of the various media tested Charcoal yeast extract medium, which is recommended for Legionella growth, does not inhibit activity of penicillin G, cefalotin, cefoxitin, amikacin, erythromycin and lincosamins, in contrast of tetracyclin, gentamicin, dibekacin, chloramphenicol, oleandomycin, pristinamycin, spiramycin and rifampicin. Charcoal, ferric pyrophosphate and L-cystein present in the culture medium are inactivating factors. However our results confirm good activity of rifampicin, erythromycin, cefoxitin, chloramphenicol, gentamicin and amikacin. Among the others macrolids tested, pristinamycin is the most active.

  9. Emergence of aminoglycoside 3-N-acetyltransferase IV in Escherichia coli and Salmonella typhimurium isolated from animals in France.

    PubMed Central

    Chaslus-Dancla, E; Martel, J L; Carlier, C; Lafont, J P; Courvalin, P

    1986-01-01

    We studied two outbreaks of calf salmonellosis caused by apramycin and gentamicin-resistant Salmonella typhimurium strains. In both cases, the responsible strains were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, tetracycline, and trimethoprim; one strain was also resistant to nalidixic acid in one outbreak. A systematic survey of the intestinal Escherichia coli strains of calves from the two affected flocks showed that 11 of 24 animals sampled were also colonized by apramycin- and gentamicin-resistant E. coli strains. These isolates belonged to four biotypes and were resistant to ampicillin, chloramphenicol, kanamycin, streptomycin, tetracycline, trimethoprim, and nalidixic acid. All of the strains were resistant to high levels of apramycin (MICs, 512 to 1,024 micrograms/ml) and to gentamicin (MICs, 8 to 32 micrograms/ml), and these resistances were always transferred en bloc. In S. typhimurium, this coresistance was borne by plasmids that were approximately 39 kilobases long (outbreak 1) or 90 kilobases long (outbreak 2), whereas in E. coli, the coresistance was due to plasmids that were approximately 110 kilobases long in both outbreaks. The two plasmids of Salmonella and four plasmids of E. coli encoded type IV aminoglycoside 3-N-acetyltransferases. The intensive use of curative and preventive treatments in calf production could be responsible for the emergence of enzymic resistance to apramycin and gentamicin. Images PMID:3521474

  10. Genes That Enhance the Ecological Fitness of Shewanella oneidensis MR-1 in Sediments Reveal the Value of Antibiotic Resistance▿ †

    PubMed Central

    Groh, Jennifer L.; Luo, Qingwei; Ballard, Jimmy D.; Krumholz, Lee R.

    2007-01-01

    Environmental bacteria persist in various habitats, yet little is known about the genes that contribute to growth and survival in their respective ecological niches. Signature-tagged mutagenesis (STM) of Shewanella oneidensis MR-1 coupled with a screen involving incubations of mutant strains in anoxic aquifer sediments allowed us to identify 47 genes that enhance fitness in sediments. Gene functions inferred from annotations provide us with insight into physiological and ecological processes that environmental bacteria use while growing in sediment ecosystems. Identification of the mexF gene and other potential membrane efflux components by STM demonstrated that homologues of multidrug resistance genes present in pathogens are required for sediment fitness of nonpathogenic bacteria. Further studies with a mexF deletion mutant demonstrated that the multidrug resistance pump encoded by mexF is required for resistance to antibiotics, including chloramphenicol and tetracycline. Chloramphenicol-adapted cultures exhibited mutations in the gene encoding a TetR family regulatory protein, indicating a role for this protein in regulating expression of the mexEF operon. The relative importance of mexF for sediment fitness suggests that antibiotic efflux may be a required process for bacteria living in sediment systems. PMID:17114320

  11. Clinical and Microbiological Aspects of Linezolid Resistance Mediated by the cfr Gene Encoding a 23S rRNA Methyltransferase▿

    PubMed Central

    Arias, Cesar A.; Vallejo, Martha; Reyes, Jinnethe; Panesso, Diana; Moreno, Jaime; Castañeda, Elizabeth; Villegas, Maria V.; Murray, Barbara E.; Quinn, John P.

    2008-01-01

    The cfr (chloramphenicol-florfenicol resistance) gene encodes a 23S rRNA methyltransferase that confers resistance to linezolid. Detection of linezolid resistance was evaluated in the first cfr-carrying human hospital isolate of linezolid and methicillin-resistant Staphylococcus aureus (designated MRSA CM-05) by dilution and diffusion methods (including Etest). The presence of cfr was investigated in isolates of staphylococci colonizing the patient's household contacts and clinical isolates recovered from patients in the same unit where MRSA CM-05 was isolated. Additionally, 68 chloramphenicol-resistant Colombian MRSA isolates recovered from hospitals between 2001 and 2004 were screened for the presence of the cfr gene. In addition to erm(B), the erm(A) gene was also detected in CM-05. The isolate belonged to sequence type 5 and carried staphylococcal chromosomal cassette mec type I. We were unable to detect the cfr gene in any of the human staphylococci screened (either clinical or colonizing isolates). Agar and broth dilution methods detected linezolid resistance in CM-05. However, the Etest and disk diffusion methods failed to detect resistance after 24 h of incubation. Oxazolidinone resistance mediated by the cfr gene is rare, and acquisition by a human isolate appears to be a recent event in Colombia. The detection of cfr-mediated linezolid resistance might be compromised by the use of the disk diffusion or Etest method. PMID:18174304

  12. Effects of dissolved organic matter from a eutrophic lake on the freely dissolved concentrations of emerging organic contaminants.

    PubMed

    Xiao, Yi-Hua; Huang, Qing-Hui; Vähätalo, Anssi V; Li, Fei-Peng; Chen, Ling

    2014-08-01

    The authors studied the effects of dissolved organic matter (DOM) on the bioavailability of bisphenol A (BPA) and chloramphenicol by measuring the freely dissolved concentrations of the contaminants in solutions containing DOM that had been isolated from a mesocosm in a eutrophic lake. The abundance and aromaticity of the chromophoric DOM increased over the 25-d mesocosm experiment. The BPA freely dissolved concentration was 72.3% lower and the chloramphenicol freely dissolved concentration was 56.2% lower using DOM collected on day 25 than using DOM collected on day 1 of the mesocosm experiment. The freely dissolved concentrations negatively correlated with the ultraviolent absorption coefficient at 254 nm and positively correlated with the spectral slope of chromophoric DOM, suggesting that the bioavailability of these emerging organic contaminants depends on the characteristics of the DOM present. The DOM-water partition coefficients (log KOC ) for the emerging organic contaminants positively correlated with the aromaticity of the DOM, measured as humic acid-like fluorescent components C1 (excitation/emission=250[313]/412 nm) and C2 (excitation/emission=268[379]/456 nm). The authors conclude that the bioavailability of emerging organic contaminants in eutrophic lakes can be affected by changes in the DOM.

  13. Role of Efflux Pumps and Topoisomerase Mutations in Fluoroquinolone Resistance in Campylobacter jejuni and Campylobacter coli

    PubMed Central

    Ge, Beilei; McDermott, Patrick F.; White, David G.; Meng, Jianghong

    2005-01-01

    Point mutations in the topoisomerase (DNA gyrase A) gene are known to be associated with fluoroquinolone resistance in Campylobacter. Recent studies have shown that an efflux pump encoded by cmeABC is also involved in decreased susceptibilities to fluoroquinolones, as well as other antimicrobials. Genome analysis suggests that Campylobacter jejuni contains at least nine other putative efflux pumps. Using insertional inactivation and site-directed mutagenesis, we investigated the potential contributions of these pumps to susceptibilities to chloramphenicol, ciprofloxacin, erythromycin, and tetracycline in C. jejuni and Campylobacter coli. Insertional inactivation of cmeB resulted in 4- to 256-fold decreases in the MICs of chloramphenicol, ciprofloxacin, erythromycin, and tetracycline, with erythromycin being the most significantly affected. In contrast, inactivation of all other putative efflux pumps had no effect on susceptibility to any of the four antimicrobials tested. Mutation of gyrA at codon 86 (Thr-Ile) caused 128- and 64-fold increases in the MICs of ciprofloxacin and nalidixic acid, respectively. The replacement of the mutated gyrA with a wild-type gyrA allele resulted in a 32-fold decrease in the ciprofloxacin MIC and no change in the nalidixic acid MIC. Our findings indicate that CmeABC is the only efflux pump among those tested that influences antimicrobial resistance in Campylobacter and that a point mutation (Thr-86-Ile) in gyrA directly causes fluoroquinolone resistance in Campylobacter. These two mechanisms work synergistically in acquiring and maintaining fluoroquinolone resistance in Campylobacter species. PMID:16048946

  14. Involvement of DNA-dependent RNA polymerase in a recA-independent pathway of genetic recombination in Escheria coli.

    PubMed

    Ikeda, H; Kobayashi, I

    1977-09-01

    Recombinant DNA molecule of phage lambda formed in Escherichia coli in the presence of chloramphenicol and/or rifampin can be assayed by their biological activity. recA- cells were found to be capable of forming recombinant lambda phage DNA in the presence of chloramphenicol. The relatively high recA-independent recombination observed in this system contrasts with the relatively low recA-independent recombination when recombinant phage particles rather than recombinant DNA are titrated. Formation of the recombinant DNA was suppressed by the the addition of rifampin. The introduction of the rif-r mutation into host bacteria made their recombination activity rifampin-resistant. These results show that DNA-dependent RNA polymerase (EC 2.7.7.6) is involved in this recA-independent pathway of recombination, which is named the "Rpo pathway." This is distinct from Red, Int, RecBC, RecE, or Der pathways of recombination. Crossover was much more frequent in the N-PL-cI and cI-PR-O regions than in the A-D and O-S regions. The crossover seems to occur in the regions that are transcribed actively. Some local change of DNA structure caused by transcription might be required for the Rpo pathway of recombination.

  15. Correlation between Infective Factors and Antibiotic Resistance in Enterococci Clinical Isolates in West of Iran

    PubMed Central

    Nasaj, Mona; Mousavi, Seyed Masoud

    2017-01-01

    The present study was done to scrutinize the possible relation between infective genes and antimicrobial resistance in Enterococcus faecalis and Enterococcus faecium. Considering the fact that the presence of recognized infective determinants among clinical isolates may promote the emergence of infections and persistence of Enterococci in hospital settings, which can lead to an increase in antimicrobial resistance. 175 E. faecalis and 67 E. faecium isolated from clinical specimens were used. The isolates were identified, and then antibiotic susceptibility testing was performed. The MIC of vancomycin and teicoplanin were determined by broth microdilution method. The presence of infective genes esp, hyl and asa1 was scrutinized using PCR. Of the 280 enterococcal isolates, 175 (62.5%) isolates were identified as E. faecalis, 67 (24%) as E. faecium and 38 (13.5%) as Enterococcus spp. The results of the antibiotic susceptibility testing showed resistance rates of 5% and 73% to vancomycin and teicoplanin in E. faecalis and E. faecium isolates, respectively. The statistical analysis showed that the esp infective gene has significant associations with ciprofloxacin, erythromycin and tetracycline in E. faecium and with chloramphenicol in E. faecalis strains; the hyl with teicoplanin and vancomycin in E. faecium strains; and also asa1 with vancomycin in E. faecium and with ampicillin and chloramphenicol in E. faecalis strains. Regarding the relationships between virulence genes and antibiotic resistance in strains of E. faecalis and E. faecium, detection of infective factors associated with invasive diseases has become a major issue of concern. PMID:28184339

  16. Isolation and characterization of the grain mold fungi Cochliobolus and Alternaria spp. from sorghum using semiselective media and DNA sequence analyses.

    PubMed

    Funnell-Harris, Deanna L; Prom, Louis K; Pedersen, Jeffrey F

    2013-02-01

    Mold diseases, caused by fungal complexes including Alternaria, Cochliobolus, and Fusarium species, limit sorghum grain production. Media were tested by plating Fusarium thapsinum, Alternaria sp., and Curvularia lunata, individually and competitively. Dichloran chloramphenicol rose bengal (DRBC) and modified V8 juice (ModV8) agars, found to be useful, were compared with commonly used agar media, dichloran chloramphenicol peptone (DCPA) and pentachloronitrobenzene (PCNB). Radial growth, starting with mycelia or single-conidia and hyphal tips, demonstrated an effect of media. For isolation of grain fungi, DRBC and ModV8 were similar or superior to DCPA and PCNB. When seedlings were inoculated with conidia of C. lunata, Alternaria sp., F. thapsinum, or mixtures, the percentage of root infection ranged from 28% to 77%. For mixed inoculations, shoot weights, lesion lengths, and percentage of root infections were similar to F. thapsinum inoculations; most colonies recovered from roots were F. thapsinum. For Alternaria grain isolates, 5 morphological types, including Alternaria alternata, were distinguished by colony morphologies and conidial dimensions. Sequence analysis using a portion of the endo-polygalacturonase gene was able to further distinguish isolates. Cochliobolus isolates were identified morphologically as C. lunata, Curvularia sorghina, and Bipolaris sorghicola. Multiple molecular genotypes were apparent from rRNA internal transcribed spacer region sequences from Cochliobolus grain isolates.

  17. Improved medium for antimicrobial susceptibility testing of Haemophilus influenzae.

    PubMed Central

    Jorgensen, J H; Redding, J S; Maher, L A; Howell, A W

    1987-01-01

    The need for complex growth media has complicated routine susceptibility testing of Haemophilus influenzae because of antagonism of certain antimicrobial agents by the medium or because of difficulties in interpretation of growth endpoints. Haemophilus test medium (HTM) is a simple, transparent medium for broth- or agar-based tests with H. influenzae. HTM incorporates Mueller-Hinton medium with additions of 15 micrograms of hematin per ml, 15 micrograms of NAD per ml, and 5 mg of yeast extract per ml as growth-promoting additives. Agar or broth microdilution MICs of 10 antimicrobial agents for a collection of 179 H. influenzae isolates determined by using HTM compared favorably with MICs determined by the conventional agar or broth dilution methods recommended by the National Committee for Clinical Laboratory Standards. Disk diffusion tests performed with HTM allowed accurate categorization of susceptible and resistant strains and were easier to interpret than tests performed with Mueller-Hinton chocolate agar. A particular advantage of HTM was the reliability of broth- or agar-based test results with trimethoprim-sulfamethoxazole. The results of the study suggest modification of current National Committee for Clinical Laboratory Standards MIC-interpretive criteria for H. influenzae with amoxicillin-clavulanate, chloramphenicol, and trimethoprim-sulfamethoxazole. Error rate-bounded analysis of MICs and disk diffusion zone sizes also suggest modified zone-interpretive criteria for ampicillin, amoxicillin-clavulanate, chloramphenicol, and tetracycline with HTM or conventional media. Interpretive zone sizes are newly proposed for cefaclor and rifampin disk diffusion tests. PMID:3500965

  18. Cloning and Analysis of the Alternative Oxidase Gene of Neurospora Crassa

    PubMed Central

    Li, Q.; Ritzel, R. G.; McLean, LLT.; McIntosh, L.; Ko, T.; Bertrand, H.; Nargang, F. E.

    1996-01-01

    Mitochondria of Neurospora crassa contain a cyanide-resistant alternative respiratory pathway in addition to the cytochrome pathway. The alternative oxidase is present only when electron flow through the cytochrome chain is restricted. Both genomic and cDNA copies for the alternative oxidase gene have been isolated and analyzed. The sequence of the predicted protein is homologous to that of other species. The mRNA for the alternative oxidase is scarce in wild-type cultures grown under normal conditions, but it is abundant in cultures grown in the presence of chloramphenicol, an inhibitor of mitochondrial protein synthesis, or in mutants deficient in mitochondrial cytochromes. Thus, induction of alternative oxidase appears to be at the transcriptional level. Restriction fragment length polymorphism mapping of the isolated gene demonstrated that it is located in a position corresponding to the aod-1 locus. Sequence analysis of mutant aod-1 alleles reveals mutations affecting the coding sequence of the alternative oxidase. The level of aod-1 mRNA in an aod-2 mutant strain that had been grown in the presence of chloramphenicol was reduced several fold relative to wild-type, supporting the hypothesis that the product of aod-2 is required for optimal expression of aod-1. PMID:8770590

  19. A prospective study on evaluation of pathogenesis, biofilm formation, antibiotic susceptibility of microbial community in urinary catheter

    NASA Astrophysics Data System (ADS)

    Younis, Khansa Mohammed; Usup, Gires; Ahmad, Asmat

    2015-09-01

    This study is aimed to isolate, detect biofilm formation ability and antibiotic susceptibility of urinary catheter adherent microorganisms from elderly hospitalized patient at the Universiti Kebangsaan Malaysia Medical Center. Microorganisms were isolated from three samples of urinary catheters (UC) surface; one of the acute vascular rejection patient (UCB) and two from benign prostate hyperplasia patients (UCC and UCD). A total of 100 isolates was isolated with 35 from UCB, 38 (UCC) and 28 (UCD). Ninety six were identified as Gram-negative bacilli, one Gram-positive bacilli and three yeasts. Results of biofilm forming on sterile foley catheter showed that all the isolates can form biofilm at different degrees; strong biofilm forming: 32% from the 35 isolates (UCB), 25% out of 38 isolates (UCC), 26% out of 28 isolates (UCD). As for moderate biofilm forming; 3% from UCB, 10% from UCC and 2% from UCD. Weak biofilm forming in UCC (3%). The antibiotic susceptibility for (UCB) isolates showed highly resistant to ampicillin, novobiocin and penicillin 100 (%), kanamycin (97%), tetracycline (94%), chloramphenicol (91%), streptomycin (77%) and showed low level of resistance to gentamycin (17%), while all the isolates from (UCC-D) showed high resistant towards ampicillin and penicillin, novobiocin (94%), tetracycline (61%), streptomycin (53%), gentamycin (50%) and low level of resistance to kanamycin (48%), chloramphenicol (47%). The findings indicate that these isolates can spread within the community on urinary catheters surface and produce strong biofilm, therefore, monitoring antibiotic susceptibility of bacteria isolated in the aggregation is recommended.

  20. Axenic culture of free-living conchocelis of Porphyra yezoensis and Porphyra haitanensis

    NASA Astrophysics Data System (ADS)

    Liu, Hui-Lian; Shuai, Li; Duan, De-Lin; Xu, Huai-Shu

    2002-03-01

    After discarding marine microorganisms from conchocelis of Porphyra yezoensis and Porphyra haitanensis, their axenic cultures were obtained through treatment with antibiotics. Antibiotic disc tests were carried out to determine the effectiveness of each antibiotic in eliminating contaminating microorganisms. Five of 12 antibiotics tested were selected and used to produce the axenic cultures in this study, which showed that 200 μg/mL streptomycin, 250 μg/mL penicillin, 252 μg/mL kanamycin, 30 μg/mL chloramphenicol were effective concentrations for eliminating microorganisms from conchocelis when antibiotics were added singly step by step; whereas simultaneous combination of 150 μg/mL streptomycin, 250 (or 350) μg/mL penicillin, 150 (or 250) μg/mL kanamycin, 70 μg/mL neomycin and 200 μg/mL chloramphenicol was also effective for producing the axenic cultures. However, it seemed that the treatments with antibiotics applied individually were more feasible than those will all antibiotics added at the same time. This may be due to the combined inhibiting effect of antibiotics on the growth and development of conchocelis.

  1. Novel bioassay using Bacillus megaterium to detect tetracycline in milk.

    PubMed

    Tumini, Melisa; Nagel, Orlando G; Molina, Pilar; Althaus, Rafael L

    2016-01-01

    Tetracyclines are used for the prevention and control of dairy cattle diseases. Residues of these drugs can be excreted into milk. Thus, the aim of this study was to develop a microbiological method using Bacillus megaterium to detect tetracyclines (chlortetracycline, oxytetracycline and tetracycline) in milk. In order to approximate the limits of detection of the bioassay to the Maximum Residue Limit (100μg/l) for milk tetracycline, different concentrations of chloramphenicol (0, 1000, 1500 and 2000μg/l) were tested. The detection limits calculated were similar to the Maximum Residue Limits when a bioassay using B. megaterium ATCC 9885 spores (2.8×10(8)spores/ml) and chloramphenicol (2000μg/l) was utilized. This bioassay detects 105μg/l of chlortetracycline, 100μg/l of oxytetracycline and 134μg/l of tetracycline in 5h. Therefore, this method is suitable to be incorporated into a microbiological multi-residue system for the identification of tetracyclines in milk.

  2. The simian virus 40 minimal origin and the 72-base-pair repeat are required simultaneously for efficient induction of late gene expression with large tumor antigen.

    PubMed

    Hartzell, S W; Byrne, B J; Subramanian, K N

    1984-10-01

    We have studied the temporal regulation of simian virus 40 (SV40) late gene expression by construction and transient expression analysis of plasmids containing the transposon Tn9 chloramphenicol acetyltransferase gene placed downstream from the late control region. The SV40 origin region in the early (but not the late) orientation promotes chloramphenicol acetyltransferase gene expression efficiently in monkey cells lacking large tumor (T) antigen. In monkey cells producing T antigen, the promoter activity of the late control region is induced by approximately 1,000-fold above the basal level. By deletion and point mutagenesis, we define two domains of the late control region required for efficient induction with T antigen. Domain I is the minimal replication origin containing T-antigen binding site II. Domain II consists of the 72-base-pair (bp) repeat and a 19-bp downstream sequence up to nucleotide 270. Domains I and II should act synergistically because the absence of either one or the other decreases induction efficiency by 2 orders of magnitude. Though a complete copy of domain II is optimal, the origin-proximal 22-bp portion of this domain is sufficient. The 21-bp repeat, located between domains I and II, is dispensable for this induction, as are sequences located downstream from nucleotide 270 in the late orientation.

  3. Modified artificial diet for rearing of tobacco budworm, Helicoverpa armigera, using the Taguchi method and Derringer's desirability function.

    PubMed

    Assemi, H; Rezapanah, M; Vafaei-Shoushtari, R; Mehrvar, A

    2012-01-01

    With the aim to improve the mass rearing feasibility of tobacco budworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), design of experimental methodology using Taguchi orthogonal array was applied. To do so, the effect of 16 ingredients of an artificial diet including bean, wheat germ powder, Nipagin, ascorbic acid, formaldehyde, oil, agar, distilled water, ascorbate, yeast, chloramphenicol, benomyl, penicillin, temperature, humidity, and container size on some biological characteristics of H. armigera was evaluated. The selected 16 factors were considered at two levels (32 experiments) in the experimental design. Among the selected factors, penicillin, container size, formaldehyde, chloramphenicol, wheat germ powder, and agar showed significant effect on the mass rearing performance. Derringer's desirability function was used for simultaneous optimization of mass rearing of tobacco budworm, H. armigera, on a modified artificial diet. Derived optimum operating conditions obtained by Derringer's desirability function and Taguchi methodology decreased larval period from 19 to 15.5 days (18.42 % improvement), decreased the pupal period from 12.29 to 11 days (10.49 % improvement), increased the longevity of adults from 14.51 to 21 days (44.72 % improvement), increased the number of eggs/female from 211.21 to 260, and increased egg hatchability from 54.2% to 72% (32.84 % improvement). The proposed method facilitated a systematic mathematical approach with a few well-defined experimental sets.

  4. Effect of mitochondrial inhibitors on metaphase-telophase progression and nuclear membrane formation in Chinese hamster cells.

    PubMed

    Chai, L S; Schumer, J M; Sandberg, A A

    1985-01-01

    Chinese hamster Don cells in log-phase were exposed to Colcemid during the G2 period with and without a combination of divalent cation chelators and mitochondrial inhibitors. Isolated metaphase cells were incubated as follows: (i) without Colcemid but with other agents and the progression was monitored from metaphase (M) to telophase (Tel) and to cell division; (ii) with Colcemid and other agents and the rate of micronuclei formation in the absence of anaphase was studied. Both EDTA and EGTA accelerated the progression from M to Tel, but did not affect the overall rate of cell division. Chloramphenicol (CAP), an inhibitor of mitochondrial protein synthesis, blocked the effect of the chelators and also retarded the progression. An inhibitor of mitochondrial respiration, Antimycin A (AA), also retarded the progression in the absence of the chelators and prevented the promoting effect of the chelators. A stimulator of ATPase for ATP breakdown. 2,4-dinitrophenol (DNP), accelerated the M to Tel progression. Chloramphenicol (CAP) and AA, as well as DNP, appeared to have little effect on the formation of micronuclei in the presence of Colcemid. EGTA, which affects cell surface Ca2+, stimulated the formation of micronuclei. This study indicates that Ca2+ ions and mitochondrial function are involved in the regulation of a certain segment of mitosis beyond metaphase, with Ca2+ sequestration in the mitochondria and chelation of Ca2+ by EGTA as dominant factors.

  5. Identification of regulatory sequences in the gene for 5-aminolevulinate synthase from rat.

    PubMed

    Braidotti, G; Borthwick, I A; May, B K

    1993-01-15

    The housekeeping enzyme 5-aminolevulinate synthase (ALAS) regulates the supply of heme for respiratory cytochromes. Here we report on the isolation of a genomic clone for the rat ALAS gene. The 5'-flanking region was fused to the chloramphenicol acetyltransferase gene and transient expression analysis revealed the presence of both positive and negative cis-acting sequences. Expression was substantially increased by the inclusion of the first intron located in the 5'-untranslated region. Sequence analysis of the promoter identified two elements at positions -59 and -88 bp with strong similarity to the binding site for nuclear respiratory factor 1 (NRF-1). Gel shift analysis revealed that both NRF-1 elements formed nucleoprotein complexes which could be abolished by an authentic NRF-1 oligomer. Mutagenesis of each NRF-1 motif in the ALAS promoter gave substantially lowered levels of chloramphenicol acetyltransferase expression, whereas mutagenesis of both NRF-1 motifs resulted in the almost complete loss of expression. These results establish that the NRF-1 motifs in the ALAS promoter are critical for promoter activity. NRF-1 binding sites have been identified in the promoters of several nuclear genes encoding mitochondrial proteins concerned with oxidative phosphorylation. The present studies suggest that NRF-1 may co-ordinate the supply of mitochondrial heme with the synthesis of respiratory cytochromes by regulating expression of ALAS. In erythroid cells, NRF-1 may be less important for controlling heme levels since an erythroid ALAS gene is strongly expressed and the promoter for this gene apparently lacks NRF-1 binding sites.

  6. Dairy farm age and resistance to antimicrobial agents in Escherichia coli isolated from dairy topsoil.

    PubMed

    Jones, Suzanna E; Burgos, Jonathan M; Lutnesky, Marvin M F; Sena, Johnny A; Kumar, Sanath; Jones, Lindsay M; Varela, Manuel F

    2011-04-01

    Antimicrobial agent usage is common in animal agriculture for therapeutic and prophylactic purposes. Selective pressure exerted by these antimicrobials on soil bacteria could result in the selection of strains that are resistant due to chromosomal- or plasmid-derived genetic components. Multiple antimicrobial resistances in Escherichia coli and the direct relationship between antimicrobial agent use over time has been extensively studied, yet the relationship between the age of an animal agriculture environment such as a dairy farm and antibiotic resistance remains unclear. Therefore, we tested the hypothesis that antimicrobial-resistance profiles of E. coli isolated from dairy farm topsoil correlate with dairy farm age. E. coli isolated from eleven dairy farms of varying ages within Roosevelt County, NM were used for MIC determinations to chloramphenicol, nalidixic acid, penicillin, tetracycline, ampicillin, amoxicillin/clavulanic acid, gentamicin, trimethoprim/sulfamethoxazole, cefotaxime, and ciprofloxacin. The minimum inhibitory concentration values of four antibiotics ranged 0.75 to >256 μg/ml, 1 to >256 μg/ml, 12 to >256 μg/ml, and 0.75 to >256 μg/ml for chloramphenicol, nalidixic acid, penicillin, and tetracycline, respectively. The study did not show a direct relationship between antibiotic resistance and the age of dairy farms.

  7. Distribution and antimicrobial resistance of enteric pathogens in Chinese paediatric diarrhoea: a multicentre retrospective study, 2008-2013.

    PubMed

    Zhang, H; Pan, F; Zhao, X; Wang, G; Tu, Y; Fu, S; Wang, J; Pan, J; Song, J; Wang, W; Jin, Z; Xu, H; Ren, Y; Li, Y; Zhong, N

    2015-09-01

    The enteric pathogens causing diarrhoea impair children's health severely. This study retrospectively analysed 1577 pathogens isolated from inpatients and outpatients in six hospitals located in Northern (Inner Mongolia), Northeastern (Hebei), Eastern (Shanghai and Jiangsu), Southern (Hainan) and Central (Hubei) China between 2008 and 2013. Of the 1577 enteric pathogens, Salmonella presented with the highest frequency (36·0%), followed by diarrhoeagenic Escherichia coli (23·7%), Staphylococcus aureus (15·0%), Shigella (13·1%), and Aeromonas (4·6%). The predominant pathogens varied in different regions of China, with Salmonella most prevalent in Shanghai and Hainan, diarrhoeagenic E. coli most prevalent in Inner Mongolia, Jiangsu and Hubei, and Shigella most prevalent in Hebei. Enteric pathogens were more frequently isolated in males (56·9%) than in females (43·1%). The highest proportion of all enteric pathogens was found in infants (67·6%) with a peak in summer and autumn (68·5%). Antimicrobial susceptibility assay demonstrated that Shigella was more resistant to ampicillin, ceftriaxone and sulfamethoxazole than Salmonella. Of the top two serotypes in Salmonella, Typhimurium was more resistant to ciprofloxacin, sulfamethoxazole and chloramphenicol than Enteritidis (P < 0·001). Meanwhile, the resistance rates of Shigella flexneri against ampicillin/sulbactam, ciprofloxacin, and chloramphenicol were significantly higher than those of Shigella sonnei (P < 0·001). Multidrug resistance was apparent in 58·2% of Shigella and 45·9% of Salmonella, and this phenomenon was more pronounced in S. flexneri.

  8. Decline in epidemic of multidrug resistant Salmonella typhi is not associated with increased incidence of antibiotic-susceptible strain in Bangladesh.

    PubMed Central

    Rahman, M.; Ahmad, A.; Shoma, S.

    2002-01-01

    Since 1987, multidrug resistant (MDR) strains of Salmonella Typhi, resistant simultaneously to ampicillin, chloramphenicol and trimethoprim-sulfamethoxazole, have caused epidemics of severe typhoid fever in Asia and Africa. A retrospective analysis of blood culture results (1989-96) in a Diarrhoea Treatment Centre in Dhaka, Bangladesh detected MDR strains in 0.3% (8 of 2793) of samples in 1990. The isolation rate peaked to 3.2% (240 of 7501) in 1994 (P < 0.01) and decreased to 1.8% (165 of 9348) in 1995 and further to 1.0% (82 of 8587) in 1996 (P < 0.01 compared to 1994) indicating the emergence and decline of MDR typhoid epidemic. Ten of 15 MDR strains tested had a 176 kb conjugative R plasmid that mediates resistance to ampicillin, chloramphenicol and trimethoprim-sulfamethoxazole to Escherichia coli K12. Unlike MDR strains, the isolation rate (approximately 3.3%) of susceptible S. Typhi remained remarkably unchanged during the study. The significant decrease in isolation of MDR strains suggests that cheaper and effective first-line antibiotics may re-emerge as drugs of choice for the treatment of typhoid fever in Bangladesh. PMID:12211593

  9. The antibiotic resistance patterns of Salmonella Typhi isolates in Italy, 1980-96. The Italian SALM-NET Working Group. Salmonella Network.

    PubMed Central

    Scuderi, G.; Fantasia, M.; Niglio, T.

    2000-01-01

    In this paper we report the distribution of Salmonella Typhi isolates in Italy and their resistance patterns to antibiotics. The data were collected by the Italian SALM-NET surveillance system in a pilot retrospective study of the period 1980-96. Data on drug-resistance were available for 82 isolates out of 176 S. Typhi isolated in Italy. Of these 82 isolates, 32 (39%) were resistant or intermediate to 1 or more antibiotics. Eight isolates were resistant and 7 intermediate to streptomycin; 4 isolates were resistant to ampicillin alone or in association with other antibiotics; only 2 strains (1 isolated in Lombardia in 1993 and the other 1 in Lazio in 1994) were resistant to chloramphenicol, and 2 (isolated in Sardegna and Piemonte in 1995 and 1996, respectively) showed intermediate resistance to chloramphenicol. The strains showing resistance to 3 or more antibiotics were very scarce: 1 (with 5 complete resistances) was isolated in Lazio in 1994, and another 1 (with complete resistance to 10 antibiotics and intermediate resistance to 2 antibiotics) was isolated in Molise in 1988. In conclusion, besides the routine activities to control typhoid fever, an accurate and continuous surveillance is necessary in order to quickly identify multidrug-resistant (MDR) S. Typhi strains and prevent their spread, even though their level, in our country, is still quite low. PMID:10722125

  10. The upstream muscle-specific enhancer of the rat muscle creatine kinase gene is composed of multiple elements.

    PubMed Central

    Horlick, R A; Benfield, P A

    1989-01-01

    A series of constructs that links the rat muscle creatine kinase promoter to the bacterial chloramphenicol acetyltransferase gene was generated. These constructs were introduced into differentiating mouse C2C12 myogenic cells to localize sequences that are important for up-regulation of the creatine kinase gene during myogenic differentiation. A muscle-specific enhancer element responsible for induction of chloramphenicol acetyltransferase expression during myogenesis was localized to a 159-base-pair region from 1,031 to 1,190 base pairs upstream of the transcription start site. Analysis of transient expression experiments using promoters mutated by deletion indicated the presence of multiple functional domains within this muscle-specific regulatory element. A DNA fragment spanning this region was used in DNase I protection experiments. Nuclear extracts derived from C2 myotubes protected three regions (designated E1, E2, and E3) on this fragment from digestion, which indicated there may be three or more trans-acting factors that interact with the creatine kinase muscle enhancer. Gel retardation assays revealed that factors able to bind specifically to E1, E2, and E3 are present in a wide variety of tissues and cell types. Transient expression assays demonstrated that elements in regions E1 and E3, but not necessarily E2, are required for full enhancer activity. Images PMID:2761536

  11. JadX is a Disparate Natural Product Binding Protein.

    PubMed

    Robertson, Andrew W; Forget, Stephanie M; Martinez-Farina, Camilo F; McCormick, Nicole E; Syvitski, Raymond T; Jakeman, David L

    2016-02-24

    We report that JadX, a protein of previously undetermined function coded for in the jadomycin biosynthetic gene cluster of Streptomyces venezuelae ISP5230, affects both chloramphenicol and jadomycin production levels in blocked mutants. Characterization of recombinant JadX through protein-ligand interactions by chemical shift perturbation and WaterLOGSY NMR spectroscopy resulted in the observation of binding between JadX and a series of jadomycins and between JadX and chloramphenicol, another natural product produced by S. venezuelae ISP5230. These results suggest JadX to be an unusual class of natural product binding protein involved in binding structurally disparate natural products. The ability for JadX to bind two different natural products in vitro and the ability to affect production of these secondary metabolites in vivo suggest a potential role in regulation or signaling. This is the first example of functional characterization of these JadX-like proteins, and provides insight into a previously unobserved regulatory process.

  12. Detection of Prohibited Fish Drugs Using Silver Nanowires as Substrate for Surface-Enhanced Raman Scattering

    PubMed Central

    Song, Jia; Huang, Yiqun; Fan, Yuxia; Zhao, Zhihui; Yu, Wansong; Rasco, Barbara A.; Lai, Keqiang

    2016-01-01

    Surface-enhanced Raman scattering or surface-enhanced Raman spectroscopy (SERS) is a promising detection technology, and has captured increasing attention. Silver nanowires were synthesized using a rapid polyol method and optimized through adjustment of the molar ratio of poly(vinyl pyrrolidone) and silver nitrate in a glycerol system. Ultraviolet-visible spectrometry, X-ray diffraction, and transmission electron microscopy were used to characterize the silver nanowires. The optimal silver nanowires were used as a SERS substrate to detect prohibited fish drugs, including malachite green, crystal violet, furazolidone, and chloramphenicol. The SERS spectra of crystal violet could be clearly identified at concentrations as low as 0.01 ng/mL. The minimum detectable concentration for malachite green was 0.05 ng/mL, and for both furazolidone and chloramphenicol were 0.1 μg/mL. The results showed that the as-prepared Ag nanowires SERS substrate exhibits high sensitivity and activity. PMID:28335303

  13. Performance of selective and differential media in the primary isolation of yeasts from different biological samples.

    PubMed

    Silva, Jaqueline Otero; Franceschini, Silvio Antônio; Lavrador, Marco Aurélio Sicchiroli; Candido, Regina Célia

    2004-01-01

    In view of the increase in yeast infections, especially polymicrobial ones, differential culture media have acquired increasing importance. The present study evaluated the Sabouraud chloramphenicol, Biggy agar, Pagano Levin agar and CHROMagar Candida media in terms of isolation, number of yeast colony forming units per plate, and inhibition of bacteria and filamentous fungi. To this end, we used 223 biological samples, including feces, and oral, vaginal and anal mucosae from 86 patients presenting or not symptoms of fungal infections. The four media did not differ significantly in terms of detection of yeast-positive cultures. The number of colony forming units per plate ranged from zero to 2.380, with a predominance of counts of 1 to 9 colonies per plate. No significant differences were observed among the four culture media in terms of number of colonies counted, for each kind of biological material. Fifteen species belonging to the genera Candida, Saccharomyces, Cryptococcus, Trichosporon and Rhodotorula were isolated, with C. albicans being the predominant species, followed by C. parapsilosis and R. rubra. CHROMagar Candida and Biggy agar were complementary in the isolation of the different species and favored a greater recovery of polymicrobial cultures. Pagano Levin agar isolated the smallest variety of species. Sabouraud chloramphenicol agar was the least effective in terms of bacterial inhibition and favored a greater development of filamentous fungi. The results suggest that more than one culture medium should be used for an adequate primary isolation.

  14. Antibiotic resistance in Staphylococci isolated from the vaginas of captive female Leontopithecus (Callitrichidae-Primates).

    PubMed

    Lilenbaum, Walter; Moraes, Ismar A; Cardoso, Verônica S; Varges, Renato G; Ferreira, Ana Maria R; Pissinatti, Alcides

    2006-08-01

    The purpose of this study was to provide current data on Staphylococcus species from the vaginas of clinically normal captive lion tamarins and to determine the antimicrobial susceptibility of these isolates. Samples were collected from 25 adult lion tamarins, processed to isolate Staphylococcus species, and tested for susceptibility to penicillin G, gentamicin, chloramphenicol, tetracycline, trimethoprim-sulfamethoxazole, streptomycin, ampicillin, and rifampicin. Isolates with the typical characteristics of the genus Staphylococcus were recovered from all 25 samples. Coagulase-negative species were the most common (68% of the isolates), and the most frequently isolated species (10 samples) was S. simulans. Other coagulase-negative species, including S. saprophyticus (n=5), S. epidermidis (n=1), and S. arlettae (n=1), were also recovered. Coagulase-positive Staphylococci were obtained from eight animals (six of from the S. aureus species and two from S. intermedius). Resistance to antibiotics was frequently observed, and 88% of the isolates (23 samples) showed resistance to at least one drug. Resistance to penicillin G was a common finding, and the most active antimicrobial agents were chloramphenicol and gentamicin. Coagulase-positive strains were more frequently resistant to antibiotics (79.7%, average=6.4 drugs) than coagulase-negative strains (38.2%, average=3.0 drugs). The high frequency of resistance observed in those isolates is surprising and very alarming. A detailed history of the use of antimicrobial drugs in these subjects did not reveal any previous exposure to any of the tested antibiotics that could justify the observed resistance rate.

  15. Evaluation of Albicans ID2 and Biggy agar for the isolation and direct identification of vaginal yeast isolates.

    PubMed

    Ilkit, Macit; Hilmioglu, Suleyha; Tasbakan, Meltem; Aydemir, Sohret

    2007-06-01

    In this study, 250 vaginal samples from patients with vulvovaginal candidosis were inoculated onto two chromogenic media, Albicans ID(2) and Biggy agar, as well as onto Sabouraud chloramphenicol agar, yielding a total of 63 yeast (25.2 %) on all three media. These strains were identified as Candida glabrata in 20 (31.8 %) samples, Candida albicans in 15 samples (23.8 %), Candida tropicalis in 10 samples (15.9 %), Candida krusei in five samples (7.9 %), Candida kefyr in five samples (7.9 %), Candida dubliniensis in four samples (6.3 %), Candida parapsilosis in two samples (3.2 %) and Candida guilliermondii in two samples (3.2 %). Mixed fungal cultures and bacterial growth or filamentous fungi were not detected on any of the selected media. The sensitivity and specificity of the Albicans ID(2) and Biggy agar with regard to the identification of C. albicans were 80.0 and 64.6 %, and 86.7 and 56.3 %, respectively. This study showed these two chromogenic media to be as effective as Sabouraud chloramphenicol agar with respect to fungal detection. However, neither Albicans ID(2) nor Biggy agar was sufficient for reliable differentiation of yeasts to the species level.

  16. Study correlating endometrial biopsies and vaginal cytology in one hundred tubectomized women.

    PubMed

    Mehta, P

    1975-01-01

    Menstrual irregularities are believed to often follow the tubectomy operation, possibly because the ovarian blood supply is altered and horm onal imbalance results. This study was designed to determine, with the help of hormonal colpocytology and endometrial histology, whether hormon al imbalance occurs after tubectomy. Subjects were 100 women attending the tubectomy follow-up clinic. All had been tubectomized 2 or more years earlier. On the 5th and 6th days of their menstrual cycles, a group was treated with an antibiotic vaginal pessary consisting of .25 gm of Chloramphenicol. From the 8th to the 24th day, serial vaginal sme ars were taken. The maturation index (MI), maturation value (MV), and k aryopyknotic index (KI) were evaluated. Endometrial biopsies were taken on Day 24 of the menstrual cycles. Of the 100 women, 98 had ovulatroy c ycles as shown by endometrial biopsy. The 2 having anovulatory cycles w ere in the treated group. Their MVs and KIs were low. None showed evid ence of hyperestrogenism either in their colpocytogram or endometrial hi stology. Disturbed ovarian function was not shown. Prior anitbiotic tr eatment is essential for hormonal cytology, as in the majority of untrea ted women the presence of Doderlein bacilli, debris, and leukocytes in the follicular phase renders preparations unsatisfactory for hormonal ev aluation. Chloramphenicol has no direct effect on the vaginal epitheliu m but tetracyclines cause massive desquamation.

  17. Subinhibitory Concentrations of Bacteriostatic Antibiotics Induce relA-Dependent and relA-Independent Tolerance to β-Lactams

    PubMed Central

    Kudrin, Pavel; Varik, Vallo; Oliveira, Sofia Raquel Alves; Beljantseva, Jelena; Del Peso Santos, Teresa; Dzhygyr, Ievgen; Rejman, Dominik; Cava, Felipe; Tenson, Tanel

    2017-01-01

    ABSTRACT The nucleotide (p)ppGpp is a key regulator of bacterial metabolism, growth, stress tolerance, and virulence. During amino acid starvation, the Escherichia coli (p)ppGpp synthetase RelA is activated by deacylated tRNA in the ribosomal A-site. An increase in (p)ppGpp is believed to drive the formation of antibiotic-tolerant persister cells, prompting the development of strategies to inhibit (p)ppGpp synthesis. We show that in a biochemical system from purified E. coli components, the antibiotic thiostrepton efficiently inhibits RelA activation by the A-site tRNA. In bacterial cultures, the ribosomal inhibitors thiostrepton, chloramphenicol, and tetracycline all efficiently abolish accumulation of (p)ppGpp induced by the Ile-tRNA synthetase inhibitor mupirocin. This abolishment, however, does not reduce the persister level. In contrast, the combination of dihydrofolate reductase inhibitor trimethoprim with mupirocin, tetracycline, or chloramphenicol leads to ampicillin tolerance. The effect is independent of RelA functionality, specific to β-lactams, and not observed with the fluoroquinolone norfloxacin. These results refine our understanding of (p)ppGpp's role in antibiotic tolerance and persistence and demonstrate unexpected drug interactions that lead to tolerance to bactericidal antibiotics. PMID:28115345

  18. In vivo analysis of the murine beta-myosin heavy chain gene promoter.

    PubMed

    Rindt, H; Gulick, J; Knotts, S; Neumann, J; Robbins, J

    1993-03-05

    The 5' upstream region of the murine beta-myosin heavy chain (MHC) gene has been isolated and tested for its ability to drive gene expression in transgenic mice. Three classes of transgenic mice were generated. The constructs contained approximately 5000, 2500, and 600 base pairs of beta-MHC upstream sequence fused to the chloramphenicol acetyltransferase gene and were termed beta 5, beta 2.5, and beta .6, respectively. Muscle-specific expression was observed with all three constructs. However, only the beta 5 lines directed high levels of muscle-specific transgene expression in both pre- and postbirth mice. Expression driven by the two shorter constructs was two to three orders of magnitude lower when the chloramphenicol acetyltransferase specific activities were compared. These data suggest that a distal-positive element directs high levels of gene expression in the ventricle and in slow skeletal muscles. Analyses of transgene expression during heart maturation revealed that some of the beta 5 lines were not able to respond in an appropriate manner to developmental transcriptional cues. Unlike the endogenous beta-MHC gene, which is down regulated in the ventricles around the time of birth, reporter gene expression in the majority of the lines generated was not shut off in the ventricles of the adult animals. These data indicate that high levels of muscle-specific beta-MHC gene expression are dependent upon the combinatorial interactions of a number of sequence elements that are distributed over a large region of the gene's upstream sequence.

  19. Methicillin-Resistant Staphylococcus aureus Keratitis after Descemet's Stripping Automated Endothelial Keratoplasty

    PubMed Central

    Miyamoto, Tatsuro; Eguchi, Hiroshi; Tserennadmid, Ehkhmaa; Mitamura-Aizawa, Sayaka; Hotta, Fumika; Mitamura, Yoshinori

    2013-01-01

    Purpose We report a case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis after Descemet's stripping automated endothelial keratoplasty (DSAEK). Case Report An 87-year-old woman who had undergone a DSAEK 4 months previously was referred to Tokushima University Hospital with a diagnosis of infectious keratitis after DSAEK. A white abscess and infiltration in the inferior cornea of the right eye were observed. We started an empiric therapy using topical levofloxacin and chloramphenicol on the basis of the microscopic findings of the corneal scraping concurrently with cultivation of the cornea. Results A strain of MRSA was isolated from the corneal sample. Although the strain was susceptible to chloramphenicol, it was resistant to quinolone. The keratitis improved rapidly due to empiric therapy, and topical steroids could be resumed 6 days after initiation of the empiric therapy. Conclusions To our knowledge, this is the first case of MRSA keratitis, and the second case of bacterial keratitis, after DSAEK. MRSA keratitis can occur following uneventful DSAEK. The empiric therapy on the basis of results from a light microscopic examination of a Gram-stained corneal scraping and restarting topical steroids in the early stages of medication contributed to the good clinical course of this case. PMID:24348415

  20. [Inactivation and reactivation of antibiotic-resistant bacteria during and after UV disinfection in reclaimed water].

    PubMed

    Huang, Jing-Jing; Tang, Fang; Xi, Jin-Ying; Pang, Yu-Chen; Hu, Hong-Ying

    2014-04-01

    Prevalence of antibiotic-resistant bacteria in wastewater effluents is concerned as an emerging contaminant. To estimate inactivation and reactivation potentials of antibiotic-resistant bacteria by UV disinfection, inactivation and reactivation of penicillin-, ampicillin-, cephalexin-, chloramphenicol-and rifampicin-resistant bacteria in the secondary effluent were studied under different UV doses. The results showed that the inactivation ratios of penicillin-, ampicillin-, cephalexin-and chloramphenicol-resistant bacteria were higher than 4-log, which was closed to that of total heterotrophic bacteria; however, the inactivation ratio of rifampicin-resistant bacteria was lower (3.7-log) under 20 mJ x cm(-2) UV exposure. After 22 h standing incubation, antibiotic-resistant bacteria widely reactivated. The colony forming ability of antibiotic-resistant bacteria was as high as 3-log when exposed to 20 mJ x cm(-2) UV light. Hence, conventional UV dose can not effectively control reactivation of antibiotic-resistant bacteria in reclaimed water by UV disinfection.

  1. Microbial selectivity of UV treatment on antibiotic-resistant heterotrophic bacteria in secondary effluents of a municipal wastewater treatment plant.

    PubMed

    Guo, Mei-Ting; Yuan, Qing-Bin; Yang, Jian

    2013-10-15

    Little is known about the microbial selectivity of UV treatment for antibiotic resistant bacteria, and the results of limited studies are conflicting. To understand the effect of UV disinfection on antibiotic resistant bacteria, both total heterotrophic bacteria and antibiotic resistant bacteria (including cephalexin-, ciprofloxacin-, erythromycin-, gentamicin-, vancomycin-, sulfadiazine-, rifampicin-, tetracycline- and chloramphenicol-resistant bacteria) were examined in secondary effluent samples from a municipal wastewater treatment plant. Bacteria resistant to both erythromycin and tetracycline were chosen as the representative of multiple-antibiotic-resistant bacteria and their characteristics after UV treatment were also investigated. UV disinfection results in effective inactivation for total heterotrophic bacteria, as well as all antibiotic resistant bacteria. After UV treatment at a fluence of 5 mJ/cm(2), the log reductions of nine types of antibiotic resistant bacteria varied from 1.0 ± 0.1 to 2.4 ± 0.1. Bacteria resistant to both erythromycin and tetracycline had a similar fluence response as did total heterotrophic bacteria. The findings suggest that UV disinfection could eliminate antibiotic resistance in wastewater treatment effluents and thus ensure public health security. Our experimental results indicated that UV disinfection led to enrichment of bacteria with resistance to sulfadiazine, vancomycin, rifampicin, tetracycline and chloramphenicol, while the proportions of cephalexin-, erythromycin-, gentamicin- and ciprofloxacin-resistant bacteria in the wastewater decreased. This reveals the microbial selectivity of UV disinfection for antibiotic resistant bacteria.

  2. Current Status of Antibiograms of Listeria ivanovii and Enterobacter cloacae Isolated from Ready-To-Eat Foods in Alice, South Africa

    PubMed Central

    Nyenje, Mirriam E.; Tanih, Nicoline F.; Green, Ezekiel; Ndip, Roland N.

    2012-01-01

    This study assessed the antimicrobial susceptibility of 51 Listeria ivanovii and 33 Enterobacter cloacae strains isolated from various ready-to-eat foods sold in Alice, South Africa. Isolates were identified using standard microbiological tests and further confirmed using API 20E and API Listeria kits. The disc diffusion technique was used to screen for antimicrobial susceptibility against 15 antimicrobials; minimum inhibitory concentration of five antibiotics was determined by the broth dilution method. All the strains of E. cloacae (100%) and 96% of L. ivanovii isolates were resistant to at least four or more of the antibiotics; nineteen antibiotypes were obtained based on the antibiotics used in the study. Antibiotype A5: AR PGR VAR ER APR was predominant in both L. ivanovii (23.5%) and E. cloacae (57.5%) isolates. Marked susceptibility of Listeriaivanovii was observed against chloramphenicol, ciprofloxacin, streptomycin and trimethoprim/sulfamethoxazole (100%) each while E. cloacae registered 100% susceptibility to ciprofloxacin only. Various percentages of susceptibility was reported to chloramphenicol and gentamicin (91%) each, nalidixic acid (97%) and streptomycin (94%). The MIC90 ranged from 0.004–7.5 µg/mL with E. cloacae being the most susceptible organism. The study demonstrated the presence of multi-resistant strains of bacteria in ready-to-eat-foods and speculates that these foods could serve as important vehicles transmitting multi-resistant bacteria to humans. PMID:23202673

  3. Sorption and degradation of wastewater-associated pharmaceuticals and personal care products in agricultural soils and sediment.

    PubMed

    Zhang, Ting; Wu, Bo; Sun, Na; Ye, Yong; Chen, Huaixia

    2013-01-01

    Pharmaceuticals and personal care products (PPCPs) have drawn popular concerns recently as an emerging class of aquatic contaminants. In this study, adsorption and degradation of four selected PPCPs, metronidazole, tinidazole, caffeine and chloramphenicol, have been investigated in the laboratory using two agricultural soils in China and sediment from Changjiang River. Adsorption tests using a batch equilibrium method demonstrated that adsorption of all tested chemicals in soils could be well described with Freundlich equation, and their adsorption affinity on soil followed the order of chloramphenicol > caffeine > tinidazole > metronidazole. Generally, higher Kf value was associated with soils which had higher organic matter contents (except for caffeine acid in this study). Degradation of selected PPCPs in soils generally followed first-order exponential decay kinetics, and half-lives ranging from 0.97 to 10.21 d. Sterilization generally decreased the degradation rates, indicating that microbial activity played a significant role in the degradation in soils. The degradation rate constant decreased with increasing initial chemical concentrations in soil, implying that the microbial activity was inhibited with high chemical loading levels.

  4. Prevalence of antimicrobial resistance in faecal enterococci from vet-visiting pets and assessment of risk factors.

    PubMed

    Leite-Martins, L; Mahú, M I; Costa, A L; Bessa, L J; Vaz-Pires, P; Loureiro, L; Niza-Ribeiro, J; de Matos, A J F; Martins da Costa, P

    2015-06-27

    The objective of this study was to determine the prevalence of antimicrobial resistance (AMR) exhibited by enterococci isolated from faeces of pets and its underlying risk factors. From September 2009 to May 2012, rectal swabs were collected from 74 dogs and 17 cats, selected from the population of animals visiting the Veterinary Hospital of University of Porto, UPVet, through a systematic random procedure. Animal owners answered a questionnaire about the risk factors that could influence the presence of AMR in faecal enterococci. Enterococci isolation, identification and antimicrobial (AM) susceptibility testing were performed. Data analyses of multilevel, univariable and multivariable generalised linear mixed models were conducted. From all enterococci isolated (n=315), 61 per cent were considered multidrug-resistant, whereas only 9.2 per cent were susceptible to all AMs tested. Highest resistance was found to tetracycline (67.0 per cent), rifampicin (60.3 per cent), azithromycin (58.4 per cent), quinupristin/dalfopristin (54.0 per cent) and erythromycin (53.0 per cent). Previous fluoroquinolone treatments and coprophagic habits were the features more consistently associated with the presence of AMR for three (chloramphenicol, ciprofloxacin and azithromycin) and seven (tetracycline, rifampicin, gentamicin, chloramphenicol, ciprofloxacin, erythromycin and azithromycin), respectively, out of nine AMs assessed. Evaluating risk factors that determine the presence of drug-resistant bacteria in pets, a possible source of resistance determinants to human beings, is crucial for the selection of appropriate treatment guidelines by veterinary practitioners.

  5. Abusive use of antibiotics in poultry farming in Cameroon and the public health implications.

    PubMed

    Guetiya Wadoum, R E; Zambou, N F; Anyangwe, F F; Njimou, J R; Coman, M M; Verdenelli, M C; Cecchini, C; Silvi, S; Orpianesi, C; Cresci, A; Colizzi, V

    2016-08-01

    The types and methods of use of antibiotics in poultry farms in Cameroon, residual levels and potential microbial resistance were determined. A questionnaire-based survey identified the different antibiotics used and high-performance liquid chromatography (HPLC) was used to determine residual levels of antibiotics. Pathogens were isolated, identified by use of commercial API kits and minimum inhibition concentration (MIC) was determined. Oxytetracyclin, tylocip and TCN (oxytetracycline, chloramphenicol and neomycin) were the most frequently used antibiotics. Antibiotics screened by HPLC were chloramphenicol, tetracycline and vancomycin. All of them except vancomycin were detected, and the concentration of these antibiotics was higher than the maximum residual limits (MRL) set by regulatory authorities. No residues of various antibiotics were found in egg albumen or yolk. The concentration of tetracycline was significantly higher in liver (150 ± 30 µg/g) than in other tissues. Foodborne pathogens, including Salmonella spp., Staphylococcus spp., Listeria spp., Clostridium spp. and Escherichia spp., were identified. Most of the pathogens were resistant to these various antibiotics tested. These findings imply the need for better management of antibiotic use to control sources of food contamination and reduce health risks associated with the presence of residues and the development of resistant pathogens by further legislation and enforcement of regulations on food hygiene and use of antibiotics.

  6. Constitutive nitrate reductase expression and inhibition in winged bean

    SciTech Connect

    Wu, Shenchuan; Harper, J.E. )

    1990-05-01

    It was found that NO{sub 3}{sup {minus}} had no effect on winged bean nitrate reductase activity (NRA). Similar NRA was expressed in plants grown on NO{sub 3}{sup {minus}}, urea, NH{sub 4}{sup +}, and nil N. This indicated that the primary NR expressed in winged bean was constitutive, rather than substrate-inducible. Maximum NRA in winged bean was obtained in the light. KClO{sub 3} was capable of inhibiting NRA of leaves if added to the root growth medium or to the NR assay medium, indicating possible competition with NO{sub 3}{sup {minus}} at the reduction site. While it has previously been shown that either cycloheximide alone, or both cycloheximide and chloramphenicol impair the synthesis of NR protein, our data unexpectedly demonstrated that cycloheximide had little effect on NRA, whereas chloramphenicol greatly inhibited the expression of NRA in winged bean. One interpretation is that chloroplasts may influence the activity and/or synthesis of constitutive NR proteins.

  7. Integrable alpha-amylase plasmid for generating random transcriptional fusions in Bacillus subtilis.

    PubMed Central

    O'Kane, C; Stephens, M A; McConnell, D

    1986-01-01

    An integrable plasmid, pOK4, which replicated independently in Escherichia coli was constructed for generating transcriptional fusions in vivo in Bacillus DNA. It did not replicate independently in Bacillus subtilis, but it could be made to integrate into the chromosome of B. subtilis if sequences homologous to chromosomal sequences were inserted into it. It had a selectable marker for chloramphenicol resistance and carried unique sites for EcoRI and SmaI just to the 5' side of a promoterless alpha-amylase gene from Bacillus licheniformis. When B. subtilis DNA fragments were ligated into one of these sites and the ligation mixture was used to transform an alpha-amylase-negative B. subtilis strain, chloramphenicol-resistant transformants could be isolated conveniently. Many of these were alpha-amylase positive, owing to the fusion of the plasmid amylase gene to chromosomal operons. In principle, because integration need not be mutagenic, it is possible to obtain fusions to any chromosomal operon. The site of each integration can be mapped, and the flanking sequences can be cloned into E. coli. The alpha-amylase gene can be used to detect regulated genes. We used it as an indicator to detect operons which are DNA-damage-inducible (din), and we identified insertions in both SP beta and PBSX prophages. Images PMID:3096966

  8. Residues of antibacterial drugs in honey from the Italian market.

    PubMed

    Baggio, A; Gallina, A; Benetti, C; Mutinelli, F

    2009-01-01

    Antibacterial drugs are used worldwide for the control of American and, less often, European foulbrood. Their administration is mostly uncontrolled and applied without approved protocols and instructions for use as well as precautionary recommendations. Consequently, this practice is responsible for the contamination of beehive products and contributes to the problem of food safety. According to this situation, 4672 analyses were carried out on 5303 honeys collected from 2001 to 2007. These samples were investigated for antibacterial residues of tetracyclines, sulphonamides, streptomycin, chloramphenicol and tylosin. Honeys were classified according to their origin: imported honey and honey from the Italian market. In the last group (only for samples collected from 2001 to 2004), another type of honey was distinguished: that of local honey. A total of 6.3% of all samples were positive for the antibacterial drugs analysed; in particular, 6.8% of imported honeys and 6.1% of honeys on the Italian market. Only 1.7% of local honey had antibacterial residues. These results are indicative of a rather frequent presence of antibacterial drug residues in both Italian and imported honeys. Furthermore, the data showed that among the active substances analysed, sulphonamides are the most used antibacterial substance followed by tetracyclines, streptomycin, tylosin, and chloramphenicol. Finally, a continuous monitoring programme is needed, accompanied by an education programme to beekeepers on proper hive management.

  9. Safety of Lactobacillus plantarum ST8Sh and Its Bacteriocin.

    PubMed

    Todorov, Svetoslav Dimitrov; Perin, Luana M; Carneiro, Bruno M; Rahal, Paula; Holzapfel, Wilhelm; Nero, Luís Augusto

    2017-02-23

    Total DNA extracted from Lb. plantarum ST8Sh was screened for the presence of more than 50 genes related to production of biogenic amines (histidine decarboxylase, tyrosine decarboxylase, and ornithine decarboxylase), virulence factors (sex pheromones, gelatinase, cytolysin, hyaluronidase, aggregation substance, enterococcal surface protein, endocarditis antigen, adhesion of collagen, integration factors), and antibiotic resistance (vancomycin, tetracycline, erythromycin, gentamicin, chloramphenicol, bacitracin). Lb. plantarum ST8Sh showed a low presence of virulence genes. Only 13 genes were detected (related to sex pheromones, aggregation substance, adhesion of collagen, tetracycline, gentamicin, chloramphenicol, erythromycin, but not to vancomycin, and bacitracin) and may be considered as indication of safety for application in fermented food products. In addition, interaction between Lb. plantarum ST8Sh and drugs from different groups were determined in order to establish possible application of the strain in combination with commercial drugs. Cytotoxicity of the semi-purified bacteriocins produced by Lb. plantarum ST8Sh was depended on applied concentration-highly cytotoxic when applied at 25 μg/mL and no cytotoxicity at 5 μg/mL.

  10. Outer Membrane Proteins form Specific Patterns in Antibiotic-Resistant Edwardsiella tarda.

    PubMed

    Peng, Bo; Wang, Chao; Li, Hui; Su, Yu-Bin; Ye, Jin-Zhou; Yang, Man-Jun; Jiang, Ming; Peng, Xuan-Xian

    2017-01-01

    Outer membrane proteins of Gram-negative bacteria play key roles in antibiotic resistance. However, it is unknown whether outer membrane proteins that respond to antibiotics behave in a specific manner. The present study specifically investigated the differentially expressed outer membrane proteins of an antibiotic-resistant bacterium, Edwardsiella tarda, a Gram-negative pathogen that can lead to unnecessary mass medication of antimicrobials and consequently resistance development in aquaculture and a spectrum of intestinal and extraintestinal diseases in humans. The comparison of a clinically isolated strain to the laboratory derived kanamycin-, tetracycline-, or chloramphenicol-resistant strains identified their respective outer membrane proteins expression patterns, which are distinct to each other. Similarly, the same approach was utilized to profile the patterns in double antibiotic-resistant bacteria. Surprisingly, one pattern is always dominant over the other as to these three antibiotics; the pattern of chloramphenicol is over tetracycline, which is over kanamycin. This type of pattern was also confirmed in clinically relevant multidrug-resistant bacteria. In addition, the presence of plasmid encoding antibiotic-resistant genes also alters the outer membrane protein profile in a similar manner. Our results demonstrate that bacteria adapt the antibiotic stress through the regulation of outer membrane proteins expression. And more importantly, different outer membrane protein profiles were required to cope with different antibiotics. This type of specific pattern provides the rationale for the development of novel strategy to design outer membrane protein arrays to identify diverse multidrug resistance profiles as biomarkers for clinical medication.

  11. Pneumococcal resistance to antibiotics.

    PubMed Central

    Klugman, K P

    1990-01-01

    The geographic distribution of pneumococci resistant to one or more of the antibiotics penicillin, erythromycin, trimethoprim-sulfamethoxazole, and tetracycline appears to be expanding, and there exist foci of resistance to chloramphenicol and rifampin. Multiply resistant pneumococci are being encountered more commonly and are more often community acquired. Factors associated with infection caused by resistant pneumococci include young age, duration of hospitalization, infection with a pneumococcus of serogroup 6, 19, or 23 or serotype 14, and exposure to antibiotics to which the strain is resistant. At present, the most useful drugs for the management of resistant pneumococcal infections are cefotaxime, ceftriaxone, vancomycin, and rifampin. If the strains are susceptible, chloramphenicol may be useful as an alternative, less expensive agent. Appropriate interventions for the control of resistant pneumococcal outbreaks include investigation of the prevalence of resistant strains, isolation of patients, possible treatment of carriers, and reduction of usage of antibiotics to which the strain is resistant. The molecular mechanisms of penicillin resistance are related to the structure and function of penicillin-binding proteins, and the mechanisms of resistance to other agents involved in multiple resistance are being elucidated. Recognition is increasing of the standard screening procedure for penicillin resistance, using a 1-microgram oxacillin disk. PMID:2187594

  12. Antibiotic susceptibility profiles of some Vibrio strains isolated from wastewater final effluents in a rural community of the Eastern Cape Province of South Africa

    PubMed Central

    2010-01-01

    Background To evaluate the antibiogram and antibiotic resistance genes of some Vibrio strains isolated from wastewater final effluents in a rural community of South Africa. V. vulnificus (18), V. metschnikovii (3), V. fluvialis (19) and V. parahaemolyticus (12) strains were isolated from final effluents of a wastewater treatment plant (WWTP) located in a rural community of South Africa. The disk diffusion method was used for the characterization of the antibiogram of the isolates. Polymerase chain reaction (PCR) was employed to evaluate the presence of established antibiotic resistance genes using specific primer sets. Results The Vibrio strains showed the typical multidrug-resistance phenotype of an SXT element. They were resistant to sulfamethoxazole (Sul), trimethoprim (Tmp), cotrimoxazole (Cot), chloramphenicol (Chl), streptomycin (Str), ampicillin (Amp), tetracycline (Tet) nalidixic acid (Nal), and gentamicin (Gen). The antibiotic resistance genes detected includes dfr18 and dfrA1 for trimethoprim; floR, tetA, strB, sul2 for chloramphenicol, tetracycline, streptomycin and sulfamethoxazole respectively. Some of these genes were only recently described from clinical isolates, demonstrating genetic exchange between clinical and environmental Vibrio species. Conclusions These results demonstrate that final effluents from wastewater treatment plants are potential reservoirs of various antibiotics resistance genes. Moreover, detection of resistance genes in Vibrio strains obtained from the wastewater final effluents suggests that these resistance determinants might be further disseminated in habitats downstream of the sewage plant, thus constituting a serious health risk to the communities reliant on the receiving waterbodies. PMID:20470419

  13. In vitro susceptibility of Burkholderia cepacia complex isolates: Comparison of disk diffusion, Etest®, agar dilution, and broth microdilution methods.

    PubMed

    Fehlberg, Lorena Cristina Corrêa; Nicoletti, Adriana Gianinni; Ramos, Ana Carolina; Rodrigues-Costa, Fernanda; de Matos, Adriana Pereira; Girardello, Raquel; Marques, Elizabeth Andrade; Gales, Ana Cristina

    2016-12-01

    Broth microdilution, agar dilution, Etest® and disk diffusion techniques were compared to evaluate the susceptibility profile of 82 Bcc clinical isolates against six antimicrobials as recommended by CLSI. Broth microdilution was considered the "gold standard" method. The regression analysis was applied to determine the essential (EA) and categorical (CA) agreement rates. STX (MIC50, 1 mg/L) was the most potent antimicrobial tested against Bcc isolates. The worst in vitro activity was observed for chloramphenicol (MIC50, 16 mg/L) and ticarcillin-clavulanic acid (MIC50, >256 mg/L). The EA among broth microdilution and agar dilution results was good for the majority of antimicrobial tested. When comparing broth microdilution and Etest®, ceftazidime, SXT and chloramphenicol exhibited EA rates below 90%. SXT showed an excellent CA (100%) when dilution methodologies were compared. However, a low CA rate was found for this agent between dilution and disk diffusion methodologies resulting in unacceptable very major and minor error rates.

  14. In Vitro Trials of Dittrichia graveolens Essential Oil Combined with Antibiotics.

    PubMed

    Miladinović, Dragoljub L; Ilić, Budimir S; Kocić, Branislava D; Marković, Marija S; Miladinović, Ljiljana C

    2016-06-01

    The chemical composition and antibacterial activity of Dittrichia graveolens (L.) Greuter essential oil were examined. Gas chromatography and gas chromatography/mass spectrometry were used to analyze the chemical composition of the essential oil. The antibacterial activity was investigated by the broth microdilution method against thirteen bacterial strains. The interactions of the essential oil and three standard antibiotics: chloramphenicol, tetracycline and streptomycin toward five selected strains were evaluated using the microdilution checkerboard assay in combination with chemometric methods: principal components analysis and hierarchical cluster analysis. Oxygenated monoterpenes were the most abundant compound class in the essential oil (40.6%), with bomyl acetate (21.7%) as the major compound. The essential oil exhibited slight antibacterial activity against the tested bacterial strains in vitro, but the combinations D. graveolens essential oil-chloramphenicol and D. graveolens-tetracycline exhibited mostly synergistic or additive interactions. These combinations reduced the minimum effective dose of the antibiotics and, consequently, minimized their adverse side effects. In contrast, the association of D. graveolens essential oil and streptomycin was characterized by strong antagonistic interactions against E. coli ATCC 25922, S. aureus ATCC 29213 and P. aeruginosa ATCC 27853. In the principal components analysis (PCA) and hierarchical cluster analysis (HCA), streptomycin against these bacterial strains stood out and formed a separate group.

  15. Selective retinol production by modulating the composition of retinoids from metabolically engineered E. coli.

    PubMed

    Jang, Hui-Jeong; Ha, Bo-Kyung; Zhou, Jia; Ahn, Jiyoon; Yoon, Sang-Hwal; Kim, Seon-Won

    2015-08-01

    Retinoids can be produced from E. coli when introduced with the β-carotene biosynthesis pathway and the BCMO gene. E. coli has no inherent metabolic pathways related to retinoids, therefore only retinal should be produced from the cleavage of β-carotene by BCMO. However, retinol and retinyl acetate were also produced in significant amounts, by the non-specific activity of inherent promiscuous enzymes or the antibiotic resistance marker of the retinal-producing plasmids. Retinol was produced by the ybbO gene of E. coli which encodes oxidoreductase and retinyl acetate was produced by the chloramphenicol resistance gene, called cat gene which encodes chloramphenicol acetyltransferase, present within the pS-NA plasmid that also contains the mevalonate pathway. The composition of retinoids could be modulated by manipulating the relevant genes. The composition of retinol, a commercially important retinoid, was significantly increased by the overexpression of ybbO gene and the removal of cat gene in the recombinant E. coli, which suggests the possibility of selective retinoid production in the future.

  16. Retinoic acid-induced down-regulation of the interleukin-2 promoter via cis-regulatory sequences containing an octamer motif.

    PubMed Central

    Felli, M P; Vacca, A; Meco, D; Screpanti, I; Farina, A R; Maroder, M; Martinotti, S; Petrangeli, E; Frati, L; Gulino, A

    1991-01-01

    Retinoic acid (RA) is known to influence the proliferation and differentiation of a wide variety of transformed and developing cells. We found that RA and the specific RA receptor (RAR) ligand Ch55 inhibited the phorbol ester and calcium ionophore-induced expression of the T-cell growth factor interleukin-2 (IL-2) gene. Expression of transiently transfected chloramphenicol acetyltransferase vectors containing the 5'-flanking region of the IL-2 gene was also inhibited by RA. RA-induced down-regulation of the IL-2 enhancer is mediated by RAR, since overexpression of transfected RARs increased RA sensitivity of the IL-2 promoter. Functional analysis of chloramphenicol acetyltransferase vectors containing either internal deletion mutants of the region from -317 to +47 bp of the IL-2 enhancer or multimerized cis-regulatory elements showed that the RA-responsive element in the IL-2 promoter mapped to sequences containing an octamer motif. RAR also inhibited the transcriptional activity of the octamer motif of the immunoglobulin heavy chain enhancer. In spite of the transcriptional inhibition of the IL-2 octamer motif, RA did not decrease the in vitro DNA-binding capability of octamer-1 protein. These results identify a regulatory pathway within the IL-2 promoter which involves the octamer motif and RAR. Images PMID:1652063

  17. Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene.

    PubMed Central

    Sohaskey, C D; Arnold, C; Barbour, A G

    1997-01-01

    A transient chloramphenicol acetyltransferase (CAT) expression system was developed for Borrelia burgdorferi. An Escherichia coli vector containing a promoterless Streptococcus agalactiae cat gene was constructed. Promoters for ospA, ospC, and flaB were placed upstream of this cat gene, and CAT assays were performed in E. coli from these stably maintained plasmids. The plasmids with putative promoters ospA and flaB were found to be approximately 20-fold more active than were the plasmids with ospC or no promoter. The level of activity correlated well with the resistance to chloramphenicol that each plasmid provided. Next, the nonreplicative plasmid constructs were transformed by electroporation into B. burgdorferi. CAT assays were performed by both thin-layer chromatography and the fluor diffusion method. Measurement of CAT activity demonstrated that the ospA promoter was again about 20-fold more active than the promoterless cat gene. The flaB and ospC promoters increased the activity seven- and threefold, respectively, over that with the promoterless construct. This simple transient-expression assay was shown to be an effective method to study promoter function in B. burgdorferi in the absence of a well-developed genetic system. PMID:9352937

  18. Untargeted metabolomic profiling of amphenicol-resistant Campylobacter jejuni by ultra-high-performance liquid chromatography-mass spectrometry.

    PubMed

    Li, Hui; Xia, Xi; Li, Xiaowei; Naren, Gaowa; Fu, Qin; Wang, Yang; Wu, Congming; Ding, Shuangyang; Zhang, Suxia; Jiang, Haiyang; Li, Jiancheng; Shen, Jianzhong

    2015-02-06

    Campylobacter jejuni, an important foodborne microorganism, poses severe and emergent threats to human health as antibiotic resistance becomes increasingly prevalent. The mechanisms of drug resistance are hard to decipher, and little is known at the metabolic level. Here we apply metabolomic profiling to discover metabolic changes associated with amphenicol (chloramphenicol and florfenicol) resistance mutations of Campylobacter jejuni. An optimized sample preparation method was combined with ultra-high-performance liquid chromatography-time-of-flight mass spectrometry (UHPLC-TOF/MS) and pattern recognition for the analysis of small-molecule biomarkers of drug resistance. UHPLC-triple quadrupole MS operated in multiple reaction monitoring mode was used for quantitative analysis of metabolic features from UHPLC-TOF/MS profiling. Up to 41 differential metabolites involved in glycerophospholipid metabolism, sphingolipid metabolism, and fatty acid metabolism were observed in a chloramphenicol-resistant mutant strain of Campylobacter jejuni. A panel of 40 features was identified in florfenicol-resistant mutants, demonstrating changes in glycerophospholipid metabolism, sphingolipid metabolism, and tryptophan metabolism. This study shows that the UHPLC-MS-based metabolomics platform is a promising and valuable tool to generate new insights into the drug-resistant mechanism of Campylobacter jejuni.

  19. Microbial strain improvement for organosulfur removal from coal. Final technical report, 1 September, 1992--31 August, 1993

    SciTech Connect

    Kilbane, J.J. II

    1993-12-31

    IGT has developed a microbial culture of Rhodococcus rhodochrous, designated as IGTS8, that is capable of specifically cleaving carbon-sulfur bonds in a range of organosulfur model compounds and is capable of removing organic sulfur from coal and petroleum without significantly sacrificing the calorific value of the fuel. Although IGTS8 possesses the ability to specifically remove organic sulfur from coal, a major research need is to develop improved strains of microorganisms that possess higher levels of desulfurization activity and therefore will permit more favorable biodesulfurization process conditions: faster rates, more complete removal, and smaller reactor size. Strain improvement is the single most important aspect to the development of a practical coal biodesulfurization process and accordingly is the focus of research in this project. During the past year, significant progress was made toward improving the biodesulfurization capabilities of Rhodococcus Rhodochrous IGTS8. The main objective was to identify and characterize strong promoters of IGTS8. The DNA sequencing of the promoter region and chloramphenicol resistance gene of pRF2, as well as six mutant promoters, was determined. The 16S structural gene of IGTS8 was isolated and used to identify the putative promoter of this gene. Four promoter probe vectors were constructed and are currently being used to analyze the strength of Rhodococcus promoters: from the IGTS8 genome, mutants of promoters from the chloramphenicol resistance gene of pRF2, the promoter from the 16S RNA gene, and various strong inducible promoters.

  20. Microbial strain improvement for organosulfur removal from coal. Technical report, December 1, 1992--February 28, 1993

    SciTech Connect

    Kilbane, J.J. II; Ho, K.

    1993-05-01

    IGT has developed a microbial culture of Rhodococcus rhodochrous, designated as IGTS8, that is capable of specifically cleaving carbon-sulfur bonds in a range of organosulfur model compounds and is capable of removing organic sulfur from coal and petroleum without significantly sacrificing the calorific value of the fuel. Although IGTSS possesses the ability to specifically remove organic sulfur from coal, a major research need is to develop strains of microorganisms that possess higher levels of desulfurization activity. During the past quarter, the DNA sequence of the promoter region of the chloramphenicol resistance gene of pRF2 was determined and tentatively localized to a 50 bp region. To further define the important sequences of this promoter mutants were generated that may have altered promoters allowing higher levels of expression of the chloramphenicol resistance gene. The promoter region from twenty such mutants has been subcloned. Additionally, promoter probe vectors were used to isolate small DNA fragments from the chromosome that possess promoters. These studies will lead to the identification and/or the construction of particularly strong Rhodococcus promoters which will subsequently be used to express the desulfurization genes.

  1. Transformation of Rhodococcus fascians by High-Voltage Electroporation and Development of R. fascians Cloning Vectors.

    PubMed

    Desomer, J; Dhaese, P; Montagu, M V

    1990-09-01

    The analysis of the virulence determinants of phytopathogenic Rhodococcus fascians has been hampered by the lack of a system for introducing exogenous DNA. We investigated the possibility of genetic transformation of R. fascians by high-voltage electroporation of intact bacterial cells in the presence of plasmid DNA. Electrotransformation in R. fascians D188 resulted in transformation frequencies ranging from 10/mug of DNA to 10/mug of DNA, depending on the DNA concentration. The effects of different electrical parameters and composition of electroporation medium on transformation efficiency are presented. By this transformation method, a cloning vector (pRF28) for R. fascians based on an indigenous 160-kilobase (chloramphenicol and cadmium resistance-encoding) plasmid pRF2 from strain NCPPB 1675 was developed. The origin of replication and the chloramphenicol resistance gene on pRF28 were used to construct cloning vectors that are capable of replication in R. fascians and Escherichia coli. The electroporation method presented was efficient enough to allow detection of the rare integration of replication-deficient pRF28 derivatives in the R. fascians D188 genome via either homologous or illegitimate recombination.

  2. Transformation of Rhodococcus fascians by high-voltage electroporation and development of R. fascians cloning vectors

    SciTech Connect

    Desomer, J.; Dhaese, P.; Montagu, M.V. )

    1990-09-01

    The analysis of the virulence determinants of phytopathogenic Rhodococcus fascians has been hampered by the lack of a system for introducing exogenous DNA. We investigated the possibility of genetic transformation of R. fascians by high-voltage electroporation of intact bacterial cells in the presence of plasmid DNA. Electrotransformation in R. fascians D188 resulted in transformation frequencies ranging from 10{sup 5}/{mu}g of DNA to 10{sup 7}/{mu}g of DNA, depending on the DNA concentration. The effects of different electrical parameters and composition of electroporation medium on transformation efficiency are present. By this transformation method, a cloning vector (pRF28) for R. fascians based on an indigenous 160-kilobase (chloramphenicol and cadmium resistance-encoding) plasmid pRF2 from strain NCPPB 1675 was developed. The origin of replication and the chloramphenicol resistance gene on pRF28 were used to construct cloning vectors that are capable of replication in R. fascians and Escherichia coli. The electroporation method presented was efficient enough to allow detection of the rare integration of replication-deficient pRF28 derivatives in the R.fascians D188 genome via either homologous or illegitimate recombination.

  3. Multiclass determination and confirmation of antibiotic residues in honey using LC-MS/MS.

    PubMed

    Lopez, Mayda I; Pettis, Jeffery S; Smith, I Barton; Chu, Pak-Sin

    2008-03-12

    A multiclass method has been 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 (lincomycin), aminoglycosides (streptomycin), sulfonamides (sulfathiazole), phenicols (chloramphenicol), and fumagillin residues using liquid chromatography tandem mass spectrometry (LC-MS/MS). Erythromycin (a macrolide) and monensin (an ionophore) can be detected and confirmed but not quantitated. Honey samples (approximately 2 g) are dissolved in 10 mL of water and centrifuged. An aliquot of the supernatant is used to determine streptomycin. The remaining supernatant is filtered through a fine-mesh nylon fabric and cleaned up by solid phase extraction. After solvent evaporation and sample reconstitution, 15 antibiotics are assayed by LC-MS/MS using electrospray ionization (ESI) in positive ion mode. Afterward, chloramphenicol is assayed using ESI in negative ion mode. The method has been validated at the low part per billion levels for most of the drugs with accuracies between 65 and 104% and coefficients of variation less than 17%. The evaluation of matrix effects caused by honey of different floral origin is presented.

  4. In vitro sensitivity of Hungarian Actinobaculum suis strains to selected antimicrobials.

    PubMed

    Biksi, I; Major, Andrea; Fodor, L; Szenci, O; Vetési, F

    2003-01-01

    In vitro antimicrobial sensitivity of 12 Hungarian isolates and the type strain ATCC 33144 of Actinobaculum suis to different antimicrobial compounds was determined both by the agar dilution and by the disc diffusion method. By agar dilution, MIC50 values in the range of 0.05-3.125 micrograms/ml were determined for penicillin, ampicillin, ceftiofur, doxycycline, tylosin, pleuromutilins, chloramphenicol, florfenicol, enrofloxacin and lincomycin. The MIC50 value of oxytetracycline and spectinomycin was 6.25 and 12.5 micrograms/ml, respectively. For ofloxacin, flumequine, neomycin, streptomycin, gentamicin, nalidixic acid, nitrofurantoin and sulphamethoxazole + trimethoprim MIC50 values were in the range of 25-100 micrograms/ml. With the disc diffusion method, all strains were sensitive to penicillin, cephalosporins examined, chloramphenicol and florfenicol, tetracyclines examined, pleuromutilins, lincomycin and tylosin. Variable sensitivity was observed for fluoroquinolones (flumequine, enrofloxacin, ofloxacin), most of the strains were susceptible to marbofloxacin. Almost all strains were resistant to aminoglycosides but most of them were sensitive to spectinomycin. A strong correlation was determined for disc diffusion and MIC results (Spearman's rho 0.789, p < 0001). MIC values of the type strain and MIC50 values of other tested strains did not differ significantly. Few strains showed a partially distinct resistance pattern for erythromycin, lincomycin and ampicillin in both methods.

  5. Detection limits of antimicrobials in ewe milk by delvotest photometric measurements.

    PubMed

    Althaus, R L; Torres, A; Montero, A; Balasch, S; Molina, M P

    2003-02-01

    The Delvotest method detection limits per manufacturer's instructions at a fixed reading time of 3 h for 24 antimicrobial agents were determined in ewe milk by photometric measurement. For each drug, eight concentrations were tested on 20 ewe milk samples from individual ewes. Detection limits, determined by means of logistic regression models, were (microg/kg): 3, amoxycillin; 2, ampicillin; 18, cloxacillin; 1, penicillin "G"; 34, cefadroxil; 430, cephalosporin "C"; 40, cephalexin; 20, cefoperazone; 33, Ceftiofur; 18, cefuroxime; 6100, streptomycin; 1200, gentamycin; 2600, neomycin; 830, erythromycin; 100, tylosin; 180, doxycycline; 320, oxytetracycline; 590, tetracycline; 88, sulfadiazine; 44, sulfamethoxazole; 140, sulfametoxypyridazine; 48, sulfaquinoxaline; 12,000, chloramphenicol; and 290, trimethoprim. Whereas the beta-lactam antibiotics, sulphonamides, and tylosin were detected by Delvotest method at levels equal to those of maximum residue limits, its sensitivity needs to be enhanced to detect aminoglycosides, tetracyclines, streptomycin, chloramphenicol, and trimethoprim residues in ewe milk or to develop an integrated residue detection system for ewe milk with different sensitive microorganisms for each group of antiinfectious agents.

  6. Mutational analysis of the lac regulatory region: second-site changes that activate mutant promoters.

    PubMed Central

    Rothmel, R K; LeClerc, J E

    1989-01-01

    Second-site mutations that restored activity to severe lacP1 down-promoter mutants were isolated. This was accomplished by using a bacteriophage f1 vector containing a fusion of the mutant E. coli lac promoters with the structural gene for chloramphenicol acetyltransferase (CAT), so that a system was provided for selecting phage revertants (or pseudorevertants) that conferred resistance of phage-infected cells to chloramphenicol. Among the second-site changes that relieved defects in mutant lac promoters, the only one that restored lacP1 activity was a T----G substitution at position -14, a weakly conserved site in E. coli promoters. Three other sequence changes, G----A at -2, A----T at +1, and C----A at +10, activated nascent promoters in the lac regulatory region. The nascent promoters conformed to the consensus rule, that activity is gained by sequence changes toward homology with consensus sequences at the -35 and -10 regions of the promoter. However, the relative activities of some promoters cannot be explained solely by consideration of their conserved sequence elements. Images PMID:2660105

  7. Prevalence, serotyping and antimicrobials resistance mechanism of Salmonella enterica isolated from clinical and environmental samples in Saudi Arabia.

    PubMed

    El-Tayeb, Mohamed A; Ibrahim, Abdelnasser S S; Al-Salamah, Ali A; Almaary, Khalid S; Elbadawi, Yahya B

    2017-02-14

    Salmonella is recognized as a common foodborne pathogen, causing major health problems in Saudi Arabia. Herein, we report epidemiology, antimicrobial susceptibility and the genetic basis of resistance among S. enterica strains isolated in Saudi Arabia. Isolation of Salmonella spp. from clinical and environmental samples resulted in isolation of 33 strains identified as S. enterica based on their biochemical characteristics and 16S-rDNA sequences. S. enterica serovar Enteritidis showed highest prevalence (39.4%), followed by S. Paratyphi (21.2%), S. Typhimurium (15.2%), S. Typhi and S. Arizona (12.1%), respectively. Most isolates were resistant to 1st and 2nd generation cephalosporin; and aminoglycosides. Moreover, several S. enterica isolates exhibited resistance to the first-line antibiotics used for Salmonellosis treatment including ampicillin, trimethoprim-sulfamethoxazole and chloramphenicol. In addition, the results revealed the emergence of two S. enterica isolates showing resistance to third-generation cephalosporin. Analysis of resistance determinants in S. enterica strains (n=33) revealed that the resistance to β-lactam antibiotics, trimethoprim-sulfamethoxazole, chloramphenicol, and tetracycline, was attributed to the presence of carb-like, dfrA1, floR, tetA gene, respectively. On the other hand, fluoroquinolone resistance was related to the presence of mutations in gyrA and parC genes. These findings improve the information about foodborne Salmonella in Saudi Arabia, alarming the emergence of multi-drug resistant S. enterica strains, and provide useful data about the resistance mechanisms.

  8. Comparing denitrification estimates for a Texas estuary by using acetylene inhibition and membrane inlet mass spectrometry.

    PubMed

    Bernot, Melody J; Dodds, Walter K; Gardner, Wayne S; McCarthy, Mark J; Sobolev, Dmitri; Tank, Jennifer L

    2003-10-01

    Characterizing denitrification rates in aquatic ecosystems is essential to understanding how systems may respond to increased nutrient loading. Thus, it is important to ensure the precision and accuracy of the methods employed for measuring denitrification rates. The acetylene (C2H2) inhibition method is a simple technique for estimating denitrification. However, potential problems, such as inhibition of nitrification and incomplete inhibition of nitrous oxide reduction, may influence rate estimates. Recently, membrane inlet mass spectrometry (MIMS) has been used to measure denitrification in aquatic systems. Comparable results were obtained with MIMS and C2H2 inhibition methods when chloramphenicol was added to C2H2 inhibition assay mixtures to inhibit new synthesis of denitrifying enzymes. Dissolved-oxygen profiles indicated that surface layers of sediment cores subjected to the MIMS flowthrough incubation remained oxic whereas cores incubated using the C2H2 inhibition methods did not. Analysis of the microbial assemblages before and after incubations indicated significant changes in the sediment surface populations during the long flowthrough incubation for MIMS analysis but not during the shorter incubation used for the C2H2 inhibition method. However, bacterial community changes were also small in MIMS cores at the oxygen transition zone where denitrification occurs. The C2H2 inhibition method with chloramphenicol addition, conducted over short incubation intervals, provides a cost-effective method for estimating denitrification, and rate estimates are comparable to those obtained by the MIMS method.

  9. Multicenter Study of Antimicrobial Susceptibility of Anaerobic Bacteria in Korea in 2012

    PubMed Central

    Lee, Yangsoon; Park, Yeon-Joon; Kim, Mi-Na; Uh, Young; Kim, Myung Sook

    2015-01-01

    Background Periodic monitoring of regional or institutional resistance trends of clinically important anaerobic bacteria is recommended, because the resistance of anaerobic pathogens to antimicrobial drugs and inappropriate therapy are associated with poor clinical outcomes. There has been no multicenter study of clinical anaerobic isolates in Korea. We aimed to determine the antimicrobial resistance patterns of clinically important anaerobes at multiple centers in Korea. Methods A total of 268 non-duplicated clinical isolates of anaerobic bacteria were collected from four large medical centers in Korea in 2012. Antimicrobial susceptibility was tested by the agar dilution method according to the CLSI guidelines. The following antimicrobials were tested: piperacillin, piperacillin-tazobactam, cefoxitin, cefotetan, imipenem, meropenem, clindamycin, moxifloxacin, chloramphenicol, metronidazole, and tigecycline. Results Organisms of the Bacteroides fragilis group were highly susceptible to piperacillin-tazobactam, imipenem, and meropenem, as their resistance rates to these three antimicrobials were lower than 6%. For B. fragilis group isolates and anaerobic gram-positive cocci, the resistance rates to moxifloxacin were 12-25% and 11-13%, respectively. Among B. fragilis group organisms, the resistance rates to tigecycline were 16-17%. Two isolates of Finegoldia magna were non-susceptible to chloramphenicol (minimum inhibitory concentrations of 16-32 mg/L). Resistance patterns were different among the different hospitals. Conclusions Piperacillin-tazobactam, cefoxitin, and carbapemems are highly active β-lactam agents against most of the anaerobes. The resistance rates to moxifloxacin and tigecycline are slightly higher than those in the previous study. PMID:26206683

  10. Differing ability to transport non-metal substrates by two RND-type metal exporters

    PubMed Central

    Conroy, Otakuye; Kim, Eun-Hae; McEvoy, Megan M.; Rensing, Christopher

    2010-01-01

    The metal-exporting systems CusCFBA of Escherichia coli and GesABC of Salmonella are RND-type multi-protein systems responsible for detoxification during metal stress. In this study the substrate range was determined for each metal transport system and possible amino acid residues important in substrate specificity identified. The Ges system, previously identified as a gold-efflux system, conferred resistance to the greatest number and variety of organic chemicals including chloramphenicol, not previously recognized as a substrate. Phylogenetic analysis showed that GesB is most closely related to a class of RND transporters including MexF that have been shown to be responsible for exporting fluoroquinolones, chloramphenicol, and biocides. However, many of the closest homologs of GesABC appear to have a role in metal resistance judging from the genetic context. In contrast, CusCFBA belongs to a distinct family of RND-type monovalent metal exporter systems containing a number of essential metal-binding methionines resulting in a much narrower substrate range. PMID:20497225

  11. Isolation and antimicrobial susceptibility of Streptococcus castoreus isolated from carcasses of European beavers (Castor fiber) in Germany.

    PubMed

    Schulze, Christoph; Kutzer, Peter; Winterhoff, Nora; Engelhardt, Andreas; Bilk, Sabine; Teubner, Jens

    2015-01-01

    Streptococcus (S.) castoreus was isolated from seven beavers (Castor fiber), which were found dead in the federal state of Brandenburg, Germany, and submitted to the Berlin-Brandenburg State laboratory for post mortem diagnostics. The isolates originated from various inflammatory processes where they were associated with other Gram negative and Gram positive aerobic and/or anaerobic bacteria (i. e. Actinobacillus sp., species of the Actinomycetaceae family, coliform bacteria, Fusobacterium sp., Prevotella sp.), but also from cloacal swabs. Testing their antimicrobial susceptibility, all S. castoreus isolates and the type strain S. castoreus (DSM 17536) were classified as being susceptible to penicillin G, ampicillin, amoxicillin-clavulanic acid, 1st generation cephalosporins, erythromycin, tetracycline, and chloramphenicole but intermediate to gentamicin. Considering both the polymicrobial flora isolated from the inflamed tissues and the fact that antimicrobials of a narrow spectrum (namely penicillins, 1st generation cephalosporins, macrolids and lincosamids) can be toxic to rodents, chloramphenicol might be a suitable drug for treatment of beavers suffering from S. castoreus (mixed)-bacterial infections.

  12. Molecular and epidemiological study of Salmonella clinical isolates.

    PubMed Central

    Rivera, M J; Rivera, N; Castillo, J; Rubio, M C; Gómez-Lus, R

    1991-01-01

    A survey of Salmonella infections was carried out over a 1-year period in the rural community covered by the Hospital Reina Sofía (Tudela, Spain). The 255 strains that were collected were studied by serotyping, antimicrobial resistance, and plasmid profile analysis. The predominant serotype was S. enteritidis (85.90%), followed by S. typhimurium (7.06%) and S. virchow (2.36%). Only 7.84% of the strains were resistant to antimicrobial agents. The most common resistance was to beta-lactam antibiotics. This resistance was due to the presence of one of two types of beta-lactamases, TEM-1 or TEM-2. Resistance to kanamycin was associated with the synthesis of a 3'-O-phosphotransferase. The resistance to streptomycin and chloramphenicol was either not enzymatic or was due to a 3"-O-phosphotransferase and a chloramphenicol acetyltransferase, respectively. Analysis of total plasmid DNA content revealed the presence of plasmids in 96.08% of the isolates. According to their plasmid profile, the strains could be classified into different groups. The three main groups, which accounted for 50.19, 20.78, and 4.70% of the isolates, respectively, corresponded to the antimicrobial-susceptible S. enteritidis serotype. These results suggested that plasmid profile analysis in conjunction with antimicrobial resistance determination can be useful for subtyping resistant Salmonella isolates. Images PMID:2056061

  13. Randomized Trial of Clarithromycin for Mediterranean Spotted Fever

    PubMed Central

    Muñoz, Tomas; Travería, Francisco Javier; Navarro, Gemma; Font, Bernat; Sanfeliu, Isabel; Segura, Ferran

    2015-01-01

    The classic antibiotic treatment for Mediterranean spotted fever (MSF) is based on tetracyclines or chloramphenicol, but chloramphenicol's bone marrow toxicity makes tetracyclines the treatment of choice. However, it is convenient to have alternatives available for patients who are allergic to tetracyclines, pregnant women, and children <8 years old. We conducted a randomized clinical trial to compare clarithromycin with doxycycline or josamycin in the treatment of MSF. Forty patients were evaluated (23 male; mean age, 39.87 years); 13 patients were aged <14 years. Seventeen patients received clarithromycin, and 23 received doxycycline or josamycin. The interval between the onset of symptoms and the start of treatment was 4.04 ± 1.70 days in the clarithromycin group versus 4.11 ± 1.60 days in the doxycycline/josamycin group (P = not significant [NS]). Time to the disappearance of fever after treatment was 2.67 ± 1.55 days in the clarithromycin group versus 2.22 ± 1.35 days in the doxycycline/josamycin (P = NS). The symptoms had disappeared at 4.70 ± 2.25 days in the clarithromycin group versus at 4.75 ± 3.08 days in the doxycycline/josamycin (P = NS). There were no adverse reactions to treatment or relapses in either group. In conclusion, clarithromycin is a good alternative to doxycycline or josamycin in the treatment of MSF. PMID:26711765

  14. Isolation and identification of zoonotic species of genus arcobacter from chicken viscera obtained from retail distributors of the metropolitan area of San José, Costa Rica.

    PubMed

    Villalobos, Edgar García; Jaramillo, Heriberto Fernández; Ulate, Carolina Chaves; Echandi, María Laura Arias

    2013-05-01

    Arcobacter is a genus of growing importance worldwide; some of its species are considered emerging enteropathogens and potential zoonotic agents. In Costa Rica, as well as in other countries, its isolation has been reported, so the objective of this project was to evaluate and identify the presence of Arcobacter in chicken viscera sold in the metropolitan area of San José, Costa Rica, as well as to determine the antimicrobial resistance patterns associated with it. One hundred fifty samples of chicken viscera including heart, liver, and other gastrointestinal organs were purchased from 15 supermarkets and 15 local retailers. De Boer and Houf broths were used as enrichment media; isolation was done with Arcobacter-selective medium and with membrane filtration with blood agar. Typical colonies were identified with genus-specific PCR, and species identification was made with multiplex PCR. Susceptibility to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, and tetracycline was done with the Epsilometer test. The isolation frequency of Arcobacter genus obtained in this study was of 17.3%. A total of 33 isolates were obtained from the poultry samples, and according to the multiplex PCR methodology, 22 (66.7%) isolates were identified as Arcobacter butzleri, 8 (24.2%) as Arcobacter cryaerophilus, and 1 (3.1%) as Arcobacter skirrowii. Two strains were not identified. No statistical significant difference was found when the source of samples was compared. Resistance toward chloramphenicol was 68.75%, followed by ampicillin (43.75%) and ciprofloxacin (18.75%); all strains were susceptible to tetracycline.

  15. Effects of antibiotics on Shiga toxin 2 production and bacteriophage induction by epidemic Escherichia coli O104:H4 strain.

    PubMed

    Bielaszewska, Martina; Idelevich, Evgeny A; Zhang, Wenlan; Bauwens, Andreas; Schaumburg, Frieder; Mellmann, Alexander; Peters, Georg; Karch, Helge

    2012-06-01

    The role of antibiotics in treatment of enterohemorrhagic Escherichia coli (EHEC) infections is controversial because of concerns about triggering hemolytic-uremic syndrome (HUS) by increasing Shiga toxin (Stx) production. During the recent large EHEC O104:H4 outbreak, antibiotic therapy was indicated for some patients. We tested a diverse panel of antibiotics to which the outbreak strain is susceptible to interrogate the effects of subinhibitory antibiotic concentrations on induction of stx(2)-harboring bacteriophages, stx(2) transcription, and Stx2 production in this emerging pathogen. Ciprofloxacin significantly increased stx(2)-harboring phage induction and Stx2 production in outbreak isolates (P values of <0.001 to <0.05), while fosfomycin, gentamicin, and kanamycin insignificantly influenced them (P > 0.1) and chloramphenicol, meropenem, azithromycin, rifaximin, and tigecycline significantly decreased them (P ≤ 0.05). Ciprofloxacin and chloramphenicol significantly upregulated and downregulated stx(2) transcription, respectively (P < 0.01); the other antibiotics had insignificant effects (P > 0.1). Meropenem, azithromycin, and rifaximin, which were used for necessary therapeutic or prophylactic interventions during the EHEC O104:H4 outbreak, as well as tigecycline, neither induced stx(2)-harboring phages nor increased stx(2) transcription or Stx2 production in the outbreak strain. These antibiotics might represent therapeutic options for patients with EHEC O104:H4 infection if antibiotic treatment is inevitable. We await further analysis of the epidemic to determine if usage of these agents was associated with an altered risk of developing HUS.

  16. Antimicrobial resistance trends among Salmonella isolates obtained from horses in the northeastern United States (2001-2013).

    PubMed

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

    2016-05-01

    OBJECTIVE To describe the antimicrobial resistance patterns of Salmonella isolates obtained from horses in the northeastern United States and to identify trends in resistance to select antimicrobials over time. SAMPLE 462 Salmonella isolates from horses. PROCEDURES Retrospective data were collected for all Salmonella isolates obtained from equine specimens that were submitted to the Cornell University Animal Health Diagnostic Center between January 1, 2001, and December 31, 2013. Temporal trends in the prevalence of resistant Salmonella isolates were investigated for each of 13 antimicrobials by use of the Cochran-Armitage trend test. RESULTS The prevalence of resistant isolates varied among antimicrobials and ranged from 0% (imipenem) to 51.5% (chloramphenicol). During the observation period, the prevalence of resistant isolates decreased significantly for amoxicillin-clavulanic acid, ampicillin, cefazolin, cefoxitin, ceftiofur, chloramphenicol, and tetracycline and remained negligible for amikacin and enrofloxacin. Of the 337 isolates for which the susceptibility to all 13 antimicrobials was determined, 138 (40.9%) were pansusceptible and 192 (57.0%) were multidrug resistant (resistant to ≥ 3 antimicrobial classes). The most common serovar isolated was Salmonella Newport, and although the annual prevalence of that serovar decreased significantly over time, that decrease had only a minimal effect on the observed antimicrobial resistance trends. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that current antimicrobial use in horses is not promoting the emergence and dissemination of antimicrobial-resistant Salmonella strains in the region served by the laboratory.

  17. Phenotypic and genetic characterization of multidrug-resistant Staphylococcus aureus in the tropics of Southeast Asia.

    PubMed

    Zulkeflle, Siti Norayuni Mohd; Yusaimi, Yus Amira; Sugiura, Norio; Iwamoto, Koji; Goto, Masafumi; Utsumi, Motoo; Othman, Nor'azizi Bin; Zakaria, Zuriati; Hara, Hirofumi

    2016-12-01

    Antibiotic resistance has become a major public health problem throughout the world. The presence of antibiotic-resistant bacteria such as Staphylococcus aureus and antibiotic resistance genes (ARGs) in hospital wastewater is a cause for great concern today. In this study, 276 Staph. aureus isolates were recovered from hospital wastewater samples in Malaysia. All of the isolates were screened for susceptibility to nine different classes of antibiotics: ampicillin, ciprofloxacin, gentamicin, kanamycin, erythromycin, vancomycin, trimethoprim and sulfamethoxazole, chloramphenicol, tetracycline and nalidixic acid. Screening tests showed that 100 % of Staph.aureus isolates exhibited resistance against kanamycin, vancomycin, trimethoprim and sulfamethoxazole and nalidixic acid. Additionally, 91, 87, 50, 43, 11 and 8.7 % of isolates showed resistance against erythromycin, gentamicin, ciprofloxacin, ampicillin, chloramphenicol and tetracycline, respectively. Based on these results, 100 % of isolates demonstrated multidrug-resistant (MDR) characteristics, displaying resistance against more than three classes of antibiotics. Of 276 isolates, nine exhibited resistance to more than nine classes of tested antibiotics; these were selected for antibiotic susceptibility testing and examined for the presence of conserved ARGs. Interestingly, a high percentage of the selected MDR Staph.aureus isolates did not contain conserved ARGs. These results indicate that non-conserved MDR gene elements may have already spread into the environment in the tropics of Southeast Asia, and unique resistance mechanisms against several antibiotics may have evolved due to stable, moderate temperatures that support growth of bacteria throughout the year.

  18. Characterization of a cryptic plasmid from a Greenland ice core Arthrobacter isolate and construction of a shuttle vector that replicates in psychrophilic high G+C Gram-positive recipients.

    PubMed

    Miteva, Vanya; Lantz, Sarah; Brenchley, Jean

    2008-05-01

    Over 60 Greenland glacial isolates were screened for plasmids and antibiotic resistance/sensitivity as the first step in establishing a genetic system. Sequence analysis of a small, cryptic, 1,950 bp plasmid, p54, from isolate GIC54, related to Arthrobacter agilis, showed a region similar to that found in theta replicating Rhodococcus plasmids. A 6,002 bp shuttle vector, pSVJ21, was constructed by ligating p54 and pUC18 and inserting a chloramphenicol acetyl transferase (CAT) cassette conferring chloramphenicol resistance. Candidate Gram-positive recipients were chosen among glacial isolates based on phylogenetic relatedness, relatively short doubling times at low temperatures, sensitivity to antibiotics, and absence of indigenous plasmids. We developed an electroporation protocol and transformed seven isolates related to members of the Arthrobacter, Microbacterium, Curtobacterium, and Rhodoglobus genera with pSVJ21. Plasmid stability was demonstrated by successive transformation into Escherichia coli and four Gram-positive isolates, growth without antibiotic, and plasmid re-isolation. This shuttle vector and our transformation protocol provide the basis for genetic experiments with different high G+C Gram-positive hosts to study cold adaptation and expression of cold-active enzymes at low temperatures.

  19. One-Pot Synthesis, Spectroscopic and Physicochemical Studies of Quinoline Based Blue Emitting Donor-Acceptor Chromophores with Their Biological Application.

    PubMed

    Asiri, Abdullah M; Khan, Salman A; Al-Thaqafya, Saad H

    2015-09-01

    Blue emitting cyano substituted isoquinoline dyes were synthesized by one-pot multicomponent reactions (MCRs) of aldehydes, malononitrile, 6-methoxy-1,2,3,4-tetrahydro-naphthalin-1-one and ammonium acetate. Results obtained from spectroscopic (FT-IR, (1)H-NMR, (13)C-NMR, EI-MS) and elemental analysis of synthesized compounds was in good agreement with their chemical structures. UV-vis and fluorescence spectroscopy measurements proved that all compounds are good absorbent and fluorescent. Fluorescence polarity study demonstrated that these compounds were sensitive to the polarity of the microenvironment provided by different solvents. In addition, spectroscopic and physicochemical parameters, including electronic absorption, excitation coefficient, stokes shift, oscillator strength, transition dipole moment and fluorescence quantum yield were investigated in order to explore the analytical potential of synthesized compounds. The anti-bacterial activity of these compounds were first studied in vitro by the disk diffusion assay against two Gram-positive and two Gram-negative bacteria then the minimum inhibitory concentration (MIC) was determined with the reference of standard drug chloramphenicol. The results displayed that compound 3 was better inhibitors of both types of the bacteria (Gram-positive and Gram-negative) than chloramphenicol. Graphical Abstract ᅟ.

  20. In vivo susceptibility of the Legionnaires disease bacterium to ten antimicrobial agents.

    PubMed

    Lewis, V J; Thacker, W L; Shepard, C C; McDade, J E

    1978-03-01

    The susceptibility of the Legionnaires disease bacterium to various antimicrobial agents was determined by inoculation of embryonated eggs via the yolk sac. When administered prophylactically, the minimal dose of drug preventing all deaths due to the infection was as follows: rifampin, 0.02 mg; gentamicin, 0.25 mg; streptomycin, 0.39 mg; erythromycin, 0.62 mg; sulfadiazine, 1.56 mg; chloramphenicol, 2.50 mg; and cephalothin, 20.0 mg. Smaller amounts delayed deaths, and larger or equal amounts rendered the embryos free of infection. Oxytetracycline in the largest tested amount, 5.0 mg, protected 80% of the embryos from death, and as little as 0.31 mg delayed death. Chlortetracycline (0.50 mg) and ampicillin (10.0 mg) were ineffective. The six most effective drugs were studied in an experiment in which they were administered at various times after infection in doses that were twice the minimal prophylactic dose preventing all deaths. In this therapeutic experiment, rifampin, and erythromycin allowed 100% survival when given even 72 h after infection; gentamicin, streptomycin, sulfadiazine, and chloramphenicol did so when given 48 h after infection. All six drugs increased mean survival time when administered 72 h after infection.