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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 toxicity in dogs.

    PubMed

    Watson, A D

    1977-07-01

    Twenty dogs were given chloramphenicol by mouth night and morning for 14 days: six dogs were dosed at 225 mg/kg/day, four each at 175 and 125 mg/kg/day and three each at 275 and 75 mg/kg/day. Six control dogs were given empty gelatin capsules twice daily for the same period. Dogs dosed at 75 mg/kg consumed more food and gained a little more weight than the control dogs, while those in the 175, 225 and 275 mg/kg groups ate less and lost weight. Four dogs dosed at 175 mg/kg or above became dull and depressed and virtually ceased to eat. No changes were observed in erythrocyte and reticulocyte counts, haemoglobin concentration, packed cell volume or total and differential leukocyte counts during the experiment. Bone marrow examination showed suppression of erythropoiesis in four of nine dogs dosed at 225 or 275 mg/kg/day. In addition, there was evidence of decreased mitotic activity and reduced rate of granulocytopoiesis in the 275 mg/kg group. Vacuolation of marrow cells was not observed. The two toxic effects observed (depression and hypophagia on the one hand, marrow suppression on the other) occurred separately or together in individual dogs.

  3. 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. PMID:22210605

  4. 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...

  5. 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...

  6. Erythromycin induces expression of the chloramphenicol acetyltransferase gene cat-86.

    PubMed Central

    Rogers, E J; Lovett, P S

    1990-01-01

    The plasmid gene cat-86 specifies chloramphenicol-inducible chloramphenicol acetyltransferase in Bacillus subtilis. This gene, like the erythromycin-inducible erm genes, is regulated by translational attenuation. Here we show that cat-86 is also inducibly regulated by erythromycin. cat-86 does not confer resistance to erythromycin. PMID:2115875

  7. 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... associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

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

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Chloramphenicol palmitate oral suspension. 520.390c Section 520.390c Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

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

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 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 § 520.390c Chloramphenicol palmitate oral suspension....

  10. 21 CFR 520.390c - Chloramphenicol palmitate oral suspension.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Chloramphenicol palmitate oral suspension. 520.390c Section 520.390c Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

  11. 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... associated with canine distemper that are caused by organisms susceptible to chloramphenicol. (3)...

  12. 21 CFR 520.390 - Chloramphenicol oral dosage forms.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Chloramphenicol oral dosage forms. 520.390 Section... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.390 Chloramphenicol oral dosage forms....

  13. 21 CFR 520.390 - Chloramphenicol oral dosage forms.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Chloramphenicol oral dosage forms. 520.390 Section... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.390 Chloramphenicol oral dosage forms....

  14. 21 CFR 520.390 - Chloramphenicol oral dosage forms.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Chloramphenicol oral dosage forms. 520.390 Section... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.390 Chloramphenicol oral dosage forms....

  15. 21 CFR 520.390 - Chloramphenicol oral dosage forms.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Chloramphenicol oral dosage forms. 520.390 Section... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.390 Chloramphenicol oral dosage forms....

  16. 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... (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS ORAL DOSAGE FORM NEW ANIMAL DRUGS § 520.390 Chloramphenicol oral dosage forms....

  17. Mechanism of Chloramphenicol-Cephaloridine Synergism on Enterobacteriaceae

    PubMed Central

    Michel, J.; Bornstein, H.; Luboshitzky, R.; Sacks, T.

    1975-01-01

    A synergistic in vitro bactericidal effect of combinations of chloramphenicol and beta-lactams on strains of Enterobacteriaceae is described. The synergism is seen with strains which are resistant to the beta-lactam and is due to chloramphenicol-induced inhibition of beta-lactamase production. PMID:1155928

  18. Antibiotics exposure and health risks: chloramphenicol.

    PubMed

    Hanekamp, Jaap C; Bast, Aalt

    2015-01-01

    The antibiotic chloramphenicol (CAP) is banned from food production. Besides being a medicinal product, CAP is also a natural product, produced by Streptomyces Venezuelae. The lack of scientific data hampers setting of an Acceptable Daily Intake (ADI). Consequently, a maximum residue limit (MRL) in food could not be established. This was then translated into a zero tolerance using the so-called Minimum Required Performance Limit (MRPL) level, viz. the achievable detection limit in food, to guide the zero tolerance policy. The MRPL is clearly not relevant to food safety and human health but is solely related to analytical technological capabilities. The increase in the latter enables detection at ever-lower levels and ignores toxicological relevance. We here provide arguments to use a Threshold of Toxicological Concern (TTC) for CAP that can accommodate developing toxicological insights. PMID:25528412

  19. 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.

  20. The effects of chloramphenicol on Ulva lactuca.

    PubMed

    Leston, Sara; Nunes, Margarida; Viegas, Ivan; Ramos, Fernando; Pardal, Miguel Ângelo

    2013-04-01

    The administration of pharmacological substances in the food producing industry is a crucial and long established practice in ensuring animal welfare. However, a very high percentage of the drugs used will directly or indirectly be present in the various compartments of natural ecosystems therefore constituting a source of pollution. The reactions that these active compounds may impose on non-target organisms are still widely unknown and further research is essential. Also, new approaches on monitoring are necessary and in this sense, the present work aimed to assess the persistence of chloramphenicol (a banned but illegally used antibiotic) in seawater, together with its effects on the growth of the green macroalgae Ulva lactuca. Moreover, the potential use of this species as a bioindicator was assessed. Results showed CAP presented an exponential degradation pattern in seawater with concentrations decreasing faster than expected. As for the effects on U. lactuca it acted as a growth promoter also contradicting the initial assumptions. Regarding the role of this species in biomonitoring it successfully took up CAP in solution while reflecting the concentrations present conferring it good characteristics as a bioindicator. On the other hand, this ability points to a possibility of CAP being accumulated and transferred along the trophic web through the consumption of U. lactuca by organisms in higher levels. PMID:23395526

  1. 21 CFR 524.390 - Chloramphenicol ophthalmic ointment.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... chloramphenicol. (b) Sponsors. See Nos. 043264 and 054771 in § 510.600(c) of this chapter. (c) Conditions of use in dogs and cats—(1) Amount. Apply every 3 hours around the clock for 48 hours, after which...

  2. 21 CFR 524.390 - Chloramphenicol ophthalmic ointment.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... chloramphenicol. (b) Sponsors. See Nos. 000856 and 043264 in § 510.600(c) of this chapter. (c) Conditions of use in dogs and cats—(1) Amount. Apply every 3 hours around the clock for 48 hours, after which...

  3. 21 CFR 524.390 - Chloramphenicol ophthalmic ointment.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... chloramphenicol. (b) Sponsors. See Nos. 000856 and 025463 in § 510.600(c) of this chapter. (c) Conditions of use in dogs and cats—(1) Amount. Apply every 3 hours around the clock for 48 hours, after which...

  4. Distribution of chloramphenicol in the genital tract of postpartum cows.

    PubMed

    Bretzlaff, K N; Ott, R S; Koritz, G D; Lock, T F; Neff-Davis, C A; Gustafsson, B K; Davis, L E

    1988-06-01

    Chloramphenicol was administered by constant IV infusion to 7 healthy postpartum cows at rates predicted to approach a steady-state plasma concentration of 5 micrograms/ml. After 8 hours of constant IV infusion, uterine tissues were removed surgically and were assayed for chloramphenicol concentrations. Mean plasma-to-tissue ratios of chloramphenicol concentrations were 3.05, 3.63 (6 cows only), and 3.22 for caruncles, endometrium, and uterine wall, respectively. Plasma-to-tissue ratios of the 3 tissues were not significantly different (P greater than 0.10). Intrauterine (IU) injections of chloramphenicol (20 mg/kg of body weight) were administered to 3 healthy post-partum cows. The mean value of the fraction of the drug absorbed from the uteri of these cows was 0.40. Mean concentrations of chloramphenicol were 43.8 micrograms/g in caruncles, 34.6 micrograms/g in endometrium, 2.8 micrograms/g in uterine wall, and 2.9 micrograms/ml in plasma 8 hours after IU injections. Chloramphenicol has now been banned for use in food-producing animals in the United States because of its potential for causing toxicosis in human beings. It is illegal to use chloramphenicol in food-producing animals in the United States and in some other countries as well. This includes use by the IU route of administration because chloramphenicol and most drugs are absorbed from the uterus into the bloodstream and are distributed to milk and tissues. PMID:3400929

  5. Bioavailability, pharmacokinetics and residues of chloramphenicol in the chicken.

    PubMed

    Anadón, A; Bringas, P; Martinez-Larrañaga, M R; Diaz, M J

    1994-02-01

    The pharmacokinetic properties of chloramphenicol were determined in broiler chickens after two single oral doses (30 and 50 mg/kg body weight) and after a single intravenous (i.v.) dose (30 mg/kg body weight). After oral and i.v. administration, the plasma concentration-time graph was characteristic of a two-compartment open model. After oral administration (30 and 50 mg/kg), chloramphenicol was absorbed rapidly (time to maximal concentration of 0.72 or 0.60 h) and eliminated with a mean half-life (t1/2 beta) of 6.87 or 7.41 h, respectively. The bioavailability was 29% at 30 mg/kg chloramphenicol and 38% at 50 mg/kg chloramphenicol. Concentrations greater than 5 micrograms/ml were achieved at 15 min and persisted up to 2 or 4 h post-administration, respectively. Statistically significant differences between the two routes of administration were found for the pharmacokinetic variables, half-lives of both distribution and elimination phases (t1/2 alpha, t1/2 beta) and apparent volume of distribution [Vd(area)]. The mean t1/2 beta of chloramphenicol and i.v. administration was 5.23 h. Chloramphenicol was extensively metabolized into dehydrochloramphenicol (DH-CAP), nitrophenylaminopropanedione (NPAP) and nitroso-chloramphenicol (NO-CAP) derivatives. Residues of chloramphenicol (CAP) and the three metabolites DH-CAP, NPAP and NO-CAP in kidney, liver and muscle were measured in chickens that received an oral dose of 50 mg/kg once daily for 4 days. The results indicate that CAP and DH-CAP residues were cleared slowly and were at or below the detection limit of 0.005 microgram/ml within 12 days after dosing. However, at the time of slaughter (12 days), the NPAP and NO-CAP residues were detected in the tissue.

  6. 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.

  7. New Insights into Chloramphenicol Biosynthesis in Streptomyces venezuelae ATCC 10712

    PubMed Central

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

    2014-01-01

    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. PMID:25267678

  8. Primary structure of a chloramphenicol acetyltransferase specified by R plasmids.

    PubMed

    Shaw, W V; Packman, L C; Burleigh, B D; Dell, A; Morris, H R; Hartley, B S

    Naturally occurring isolates of chloramphenicol-resistant bacteria commonly synthesise chloramphenicol acetyltransferase (EC 2.3.28; CAT) in amounts which are sufficient to account for the resistance phenotype and often harbour plasmids which carry the structural gene for CAT. The findings of CAT in such diverse prokaryotes as Proteus mirabilis, Agrobacterium tumefaciens, Streptomyces sp., and a soil Flavobacterium has led to speculation concerning the origin and evolution of the more commonly observed CAT variants specified by plasmids in clinically important bacteria. To provide a more solid basis for studying the evolution and spread of CAT within prokaryotes we chose to determine the complete amino acid sequence of a type I variant of CAT, the variant known to be associated with most F-like plasmids conferring chloramphenicol resistance. The sequence has been determined by combining the results obtained from manual and automated sequential degradation with those obtained by mass spectrometry of peptides generated by enzymatic digestion. The directly determined primary structure is identical with that predicted by the DNA sequence analysis of the chloramphenicol resistance transponson Tn9 known to specify a type I variant of chloramphenicol acetyltransferase.

  9. Induction of chloramphenicol and tetracycline resistance in Flexibacter sp. strain FS-1.

    PubMed Central

    Barcak, G J; Burchard, R P

    1985-01-01

    The gliding bacterium Flexibacter sp. strain FS-1 exhibits inducible resistance to chloramphenicol (Cmr) and tetracycline (Tcr). Either chloramphenicol or tetracycline alone induced a Cmr Tcr phenotype. The resistance is apparently not plasmid encoded. PMID:3855409

  10. 21 CFR 520.390b - Chloramphenicol capsules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Chloramphenicol capsules. 520.390b Section 520.390b Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED..., and bacterial infections associated with canine distemper caused by susceptible organisms....

  11. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... vitro culturing and susceptibility tests on samples collected prior to treatment. If no response to... which are raised for food production. Chloramphenicol products must not be used in meat-, egg-, or milk... be conducted, including in vitro culturing and susceptibility tests on samples collected prior...

  12. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... vitro culturing and susceptibility tests on samples collected prior to treatment. If no response to... which are raised for food production. Chloramphenicol products must not be used in meat-, egg-, or milk... be conducted, including in vitro culturing and susceptibility tests on samples collected prior...

  13. [Physiological Effect of Vallisneria natans Under Different Concentrations of Nitrogen, Phosphorus and Chloramphenicol].

    PubMed

    Hu, Zhen-zhen; Cui, Yi-bin; Li, Mei; Yu, Jing

    2015-09-01

    The effects of peroxidase(POD), superoxide dismutase(SOD) activities and malondialdehyde(MDA), soluble proteins and chlorophyll in the leaves of Vallisneria natans exposed to different concentrations of nitrogen and phosphorus in the eutrophication water body and chloramphenicol after 7 days were investigated in the study. The soluble protein content increased significantly in group eutrophic water and 0.2 µg.L-1 chloramphenicol, and the concentration of protein was 2.38 times of that in group 0 µg.L-1 chloramphenicol. In group of eutrophic water and 0. 2 µg.L-1 chloramphenicol, POD activities decreased significantly to 33. 84% of that in group 0 µg.L-1 chloramphenicol. With the increasing of the joint concentration, SOD activities decreased. SOD activities in group of mesotrophic and 0. 2 µg.L-1 chloramphenicol was 28. 59% of that in group of 0 µg.L-1 chloramphenicol. PMID:26717684

  14. A competitive enzyme-linked immunosorbent assay for determination of chloramphenicol.

    PubMed

    Wesongah, J O; Murilla, G A; Guantai, A N; Elliot, C; Fodey, T; Cannavan, A

    2007-02-01

    Chloramphenicol is a broad-spectrum antibiotic shown to have specific activity against a wide variety of organisms that are causative agents of several disease conditions in domestic animals. Chloramphenicol has been banned for use in food-producing animals for its serious adverse toxic effects in humans. Due to the harmful effects of chloramphenicol residues livestock products should be free of any traces of these residues. Several analytical methods are available for chloramphenicol analysis but sensitive methods are required in order to ensure that no traces of chloramphenicol residues are present in edible animal products. In order to prevent the illegal use of chloramphenicol, regulatory control of its residues in food of animal origin is essential. A competitive enzyme-linked immunosorbent assay for chloramphenicol has been locally developed and optimized for the detection of chloramphenicol in sheep serum. In the assay, chloramphenicol in the test samples and that in chloramphenicol-horseradish peroxidase conjugate compete for antibodies raised against the drug in camels and immobilized on a microtitre plate. Tetramethylbenzidine-hydrogen peroxide (TMB/H2O2) is used as chromogen-substrate system. The assay has a detection limit of 0.1 ng/mL of serum with a high specificity for chloramphenicol. Cross-reactivity with florfenicol, thiamphenicol, penicillin, tetracyclines and sulfamethazine was not observed. The assay was able to detect chloramphenicol concentrations in normal sheep serum for at least 1 week after intramuscular injection with the drug at a dose of 25 mg/kg body weight (b.w.). The assay can be used as a screening tool for chloramphenicol use in animals. PMID:17217404

  15. Occurrence of chloramphenicol-acetylating enzymes in various gram-negative bacilli.

    PubMed

    Okamoto, S; Suzuki, Y; Mise, K; Nakaya, R

    1967-11-01

    The occurrence of a chloramphenicol-acetylating enzyme, similar to that found in Escherichia coli, carrying an R factor was investigated in various gram-negative bacilli. The acetylated products of chloramphenicol were identified by chromatography and quantitatively assayed after benzene extraction. The investigated strains were of the Salmonella-Arizona group, the Klebsiella-Aerobacter group, Serratia marcescens, the Proteus group, and Pseudomonas aeruginosa, most of which were isolated from 1947 to 1957. Both chloramphenicol-sensitive and -resistant strains were included, but none of them was able to transfer chloramphenicol resistance by conjugation. In the Proteus group, a significant level of a chloramphenicol-acetylating enzyme was found in most strains, whether they were sensitive or resistant to chloramphenicol; the resistant strains showed higher levels of the enzyme. Some chloramphenicol-sensitive strains lacked this enzyme. Only the sensitive strains containing the enzyme could easily produce chloramphenicol-resistant mutants with higher enzyme activity. Thus, the chloramphenicol resistance of this group can be reasonably explained on the basis of the chloramphenicol-acetylating enzyme. All of the Pseudomonas aeruginosa strains were resistant to chloramphenicol, and most strains showed low levels of the enzyme (which, however, did not appear sufficient to explain their resistance). All of the strains of the other groups (except one strain of Enterobacter cloacae) lacked the enzyme, although most strains of the Klebsiella-Aerobacter group and of S. marcescens were resistant to chloramphenicol. With respect to the origin of the resistance gene of the R factor, it is noteworthy that the strains of Proteus mirabilis isolated in 1947 possessed this enzyme before the discovery of chloramphenicol.

  16. 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.

  17. Demonstration of chloramphenicol hypersensitivity by measurement of histone methylation in lymphocyte cultures

    PubMed Central

    Dobozy, A.; Hunyadi, J.; Simon, N.

    1971-01-01

    In PHA-stimulated lymphocyte cultures, chloramphenicol considerably inhibited the incorporation of thymidine, but had little influence on the methylation of histone. In lymphocyte cultures from chloramphenicol-sensitive patients, the antibiotic enhanced histone methylation, but reduced the incorporation of thymidine. The measurement of the increase of histone methylation seems to be a suitable method for the in vitro demonstration of hypersensitivity to chloramphenicol. PMID:5091617

  18. Effects of ionizing radiations on a pharmaceutical compound, chloramphenicol

    NASA Astrophysics Data System (ADS)

    Varshney, L.; Patel, K. M.

    1994-05-01

    Chloramphenicol, a broad spectrum antibiotic, has been irradiated using Cobalt-60 γ radiation and electron beam at graded radiation doses upto 100 kGy. Several degradation products and free radicals are formed on irradiation. Purity, degradation products, free radicals, discolouration, crystallinity, solubility and entropy of radiation processing have been investigated. Aqueous solutions undergo extensive radiolysis even at low doses. Physico-chemical, microbiological and toxicological tests do not show significant degradation at sterilization dose. High performance liquid chromatography (HPLC), differential scanning calorimetry (DSC), UV-spectrophotometry, diffuse reflectance spectroscopy (DRS) and electron spin resonance spectroscopy (ESR) techniques were employed for the investigations.

  19. Occurrence of chloramphenicol in crops through natural production by bacteria in soil.

    PubMed

    Berendsen, Bjorn; Pikkemaat, Mariel; Römkens, Paul; Wegh, Robin; van Sisseren, Maarten; Stolker, Linda; Nielen, Michel

    2013-05-01

    Due to the unexpected findings of the banned antibiotic chloramphenicol in products of animal origin, feed, and straw, the hypothesis was studied that the drug is naturally present in soil, through production by soil bacteria, and subsequently can be transferred to crops. First, the stability of chloramphenicol in soil was studied. The fate of chloramphenicol highly depends on soil type and showed a half-life of approximately one day in nonsterile topsoil. It was found to be more stable in subsoil and sterile soils. Second, the production of chloramphenicol in soil was studied, and it was confirmed that Streptomyces venezuelae can produce chloramphenicol at appreciable amounts in nonsterile soil. Third, a transfer study was carried out using wheat and maize grown on three different soils that were weekly exposed to aqueous chloramphenicol solutions at different levels. Chloramphenicol was taken up by crops as determined by chiral liquid chromatography coupled to tandem mass spectrometric analysis, and the levels in crops were found to be bioavailability related. It was concluded that chloramphenicol residues can occur naturally in crops as a result of the production of chloramphenicol by soil bacteria in their natural environment and subsequent uptake by crops. PMID:23570678

  20. [Influence of centrophenoxine on the effect of cyclophosphamide and chloramphenicol in teratological experiment].

    PubMed

    Neumann, H J

    1977-01-15

    Cyclophosphamide and chloramphenicol underwent a teralogical test in the animal experiment. The prenatal changes in the application of cyclophosphamide reveal the cytostatic remedy as a strong teratogenic remedy, whereas chloramphenicol in the first place has an embryotoxic effect and only a weak teratogenic one. The possibility of an influence on the prenatal changes caused by cyclophosphamide and chloramphenicol was tried with centrophenoxin. Apart from the weight of the foetuses the induced changes remained without influence in cyclophosphamide, whereas in chloramphenicol even in increase of the toxic effect could be observed.

  1. Chloramphenicol Resistance Plasmids in Escherichia coli Isolated from Diseased Piglets

    PubMed Central

    Jørgensen, Sigrid Tue

    1978-01-01

    The plasmids in 19 chloramphenicol-resistant Escherichia coli strains of three pig pathogenic antigen types were studied in conjugation and transduction experiments. The plasmids had identical resistance patterns: streptomycin, spectinomycin, sulfonamides, and chloramphenicol (Sm, Sp, Su, Cm) and belonged to IncFII. One plasmid carried ampicillin resistance in addition. Restriction enzyme analysis of the deoxyribonucleic acid from five of the plasmids originating from the same herd showed that their digestion patterns with EcoRI were indistinguishable. EcoRI cleaved the deoxyribonucleic acid of a sixth plasmid from the same herd and displayed nine of the ten bands of the other five plasmids plus an additional six. It appears that the five plasmids with identical restriction patterns have a common origin and may be copies of the same plasmid from which the sixth may have developed. Four strains carried two plasmids each. In two of these strains, a plasmid with a tetracycline marker (Tc), or possibly the tetracycline marker alone, recombined frequently with the Sm Sp Su Cm plasmid without destroying any known function of the latter. The possibility that Tc is carried on a translocation sequence is discussed. Images PMID:352263

  2. Clinical isolate of a porinless Salmonella typhi resistant to high levels of chloramphenicol.

    PubMed Central

    Toro, C S; Lobos, S R; Calderón, I; Rodríguez, M; Mora, G C

    1990-01-01

    We studied a clinical isolate of Salmonella typhi (strain 1895) characterized by resistance to 200 micrograms of chloramphenicol per ml despite the absence of chloramphenicol-inactivating activity. The outer membrane protein profile analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated a deficiency of one of the major protein species which may serve as a porin for entry of chloramphenicol. When the strain was transformed with a plasmid encoding chloramphenicol acetyltransferase, chloramphenicol added to the culture was not inactivated, suggesting a drastic reduction of permeability towards the drug. Moreover, transformants bearing a plasmid coding for the Escherichia coli OmpF porin became considerably more susceptible to chloramphenicol (40 micrograms/ml). On the other hand, transformants carrying a plasmid encoding the Salmonella typhi ompC gene remained as resistant to the drug as the parental strain, even though they overexpressed OmpC. These findings indicate that the lack of OmpF plays a major role in the resistance to chloramphenicol in strain 1895. Images PMID:2285283

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

    PubMed

    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 R(2)=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. PMID:25754387

  4. Photocatalytic degradation of the antibiotic chloramphenicol and effluent toxicity effects.

    PubMed

    Lofrano, Giusy; Libralato, Giovanni; Adinolfi, Roberta; Siciliano, Antonietta; Iannece, Patrizia; Guida, Marco; Giugni, Maurizio; Volpi Ghirardini, Annamaria; Carotenuto, Maurizio

    2016-01-01

    Chloramphenicol sodium succinate (CAP, C15H15Cl2N2 Na2O8) is a broad-spectrum antibiotic exhibiting activity against both Gram-positive and Gram-negative bacteria as well as other groups of microorganisms only partially removed by conventional activated sludge wastewater treatment plants. Thus, CAP and its metabolites can be found in effluents. The present work deals with the photocatalytic degradation of CAP using TiO2 as photocatalyst. We investigated the optimization of reaction contact time and concentration of TiO2 considering CAP and its by-products removal as well as effluent ecotoxicity elimination. Considering a CAP real concentration of 25mgL(-1), kinetic degradation curves were determined at 0.1, 0.2, 0.4, 0.8, 1.6 and 3.2gL(-1) TiO2 after 5, 10, 30, 60 and 120min reaction time. Treated samples were checked for the presence of by-products and residual toxicity (V. fischeri, P. subcapitata, L. sativum and D. magna). Results evidenced that the best combination for CAP and its by-products removal could be set at 1.6gL(-1) of TiO2 for 120min with an average residual toxicity of approximately 10%, that is the threshold set for negative controls in most toxicity tests for blank and general toxicity test acceptability.

  5. 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.

  6. 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. PMID:26452956

  7. 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

  8. 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.

  9. 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.

  10. Stability-Indicating HPTLC Method for Simultaneous Estimation of Flurbiprofen and Chloramphenicol in Ophthalmic Solution.

    PubMed

    Sadakwala, Vaishnavi M; Chauhan, Renu S; Shah, Shailesh A; Shah, Dinesh R

    2016-01-01

    A specific, accurate and reproducible stability-indicating high performance thin layer chromatography (HPTLC) method was developed for the estimation of flurbiprofen and chloramphenicol in the presence of their degradation products. Degradation studies of both the drugs were carried out in acidic, alkaline, neutral, oxidative, photolytic and thermal stress conditions. Separation was performed on thin layer chromatography plate precoated with silica gel 60 F254 using ethyl acetate : n-hexane : methanol : tri-ethyl amine (5 : 4 : 2 : 0.5, v/v/v/v). Spots at retention factor 0.29 and 0.62 were recognized as flurbiprofen and chloramphenicol, respectively, and were quantified through densitometric measurements at wavelength 267 nm. Method was found to be linear over the concentration range 12-60 ng/spot with correlation coefficient of 0.9997 for flurbiprofen and 200-1,000 ng/spot with correlation coefficient of 0.9977 for chloramphenicol. The proposed method was applied to the estimation of flurbiprofen and chloramphenicol in commercial ophthalmic formulation. The developed HPTLC method can be applied for routine analysis of flurbiprofen and chloramphenicol in the presence of their degradation products in their individual as well as combined pharmaceutical formulations.

  11. 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. PMID:19459958

  12. Treatment of acne vulgaris comparing two similar lotion formulations, one with ('Actinac') and one without chloramphenicol.

    PubMed

    Fraser, N B; Main, R A; Stewart, T W; Thornton, E J

    1980-01-01

    A double-blind, randomized study was carried out in 37 patients with acne vulgaris to compare the efficacy of two similar lotion formulations, one with ('Actinac') and the other without chloramphenicol (4%). Patients used one or other of the two formulations twice daily for the first 4 days and then at night only for the remainder of the 3-month trial period. At the end of the first month's treatment, the group treated with the full formulation showed a statistically significant greater reduction in lesion count score than in the group receiving the formulation without chloramphenicol. Both treatments produced statistically significant reductions in lesion counts at all monthly follow-up visits. After 3-months' treatment, the mean lesion count score was reduced from 28.8 to 9.3 in the full formulation-treated group and from 39.5 to 14.3 in the formulation minus chloramphenicol group, but this difference did not reach statistical significance.

  13. Modifications and adaptations of the Charm II rapid antibody assay for chloramphenicol in honey.

    PubMed

    McMullen, Sarah E; Lansden, John A; Schenck, Frank J

    2004-07-01

    The Charm II screening method for the presence of chloramphenicol in honey has a sensitivity of 0.3 ppb. This screening method is a simple, rapid antibody assay using [3H]chloramphenicol and a binding reagent. Analysis of different types of honey revealed considerable differences in results. Honey can be liquid, crystallized (creamed), or partially crystallized and is classified by the U.S. Department of Agriculture into seven color categories: water white, extra white, white, extra light amber, light amber, amber, and dark amber. Fortified and nonfortified liquid amber honey tested appropriately with the Charm II unit and the negative control provided with the unit after slight modifications were made. However, approximately 70% of creamed honey samples fortified at 0.6 ppb did not test positive for the presence of chloramphenicol using the provided negative control. Matrix quenching effects were evaluated, and these effects were accounted for by establishing different assay conditions for different honey types.

  14. Measurement of Three Antibiotics (Penicillin, Cephalothin, and Chloramphenicol) When Present Together in Mixtures

    PubMed Central

    Sabath, Leon D.; Loder, P. Bronwen; Gerstein, Deborah A.; Finland, Maxwell

    1968-01-01

    Benzylpenicillin, cephalothin, and chloramphenicol were measured individually and quantitatively when present together in mixtures at concentrations found in patients. The assay depended on the selective inactivation of two of the antibiotics, permitting the third to be assayed as if present alone. Agar seeded with a chloramphenicol-resistant, penicillinase-negative Staphylococcus aureus was used for a cylinder-plate assay of appropriately diluted fluid to determine the activity of either benzylpenicillin or cephalothin after either β-lactamase inactivation of the former or ultraviolet light inactivation of the latter. Chloramphenicol was assayed with Sarcina lutea after appropriate β-lactamase inactivation of both cephalothin and benzylpenicillin. Fluids containing various amounts of the three antibiotics were assayed by this method with less than 8% error. Procedures that fail to measure each antibiotic individually may give larger errors. PMID:5187067

  15. Effect of chloramphenicol and ethidium bromide on the level of ornithine carbamoyltransferase in Neurospora crassa

    SciTech Connect

    Zerez, C.R.; Weiss, R.L.

    1986-05-01

    The specific activity of the nuclear-gene-encoded, mitochondrial arginine biosynthetic enzyme ornithine carbamoyltransferase in Neurospora crassa was elevated in mycelia treated with chloramphenicol or ethidium bromide. The increase in specific activity was caused by an increase in the number of mature enzyme molecules rather than by the activation of a preexisting enzyme. Chloramphenicol and ethidium bromide appeared to act indirectly via arginine-mediated derepression. However, derepression did not appear to result from a drug-mediated decrease in the arginine pool.

  16. Preparation and characterization of chloramphenicol niosomes and comparison with chloramphenicol eye drops (0.5%w/v) in experimental conjunctivitis in albino rabbits.

    PubMed

    Yasin, Muhammad Naveed; Hussain, Shahzad; Malik, Farnaz; Hameed, Abdul; Sultan, Tipu; Qureshi, Fahim; Riaz, Humayun; Perveen, Ghazala; Wajid, Amina

    2012-01-01

    Niosomes has gained tremendous popularity as ultimate drug carrier. Lot of research work is being carried out on preparation of niosomes for ophthalmic use having no significant effect on vision and its sustained release pattern. Chloramphenicol niosomes were prepared using two different ratios of cholesterol, drug and surfactant, termed as EIN-1, EIN-2 by ether injection method and their entrapment efficiency, particle size. The in vitro drug release pattern was observed for ten hours. The EIN-2 showed 90% entrapment and released 81% of entrapped drug after 10 hours. Zeta potential & viscosity were determined and in-vivo comparison was made with Chloramphenicol eye drops where it exhibited Cmax of 15 μ g/ml. Stability studies were done to determine shelf life. MIC of selected strain of S. aureus was also determined. EIN 2 niosomal suspension was compared with Chloramphenicol eye drops in experimental conjunctivitis in albino rabbits. In-vitro studies are encouraging as niosomes released about 75% of total entrapped drug by EIN-1 and 81% of total entrapped drug by EIN 2. In vivo study shows that niosomes released the drug in eye in acceptable range and showed a sustained release pattern without affecting the vision. Niosomes were found ultimate ophthalmic drug carriers capable to release drug in sustained and determined pattern.

  17. Responses of Wild-Type and Resistant Strains of the Hyperthermophilic Bacterium Thermotoga maritima to Chloramphenicol Challenge▿ †

    PubMed Central

    Montero, Clemente I.; Johnson, Matthew R.; Chou, Chung-Jung; Conners, Shannon B.; Geouge, Sarah G.; Tachdjian, Sabrina; Nichols, Jason D.; Kelly, Robert M.

    2007-01-01

    Transcriptomes and growth physiologies of the hyperthermophile Thermotoga maritima and an antibiotic-resistant spontaneous mutant were compared prior to and following exposure to chloramphenicol. While the wild-type response was similar to that of mesophilic bacteria, reduced susceptibility of the mutant was attributed to five mutations in 23S rRNA and phenotypic preconditioning to chloramphenicol. PMID:17557852

  18. An in vitro synergistic interaction of combinations of Thymus glabrescens essential oil and its main constituents with chloramphenicol.

    PubMed

    Ilić, Budimir S; Kocić, Branislava D; Cirić, Vojislav M; Ćvetković, Olga G; Miladinović, Dragoljub L

    2014-01-01

    The chemical composition and antibacterial activity of Thymus glabrescens Willd. (Lamiaceae) essential oil were examined, as well as the association between it and chloramphenicol. The antibacterial activities of geraniol and thymol, the main constituents of T. glabrescens oil, individually and in combination with chloramphenicol, were also determined. The interactions of the essential oil, geraniol, and thymol with chloramphenicol toward five selected strains were evaluated using the microdilution checkerboard assay in combination with chemometric methods. Oxygenated monoterpenes were the most abundant compound class in the oil, with geraniol (22.33%) as the major compound. The essential oil exhibited in vitro antibacterial activity against all tested bacterial strains, but the activities were lower than those of the standard antibiotic and thymol. A combination of T. glabrescens oil and chloramphenicol produced a strong synergistic interaction (FIC indices in the range 0.21-0.87) and a substantial reduction of the MIC value of chloramphenicol, thus minimizing its adverse side effects. The combinations geraniol-chloramphenicol and thymol-chloramphenicol produced synergistic interaction to a greater extent, compared with essential oil-chloramphenicol association, which may indicate that the activity of the thyme oil could be attributed to the presence of significant concentrations of geraniol and thymol. PMID:24616649

  19. The chloramphenicol acetyltransferase gene of Tn2424: a new breed of cat.

    PubMed

    Parent, R; Roy, P H

    1992-05-01

    We have sequenced the gene coding for the chloramphenicol acetyltransferase of Tn2424 of plasmid NR79. This gene codes for a protein of 23,500 Da, and the derived protein sequence is similar to those of the chromosomal chloramphenicol acetyltransferases of Agrobacterium tumefaciens and Pseudomonas aeruginosa and of unidentified open reading frames, which may encode chloramphenicol acetyltransferases, adjacent to the ermG macrolide-lincosamide-streptogramin resistance gene of Bacillus sphaericus and the vgb virginiamycin resistance gene of Staphylococcus aureus. Weaker similarity to the LacA (thiogalactoside acetyltransferase) and CysE (serine acetyltransferase) proteins of Escherichia coli and the NodL protein of Rhizobium leguminosarum is also observed. There is no significant similarity to any other chloramphenicol acetyltransferase genes, such as that of Tn9. The Tn2424 cat gene is part of a 4.5-kb region which also contains the aacA1a aminoglycoside-6'-N-acetyltransferase gene; Tn2424 is similar to Tn21 except for the presence of this region. Sequences flanking the cat gene are typical of those flanking other genes inserted into pVS1-derived "integrons" by a site-specific recombinational mechanism.

  20. 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

  1. 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.

  2. Antagonism of ampicillin and chloramphenicol for meningeal isolates of group B streptococci.

    PubMed Central

    Weeks, J L; Mason, E O; Baker, C J

    1981-01-01

    The increasing prevalence of ampicillin-resistant Haemophilus influenzae type b has led to the recommendation that ampicillin and chloramphenicol be given as the initial therapy for suspected bacterial meningitis in infants and children. However, during the first 2 months of life, H. influenzae type b is a rare cause of meningitis, whereas group B streptococcus is the most frequently isolated agent. Since ampicillin and chloramphenicol have been shown to be antagonistic for other streptococci, an in vitro study of their effect on group B streptococci was performed. The effect of ampicillin and chloramphenicol, alone and in combination, on 18 meningeal isolates was determined for 2 different inocula of group B streptococci, using microtiter broth dilution and growth kinetic assays. Isoboles, fractional lethal concentration indices, or both indicated antagonism for all strains. Growth kinetic assays for two representative strains demonstrated inhibition of the early bactericidal activity of ampicillin by chloramphenicol. These findings of in vitro antagonism suggest that this combination may be contraindicated for the treatment of infants with group B streptococcal meningitis. PMID:7030197

  3. Eriobotrya japonica counteracts reactive oxygen species and nitric oxide stimulated by chloramphenicol.

    PubMed

    Eraso, Alberto Jorge; Albesa, Inés

    2007-01-01

    Chloramphenicol is a toxic antibiotic used for certain infections, though aplastic anaemia is one of its side-effects. The results of our experiments showed that blood cells suffered oxidative stress in the presence of chloramphenicol, with a significant increase in reactive oxygen species (ROS) detected by luminol-chemiluminescence (CL). The extract of fruits of Eriobotrya japonica markedly decreased ROS in leukocytes and erythrocytes, the oxidative stress caused by this antibiotic. Nitro Blue Tetrazolium (NBT) assay with purified leukocytes demonstrated that the antioxidant action of E. japonica caused an intracellular reduction in ROS, and that the extracts decreased these promoters of oxidative stress to normal levels in the cytoplasm. Determinations of nitric oxide (NO) generation indicated that E. japonica extracts also inhibited the stimuli of NO provoked by chloramphenicol. This study showed that the immediate antioxidant effect of E. japonica could be associated with the action of vitamin A. The protective action of this fruit was seen on mature leukocytes and erythrocytes, beneficial effect on blood cells suggest that its extract could be used as an antioxidant agent complementing the administration of chloramphenicol, as a modern-day extension to its traditional use in Chinese medicine.

  4. Chloramphenicol- and tetracycline-resistant uropathogenic Escherichia coli (UPEC) exhibit reduced virulence potential.

    PubMed

    Starcic Erjavec, Marjanca; Rijavec, Matija; Krizan-Hergouth, Veronika; Fruth, Angelika; Zgur-Bertok, Darja

    2007-11-01

    It is well documented that uropathogenic Escherichia coli (UPEC) isolates resistant to nalidixic acid have reduced virulence potential. Our goal was to assess whether UPEC isolates resistant to chloramphenicol, tetracycline and streptomycin also exhibit reduced virulence potential. Among 110 human UPEC isolates, the prevalences of the virulence factors fimH, papC, papGII, papGIII, sfa/focDE, afa, hlyA, cnf1, usp, ibeA, fyuA, iroN, iucD, ireA, and K1 and K5 capsules as well as of pathotypes, phylogenetic groups, O antigens and a pathogenicity island (PAI) marker were compared between chloramphenicol-, tetracycline-, streptomycin- and, as a control, nalidixic acid-resistant and -susceptible strains. Our findings show that among human UPEC isolates, not only nalidixic acid-resistant but also chloramphenicol- and tetracycline-resistant isolates have reduced virulence potential compared with susceptible strains. To our knowledge, this is the first report of a statistically significant reduction in virulence traits among chloramphenicol- and tetracycline-resistant isolates.

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

    PubMed

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

    2003-07-01

    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).

  6. Highly sensitive determination of chloramphenicol based on thin-layered MoS2/polyaniline nanocomposite.

    PubMed

    Yang, Tao; Chen, Huaiyin; Ge, Tong; Wang, Jin; Li, Weihua; Jiao, Kui

    2015-11-01

    The nanocomposite of molybdenum disulfide (MoS2) and polyaniline (PANI) was prepared through in situ polymerization of aniline on the surface and interlayer of thin-layered MoS2. Owing to the physisorption of aromatic aniline onto the basal plane of MoS2, the electrochemical properties of MoS2/PANI nanocomposite were improved. And a novel electrochemical sensor based on MoS2/PANI nanocomposite was used to determine chloramphenicol by differential pulse voltammetry, exhibiting excellent performance. The detection range was from 1×10(-7) mol L(-1) to 1×10(-4) mol L(-1), with a high sensitivity and a low detection limit of 6.9×10(-8) mol L(-1). In addition, this sensor can be used for the determination of chloramphenicol in real samples. PMID:26452965

  7. 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.

  8. Exocrine pancreatic function in man after treatment with oxytetracycline and chloramphenicol.

    PubMed

    Fleischer, K

    1976-01-01

    The exocrine pancreatic function was studied in humans by performing a secretin-cholecystokinin test before and after treatment with oxytetracycline or chloramphenicol. In the oxytetracycline-treated patients there was a depression of the amylase and lipase outputs in the duodenal secretion, chymotrypsin decreasing only slightly. After treatment with the two antibiotics the calcium secretion was reduced. The other parameters measured in the duodenal secretion remained essentially unchanged. The enzyme dissociation observed in the present studies is considered to reflect the onset of pancreatic dysfunction due to antibiotic administration. As in the previous animal onset of pancreatic dysfunction due to antibiotic administration. As in the previous animal experiments, the suggested explanation for the changes in enzyme secretion is an inhibition of protein synthesis in the exocrine pancreas due to oxytetracycline and chloramphenicol. PMID:950076

  9. Highly sensitive determination of chloramphenicol based on thin-layered MoS2/polyaniline nanocomposite.

    PubMed

    Yang, Tao; Chen, Huaiyin; Ge, Tong; Wang, Jin; Li, Weihua; Jiao, Kui

    2015-11-01

    The nanocomposite of molybdenum disulfide (MoS2) and polyaniline (PANI) was prepared through in situ polymerization of aniline on the surface and interlayer of thin-layered MoS2. Owing to the physisorption of aromatic aniline onto the basal plane of MoS2, the electrochemical properties of MoS2/PANI nanocomposite were improved. And a novel electrochemical sensor based on MoS2/PANI nanocomposite was used to determine chloramphenicol by differential pulse voltammetry, exhibiting excellent performance. The detection range was from 1×10(-7) mol L(-1) to 1×10(-4) mol L(-1), with a high sensitivity and a low detection limit of 6.9×10(-8) mol L(-1). In addition, this sensor can be used for the determination of chloramphenicol in real samples.

  10. Induction of cat-86 by chloramphenicol and amino acid starvation in relaxed mutants of Bacillus subtilis.

    PubMed Central

    Ambulos, N P; Rogers, E J; Alexieva, Z; Lovett, P S

    1988-01-01

    The chloramphenicol acetyltransferase gene cat-86 is induced through a mechanism that is a variation of classical attenuation. Induction results from the destabilization of an RNA stem-loop that normally sequesters the cat-86 ribosome-binding site. Destabilization of the stem-loop is due to the stalling of a ribosome in the leader region of cat-86 mRNA at a position that places the A site of the stalled ribosome at leader codon 6. Two events can stall ribosomes at the correct location to induce cat-86 translation: addition of chloramphenicol to cells and starvation of cells for the amino acid specified by leader codon 6. Induction by amino acid starvation is an anomaly because translation of the cat-86 coding sequence requires all 20 amino acids. To explain this apparent contradiction we postulated that amino acid starvation triggers intracellular proteolysis, thereby providing levels of the deprived amino acid sufficient for cat-86 translation. Here we show that a mutation in relA, the structural gene for stringent factor, blocks intracellular proteolysis that is normally triggered by amino acid starvation. The relA mutation also blocks induction of cat-86 by amino acid starvation, but the mutation does not interfere with chloramphenicol induction. Induction by amino acid starvation can be demonstrated in relA mutant cells if the depleted amino acid is restored at very low levels (e.g., 2 micrograms/ml). A mutation in relC, which may be the gene for ribosomal protein L11, blocks induction of cat-86 by either chloramphenicol or amino acid starvation. We believe this effect is due to a structural alteration of the ribosome resulting from the relC mutation and not to the relaxed phenotype of the cells. PMID:3142854

  11. Chloramphenicol induction of cat-86 requires ribosome stalling at a specific site in the leader.

    PubMed

    Alexieva, Z; Duvall, E J; Ambulos, N P; Kim, U J; Lovett, P S

    1988-05-01

    The plasmid gene cat-86 specifies chloramphenicol-inducible chloramphenicol acetyltransferase in Bacillus subtilis. Induction by the antibiotic is primarily due to activation of the translation of cat-86-encoded mRNA. It has been suggested that the inducer stalls ribosomes at a discrete location in the leader region of cat-86 mRNA, which causes the destabilization of a downstream RNA secondary structure that normally sequesters the cat-86 ribosome binding site. It is the destabilization of this RNA secondary structure that permits translation of the cat-86 coding sequence. In the present report, we show that ribosomes that were stalled in the cat-86 leader by starvation of host cells for the amino acid specified by leader codon 6 induced gene expression to a level above that detected when cells were starved for the amino acids specified by leader codons 7 and 8. Starvation for amino acids specified by leader codons 3, 4, or 5 failed to activate cat-86 expression. These results indicate that the stalled ribosome that is most active in cat-86 induction has its aminoacyl site occupied by leader codon 6. To determine if chloramphenicol also stalled ribosomes in the cat-86 regulatory leader such that the aminoacyl site was occupied by codon 6, we separately changed leader codons 3, 4, 5, and 6 to the translation termination (ochre) codon TAA. Each of the mutated genes was tested for its ability to be induced by chloramphenicol. The results show that replacement of leader codons 3, 4, or 5 by the ochre codon blocked induction, whereas replacement of leader codon 6 by the ochre codon permitted induction. Collectively, these observations lead to the conclusion that cat-86 induction requires ribosome stalling in leader mRNA, and they identify leader codon 6 as the codon most likely to be occupied by the aminoacyl site of a stalled ribosome that is active in the induction.

  12. Induction of cat-86 by chloramphenicol and amino acid starvation in relaxed mutants of Bacillus subtilis.

    PubMed

    Ambulos, N P; Rogers, E J; Alexieva, Z; Lovett, P S

    1988-12-01

    The chloramphenicol acetyltransferase gene cat-86 is induced through a mechanism that is a variation of classical attenuation. Induction results from the destabilization of an RNA stem-loop that normally sequesters the cat-86 ribosome-binding site. Destabilization of the stem-loop is due to the stalling of a ribosome in the leader region of cat-86 mRNA at a position that places the A site of the stalled ribosome at leader codon 6. Two events can stall ribosomes at the correct location to induce cat-86 translation: addition of chloramphenicol to cells and starvation of cells for the amino acid specified by leader codon 6. Induction by amino acid starvation is an anomaly because translation of the cat-86 coding sequence requires all 20 amino acids. To explain this apparent contradiction we postulated that amino acid starvation triggers intracellular proteolysis, thereby providing levels of the deprived amino acid sufficient for cat-86 translation. Here we show that a mutation in relA, the structural gene for stringent factor, blocks intracellular proteolysis that is normally triggered by amino acid starvation. The relA mutation also blocks induction of cat-86 by amino acid starvation, but the mutation does not interfere with chloramphenicol induction. Induction by amino acid starvation can be demonstrated in relA mutant cells if the depleted amino acid is restored at very low levels (e.g., 2 micrograms/ml). A mutation in relC, which may be the gene for ribosomal protein L11, blocks induction of cat-86 by either chloramphenicol or amino acid starvation. We believe this effect is due to a structural alteration of the ribosome resulting from the relC mutation and not to the relaxed phenotype of the cells.

  13. Chloramphenicol induction of cat-86 requires ribosome stalling at a specific site in the leader.

    PubMed Central

    Alexieva, Z; Duvall, E J; Ambulos, N P; Kim, U J; Lovett, P S

    1988-01-01

    The plasmid gene cat-86 specifies chloramphenicol-inducible chloramphenicol acetyltransferase in Bacillus subtilis. Induction by the antibiotic is primarily due to activation of the translation of cat-86-encoded mRNA. It has been suggested that the inducer stalls ribosomes at a discrete location in the leader region of cat-86 mRNA, which causes the destabilization of a downstream RNA secondary structure that normally sequesters the cat-86 ribosome binding site. It is the destabilization of this RNA secondary structure that permits translation of the cat-86 coding sequence. In the present report, we show that ribosomes that were stalled in the cat-86 leader by starvation of host cells for the amino acid specified by leader codon 6 induced gene expression to a level above that detected when cells were starved for the amino acids specified by leader codons 7 and 8. Starvation for amino acids specified by leader codons 3, 4, or 5 failed to activate cat-86 expression. These results indicate that the stalled ribosome that is most active in cat-86 induction has its aminoacyl site occupied by leader codon 6. To determine if chloramphenicol also stalled ribosomes in the cat-86 regulatory leader such that the aminoacyl site was occupied by codon 6, we separately changed leader codons 3, 4, 5, and 6 to the translation termination (ochre) codon TAA. Each of the mutated genes was tested for its ability to be induced by chloramphenicol. The results show that replacement of leader codons 3, 4, or 5 by the ochre codon blocked induction, whereas replacement of leader codon 6 by the ochre codon permitted induction. Collectively, these observations lead to the conclusion that cat-86 induction requires ribosome stalling in leader mRNA, and they identify leader codon 6 as the codon most likely to be occupied by the aminoacyl site of a stalled ribosome that is active in the induction. PMID:3129723

  14. Transmissible Resistance to Penicillin G, Neomycin, and Chloramphenicol in Rhizobium japonicum1

    PubMed Central

    Cole, Michael A.; Elkan, Gerald H.

    1973-01-01

    The genetic basis for resistance to a number of antibiotics was examined in Rhizobium japonicum. Resistance to penicillin G, neomycin, and chloramphenicol appears to be mediated by an extrachromosomal element similar to that found in the Enterobacteriaceae. Resistance to these antibiotics was eliminated from cells by treatment with acridine orange, and resistance to all three antibiotics could be transferred en bloc to Agrobacterium tumefaciens under conditions excluding transformation or transduction as possible genetic mechanisms. PMID:4491197

  15. 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.

  16. In vitro assessment of chloramphenicol and florfenicol as second-line antimicrobial agents in dogs.

    PubMed

    Maaland, M G; Mo, S S; Schwarz, S; Guardabassi, L

    2015-10-01

    The aim of this study was to evaluate the potential of chloramphenicol and florfenicol as second-line antimicrobial agents for treatment of infections caused by methicillin-resistant Staphylococcus pseudintermedius (MRSP) and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in dogs, through a systematic in vitro assessment of the pharmacodynamic properties of the two drugs. Minimum inhibitory concentrations (MIC) and phenicol resistance genes were determined for 169 S. pseudintermedius and 167 E. coli isolates. Minimum bactericidal concentrations (MBC), time-killing kinetics, and postantibiotic effect (PAE) of both agents against wild-type isolates of each species were assessed. For S. pseudintermedius, the chloramphenicol MIC90 was 32 μg/mL. No florfenicol resistance was detected in this species (MIC90 = 4 μg/mL). The MIC90 of both agents against E. coli was 8 μg/mL. Resistance genes found were catpC221 in S. pseudintermedius and catA1 and/or floR in E. coli. The phenicols displayed a time-dependent, mainly, bacteriostatic effect on both species. Prolonged PAEs were observed for S. pseudintermedius, and no PAEs were detected for E. coli. More research into determination of PK/PD targets of efficacy is needed to further assess the clinical use of chloramphenicol and florfenicol as second-line agents in dogs, optimize dosage regimens, and set up species-specific clinical break points. PMID:25623169

  17. The Circadian Rhythm in Photosynthesis in Acetabularia in the Presence of Actinomycin D, Puromycin, and Chloramphenicol

    PubMed Central

    Sweeney, Beatrice M.; Tuffli, Charles F.; Rubin, Richard H.

    1967-01-01

    Anucleate Acetabularia crenulata shows a circadian rhythm in photosynthesis. In this study, an oxygen electrode was employed to measure this photosynthetic rhythm in the presence and absence of the inhibitors, actinomycin D, chloramphenicol, and puromycin. High concentrations of the inhibitors were used: actinomycin D, 20–40 µg ml-1 puromycin, 30 and 100 µg ml-1; and chloramphenicol, 250 µg ml-1. The effectiveness of these inhibitors on protein synthesis was also measured under the same conditions used for the determination of rhythmicity. In spite of large effects of all three inhibitors on the incorporation of 14C leucine, no effect on the period or the phase of the photosynthetic rhythm was observed. The higher concentration of puromycin and chloramphenicol produced toxic effects which were expressed as a reduction in the amount of photosynthesis, but rhythmicity was still apparent. After 3 or 4 days' exposure to actinomycin, Acetabularia became resistant to its effect. Recovery was also observed in the ability to incorporate leucine. The implications of these results for theories of the basic oscillator responsible for circadian rhythmicity are discussed. PMID:11526851

  18. The circadian rhythm in photosynthesis in Acetabularia in the presence of actinomycin D, puromycin, and chloramphenicol.

    PubMed

    Sweeney, B M; Tuffli, C F; Rubin, R H

    1967-01-01

    Anucleate Acetabularia crenulata shows a circadian rhythm in photosynthesis. In this study, an oxygen electrode was employed to measure this photosynthetic rhythm in the presence and absence of the inhibitors, actinomycin D, chloramphenicol, and puromycin. High concentrations of the inhibitors were used: actinomycin D, 20-40 micrograms ml-1; puromycin, 30 and 100 micrograms ml-1; and chloramphenicol, 250 micrograms ml-1. The effectiveness of these inhibitors on protein synthesis was also measured under the same conditions used for the determination of rhythmicity. In spite of large effects of all three inhibitors on the incorporation of 14C leucine, no effect on the period or the phase of the photosynthetic rhythm was observed. The higher concentration of puromycin and chloramphenicol produced toxic effects which were expressed as a reduction in the amount of photosynthesis, but rhythmicity was still apparent. After 3 or 4 days' exposure to actinomycin, Acetabularia became resistant to its effect. Recovery was also observed in the ability to incorporate leucine. The implications of these results for theories of the basic oscillator responsible for circadian rhythmicity are discussed.

  19. 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

  20. 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

  1. Evidence of natural occurrence of the banned antibiotic chloramphenicol in herbs and grass.

    PubMed

    Berendsen, Bjorn; Stolker, Linda; de Jong, Jacob; Nielen, Michel; Tserendorj, Enkhtuya; Sodnomdarjaa, Ruuragchas; Cannavan, Andrew; Elliott, Christopher

    2010-07-01

    Chloramphenicol (CAP), a broad-spectrum antibiotic, was detected in several herb and grass samples from different geographic origins. Due to its suspected carcinogenicity and linkages with the development of aplastic anemia in humans, CAP is banned for use in food-producing animals in the European Union (EU) and many other countries. However, products of animal origin originating from Asian countries entering the European market are still found noncompliant (containing CAP) on a regular basis, even when there is no history of chloramphenicol use in these countries. A possible explanation for the continued detection of these residues is the natural occurrence of CAP in plant material which is used as animal feed, with the consequent transfer of the substance to the animal tissues. Approximately 110 samples were analyzed using liquid chromatography coupled with mass spectrometric detection. In 26 samples, the presence of CAP was confirmed using the criteria for banned substances defined by the EU. Among other plant materials, samples of the Artemisia family retrieved from Mongolia and from Utah, USA, and a therapeutic herb mixture obtained from local stores in the Netherlands proved to contain CAP at levels ranging from 0.1 to 450 microg/kg. These findings may have a major impact in relation to international trade and safety to the consumer. The results of this study demonstrate that noncompliant findings in animal-derived food products may in part be due to the natural occurrence of chloramphenicol in plant material. This has implications for the application of current EU, USA, and other legislation and the interpretation of analytical results with respect to the consideration of CAP as a xenobiotic veterinary drug residue and the regulatory actions taken upon its detection in food. PMID:20431869

  2. 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.

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

    PubMed Central

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

    1984-01-01

    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. Images PMID:6327638

  4. The leader peptides of attenuation-regulated chloramphenicol resistance genes inhibit translational termination.

    PubMed Central

    Moffat, J G; Tate, W P; Lovett, P S

    1994-01-01

    Placing a translation stop codon at the ribosomal pause site in the leader of the attenuation-regulated cat-86 gene activates cat expression in the absence of the inducer, chloramphenicol. Genetic experiments have shown that this phenomenon depends on the amino acid sequence of the leader-encoded peptide and could readily be explained if the peptide was an inhibitor of translation termination. Here we demonstrate that the cat-86 leader pentapeptide is an in vitro inhibitor of translation termination in addition to its previously described antipeptidyltransferase activity. PMID:7961481

  5. 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.

  6. 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

    2015-03-01

    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. PMID:25682427

  7. Pharmacokinetics of a long-acting chloramphenicol formulation administered by intramuscular and subcutaneous routes in cattle.

    PubMed

    Sanders, P; Guillot, P; Mourot, D

    1988-06-01

    The pharmacokinetic properties of a long-acting formulation of chloramphenicol were determined in six yearling cattle after a single intravenous (i.v.) administration (40 mg/kg body weight) and after two sequential subcutaneous (s.c.) or intramuscular (i.m.) administrations (90 mg/kg/48 h). The two extravascular routes were studied during a crossover trial for a bioequivalence test. After i.v. administration, the plasma concentration-time graph was characteristic of a two-compartment open model. Mean values were a half-life of 4.1 h, a volume of distribution of 0.86 l/kg and a body clearance of 0.128 l/kg/h. Plasma concentrations of chloramphenicol following i.m. and s.c. administrations increased slowly to a broad peak at 10-15 micrograms/ml between 9 and 12 h. Bioavailability was 19.1% after i.m. injection and 12.4% after s.c. administration. The extent of absorption from the two routes did not differ significantly. The rate of absorption was significantly lower after s.c. application than it was after i.m. injection. The time necessary for the plasma concentration to exceed 5 micrograms/ml was the same for the two routes. Thus, i.m. and s.c. routes are bioequivalent.

  8. 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.

  9. 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

  10. 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.

  11. Isocratic liquid chromatographic determination of theophylline, acetaminophen, chloramphenicol, caffeine, anticonvulsants, and barbiturates in serum.

    PubMed

    Meatherall, R; Ford, D

    1988-01-01

    An isocratic reverse-phase high-performance liquid chromatographic system is described for resolving theophylline, acetaminophen, caffeine, chloramphenicol, ethosuximide, primidone, phenobarbital, phenytoin, and carbamazepine within 7 min. A procedure for routinely measuring these drugs in serum is validated and has been extended to include the quantification of N-desmethylmethsuximide, barbital, amobarbital, secobarbital, pentobarbital, mephobarbital, and thiopental. A 250 x 4.6 mm column packed with trimethylsilyl (C-1)-coated 5-microns particles is used. The mobile phase is phosphate buffer (10 mmol/L, pH 6.3):methanol:acetonitrile, 65:17.5:17.5. A common solvent extraction procedure is used for all of these drugs. The extractant is chloroform:isopropanol (95:5), containing three internal standards: 3-isobutyl-l-methylxanthine (IMX), tolybarb, and methsuximide. Theophylline, acetaminophen, caffeine, and chloramphenicol are quantified at 273 nm with IMX as the internal standard. With two exceptions, the rest of the drugs are quantified at 204 nm using tolybarb as the internal standard; ethosuximide is quantified at 204 nm using methsuximide as the internal standard, and thiopental is quantified at 285 nm using IMX as the internal standard.

  12. 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

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

    PubMed

    Yang, Mengrui; Wang, Min; 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.

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

    PubMed

    Yang, Mengrui; Wang, Min; 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

  15. 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

  16. Synergistic killing of NDM-producing MDR Klebsiella pneumoniae by two ‘old’ antibiotics—polymyxin B and chloramphenicol

    PubMed Central

    Abdul Rahim, Nusaibah; Cheah, Soon-Ee; Johnson, Matthew D.; Yu, Heidi; Sidjabat, Hanna E.; Boyce, John; Butler, Mark S.; Cooper, Matthew A.; Fu, Jing; Paterson, David L.; Nation, Roger L.; Bergen, Phillip J.; Velkov, Tony; Li, Jian

    2015-01-01

    Objectives Combination therapy is an important option in the fight against Gram-negative ‘superbugs’. This study systematically investigated bacterial killing and the emergence of polymyxin resistance with polymyxin B and chloramphenicol combinations used against New Delhi metallo-β-lactamase (NDM)-producing MDR Klebsiella pneumoniae. Methods Four NDM-producing K. pneumoniae strains were employed. The presence of genes conferring resistance to chloramphenicol was examined by PCR. Time–kill studies (inocula ∼106 cfu/mL) were conducted using various clinically achievable concentrations of each antibiotic (range: polymyxin B, 0.5–2 mg/L; chloramphenicol, 4–32 mg/L), with real-time population analysis profiles documented at baseline and 24 h. The microbiological response was examined using the log change method and pharmacodynamic modelling in conjunction with scanning electron microscopy (SEM). Results Multiple genes coding for efflux pumps involved in chloramphenicol resistance were present in all strains. Polymyxin B monotherapy at all concentrations produced rapid bacterial killing followed by rapid regrowth with the emergence of polymyxin resistance; chloramphenicol monotherapy was largely ineffective. Combination therapy significantly delayed regrowth, with synergy observed in 25 out of 28 cases at both 6 and 24 h; at 24 h, no viable bacterial cells were detected in 15 out of 28 cases with various combinations across all strains. No polymyxin-resistant bacteria were detected with combination therapy. These results were supported by pharmacodynamic modelling. SEM revealed significant morphological changes following treatment with polymyxin B both alone and in combination. Conclusions The combination of polymyxin B and chloramphenicol used against NDM-producing MDR K. pneumoniae substantially enhanced bacterial killing and suppressed the emergence of polymyxin resistance. PMID:26023209

  17. 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

  18. 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.

  19. 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. PMID:26496159

  20. Chloramphenicol Mediates Superoxide Production in Photosystem II and Enhances Its Photodamage in Isolated Membrane Particles

    PubMed Central

    Rehman, Ateeq Ur; Kodru, Sandeesha; Vass, Imre

    2016-01-01

    Chloramphenicol (CAP) is an inhibitor of protein synthesis, which is frequently used to decouple photodamage and protein synthesis dependent repair of Photosystem II during the process of photoinhibition. It has been reported earlier that CAP is able to mediate superoxide production by transferring electrons from the acceptor side of Photosystem I to oxygen. Here we investigated the interaction of CAP with Photosystem II electron transport processes by oxygen uptake and variable chlorophyll fluorescence measurements. Our data show that CAP can accept electrons at the acceptor side of Photosystem II, most likely from Pheophytin, and deliver them to molecular oxygen leading to superoxide production. In addition, the presence of CAP enhances photodamage of Photosystem II electron transport in isolated membrane particles, which effect is reversible by superoxide dismutase. It is concluded that CAP acts as electron acceptor in Photosystem II and mediates its superoxide dependent photodamage. This effect has potential implications for the application of CAP in photoinhibitory studies in intact systems. PMID:27092170

  1. 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

  2. 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.

  3. Comparative efficacy of chloramphenicol loaded chondroitin sulfate and dextran sulfate nanoparticles to treat intracellular Salmonella infections.

    PubMed

    Kiruthika, V; Maya, S; Suresh, Maneesha K; Kumar, V Anil; Jayakumar, R; Biswas, Raja

    2015-03-01

    Salmonella Paratyphi A is a food-borne Gram-negative pathogen and a major public health challenge in the developing world. Upon reaching the intestine, S. Paratyphi A penetrates the intestinal epithelial barrier; and infects phagocytes such as macrophages and dendritic cells. S. Paratyphi A surviving within macrophages is protected from the lethal action of antibiotics due to their poor penetration into the intracellular compartments. Hence we have developed chloramphenicol loaded chondroitin sulfate (CS-Cm Nps) and dextran sulfate (DS-Cm Nps) nanoparticles through ionotropic-gelation method for the intracellular delivery of chloramphenicol. The size of these nanoparticles ranged between 100 and 200 nm in diameter. The encapsulation efficiency of both the nanoparticles was found to be around 65%. Both the nanoparticles are found to be non-hemolytic and non-toxic to fibroblast and epithelial cells. The prepared nanoparticles exhibited sustained release of the drug of up to 40% at pH 5 and 20-25% at pH 7.0 after 168 h. The anti-microbial activities of both nanoparticles were tested under in vitro and ex vivo conditions. The delivery of DS-Cm Nps into the intracellular compartments of the macrophages was 4 fold more compared to the CS-Cm Nps which lead to the enhanced intracellular antimicrobial activity of Ds-Cm Nps. Enhanced anti-microbial activity of Ds-Cm Nps was further confirmed in an ex vivo chicken intestine infection model. Our results showed that Cm loaded DS Nps can be used to treat intracellular Salmonella infections.

  4. Comparative efficacy of chloramphenicol loaded chondroitin sulfate and dextran sulfate nanoparticles to treat intracellular Salmonella infections.

    PubMed

    Kiruthika, V; Maya, S; Suresh, Maneesha K; Kumar, V Anil; Jayakumar, R; Biswas, Raja

    2015-03-01

    Salmonella Paratyphi A is a food-borne Gram-negative pathogen and a major public health challenge in the developing world. Upon reaching the intestine, S. Paratyphi A penetrates the intestinal epithelial barrier; and infects phagocytes such as macrophages and dendritic cells. S. Paratyphi A surviving within macrophages is protected from the lethal action of antibiotics due to their poor penetration into the intracellular compartments. Hence we have developed chloramphenicol loaded chondroitin sulfate (CS-Cm Nps) and dextran sulfate (DS-Cm Nps) nanoparticles through ionotropic-gelation method for the intracellular delivery of chloramphenicol. The size of these nanoparticles ranged between 100 and 200 nm in diameter. The encapsulation efficiency of both the nanoparticles was found to be around 65%. Both the nanoparticles are found to be non-hemolytic and non-toxic to fibroblast and epithelial cells. The prepared nanoparticles exhibited sustained release of the drug of up to 40% at pH 5 and 20-25% at pH 7.0 after 168 h. The anti-microbial activities of both nanoparticles were tested under in vitro and ex vivo conditions. The delivery of DS-Cm Nps into the intracellular compartments of the macrophages was 4 fold more compared to the CS-Cm Nps which lead to the enhanced intracellular antimicrobial activity of Ds-Cm Nps. Enhanced anti-microbial activity of Ds-Cm Nps was further confirmed in an ex vivo chicken intestine infection model. Our results showed that Cm loaded DS Nps can be used to treat intracellular Salmonella infections. PMID:25645750

  5. An Unusual Peroxo Intermediate of the Arylamine Oxygenase of the Chloramphenicol Biosynthetic Pathway

    PubMed Central

    Makris, Thomas M.; Vu, Van V.; Meier, Katlyn K.; Komor, Anna J.; Rivard, Brent S.; Münck, Eckard; Que, Lawrence; Lipscomb, John D.

    2015-01-01

    Streptomyces venezuelae CmlI catalyzes the 6-electron oxygenation of the arylamine precursor of chloramphenicol in a nonribosomal peptide synthetase (NRPS)-based pathway to yield the nitroaryl group of the antibiotic. Optical, EPR, and Mössbauer studies show that the enzyme contains a nonheme dinuclear iron cluster. Addition of O2 to the diferrous state of the cluster results in an exceptionally long-lived intermediate (t1/2 = 3 h at 4 °C) that is assigned as a peroxodiferric species (CmlI-peroxo) based upon the observation of an 18O2-sensitive resonance Raman (rR) vibration. CmlI-peroxo is spectroscopically distinct from the well characterized and commonly observed cis-μ-1,2-peroxo (μ-η1:η1) intermediates of nonheme diiron enzymes. Specifically, it exhibits a blue-shifted broad absorption band around 500 nm and a rR spectrum with a ν(O–O) that is at least 60 cm−1 lower in energy. Mössbauer studies of the peroxo state reveal a diferric cluster having iron sites with small quadrupole splittings and distinct isomer shifts (0.54 and 0.62 mm/s). Taken together, the spectroscopic comparisons clearly indicate that CmlI-peroxo does not have a μ-η1:η1-peroxo ligand; we propose that a μ-η1:η2-peroxo ligand accounts for its distinct spectroscopic properties. CmlI-peroxo reacts with a range of arylamine substrates by an apparent second order process, indicating that CmlI-peroxo is the reactive species of the catalytic cycle. Efficient production of chloramphenicol from the free arylamine precursor suggests that CmlI catalyzes the ultimate step in the biosynthetic pathway, and that the precursor is not bound to the NRPS during this step. PMID:25564306

  6. 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

  7. Construction and Use of a Replication-Competent Human Immunodeficiency Virus (HIV-1) that Expresses the Chloramphenicol Acetyltransferase Enzyme

    NASA Astrophysics Data System (ADS)

    Terwilliger, E. F.; Godin, B.; Sodroski, J. G.; Haseltine, W. A.

    1989-05-01

    The construction and properties of an infectious human immunodeficiency virus (HIV) that expresses the bacterial gene chloramphenicol acetyltransferase are described. This virus can be used in vitro to screen for drugs that inhibit HIV infection. The marked virus may also be used to trace the routes of infection from the site of inoculation in animal experiments.

  8. Anti-peptidyl transferase leader peptides of attenuation-regulated chloramphenicol-resistance genes.

    PubMed Central

    Gu, Z; Harrod, R; Rogers, E J; Lovett, P S

    1994-01-01

    The chloramphenicol (Cm)-inducible cmlA gene of Tn1696 specifies nonenzymatic resistance to Cm and is regulated by attenuation. The first eight codons of the leader specify a peptide that inhibits peptidyl transferase in vitro. Functionally similar, but less inhibitory, peptides are encoded by the leaders of Cm-inducible cat genes. However, the cat and cmlA coding sequences are unrelated and specify proteins of unrelated function. The inhibition of peptidyl transferase by the leader peptides is additive with that of Cm. Erythromycin competes with the inhibitory action of the peptides, and erythromycin and the peptides footprint to overlapping sites at the peptidyl transferase center of 23S rRNA. It is proposed that translation of the cmlA and cat leaders transiently pauses upon synthesis of the inhibitor peptides. The predicted site of pausing is identical to the leader site where long-term occupancy by a ribosome (ribosome stalling) will activate downstream gene expression. We therefore propose the inducer, Cm, converts a peptide-paused ribosome to the stalled state. We discuss the idea that cooperativity between leader peptide and inducer is necessary for ribosome stalling and may link the activation of a specific drug-resistance gene with a particular antibiotic. Images PMID:7515506

  9. 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.

  10. 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. PMID:27101357

  11. Determination of chloramphenicol residues in crustaceans: preparation and evaluation of a proficiency test in Germany.

    PubMed

    Polzer, J; Hackenberg, R; Stachel, C; Gowik, P

    2006-11-01

    Chloramphenicol (CAP) is banned for use in food-producing animals and is, thus, controlled on the basis of the National Residue Control Plans in the European Union. Due to current problems with residues of CAP in shrimp, crayfish and prawns, a sensitive GC/NCI/MS method was optimised and in-house validated. The validation study resulted in a decision limit (CCalpha) of 0.07 microg kg-1, a recovery of 95% and a within-laboratory reproducibility of 9%. The method was used for preparing a proficiency test to assess the quality of residue control in Germany. The proficiency test involved analysis of five samples and the results were very satisfactory. The reproducibility standard deviation for five samples ranged from 17 to 24%, and the median concentrations lay between 0.43 and 0.51 microg kg-1 CAP. These values are clearly below the corresponding Horwitz standard deviation of about 50%. From the study, it can be concluded that there are, irrespective of the method applied, well-established and proper working analytical procedures for the control of CAP around the minimum required performance limit (MRPL) of 0.3 microg kg-1. PMID:17071515

  12. 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.

  13. 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.

  14. Development and physical characterization of chloramphenicol loaded biodegradable nanoparticles for prolonged release.

    PubMed

    Mandal, B; Halder, K K; Dey, S K; Bhoumik, M; Debnath, M C; Ghosh, L K

    2009-07-01

    The objectives of our study were to prepare a biodegradable nanoparticulate system of chloramphenicol (CHL) and to evaluate its ability to prolong in vitro release of CHL compared to free drug suspension (FDS). CHL-loaded polylactide-co-glycolide nanoparticles (CHL-PLGA-NPs) were prepared by an emulsion/solvent evaporation method using ethyl acetate and polyvinyl alcohol. CHL-PLGA-NPs were characterized by particle size, zeta potential, infrared spectra, drug entrapment efficiency and in vitro release kinetics measurement. Sonication was done with an ultrasound pulse sonicator at 70 W, 30 kHz for 60 s to produce stable NPs of mean size range from 277 nm to 433 nm. Drug to polymer ratio (D:P) was selected as formulation variable and significantly influenced entrapment efficiency (approximately 30% to 66%) and release (p < 0.05). Entrapment of CHL in biodegradable NPs significantly prolonged drug release compared to FDS and thus implies potential antibiotic delivery system for ocular application.

  15. 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. PMID:26031849

  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. Direct detection of chloramphenicol in honey by neutral desorption-extractive electrospray ionization mass spectrometry.

    PubMed

    Huang, X Y; Fang, X W; Zhang, X; Dai, X M; Guo, X L; Chen, H W; Luo, L P

    2014-11-01

    Herein, we constructed a platform of neutral desorption-extractive electrospray ionization mass spectrometry (ND-EESI-MS) for direct and rapid detection of chloramphenicol (CAP) in honey samples diluted with methanol. Under the optimized working conditions, the quantitative information of CAP residues was acquired effectively by EESI-Ion Trap MS (n) . Using heated methanol-N2 as spray reagent, we reduced the limit of determination (LOD) from 73.3 ng/mL to 0.3 ng/mL, and the CAP detection is linear in the range of 1-5000 ng/mL (R = 0.9947). For the honey samples with CAP of 10, 100, and 1000 ng/mL, the recoveries were 133.0, 80.6, and 101.1%, and the relative standard deviations were 5.96, 8.82, and 8.71%, respectively. The reproducibility assays showed the stability of this method. Therefore, this ND-EESI-MS method is powerful for direct, rapid, and quantitative CAP analysis in honey samples with high sensitivity, precision, and specificity. PMID:25277102

  18. Electrochemical sensor for chloramphenicol based on novel multiwalled carbon nanotubes@molecularly imprinted polymer.

    PubMed

    Yang, Guangming; Zhao, Faqiong

    2015-02-15

    Herein, we present a novel electrochemical sensor for the determination of chloramphenicol (CAP), which is based on multiwalled carbon nanotubes@molecularly imprinted polymer (MWCNTs@MIP), mesoporous carbon (CKM-3) and three-dimensional porous graphene (P-r-GO). Firstly, 3-hexadecyl-1-vinylimidazolium chloride (C16VimCl) was synthetized by using 1-vinylimidazole and 1-chlorohexadecane as precursors. Then, C16VImCl was used to improve the dispersion of MWCNT and as monomer to prepare MIP on MWCNT surface to obtain MWCNTs@MIP. After that, the obtained MWCNTs@MIP was coated on the CKM-3 and P-r-GO modified glassy carbon electrode to construct an electrochemical sensor for the determination of CAP. The parameters concerning this assay strategy were carefully considered. Under the optimal conditions, the electrochemical sensor offered an excellent response for CAP. The linear response ranges were 5.0 × 10(-9)-5 × 10(-7)mol L(-1) and 5.0 × 10(-7)-4.0 × 10(-6), respectively, and the detection limit was 1.0 × 10(-10)mol L(-1). The electrochemical sensor was applied to determine CAP in real samples with satisfactory results.

  19. Fluorescent aptasensor for chloramphenicol detection using DIL-encapsulated liposome as nanotracer.

    PubMed

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

    2016-07-15

    A novel fluorescence aptasensor was successfully developed to respond to chloramphenicol (CAP) in food based on magnetic aptamer-liposome vesicle probe. In order to fabricate it, aptamer labeled on functionalized magnetic beads (MB) was firstly employed as capture adsorbent (MB-Apt), then SSB (single-stranded DNA binding protein) and DIL (1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanineperchlorate) coimmobilized liposomes (SSB/DIL-Lip) was employed as vesicle signal tracer. The composite vesicle probe is formed between SSB/DIL-Lip and MB-Apt based on SSB's specific recognition towards aptamer on vesicle signal tracer. Upon the vesicle probe solution reacted with CAP, the aptamer on the magnetic beads preferentially bounded with CAP, and then released SSB/DIL-Lip vesicle signal tracer in the supernatant after magnetic separation. The released tracer can emit fluorescence which was correspondence with the concentration of the analyte. At the optimum conditions, the aptasensor exhibited a good linear response for CAP detection in the range of 0.003-10nM with a detection limit of 1pM. Importantly, the methodology was further validated for analyzing CAP in fish samples with consistent results obtained by ELISA kit, thus providing a promising approach for quantitative monitoring of CAP and significant anti-interference ability in food safety. PMID:27015148

  20. 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

  1. Simultaneous determination of chloramphenicol, florfenicol and florfenicol amine in ham sausage with a hybrid chemiluminescent immunoassay.

    PubMed

    Tao, Xiaoqi; Jiang, Haiyang; Yu, Xuezhi; Zhu, Jinghui; Wang, Xia; Wang, Zhanhui; Niu, Lanlan; Wu, Xiaoping; Shen, Jianzhong

    2013-01-01

    A novel chemiluminescent immunoassay utilising two types of primary antibodies (murine monoclonal antibody and rabbit polyclonal antibody) and two types of horseradish peroxidase-labelled secondary antibodies was established for simultaneously detecting multiple amphenicol residues in ham sausage. After combining the extract procedure of the target amphenicol into one simplified method, this hybrid chemiluminescent immunoassay could screen chloramphenicol (CAP), florfenicol (FF) and its metabolite florfenicol amine (FFA) at the same time by adding the corresponding secondary antibody. Ham sausage samples were analysed by using this hybrid immunoassay, with LODs of CAP being 0.01 μg kg⁻¹, of FF being 2.8 μg kg⁻¹ and of FFA being 3.0 μg kg⁻¹. The applicability of the proposed method has been validated by determining CAP, FF and FFA in ham sausage samples with satisfactory results. Good recoveries and high correlation with traditional enzyme-linked immunosorbent assay and LC-MS/MS results illustrated that the developed hybrid chemiluminescent immunoassay could screen high-throughput ultra-trace amphenicol residues effectively at one time.

  2. 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

    2012-11-01

    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.

  3. Possible protective role of chloramphenicol in TSST-1 and coagulase-positive Staphylococcus aureus-induced septic arthritis with altered levels of inflammatory mediators.

    PubMed

    Majumdar, Sayantani; Dutta, Kallol; Manna, Sunil K; Basu, Anirban; Bishayi, Biswadev

    2011-08-01

    Chloramphenicol is mostly used against coagulase-negative Staphylococcus aureus, and its protective role against coagulase-positive S. aureus is not well studied. In our study, arthritis was induced in mice by S. aureus (Apollo Gleneagles 33 (AG-33) or American Type Culture Collection 25923 (ATCC-25923)) infection. Chloramphenicol was administered after 2 h of infection. Mice were killed at 1, 3, 5 days post-infection. Mice inoculated with pathogenic Staphylococci (AG-33) expressing coagulase and Toxic shock syndrome toxin-1 (TSST-1), displayed severe arthritis with enhanced bacterial burden in the spleen, cytokine production in serum and synovial tissue, neutrophil recruitment, and cyclooxegenase-2 expression in synovial tissue compared with ATCC-25923-infected groups. Severity of arthritis was regulated by chloramphenicol treatment. Our study suggests that alteration in the inflammatory cytokine levels and pronounced production of cyclooxygenase-2 play important roles in progression of arthritis which is regulated by application of chloramphenicol.

  4. Changing antibiotic sensitivity pattern and scope of chloramphenicol in the management of hospitalised patients of typhoid fever.

    PubMed

    Beig, Farzana K; Ahmad, Farida; Abqari, Shaad

    2009-12-01

    Enteric fever is a global health problem and there is emerging drug resistance with some reports of re-emerging sensitivity to previously used antibiotics eg, chloramphenicol. This study was done to compare the drug sensitivity pattern of enteric fever over a decade period. Twenty-five culture positive patients for S typhi from one study done between October 1993 to February 1995 and 35 positive patients from another study done between September 2005 and August 2006 in the same hospital were taken into account and their sensitivity pattern was compared. A total of 36% of cases were sensitive to all the drugs and equal number were multidrug resistant in the study in 1995, while in 2006 only 14.2% (p<0.05) were sensitive to all drugs and percentage of multidrug resistance has increased to 42% (p>0.05). Sensitivity to chloramphenicol had decreased from 68.00% to 54.30% (p>0.05). With the emergence of resistant strains there is significant decline in overall sensitivity to all first line drugs. However there is no change in chloramphenicol sensitivity of Salmonella enterica serovar typhi 10 years back and now, this alone is not the sufficient reason for reintroduction of this drug especially in younger patients. PMID:20509470

  5. Validation of a rapid and sensitive routine method for determination of chloramphenicol in honey by LC-MS/MS.

    PubMed

    Taka, Tsuyoshi; Baras, Marina C; Chaudhry Bet, Zahra F

    2012-01-01

    Chloramphenicol (CAP) is a broad spectrum antibiotic used in the treatment of human and animal diseases. However, CAP can exhibit toxic effects in certain susceptible individuals, causing bone marrow depression, including fatal aplastic anemia. As this condition is dose-independent, CAP has been banned for use in food-producing animals, including honeybees. In this study, a quick, simple and low-cost routine analytical method was developed for the screening and confirmation of chloramphenicol in honey by LC-MS/MS. Sample clean-up takes only two steps without SPE procedure and with recoveries >97%. Honey samples were selected from several producers in Brazil and diluted in a small amount of water. After fortification and addition of d (s)-chloramphenicol as internal standard, the samples were extracted with ethyl acetate. Complete validation of the method was performed on the basis of EU decision 2002/657. Within-laboratory CV reproducibility at the lowest concentration was <10%. An evaluation of two different methods to calculate the decision limit and detection capability gave 0.08 µg kg(-1) for CCα and 0.12 µg kg(-1) for CCβ. PMID:22088167

  6. Misfolding of chloramphenicol acetyltransferase due to carboxy-terminal truncation can be corrected by second-site mutations.

    PubMed

    Van der Schueren, J; Robben, J; Volckaert, G

    1998-12-01

    Folding of chloramphenicol acetyltransferase (CAT) in Escherichia coli is hampered by deletion of the carboxy-terminal tail including the last residue of the carboxy-terminal alpha-helix. Such truncated CAT polypeptides quantitatively aggregate into cytoplasmic inclusion bodies, which results in absence of a chloramphenicol-resistant phenotype for the producing host. In this paper, a genetic approach is presented to examine this aggregation process in more detail. Random mutagenesis of inactive CAT followed by direct phenotypic selection for revertants with restored chloramphenicol resistance was used to isolate second-site suppressors of inactive truncation mutants of CAT. Two random mutagenesis procedures, independently of each other, yielded a unique substitution of Phe for Leu at amino acid position 145. This second-site mutation does not drastically affect the proteins' stability under normal growth conditions of E. coli. Hence, the introduction of Phe at amino acid position 145 improves the ability of the protein to fold into a soluble, enzymatically active conformation. The conservative character of the Leu145Phe replacement indicates that limited changes at crucial positions can have important effects on protein folding in vivo.

  7. 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.

  8. Chloramphenicol-resistant Salmonella newport traced through hamburger to dairy farms. A major persisting source of human salmonellosis in California.

    PubMed

    Spika, J S; Waterman, S H; Hoo, G W; St Louis, M E; Pacer, R E; James, S M; Bissett, M L; Mayer, L W; Chiu, J Y; Hall, B

    1987-03-01

    Animal-to-human transmission of drug-resistant salmonella and the role of antimicrobial use in food animals in the emergence of these bacteria are controversial subjects. Investigation of a 4.9-fold increase in Salmonella newport isolations from Californians in 1985 showed that 87 percent of the isolates had an unusual antimicrobial-resistance pattern (including chloramphenicol resistance) and a single, identical plasmid. Interviews of 45 patients and 89 matched controls in Los Angeles County showed that illness was associated with penicillin or tetracycline use during the month before onset (P less than 0.001) and with eating ground beef during the week before onset (P = 0.052). The epidemic strain was isolated from hamburger products eaten by cases, abattoirs where the animals from which the meat came were slaughtered, dairies that sent cows for slaughter on days when culture-positive products were processed, and ill dairy cows. Isolation of salmonella from beef carcasses in abattoirs correlated with the proportion of dead or moribund animals received for slaughter (r = 0.60, P less than 0.05). Isolation of chloramphenicol-resistant salmonella from dairy farms was associated with the use of chloramphenicol at those dairies. We conclude that food animals are a major source of antimicrobial-resistant salmonella infections in humans and that these infections are associated with antimicrobial use on farms. PMID:3807951

  9. 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

    2016-06-01

    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.

  10. Performance characteristics of an analytical procedure for determining chloramphenicol residues in muscle tissue by gas chromatography-electron capture detection.

    PubMed

    Cerkvenik-Flajs, Vesna

    2006-10-01

    Validation of an analytical method for determining chloramphenicol residues in muscle tissue by gas chromatography-electron capture detection (GC-ECD) was performed according to the latest European Union criteria for the analysis of veterinary drugs in food, laid down by Commission Decision 2002/657/EC. The method using the meta isomer of chloramphenicol as an internal standard proved to be very selective, specific to other related phenicols and accurate to within +3.6% at a concentration level of 8.9 microg/kg, as present in the certified reference material available. The correlation coefficient of the calibration curve was 0.9991. At all three fortification levels studied (0.3, 0.45 and 0.6 microg/kg), repeatability and intra-laboratory reproducibility were <8 and < or =9%, respectively. The decision limit (CCalpha) and detection capability (CCbeta) were 0.07 and 0.12 microg/kg, respectively. The validation results and the results of participation in an international inter-laboratory proficiency test indicate that the method presented is completely suited for regulatory control to screen and quantify chloramphenicol residues in various muscle tissues on a routine basis.

  11. 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.

  12. Assessment of commutability for candidate certified reference material ERM-BB130 "chloramphenicol in pork".

    PubMed

    Zeleny, Reinhard; Emteborg, Håkan; Schimmel, Heinz

    2010-10-01

    Chloramphenicol (CAP), an effective antibiotic against many microorganisms, is meanwhile banned in the EU for treatment of food-producing animals due to adverse health effects. The Institute for Reference Materials and Measurements (IRMM) is currently developing a certified reference material (CRM) for CAP in pork, intended for validation and method performance verifications of analytical methods. The material will be certified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-mass spectrometry (GC-MS) methods and has a target CAP level around the minimum required performance limit (MRPL) of 0.3 microg/kg. To prove that the material can be applied as a quality control tool for screening methods, a commutability study was conducted, involving five commercially available enzyme-linked immunosorbent assay kits and one biosensor assay (BiaCore kit). Meat homogenates (cryo-milled wet tissue) with CAP concentrations around the MRPL and the candidate CRM (lyophilised powder) were measured by LC-MS/MS and GC-MS as well as the six screening methods. Pairwise method comparisons of results obtained for the two sample types showed that the CRM can successfully be applied as quality control (QC) sample to all six screening methods. The study suggests that ERM-BB130 is sufficiently commutable with the investigated assays and that laboratories applying one of the investigated kits therefore benefit from using ERM-BB130 to demonstrate the correctness of their results. However, differences among the assays were observed, either in the abundance of bias between screening and confirmatory LC and GC methods, the repeatability of test results, or goodness of fit between the methods. PMID:20665007

  13. Crystal structure of CmlI, the arylamine oxygenase from the chloramphenicol biosynthetic pathway.

    PubMed

    Knoot, Cory J; Kovaleva, Elena G; Lipscomb, John D

    2016-09-01

    The diiron cluster-containing oxygenase CmlI catalyzes the conversion of the aromatic amine precursor of chloramphenicol to the nitroaromatic moiety of the active antibiotic. The X-ray crystal structures of the fully active, N-terminally truncated CmlIΔ33 in the chemically reduced Fe(2+)/Fe(2+) state and a cis μ-1,2(η (1):η (1))-peroxo complex are presented. These structures allow comparison with the homologous arylamine oxygenase AurF as well as other types of diiron cluster-containing oxygenases. The structural model of CmlIΔ33 crystallized at pH 6.8 lacks the oxo-bridge apparent from the enzyme optical spectrum in solution at higher pH. In its place, residue E236 forms a μ-1,3(η (1):η (2)) bridge between the irons in both models. This orientation of E236 stabilizes a helical region near the cluster which closes the active site to substrate binding in contrast to the open site found for AurF. A very similar closed structure was observed for the inactive dimanganese form of AurF. The observation of this same structure in different arylamine oxygenases may indicate that there are two structural states that are involved in regulation of the catalytic cycle. Both the structural studies and single crystal optical spectra indicate that the observed cis μ-1,2(η (1):η (1))-peroxo complex differs from the μ-η (1):η (2)-peroxo proposed from spectroscopic studies of a reactive intermediate formed in solution by addition of O2 to diferrous CmlI. It is proposed that the structural changes required to open the active site also drive conversion of the µ-1,2-peroxo species to the reactive form. PMID:27229511

  14. 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.

  15. 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

  16. 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

  17. Sensitive voltammetric determination of chloramphenicol by using single-wall carbon nanotube-gold nanoparticle-ionic liquid composite film modified glassy carbon electrodes.

    PubMed

    Xiao, Fei; Zhao, Faqiong; Li, Jiangwen; Yan, Rui; Yu, Jingjing; Zeng, Baizhao

    2007-07-16

    A novel composite film modified glassy carbon electrode has been fabricated and characterized by scanning electron microscope (SEM) and voltammetry. The composite film comprises of single-wall carbon nanotube (SWNT), gold nanoparticle (GNP) and ionic liquid (i.e. 1-octyl-3-methylimidazolium hexafluorophosphate), thus has the characteristics of them. The resulting electrode shows good stability, high accumulation efficiency and strong promotion to electron transfer. On it, chloramphenicol can produce a sensitive cathodic peak at -0.66 V (versus SCE) in pH 7.0 phosphate buffer solutions. Parameters influencing the voltammetric response of chloramphenicol are optimized, which include the composition of the film and the operation conditions. Under the optimized conditions, the peak current is linear to chloramphenicol concentration in the range of 1.0x10(-8)-6.0x10(-6) M, and the detection limit is estimated to be 5.0x10(-9) M after an accumulation for 150 s on open circuit. The electrode is applied to the determination of chloramphenicol in milk samples, and the recoveries for the standards added are 97.0% and 100.3%. In addition, the electrochemical reaction of chloramphenicol and the effect of single-wall carbon nanotube, gold nanoparticle and ionic liquid are discussed.

  18. 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.

  19. 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

  20. 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.

  1. Characterization of the myelotoxicity of chloramphenicol succinate in the B6C3F1 mouse.

    PubMed

    Turton, John A; Fagg, Rajni; Sones, William R; Williams, Thomas C; Andrews, C Michael

    2006-04-01

    Chloramphenicol (CAP) is haemotoxic in man, inducing two types of toxicity. First, a dose-related, reversible anaemia with reticulocytopenia, sometimes seen in conjunction with leucopenia and thrombocytopenia; this form of toxicity develops during drug treatment. The second haemotoxicity is aplastic anaemia (AA) which is evident in the blood as severe pancytopenia. AA development is not dose-related and occurs weeks or months after treatment. We wish, in the longer term, to investigate CAP-induced AA in the busulphan-pretreated mouse. However, as a prelude to that study, we wanted to characterize in detail the reversible haemotoxicity of CAP succinate (CAPS), administered at high dose levels in the mouse, and follow the recovery of the bone marrow in the post-dosing period. Female B6C3F1 mice were gavaged with CAPS at 0, 2500 and 3500 mg/kg, daily, for 5 days and sampled (n = 5) at 1, 7, 14 and 21 days post-dosing. Blood, bone marrow and spleen samples were analysed and clonogenic assays carried out. At day 1 post-dosing, at both CAPS dose levels, decreases were seen in erythrocytes and erythrocyte precursors; marrow erythroid cells were reduced. Reductions were also evident in splenic nucleated cell counts, blood high fluorescence ratio (HFR) reticulocyte counts and total reticulocyte counts; burst-forming units-erythroid and colony-forming units-erythroid showed decreases. At day 7 post-dosing (2500 mg/kg CAPS), there was regeneration of erythrocyte production, with marked splenic erythropoietic activity, and raised blood HFR reticulocytes. At day 7, at 3500 mg/kg CAPS, erythrocyte and reticulocyte parameters remained depressed. At 14 days post-dosing (2500 mg/kg CAPS), many erythrocyte parameters had returned to normal; at 3500 mg/kg CAPS, there was erythroid regeneration. By 21 days post-dosing, at both CAPS dose levels, most erythrocytic parameters were equivalent to control values. For leucocyte parameters, there was some depression at day 1 post-dosing (at

  2. 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

    2016-11-01

    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. PMID:27516318

  3. 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

    2016-11-01

    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.

  4. 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.

  5. 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

  6. Validation of a method for the determination of chloramphenicol in poultry and swine liver by ultra-performance liquid chromatography coupled with tandem mass spectrometry.

    PubMed

    Xia, Xi; Li, Xiaowei; Ding, Shuangyang; Shen, Jianzhong

    2010-01-01

    A sensitive and reliable method has been developed and validated for the determination of chloramphenicol in poultry and swine liver using SPE and ultra-performance liquid chromatography (UPLC)/MS/MS. The liver samples were extracted with ethyl acetate, defatted with n-hexane, and further cleaned up using SPE cartridges with polymeric sorbent. An Acquity BEH C18 column was used for gradient UPLC separation, with water and acetonitrile as the mobile phase. The multiple reaction monitoring mode was used for two precursor-product ion transitions for chloramphenicol and one for the internal standard. The method was validated at 0.1, 0.3, and 1.0 microg/kg. Mean recoveries from fortified samples ranged from 95.5 to 106.7% with an RSD of 12.2%. The method LOD was < 0.02 microg/kg. PMID:21140679

  7. Label-free and sensitive aptasensor based on dendritic gold nanostructures on functionalized SBA-15 for determination of chloramphenicol.

    PubMed

    Bagheri Hashkavayi, Ayemeh; Raoof, Jahan Bakhsh; Azimi, Razieh; Ojani, Reza

    2016-04-01

    A highly sensitive and low-cost electrochemical aptasensor was developed for the determination of chloramphenicol (CAP). The system was based on a CAP-binding aptamer, a molecular recognition element, and 1,4-diazabicyclo[2.2.2]octane (DABCO)-supported mesoporous silica SBA-15 on the surface of a screen-printed graphite electrode for formation of dendritic gold nanostructures and improving the performance and conductivity of the biosensor. Hemin has been applied as an electrochemical indicator which interacted with the guanine bases of the aptamer. In the absence of CAP, hemin binds to the aptamer and produces a weak differential pulse voltammetric (DPV) signal. The presence of CAP led to stabilization of the folded aptamer, which generated an amplified DPV signal. The peak current of hemin increased linearly with the concentration of CAP. Under optimal conditions, two linear ranges were obtained from 0.03 to 0.15 μM and 0.15 to 7.0 μM, respectively, and the detection limit was 4.0 nM. The prepared biosensor has good selectivity against other non-target drugs. Thus, the sensor could provide a promising platform for the fabrication of aptasensors. The feasibility of using this aptasensor was demonstrated by determination of CAP in a human blood serum sample. PMID:26879648

  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. PMID:26264269

  9. Comparative inhibition of chloramphenicol acetyltransferase gene expression by antisense oligonucleotide analogues having alkyl phosphotriester, methylphosphonate and phosphorothioate linkages.

    PubMed Central

    Marcus-Sekura, C J; Woerner, A M; Shinozuka, K; Zon, G; Quinnan, G V

    1987-01-01

    Several classes of oligonucleotide antisense compounds of sequence complementary to the start of the mRNA coding sequence for chloramphenicol acetyl transferase (CAT), including methylphosphonate, alkyltriester, and phosphorothioate analogues of DNA, have been compared to "normal" phosphodiester oligonucleotides for their ability to inhibit expression of plasmid-directed CAT gene activity in CV-1 cells. CAT gene expression was inhibited when transfection with plasmid DNA containing the gene for CAT coupled to simian virus 40 regulatory sequences (pSV2CAT) or the human immunodeficiency virus enhancer (pHIVCAT) was carried out in the presence of 30 microM concentrations of analogue. For the oligo-methylphosphonate analogue, inhibition was dependent on both oligomer concentration and chain length. Analogues with phosphodiester linkages that alternated with either methylphosphonate, ethyl phosphotriester, or isopropyl phosphotriester linkages were less effective inhibitors, in that order. The phosphorothioate analogue was about two-times more potent than the oligo-methylphosphonate, which was in turn approximately twice as potent as the normal oligonucleotide. Images PMID:3475677

  10. Development of a subcritical water extraction approach for trace analysis of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry tissues.

    PubMed

    Xiao, Zhiming; Song, Rong; Rao, Zhenghua; Wei, Shulin; Jia, Zheng; Suo, Decheng; Fan, Xia

    2015-10-30

    Subcritical water extraction was investigated as a novel and alternative technology for the separation of trace amounts of chloramphenicol, thiamphenicol, florfenicol and its major metabolite florfenicol amine from poultry tissues and its results were compared with those of conventional shaking extraction, ultrasonic extraction, and pressurized liquid extraction. Decreasing the polarity of water by successively increasing the extraction temperature from 50°C to 200°C at the moderate pressure enabled selective, highly effective extractions to be performed. Rapid quantification of the target compounds was carried out by ultra-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). The critical parameters of subcritical water extraction such as solvent modifier, temperature, pressure, extraction time, and static cycles were varied with control. The optimized extraction procedures using subcritical water as extraction solvent, were carried out on a pressurized liquid extractor operated at 150°C and 100bar, applying two static cycles for 3min. Average recoveries of the four analytes from fortified samples ranged between 86.8% and 101.5%, with relative standard deviations (RSDs) lower than 7.7%. The limits of detection (LODs) and quantification (LOQs) for the target compounds were in the ranges of 0.03-0.5μgkg(-1) and 0.1-2.0μgkg(-1), respectively. The proposed method is fast, sensitive, water-based thus more environmental acceptable, making it a suitable replacement for conventional organic solvent extraction in veterinary drug residue analysis.

  11. Development of Suberin Fatty Acids and Chloramphenicol-Loaded Antimicrobial Electrospun Nanofibrous Mats Intended for Wound Therapy.

    PubMed

    Tamm, Ingrid; Heinämäki, Jyrki; Laidmäe, Ivo; Rammo, Liisi; Paaver, Urve; Ingebrigtsen, Sveinung G; Škalko-Basnet, Nataša; Halenius, Anna; Yliruusi, Jouko; Pitkänen, Pauliina; Alakurtti, Sami; Kogermann, Karin

    2016-03-01

    Suberin fatty acids (SFAs) isolated from outer birch bark were investigated as an antimicrobial agent and biomaterial in nanofibrous mats intended for wound treatment. Electrospinning (ES) was used in preparing the composite nonwoven nanomats containing chloramphenicol (CAM; as a primary antimicrobial drug), SFAs, and polyvinylpyrrolidone (as a carrier polymer for ES). The X-ray powder diffraction, differential scanning calorimetry, scanning electron microscopy, atomic force microscopy, and texture analysis were used for the physicochemical and mechanical characterization of the nanomats. ES produced nanofibrous mats with uniform structure and with an average fiber diameter ranging from 370 to 425 nm. Microcrystalline SFAs and crystalline CAM were found to undergo a solid-state transformation during ES processing. The ES process caused also the loss of CAM in the final nanofibers. In the texture analysis, the SFAs containing nanofibers exhibited significantly higher maximum detachment force to an isolated pig skin (p < 0.05) than that obtained with the reference nanofibers. CAM exists in an amorphous form in the nanofibers which needs to be taken into account in controlling the physical storage stability. In conclusion, homogeneous composite nanofibrous mats for wound healing can be electrospun from the ternary mixture(s) of CAM, SFAs, and polyvinylpyrrolidone. PMID:26886306

  12. 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

    2016-01-01

    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.

  13. 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.

  14. Analysis and monitoring of chloramphenicol residues in food of animal origin in Slovenia from 1991 to 2000.

    PubMed

    Cerkvenik, V

    2002-04-01

    To prevent the illegal use of chloramphenicol (CAP), regulatory control of its residues in food of animal origin is essential. In Slovenia, the monitoring of CAP residues for statutory purpose started in 1991. The results of a 10-year period are presented. CAP residues were determined by capillary gas chromatography (GC) with electron capture detection (ECD) using meta-CAP as an internal standard (ISTD). Before chromatographic determination, analytes were derivatized by silylation. Overall, CAP recovery, adjusted for ISTD, was for bovine muscle tissue and raw cow's milk (in the region of 2-10 microg kg(-1)) 89 and 102%, respectively, and for whole eggs, 87% (in the region of 1-10 microg kg(-1)). The use of meta-CAP improved significantly the precision of the method. The detection limit for CAP was 1 microg kg(-1), which was sufficiently sensitive for routine use. A total of 1308 random samples of Slovenian origin were analysed from 1991 to 2000, covering all parts of the country. CAP was found only in one milk sample in 1997 at a concentration of 4.6 microg kg(-1).

  15. 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. PMID:27037966

  16. Heavy metal (lead, Cadmium) and antibiotic (Tetracycline and Chloramphenicol) residues in fresh and frozen fish types (Clarias gariepinus, Oreochromis niloticus) in Ibadan, Oyo State, Nigeria.

    PubMed

    Olusola, Adetunji Victoria; Folashade, Popoola Amirah; Ayoade, Odetokun Ismail

    2012-09-15

    This study was carried out to assess the level of heavy metals (lead (Pb) and cadmium (Cd)) and antibiotics (tetracycline and chloramphenicol) residues in frozen and fresh fish types obtained from Eleyele river, Officer's mess, Alfa farm and a major frozen meat outlet in Ibadan, Oyo State Nigeria. The Atomic Absorption Spectrophotometer (AAS) and High Performance Liquid Chromatography (HPLC) were used to analyze the heavy metals and antibiotics residue levels in fresh and frozen fish, respectively. The results showed mean concentrations of antibiotics was higher (p<0.05) in fresh than in frozen fish samples while there were no significant differences in the mean concentrations of heavy metal residue. The differences of mean residue levels in both antibiotics and heavy metals tested in the cranial and caudal parts of the fish samples were not significant (p<0.05). However, there was a significant difference between species (tilapia and catfish) and sources. The highest heavy metals and tetraxycline residues were observed in Alfa's farm (Pb: 0.039+/-0.004 ppm; Cd: 0.020+/-0.006 ppm; tetraxycline: 2.185+/-0.412). Chloramphenicol was highest in Officers mess (0.837+/-0.165 ppm). The heavy metals (Pb and Cd) concentrations determined were below the maximum permissible limits set by both local and international safety agencies. Tetraxcycline exceeded international limits of 0.2 ppm while Chloramphenicol which has a zero tolerance level was also detected from all sources. This study accentuates the need for control of heavy metals and antibiotics in fish sold for human consumption in Ibadan, Nigeria. The need to ban chloramphenicol in treatment of fish and other animals is emphasised. PMID:24205760

  17. 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). PMID:27112147

  18. [Treatment of puerperal endometritis. Evaluation of the efficacy and safety of clindamycin + gentamycin vs. penicillin + chloramphenicol + gentamycin].

    PubMed

    Gutiérrez, C; Carrillo, C; Escudero, F; Caciano, S; García Hjarles, M

    1994-11-01

    This was a prospective, single-blind, comparative study in patients with diagnosis of puerperal endometritis, carried out at the Loayza Hospital in Lima, Peru. The objective of this study was to evaluate the efficacy and safety of clindamycin and gentamicin in the management of endometritis vs. penicillin, chloramphenicol and gentamicin for 10 days. Sixty-five patients were enrolled and 62 were evaluable for efficacy. Both treatment groups were comparable in the pre-treatment period in terms of age, history of pregnancies, controls by gynecologist, days of disease and fever, clinical symptoms like fever, pelvic pain, pulse, uterine size and in laboratory, in hematocrit and leukocytes count. In the culture of endometrium tissue, 27/32 patients (84.4%) in Group A (penicillin + CAF + gentamicin) and 27/30 patients (90%) in Group B (clindamycin + gentamicin) had positive cultures at baseline; 18 and 22 patients showed anaerobes; 8 and 4 patients showed anaerobes plus aerobes and, one patient in each treatment group showed aerobes only. Peptostreptococcus and Bacteroides fragilis were the most frequently isolated pathogens. Improvement in lochia fetidity was more rapid in Group B, it turned transparent and not fetid since day 3. Complete cure was significantly better in Group B 24/30 (80%) in comparison with Group A 16/32 (50%) (p = 0.02). Partial response was found in 15 patients (43.3%) in Group A and 5 patients (16.6%) in Group B. Only one case was considered as bacteriological failure in Group A and only one patient in Group B was considered as failure and required an additional operation due to residual abscess.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7821833

  19. Comparsion of an immunochromatographic strip with ELISA for simultaneous detection of thiamphenicol, florfenicol and chloramphenicol in food samples.

    PubMed

    Guo, Lingling; Song, Shanshan; Liu, Liqiang; Peng, Juan; Kuang, Hua; Xu, Chuanlai

    2015-09-01

    Rapid and sensitive indirect competitive enzyme-linked immunosorbent assays (ic-ELISA) and gold nanoparticle immunochromatographic strip tests were developed to detect thiamphenicol (TAP), florfenicol (FF) and chloramphenicol (CAP) in milk and honey samples. The generic monoclonal antibody for TAP, FF and CAP was prepared based on a hapten [D-threo-1-(4-aminophenyl)-2- dichloroacetylamino-1,3-propanediol], and the haptenwas linked to a carrier protein using the diazotization method. After the optimization of several parameters (coating, pH, sodium chloride content and methanol content), the ic-ELISA was established. The quantitative working range for TAP was 0.11-1.36 ng/mL, with an IC50 of 0.39 ng/mL. The optimized ELISA showed cross-reactivity to CAP (300%) and FF (15.6%), with IC50 values of 0.13 and 2.5 ng/mL, respectively. The analytical recovery of TAP, FF and CAP in milk and honey samples in the ic-ELISA ranged from 81.2 to 112.9%. Based on this monoclonal antibody, a rapid and sensitive immunochromatographic test strip was also developed. This strip had a detection limit of 1 ng/mL for TAP, FF and CAP in milk and honey samples. Moreover, the test was completed within 10 min. Our results showed that the proposed ic-ELISA and immunochromatographic test strip method are highly useful screening tools for TAP, FF and CAP detection in milk and honey samples. PMID:25675893

  20. Leader peptides of inducible chloramphenicol resistance genes from gram-positive and gram-negative bacteria bind to yeast and Archaea large subunit rRNA.

    PubMed Central

    Harrod, R; Lovett, P S

    1997-01-01

    catA86 is the second gene in a constitutively transcribed, two-gene operon cloned from Bacillus pumilus . The region that intervenes between the upstream gene, termed the leader, and the catA86 coding sequence contains a pair of inverted repeat sequences which cause sequestration of the catA86 ribosome binding site in mRNA secondary structure. As a consequence, the catA86 coding sequence is untranslatable in the absence of inducer. Translation of the catA86 coding sequence is induced by chloramphenicol in Gram-positives and induction requires a function of the leader coding sequence. The leader-encoded peptide has been proposed to instruct its translating ribosome to pause at leader codon 6, enabling chloramphenicol to stall the ribosome at that site. Ribosome stalling causes destabilization of the RNA secondary structure, exposing the catA86 ribosome binding site, allowing activation of its translation. A comparable mechanism of induction by chloramphenicol has been proposed for the regulated cmlA gene from Gram-negative bacteria. The catA86 and cmlA leader-encoded peptides are in vitro inhibitors of peptidyl transferase, which is thought to be the basis for selection of the site of ribosome stalling. Both leader-encoded peptides have been shown to alter the secondary structure of Escherichia coli 23S rRNA in vitro. All peptide-induced changes in rRNA conformation are within domains IV and V, which contains the peptidyl transferase center. Here we demonstrate that the leader peptides alter the conformation of domains IV and V of large subunit rRNA from yeast and a representative of the Archaea. The rRNA target for binding the leader peptides is therefore conserved across kingdoms. PMID:9108153

  1. 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

  2. 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%).

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

    PubMed

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

    2015-03-15

    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 (R(2)>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%).

  4. [Simultaneous determination of zeranols and chloramphenicol in foodstuffs of animal origin by combination immunoaffinity column clean-up and liquid chromatography-tandem mass spectrometry].

    PubMed

    Wang, Qing; Wang, Guomin; Xi, Cunxian; Li, Xianliang; Chen, Dongdong; Tang, Bobin; Zhang, Lei; Zhao, Hua

    2014-06-01

    A combination immunoaffinity column (IAC-CZ) clean-up and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method was successfully developed for zearalenol, beta-zearalenol and zearalenone) and chloramphenicol (CAP) in foodstuffs of animal origin. The samples (fish, liver, milk and honey) were enzymatically digested by beta-glucuronidase/sulfatase for about 16 h and then extracted with ether. The extracts were evaporated to dryness and then the residues were dissolved by 1.0 mL of 50% acetonitrile solution. After filtered and diluted with PBS buffer, the reconstituted solution were cleaned-up with a IAC-CZ and then analyzed by LC-MS/MS in multiple reaction monitoring (MRM) mode. The chromatographic separation was performed on a Shimadzu Shim-pack VP-ODS column with gradient elution by acetonitrile and 2 mmol/L ammonium acetate solution. The detection was carried out by electrospray negative ionization mass spectrometry in MRM mode. The proposed method was validated by the limit of detection (0.04-0.10 microg/kg), linearity (R2 > or = 0.999 0), average recoveries (70.9%-95.6%) and precisions (2.0% - 11.8%). The developed method is reliable, sensitive and has good applicability. The combination immunoaffinity column was proved to be an effective pretreatment technique to decrease the matrix effect, and it met the requirements of residue analysis of co-occurring zeranols and chloramphenicol. PMID:25269264

  5. Dissemination of a chloramphenicol- and tetracycline-resistant but penicillin-susceptible invasive clone of serotype 5 Streptococcus pneumoniae in Colombia.

    PubMed

    Tamayo, M; Sá-Leão, R; Santos Sanches, I; Castañeda, E; de Lencastre, H

    1999-07-01

    A national surveillance conducted in Colombia between 1994 and 1996 identified serotype 5 Streptococcus pneumoniae as the second most frequent cause of invasive disease in children younger than 5 years of age. All 43 serotype 5 isolates collected during this period were shown to be susceptible to penicillin, erythromycin, cefotaxime, and vancomycin, but most (38 of 43, or 88%) were highly resistant to chloramphenicol. In order to clarify a possible genetic relatedness among these isolates, additional microbiological and molecular characterizations were performed. Most (40 of 43, or 93%) of the isolates were found to be resistant to tetracycline. Pulsed-field gel electrophoresis (PFGE) patterns of chromosomal DNAs revealed that all the 43 isolates were closely related and that 38 of the 43 isolates were representatives of a "Colombian clone" of S. pneumoniae isolates which were recovered throughout the 3-year surveillance period from patients in 13 hospitals located in five Colombian cities. Isolates belonging to this Colombian clone were resistant to chloramphenicol and tetracycline, hybridized with the cat and tetM DNA probes in the same 340-kb SmaI fragment, and had identical PFGE patterns after both SmaI and ApaI digestions.

  6. [Simultaneous determination of zeranols and chloramphenicol in foodstuffs of animal origin by combination immunoaffinity column clean-up and liquid chromatography-tandem mass spectrometry].

    PubMed

    Wang, Qing; Wang, Guomin; Xi, Cunxian; Li, Xianliang; Chen, Dongdong; Tang, Bobin; Zhang, Lei; Zhao, Hua

    2014-06-01

    A combination immunoaffinity column (IAC-CZ) clean-up and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method was successfully developed for zearalenol, beta-zearalenol and zearalenone) and chloramphenicol (CAP) in foodstuffs of animal origin. The samples (fish, liver, milk and honey) were enzymatically digested by beta-glucuronidase/sulfatase for about 16 h and then extracted with ether. The extracts were evaporated to dryness and then the residues were dissolved by 1.0 mL of 50% acetonitrile solution. After filtered and diluted with PBS buffer, the reconstituted solution were cleaned-up with a IAC-CZ and then analyzed by LC-MS/MS in multiple reaction monitoring (MRM) mode. The chromatographic separation was performed on a Shimadzu Shim-pack VP-ODS column with gradient elution by acetonitrile and 2 mmol/L ammonium acetate solution. The detection was carried out by electrospray negative ionization mass spectrometry in MRM mode. The proposed method was validated by the limit of detection (0.04-0.10 microg/kg), linearity (R2 > or = 0.999 0), average recoveries (70.9%-95.6%) and precisions (2.0% - 11.8%). The developed method is reliable, sensitive and has good applicability. The combination immunoaffinity column was proved to be an effective pretreatment technique to decrease the matrix effect, and it met the requirements of residue analysis of co-occurring zeranols and chloramphenicol.

  7. 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.

  8. 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

    2015-01-01

    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).

  9. Validation of a method for simultaneous determination of nitroimidazoles, benzimidazoles and chloramphenicols in swine tissues by ultra-high performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Xia, Xi; Wang, Yuanyuan; Wang, Xia; Li, Yun; Zhong, Feng; Li, Xiaowei; Huang, Yaoling; Ding, Shuangyang; Shen, Jianzhong

    2013-05-31

    This paper presents a sensitive and confirmatory multi-residue method for the analysis of 23 veterinary drugs and metabolites belonging to three classes (nitroimidazoles, benzimidazoles, and chloramphenicols) in porcine muscle, liver, and kidney. After extracted with ethyl acetate and basic ethyl acetate sequentially, the crude extracts were defatted with hexane and further purified using Oasis MCX solid-phase extraction cartridges. Rapid determination was carried out by ultra-high performance liquid chromatography-electrospray ionization tandem mass spectrometry. Data acquisition was performed under positive and negative mode simultaneously. Recoveries based on matrix-matched calibrations for meat, liver, and kidney ranged from 50.6 to 108.1%. The method quantification limits were in the range of 3-100ng/kg. PMID:23017446

  10. 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.

  11. Screening for chloramphenicol residues in the tissues and fluids of treated cattle by the four plate test, Charm II radioimmunoassay and Ridascreen CAP-Glucuronid enzyme immunoassay.

    PubMed

    Lynas, L; Currie, D; Elliott, C T; McEvoy, J D; Hewitt, S A

    1998-12-01

    The administration of chloramphenicol (CAP) is banned in food animals in the European Union (EU). It is, therefore, important to have adequate screening methods to determine if residues of CAP and its major metabolite, chloramphenicol-glucuronide (CAP-Gluc), are present in samples taken for monitoring purposes. Six castrated male cattle were treated with a single intramuscular injection of 10 mg kg-1 CAP. Animals were sampled once daily for urine and were slaughtered at 3 and 6 d post-injection. Samples of bile, kidney, liver and diaphragmatic muscle were removed at slaughter. All matrices were analysed using the four plate test (FPT) bioassay, the Charm II radioimmunoassay and a Ridascreen CAP-Glucuronid competitive enzyme immunoassay (EIA). The FPT detected CAP residues in urine samples taken up to 2 d post-treatment. The Charm assay detected CAP in the urine for up to 4 d post-treatment. The EIA detected CAP throughout the 6 d sampling period. Samples of bile were positive by both the EIA and the Charm assay at day 3 and day 6. No zones of inhibition were obtained using the FPT in bile or diaphragm either with or without sample pre-treatment with beta-glucuronidase. However, the kidney and the liver from one animal killed at day 6 gave larger zones of inhibition after treatment with beta-glucuronidase, indicating the presence of CAP. The kidneys of all treated animals slaughtered at day 3 were positive by both the EIA and the Charm assay but none of the kidneys at day 6 tested positive by either method. Owing to technical difficulties, the Charm assay was not suitable for the analysis of liver. The EIA failed to detect CAP in the liver of any treated animal. It is concluded that urine appears to be the best matrix for screening purposes. The sensitivity of the FPT is inadequate for the determination of CAP residues were minimal withdrawal periods have been observed. The Charm assay and the EIA were suitable for the detection of both CAP and CAP-Gluc in tissues

  12. 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

    2016-01-01

    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%. PMID:26686562

  13. Identification of the O antigen polymerase (rfc) gene in Escherichia coli O4 by insertional mutagenesis using a nonpolar chloramphenicol resistance cassette.

    PubMed Central

    Lukomski, S; Hull, R A; Hull, S I

    1996-01-01

    Computer analysis of the O4 polysaccharide gene cluster of Escherichia coli revealed the presence of two open reading frames (ORFs) encoding strongly hydrophobic polypeptides. O antigen polymerase, which is encoded by the rfc gene, is a potential membrane protein and therefore should be hydrophobic. To identify the rfc gene, these two ORFs were subjected to insertional mutagenesis. A chloramphenicol resistance cassette was designed which, when properly inserted, does not cause a polar effect in downstream genes. Each of two ORFs, cloned into a plasmid vector, was inactivated with this cassette. Two types of mutants bearing chromosomal insertions of the cassettes in each ORF were constructed by homologous recombination. These mutants were characterized by PCR, Southern blotting, and transverse-alternating-field electrophoresis. Only one class of mutants exhibited the expected O polymerase-deficient phenotype; they produced O4-specific, semirough lipopolysaccharide. Therefore, this ORF was identified as the rfc gene. The chromosomal rfc mutation was complemented in trans by the rfc gene expressed from a plasmid vector. PMID:8550424

  14. Detection and quantification of chloramphenicol in milk and honey using molecularly imprinted polymers: Canadian penny-based SERS nano-biosensor.

    PubMed

    Gao, Fang; Feng, Shaolong; Chen, Zhiwen; Li-Chan, Eunice C Y; Grant, Edward; Lu, Xiaonan

    2014-12-01

    We integrated molecularly imprinted polymers with surface-enhanced Raman spectroscopy (MIPs-SERS) to develop an innovative nano-biosensor for the determination of chloramphenicol (CAP) in milk and honey products. Template molecule (CAP), functional monomer (acrylamide), cross-linking agent (ethylene glycol dimethacrylate), initiator (2,2'-azobis(isobutyronitrile)), and porogen (methanol) were employed to form MIPs via "dummy" precipitation polymerization. Static and kinetic studies validated the specific selectivity of MIPs toward CAP over nonimprinted polymers (imprinting factor >4). Canadian penny-based silver nano-structure was synthesized as SERS-active substrate for determination of CAP in food matrices. Collected spectra were processed by principal component analysis to differentiate various concentrations of CAP in foods. Partial least squares regression models showed good prediction values (R > 0.9) of actual spiked contents (0, 0.1, 0.5, 1, 5 ppm) of CAP in milk and honey. This developed nano-biosensor is low cost, requires little sample pretreatment, and can provide reliable detection of trace level of chemical hazards in food systems within a total of 15 min. PMID:25393060

  15. Stimulation and inhibition of bacterial growth by caffeine dependent on chloramphenicol and a phenolic uncoupler--a ternary toxicity study using microfluid segment technique.

    PubMed

    Cao, Jialan; Kürsten, Dana; Schneider, Steffen; Köhler, J Michael

    2012-10-01

    A droplet-based microfluidic technique for the fast generation of three dimensional concentration spaces within nanoliter segments was introduced. The technique was applied for the evaluation of the effect of two selected antibiotic substances on the toxicity and activation of bacterial growth by caffeine. Therefore a three-dimensional concentration space was completely addressed by generating large sequences with about 1150 well separated microdroplets containing 216 different combinations of concentrations. To evaluate the toxicity of the ternary mixtures a time-resolved miniaturized optical double endpoint detection unit using a microflow-through fluorimeter and a two channel microflow-through photometer was used for the simultaneous analysis of changes on the endogenous cellular fluorescence signal and on the cell density of E. coli cultivated inside 500 nL microfluid segments. Both endpoints supplied similar results for the dose related cellular response. Strong non-linear combination effects, concentration dependent stimulation and the formation of activity summits on bolographic maps were determined. The results reflect a complex response of growing bacterial cultures in dependence on the combined effectors. A strong caffeine induced enhancement of bacterial growth was found at sublethal chloramphenicol and sublethal 2,4-dinitrophenol concentrations. The reliability of the method was proved by a high redundancy of fluidic experiments. The results indicate the importance of multi-parameter investigations for toxicological studies and prove the potential of the microsegmented flow technique for such requirements. PMID:22888747

  16. 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.

  17. 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

    2015-04-01

    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.

  18. 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.

  19. Determination of chloramphenicol, thiamphenicol, florfenicol, and florfenicol amine in poultry and porcine muscle and liver by gas chromatography-negative chemical ionization mass spectrometry.

    PubMed

    Shen, Jianzhong; Xia, Xi; Jiang, Haiyang; Li, Cun; Li, Jiancheng; Li, Xiaowei; Ding, Shuangyang

    2009-05-15

    A sensitive and reliable method using gas chromatography-negative chemical ionization mass spectrometry (GC-NCI/MS) was developed for the simultaneous determination of chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF), and florfenicol amine (FFA) at trace levels in muscle and liver. Before extraction with ethyl acetate, CAP-d(5) was added to tissue samples as internal standard. The organic extracts were frozen to remove lipid and further purified by liquid-liquid extraction (LLE) with hexane and solid-phase extraction (SPE) using Oasis HLB cartridges. The target compounds were derivatized with BSTFA+1% TMCS prior to GC-NCI/MS determination in selected ion monitoring mode (SIM). The recovery values ranged from 78.5 to 105.5%, with relative standard deviations (RSD) <17%. The limits of detections (LODs) of 0.1 microg/kg for CAP and 0.5 microg/kg for TAP, FF, and FFA were obtain. Incurred sample and samples from local market were successfully analyzed using this method. PMID:19395324

  20. Detection and quantification of chloramphenicol in milk and honey using molecularly imprinted polymers: Canadian penny-based SERS nano-biosensor.

    PubMed

    Gao, Fang; Feng, Shaolong; Chen, Zhiwen; Li-Chan, Eunice C Y; Grant, Edward; Lu, Xiaonan

    2014-12-01

    We integrated molecularly imprinted polymers with surface-enhanced Raman spectroscopy (MIPs-SERS) to develop an innovative nano-biosensor for the determination of chloramphenicol (CAP) in milk and honey products. Template molecule (CAP), functional monomer (acrylamide), cross-linking agent (ethylene glycol dimethacrylate), initiator (2,2'-azobis(isobutyronitrile)), and porogen (methanol) were employed to form MIPs via "dummy" precipitation polymerization. Static and kinetic studies validated the specific selectivity of MIPs toward CAP over nonimprinted polymers (imprinting factor >4). Canadian penny-based silver nano-structure was synthesized as SERS-active substrate for determination of CAP in food matrices. Collected spectra were processed by principal component analysis to differentiate various concentrations of CAP in foods. Partial least squares regression models showed good prediction values (R > 0.9) of actual spiked contents (0, 0.1, 0.5, 1, 5 ppm) of CAP in milk and honey. This developed nano-biosensor is low cost, requires little sample pretreatment, and can provide reliable detection of trace level of chemical hazards in food systems within a total of 15 min.

  1. Separation, concentration and determination of chloramphenicol in environment and food using an ionic liquid/salt aqueous two-phase flotation system coupled with high-performance liquid chromatography.

    PubMed

    Han, Juan; Wang, Yun; Yu, Cuilan; Li, Chunxiang; Yan, Yongsheng; Liu, Yan; Wang, Liang

    2011-01-31

    Ionic liquid-salt aqueous two-phase flotation (ILATPF) is a novel, green, non-toxic and sensitive samples pretreatment technique. ILATPF coupled with high-performance liquid chromatography (HPLC) was developed for the analysis of chloramphenicol, which combines ionic liquid aqueous two-phase system (ILATPS) based on imidazolium ionic liquid (1-butyl-3-methylimidazolium chloride, [C(4)mim]Cl) and inorganic salt (K(2)HPO(4)) with solvent sublation. In ILATPF systems, phase behaviors of the ILATPF were studied for different types of ionic liquids and salts. The sublation efficiency of chloramphenicol in [C(4)mim]Cl-K(2)HPO(4) ILATPF was influenced by the types of salts, concentration of K(2)HPO(4) in aqueous solution, solution pH, nitrogen flow rate, sublation time and the amount of [C(4)mim]Cl. Under the optimum conditions, the average sublation efficiency is up to 98.5%. The mechanism of ILATPF contains two principal processes. One is the mechanism of IL-salt ILATPS formation, the other is solvent sublation. This method was practical when applied to the analysis of chloramphenicol in lake water, feed water, milk, and honey samples with the linear range of 0.5-500 ng mL(-1). The method yielded limit of detection (LOD) of 0.1 ng mL(-1) and limit of quantification (LOQ) of 0.3 ng mL(-1). The recovery of CAP was 97.1-101.9% from aqueous samples of environmental and food samples by the proposed method. Compared with liquid-liquid extraction, solvent sublation and ionic liquid aqueous two-phase extraction, ILATPF can not only separate and concentrate chloramphenicol with high sublation efficiency, but also efficiently reduce the wastage of IL. This novel technique is much simpler and more environmentally friendly and is suggested to have important applications for the concentration and separation of other small biomolecules. PMID:21168562

  2. Performance Assessment and Comparability of a Commercial Enzyme-Linked Immunosorbent Assay Kit with Liquid Chromatography-Tandem Mass Spectrometry for Chloramphenicol Residues in Crab and Shrimp.

    PubMed

    Jester, Edward L E; Loader, Jared I; El Said, Kathleen R; Abraham, Ann; Flores Quintana, Harold A; Plakas, Steven M

    2016-01-01

    Monitoring for chloramphenicol (CAP) in aquaculture products is primarily performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), which requires expensive equipment and specialized training. Many laboratories prefer to screen samples with facile and high-throughput enzyme-linked immunosorbent assay (ELISA) kits for CAP residues before submitting samples for LC-MS/MS quantification and confirmation. We evaluated the performance of a Ridascreen (R-Biopharm) ELISA kit for CAP in spiked and incurred crab and shrimp muscle at levels bracketing the minimum required performance level for analysis (0.3 ng/g). The Ridascreen ELISA kit incorporates antibody directed against CAP. Incurred CAP levels in crab and shrimp muscle were verified using LC-MS/MS. We found good repeatability (relative standard deviation) of the ELISA in spiked and incurred crab and shrimp muscle samples, with values ranging from 6.8 to 21.7%. Recoveries of CAP from tissues spiked at 0.15 to 0.60 ng/g ranged from 102 to 107%. Minimal cross-reactivity with blank crab and shrimp muscle matrix components was observed. ELISA data were highly correlated with those of LC-MS/MS for CAP in incurred muscle tissue. We believe this study to be the first evaluation of the performance and comparability of a CAP ELISA kit and LC-MS/MS for determination of CAP residues, as well as their elimination, in crab muscle. Our findings support the use of this ELISA kit for screening purposes and, when used in conjunction with validated instrumental methods, for regulatory monitoring of CAP in these species.

  3. A triple-amplification SPR electrochemiluminescence assay for chloramphenicol based on polymer enzyme-linked nanotracers and exonuclease-assisted target recycling.

    PubMed

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

    2016-12-15

    The present study aimed to explore a novel triple-amplification electrochemiluminescence (ECL) assay for detecting of chloramphenicol (CAP). This strategy was based on single-stranded DNA-binding protein (SSB) and horseradish peroxidase (HRP) enzyme-linked polymer (EnVision reagent, EV) labeled on Au nanoparticles (EV-Au-SSB) as nanotracer and exonuclease-assisted target recycling. The composite probes were prepared via immunoreactions between the CdS nanocrystal (CdS NC)-functionalized partial complementary DNA and aptamer (CdSNCs/Apt-ssDNA1) as capture probes, and EV-Au-SSB as nanotracer. When the composite probe solution co-existed with CAP and Exo I, the aptamer on the capture probes preferentially combined with CAP, and then CAP-Apt and nanotracer complex were released into the solution. Subsequently, Exo I in the solution could further digest the CAP-Apt from the 3'-end of the aptamer and release CAP, which could participate in further reaction with the probes. It was worth mentioning that EV contained a large number of HRPs on its dendritic chain. In the EV-Au-SSB, Au could enhance ECL intensity of CdS NCs by surface plasmon resonance. What's more, HRPs on EV could catalyze the reaction of H2O2, which could obviously enhance ECL intensity of CdS NCs. This study demonstrated excellent performance of the triple-amplification ECL assay, which makes this aptasensor system suitable and promising for the practical application of CAP residues in fish samples. Moreover, the assay might provide a promising avenue to develop efficient aptasensors to determine small-molecule harmful substances in environmental monitoring and food safety. PMID:27434234

  4. 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

  5. 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.

  6. 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. PMID:27251948

  7. Quantitative liquid chromatography/tandem mass spectrometry determination of chloramphenicol residues in food using sub-2 microm particulate high-performance liquid chromatography columns for sensitivity and speed.

    PubMed

    Kaufmann, Anton; Butcher, Patrick

    2005-01-01

    The use of chloramphenicol (CAP)--a highly effective broad-spectrum antibiotic used in animal husbandry--is banned in many countries. Therefore, a very low minimum required performance limit (MRPL) of 0.3 microg/kg CAP in meat for human consumption has been defined. Analytical methods capable of quantifying and confirming such low residue levels require sophisticated instrumentation. Preferably sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) or gas chromatography/mass spectrometry (GC/MS) methods have been used. This paper suggests the use of sub-2 microm particulate high-performance liquid chromatography (HPLC) columns to gain additional sensitivity and improve resolution as well as speed. Depending on the operating conditions, higher chromatographic resolution and speed can be obtained at the price of a significantly increased operating pressure, requiring dedicated LC equipment. A 3-4-fold overall improvement of the signal-to-noise ratio for CAP was obtained compared to more classical 5 microm particulate HPLC columns. The proposed analytical methodology includes an enzymatic digestion, which liberates glucuronide-bound CAP from kidney tissue. The extracts obtained after an Extrelut clean-up are sufficiently pure to permit routine injection of biological samples into the sub-2 microm particulate HPLC column, without observing rapid deterioration of peak shape or column clogging problems. The time for one chromatographic run was 4.2 min. The described method was validated for two particularly difficult matrices (kidney and honey). Decision limits (CC alpha) were 0.007 microg/kg (honey) und 0.011 microg/kg (kidney), which are significantly below the current MRPL. PMID:16299695

  8. Role of the Two Component Signal Transduction System CpxAR in Conferring Cefepime and Chloramphenicol Resistance in Klebsiella pneumoniae NTUH-K2044

    PubMed Central

    Srinivasan, Vijaya Bharathi; Vaidyanathan, Vasanth; Mondal, Amitabha; Rajamohan, Govindan

    2012-01-01

    Background Klebsiella pneumoniae is a Gram-negative, non-motile, facultative anaerobe belonging to the Enterobacteriaceae family of the γ-Proteobacteria class in the phylum Proteobacteria. Multidrug resistant K. pneumoniae have caused major therapeutic problems worldwide due to emergence of extended-spectrum β-lactamase producing strains. Two-component systems serve as a basic stimulus-response coupling mechanism to allow organisms to sense and respond to changes in many different environmental conditions including antibiotic stress. Principal Findings In the present study, we investigated the role of an uncharacterized cpxAR operon in bacterial physiology and antimicrobial resistance by generating isogenic mutant (ΔcpxAR) deficient in the CpxA/CpxR component derived from the hyper mucoidal K1 strain K. pneumoniae NTUH-K2044. The behaviour of ΔcpxAR was determined under hostile conditions, reproducing stresses encountered in the gastrointestinal environment and deletion resulted in higher sensitivity to bile, osmotic and acid stresses. The ΔcpxAR was more susceptible to β-lactams and chloramphenicol than the wild-type strain, and complementation restored the altered phenotypes. The relative change in expression of acrB, acrD, eefB efflux genes were decreased in cpxAR mutant as evidenced by qRT-PCR. Comparison of outer membrane protein profiles indicated a conspicuous difference in the knock out background. Gel shift assays demonstrated direct binding of CpxRKP to promoter region of ompCKP in a concentration dependent manner. Conclusions and Significance The Cpx envelope stress response system is known to be activated by alterations in pH, membrane composition and misfolded proteins, and this systematic investigation reveals its direct involvement in conferring antimicrobial resistance against clinically significant antibiotics for the very first time. Overall results displayed in this report reflect the pleiotropic role of the CpxAR signaling system and

  9. 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. PMID:26838404

  10. Introduction of UAG, UAA, and UGA nonsense mutations at a specific site in the Escherichia coli chloramphenicol acetyltransferase gene: use in measurement of amber, ochre, and opal suppression in mammalian cells.

    PubMed Central

    Capone, J P; Sedivy, J M; Sharp, P A; RajBhandary, U L

    1986-01-01

    We have used oligonucleotide-directed site-specific mutagenesis to convert serine codon 27 of the Escherichia coli chloramphenicol acetyltransferase (cat) gene to UAG, UAA, and UGA nonsense codons. The mutant cat genes, under transcriptional control of the Rous sarcoma virus long terminal repeat, were then introduced into mammalian cells by DNA transfection along with UAG, UAA, and UGA suppressor tRNA genes derived from a human serine tRNA. Assay for CAT enzymatic activity in extracts from such cells allowed us to detect and quantitate nonsense suppression in monkey CV-1 cells and mouse NIH3T3 cells. Using such an assay, we provide the first direct evidence that an opal suppressor tRNA gene is functional in mammalian cells. The pattern of suppression of the three cat nonsense mutations in bacteria suggests that the serine at position 27 of CAT can be replaced by a wide variety of amino acids without loss of enzymatic activity. Thus, these mutant cat genes should be generally useful for the quantitation of suppressor activity of suppressor tRNA genes introduced into cells and possibly for the detection of naturally occurring nonsense suppressors. Images PMID:3023959

  11. 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.

  12. 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. PMID:27664624

  13. Isolation of Escherichia coli strains with AcrAB-TolC efflux pump-associated intermediate interpretation or resistance to fluoroquinolone, chloramphenicol and aminopenicillin from dogs admitted to a university veterinary hospital.

    PubMed

    Sato, Toyotaka; Yokota, Shin-ichi; Ichihashi, Risa; Miyauchi, Tomoka; Okubo, Torahiko; Usui, Masaru; Fujii, Nobuhiro; Tamura, Yutaka

    2014-07-01

    Understanding the prevalence of antimicrobial-resistance and the relationship between emergence of resistant bacteria and clinical treatment can facilitate design of effective treatment strategies. We here examined antimicrobial susceptibilities of Escherichia coli isolated from dogs admitted to a university hospital (University hospital) and companion animal clinics (Community clinics) in the same city and investigated underlying multidrug-resistance mechanisms. The prevalence of E. coli with intermediate and resistant interpretations to ampicillin (AMP), enrofloxacin (ENR) and chloramphenicol (CHL) was higher in the University hospital than in the Community clinics cases. Use of antimicrobials, including fluoroquinolone, was also significantly higher in the University hospital than in the Community clinics cases. Upon isolation using ENR-supplemented agar plates, all ENR-resistant isolates had 3-4 nucleotide mutations that accompanied by amino acid substitutions in the quinolone-resistance-determining regions of gyrA, parC and parE, and 94.7% of all isolates derived from the University hospital showed AMP and/or CHL resistance and possessed blaTEM and/or catA1. The average mRNA expression levels of acrA, acrB and tolC and the prevalence of organic solvent tolerance, in isolates derived from ENR-supplemented agar plates were significantly higher in the University hospital than in the Community clinics isolates. Thus, E. coli derived from the University hospital cases more often showed concomitant decreased susceptibilities to aminopenicillins, fluoroquinolones and CHL than did those derived from the Community clinics; this was related to an active AcrAB-TolC efflux pump, in addition to acquisition of specific resistance genes and genetic mutations.

  14. Extraction and determination of chloramphenicol in feed water, milk, and honey samples using an ionic liquid/sodium citrate aqueous two-phase system coupled with high-performance liquid chromatography.

    PubMed

    Han, Juan; Wang, Yun; Yu, Cui-lan; Yan, Yong-sheng; Xie, Xue-qiao

    2011-01-01

    A green, simple, non-toxic, and sensitive sample pretreatment procedure coupled with high-performance liquid chromatography (HPLC) was developed for the analysis of chloramphenicol (CAP) that exploits an aqueous two-phase system based on imidazolium ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate, [Bmim]BF(4)) and organic salt (Na(3)C(6)H(5)O(7)) using a liquid-liquid extraction technique. The influence factors on partition behaviors of CAP were studied, including the type and amount of salts, the pH value, the volume of [Bmim]BF(4), and the extraction temperature. Extraction efficiency of the CAP was found to increase with increasing temperature and the volume of [Bmim]BF(4). Thermodynamic studies indicated that hydrophobic interactions were the main driving force, although electrostatic interactions and salting-out effects were also important for the transfer of the CAP. Under the optimal conditions, 90.1% of the CAP could be extracted into the ionic liquid-rich phase in a single-step extraction. This method was practical when applied to the analysis of CAP in feed water, milk, and honey samples with a linear range of 2~1,000 ng mL(-1). The method yielded a limit of detection of 0.3 ng mL(-1) and a limit of quantification of 1.0 ng mL(-1). The recovery of CAP was 90.4-102.7% from aqueous samples of real feed water, milk, and honey samples by the proposed method. This novel process is much simpler and more environmentally friendly and is suggested to have important applications for the separation of antibiotics. PMID:21063686

  15. Isolation of Escherichia coli Strains with AcrAB–TolC Efflux Pump-Associated Intermediate Interpretation or Resistance to Fluoroquinolone, Chloramphenicol and Aminopenicillin from Dogs Admitted to a University Veterinary Hospital

    PubMed Central

    SATO, Toyotaka; YOKOTA, Shin-ichi; ICHIHASHI, Risa; MIYAUCHI, Tomoka; OKUBO, Torahiko; USUI, Masaru; FUJII, Nobuhiro; TAMURA, Yutaka

    2014-01-01

    ABSTRACT Understanding the prevalence of antimicrobial-resistance and the relationship between emergence of resistant bacteria and clinical treatment can facilitate design of effective treatment strategies. We here examined antimicrobial susceptibilities of Escherichia coli isolated from dogs admitted to a university hospital (University hospital) and companion animal clinics (Community clinics) in the same city and investigated underlying multidrug-resistance mechanisms. The prevalence of E. coli with intermediate and resistant interpretations to ampicillin (AMP), enrofloxacin (ENR) and chloramphenicol (CHL) was higher in the University hospital than in the Community clinics cases. Use of antimicrobials, including fluoroquinolone, was also significantly higher in the University hospital than in the Community clinics cases. Upon isolation using ENR-supplemented agar plates, all ENR-resistant isolates had 3–4 nucleotide mutations that accompanied by amino acid substitutions in the quinolone-resistance-determining regions of gyrA, parC and parE, and 94.7% of all isolates derived from the University hospital showed AMP and/or CHL resistance and possessed blaTEM and/or catA1. The average mRNA expression levels of acrA, acrB and tolC and the prevalence of organic solvent tolerance, in isolates derived from ENR-supplemented agar plates were significantly higher in the University hospital than in the Community clinics isolates. Thus, E. coli derived from the University hospital cases more often showed concomitant decreased susceptibilities to aminopenicillins, fluoroquinolones and CHL than did those derived from the Community clinics; this was related to an active AcrAB–TolC efflux pump, in addition to acquisition of specific resistance genes and genetic mutations. PMID:24646457

  16. 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. PMID:26190470

  17. 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.

  18. 21 CFR 522.390 - Chloramphenicol injection.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... severe infections, use 4 to 6 hour treatment intervals the first day. If no response is obtained in 3 to 5 days, discontinue use and reevaluate diagnosis. (2) Indications for use. Treatment of infections of the respiratory tract, the urinary tract, and enteritis and tonsillitis caused by...

  19. 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...

  20. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial infections...

  1. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial infections...

  2. 21 CFR 520.390a - Chloramphenicol tablets.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... infections, bacterial infections of the urinary tract, bacterial enteritis, and bacterial infections... gastroenteritis associated with bacterial diarrhea, bacterial pulmonary infections, and bacterial infections...

  3. 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...

  4. 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...

  5. 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...

  6. 21 CFR 524.390b - Chloramphenicol ophthalmic solution.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Section 524.390b Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... use. Dogs and Cats—(1) Amount. Apply one or two drops, 4 to 6 times a day for the first 72 hours...) Limitations. Therapy for cats should not exceed 7 days. As with other antibiotics, prolonged use may result...

  7. 21 CFR 524.390b - Chloramphenicol ophthalmic solution.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Section 524.390b Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES... use. Dogs and Cats—(1) Amount. Apply one or two drops, 4 to 6 times a day for the first 72 hours...) Limitations. Therapy for cats should not exceed 7 days. As with other antibiotics, prolonged use may result...

  8. 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

    2015-08-01

    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. PMID:26049592

  9. 77 FR 41412 - Determination That CHLOROMYCETIN (Chloramphenicol) Capsules, 250 Milligrams, Were Withdrawn From...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-13

    .... In the Federal Register of February 11, 2009 (74 FR 6896), FDA announced that it was withdrawing... evaluated relevant literature and data for possible postmarketing adverse events. At the time of the... later terminated in leukemia. There is published literature which suggests that the risk of...

  10. 77 FR 4895 - New Animal Drugs; Chloramphenicol, Diethylcarbamazine Citrate, Hygromycin B, Methoxyflurane...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-01

    ... Nos. 000856 and 025463 in Sec. 510.600(c) of this chapter. (c) Conditions of use in dogs and cats--(1...: (i) No. 061623 for 50-, 100-, 250-, and 500-mg; and 1-g tablets; (ii) (c) Conditions of use in...

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

    PubMed Central

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

    2015-01-01

    The complete nucleotide sequence of a 7.7 kb 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. PMID:26049592

  12. 75 FR 55810 - Withdrawal of Approval of New Animal Drug Applications; Chloramphenicol, Lincomycin, Pyrantel...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-14

    ... four new animal drug applications (NADAs). In a final rule published elsewhere in this issue of the Federal Register, FDA is amending the regulations to remove portions reflecting approval of these NADAs... FDA withdraw approval of the four NADAs listed in table 1 because they are no longer manufactured...

  13. 21 CFR 524.390d - Chloramphenicol-prednisolone ophthalmic ointment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    .... 524.390d Section 524.390d Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN.... 017030 in § 510.600(c) of this chapter. (c) Conditions of use. Dogs and cats—(1) Amount. Apply 4 to 6...) Limitations. Therapy for cats should not exceed 7 days, prolonged use in cats may produce blood dyscrasia....

  14. 21 CFR 524.390d - Chloramphenicol-prednisolone ophthalmic ointment.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... 524.390d Section 524.390d Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN.... 017030 in § 510.600(c) of this chapter. (c) Conditions of use. Dogs and cats—(1) Amount. Apply 4 to 6...) Limitations. Therapy for cats should not exceed 7 days, prolonged use in cats may produce blood dyscrasia....

  15. In vitro antimicrobial production of beta-lactamases, aminoglycoside-modifying enzymes, and chloramphenicol acetyltransferase by and susceptibility of clinical isolates of Acinetobacter baumannii.

    PubMed Central

    Vila, J; Marcos, A; Marco, F; Abdalla, S; Vergara, Y; Reig, R; Gomez-Lus, R; Jimenez de Anta, T

    1993-01-01

    Antimicrobial susceptibility testing was performed on 54 epidemiologically unrelated clinical isolates of Acinetobacter baumannii by using a standard agar dilution technique. On the basis of the in vitro activities, imipenem and doxycycline were the most active agents, whereas amikacin, isepamicin, and the new fluorquinolones ciprofloxacin and ofloxacin presented moderate activity. Cephalosporinase activity was found in 98% of the strains, whereas lactamases of TEM type 1 and one with a pI of 7 to 7.5 were present in 16 and 11% of the strains, respectively. Resistance to aminoglycosides was explained by the production of the three classes of aminoglycoside-modifying enzymes, with predominance of aminoglycoside-3'-phosphotransferase VI in 28% of the strains. PMID:8431011

  16. 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...

  17. Synergistic Interactions of Methanolic Extract of Acacia mearnsii De Wild. with Antibiotics against Bacteria of Clinical Relevance

    PubMed Central

    Olajuyigbe, Olufunmiso O.; Afolayan, Anthony J.

    2012-01-01

    With the emergence of multidrug-resistant organisms, combining medicinal plants with synthetic or orthodox medicines against resistant bacteria becomes necessary. In this study, interactions between methanolic extract of Acacia mearnsii and eight antibiotics were investigated by agar diffusion and checkerboard assays. The minimum inhibitory concentrations (MICs) of all the antibiotics ranged between 0.020 and 500 μg/mL while that of the crude extract varied between 0.156 and 1.25 mg/mL. The agar diffusion assay showed that extract-kanamycin combination had zones of inhibition ≥20 ± 1.0 mm in all the bacteria tested (100%), followed by extract-chloramphenicol (90%) > extract-ciprofloxacin = extract-tetracycline (70%) > extract-amoxicillin (60%) > extract-nalidixic acid (50%) > extract-erythromycin (40%) > extract-metronidazole (20%). The checkerboard showed synergistic interaction (61.25%), additivity/indifference (23.75%) and antagonistic (15%) effects. The synergistic interaction was most expressed by combining the extract with tetracycline, metronidazole, amoxicillin, ciprofloxacin, chloramphenicol and nalidixic acid against E. coli (ATCC 25922), erythromycin, metronidazole, amoxicillin, chloramphenicol and kanamycin against S. aureus (ATCC 6538), erythromycin, tetracycline, amoxicillin, nalidixic acid and chloramphenicol against B. subtilis KZN, erythromycin, metronidazole and amoxicillin against E. faecalis KZN, erythromycin, tetracycline, nalidixic acid and chloramphenicol against K. pneumoniae (ATCC 10031), erythromycin, tetracycline, metronidazole and chloramphenicol against P. vulgaris (ATCC 6830), erythromycin, tetracycline, amoxicillin and chloramphenicol against S. sonnei (ATCC 29930), metronidazole, amoxicillin and chloramphenicol against E. faecalis (ATCC 29212) and ciprofloxacin and chloramphenicol against Proteus vulgaris KZN. The synergistic interactions indicated that the bactericidal potentials of the antibacterial agents were improved and

  18. 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...

  19. Meningitis - meningococcal

    MedlinePlus

    ... the most commonly used antibiotics for meningococcal meningitis. Penicillin in high doses is almost always effective, too. If the patient is allergic to penicillin, chloramphenicol may be used. Sometimes corticosteroids may be ...

  20. 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,...

  1. Meningococcal Meningitis

    MedlinePlus

    ... cord. The extended meningitis belt of sub-Saharan Africa, stretching from Senegal in the west to Ethiopia ... ampicillin, chloramphenicol and ceftriaxone. Under epidemic conditions in Africa in areas with limited health infrastructure and resources, ...

  2. Shuttle mutagenesis of Neisseria gonorrhoeae: pilin null mutations lower DNA transformation competence.

    PubMed Central

    Seifert, H S; Ajioka, R S; Paruchuri, D; Heffron, F; So, M

    1990-01-01

    The method of shuttle mutagenesis has been extended to Neisseria gonorrhoeae. We have constructed a defective mini-Tn3 derivative that encodes chloramphenicol resistance in both N. gonorrhoeae and Escherichia coli and selected for mutations in the chloramphenicol resistance gene that express higher levels of antibiotic resistance in N. gonorrhoeae. Isogenic N. gonorrhoeae strains that differ only in pilin expression were constructed and used to test the effect of pilin null mutations on DNA transformation competence. PMID:2152910

  3. 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

  4. [Change of Electrophysical Properties of Escherichia coli Cells Due to Levomycetin and Tetracycline Action].

    PubMed

    Guliy, O I; Bunin, V D; Larionova, O S; Zhnichkova, E G; Balkos, A B; Ignatov, O V

    2016-01-01

    The effect of chloramphenicol and tetracycline, as inhibitors of protein synthesis, on electrophysical properties of Escherichia coli K-12 cells was investigated. Significant changes in the orientation spectra (OS) of the cell suspensions incubated with various concentrations of chloramphenicol were observed only at the first five frequencies of the electric field (10-1000 kHz). When the cells were exposed to chloramphenicol (1.5 mcg/ml) or tetracycline (1.7 mcg/ml), no changes in the OS were recorded. Significant changes in the electrooptic signal were observed, when the K-12 cells were simultaneously incubated with chloramphenicol (1.5 mcg/ml) and tetracycline (1.7 mcg/ml), that could be due to the synergistic action of the antibiotics. Therefore, the electrooptic analysis provided registration of higher antibacterial effect with the simultaneous use of chloramphenicol and tetracycline. Additional control experiments with the cell culture on the LB nutrient medium containing chloramphenicol and tetracycline were performed. The results suggested that the use of electrophysical methods for investigation of antibiotics effect on microorganisms was rather efficient. PMID:27337862

  5. Events Surrounding the Early Development of Euglena Chloroplasts

    PubMed Central

    Vaisberg, Abraham J.; Schiff, Jerome A.; Li, Lynn; Freedman, Zachary

    1976-01-01

    d(−)threo-Chloramphenicol blocks chlorophyll and plastid protein synthesis in Euglena. During chloroplast development in white light, but not in red, the cells escape from chloramphenicol inhibition and chlorophyll formation is restored. Concomitantly, chloramphenicol is reduced. Reduction of chloramphenicol in an enzyme extract from Euglena requires NADPH and ferredoxin for maximal activity. Methyl viologen replaces ferredoxin, and when chemically reduced, ferredoxin or methyl viologen reduces chloramphenicol directly. This suggests that the enzyme involved is ferredoxin-NADP reductase. In agreement, crude extracts from wild type and W3BUL, a mutant lacking detectable plastids and plastid DNA, when separated on acrylamide gels, show a single band which reduces methyl viologen with NADPH, and its mobility is similar in wild type and in mutant W3BUL. The reductase is inducible by light and increases 3-fold in wild type in white or red light and 1.5-fold in W3BUL in white light. DCMU does not block chloramphenicol reduction in vivo indicating that electrons originate from sources other than photosynthetic electron transport. We infer that chloramphenicol is reduced by ferredoxin which receives electrons via ferredoxin-NADP reductase. The limiting step is not the enzyme but the source of reducing power which can be supplied from the cytoplasm, probably under control of the blue light receptor. Ferredoxin and ferredoxin NADP reductase appear to be coded in the nuclear genome, synthesized on cytoplasmic ribosomes, and join a group of enzymes which cannot be precisely localized, since they may be active anywhere from their site of synthesis in the cytoplasm to their place of deposition in the chloroplast. PMID:16659534

  6. Reduced plasma membrane permeability in a multiple cross-resistant strain of Saccharomyces cerevisiae.

    PubMed Central

    Rank, G H; Robertson, A; Phillips, K

    1975-01-01

    Single nuclear gene inheritance was shown to be responsible for increased resistance to: eight diverse inhibitors of mitochondrial function (antimycin, carbonylcyanide-m-chlorophenylhydrazone, chloramphenicol, oligomycin, tetracycline, triethyltin bromide, triphenylmethylphosphonium bromide and triton-X-165); and an inhibitor of cytoplasmic protein synthesis (cycloheximide). Continuous monitoring of oxygen uptake during respiratory adaptation showed that anerobic pretreatment of resistant cells sensitized respiratory adaptation to chloramphenicol and antimycin. However, since a depression of mitochondrial function by catabolite repression did not result in sensitization to antimycin, alteration of the mitochondrial membrane does not appear to be responsible for resistance to mitochondrial inhibition. Alteration of cellular binding sites was not responsible for resistance since in vitro mitochondrial protein synthesis was sensitive to chloramphenicol and in vitro mitochondrial respiration was sensitive to oligomycin, carbonylcyanide-m-chlorophenylhydrazone, and antimycin. Autoradiography of an ethylacetate-ethanol extract of [14C]chloramphenicol-treated resistant cells indicated that resistance was not due to enzymatic modification of inhibitors. The maintenance of an antimycin-resistant respiration by protoplasts of resistant cells ruled out the involvement of the cell wall in cellular resistance. The reduced transport of [14C]chloramphenicol by resistant cells (1% of normal cells) indicated that a single nuclear gene mutation can alter the permeability of the plasma membrane to many diverse inhibitors. PMID:1092646

  7. Antimicrobial susceptibility testing of Haemophilus parainfluenzae by a kinetic killing-curve method.

    PubMed

    Jemsek, J G; Martin, R R; Greenberg, S B; Gentry, L O

    1980-03-01

    A kinetic killing-curve method, designed to mimic several aspects of clinical therapy in endocarditis, was used to test 10 strains of Haemophilus parainfluenzae against 28 antibiotic regimens. In an effort to simulate changing in vivo levels of antibiotic in serum, concentrations of three penicillins, three cephalosporins, gentamicin, and chloramphenicol were sequentially adjusted over a 12-hr period. Against six beta-lactamase-negative strains, gentamicin in combination with penicillin or cephalosporin invariably resulted in an additive or synergistic effect. Chloramphenicol and a penicillin or cephalosporin usually displayed an indifferent effect, but chloramphenicol was often antagonistic when combined with gentamicin. With four beta-lactamase-positive strains, variable responses were noted to penicillin-aminoglycoside combinations; cephalosporin-aminoglycoside combinations were usually synergistic. This dynamic approach to killing-curve studies may be more appropriate than a static system for in vitro examination of the effect of antimicrobial combinations against selected organisms.

  8. 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.

  9. 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

  10. 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.

  11. Multiple antibiotic resistance in Rhizobium japonicum.

    PubMed

    Cole, M A; Elkan, G H

    1979-05-01

    A total of 48 strains of the soil bacterium Rhizobium japonicum were screened for their response to several widely used antibiotics. Over 60% of the strains were resistant to chloramphenicol, polymyxin B, and erythromycin, and 47% or more of the strains were resistant to neomycin and penicillin G, when tested by disk assay procedures. The most common grouping of resistances in strains was simultaneous resistance to tetracycline, penicillin G, neomycin, chloramphenicol, and streptomycin (25% of all strains tested). The occurrence of multiple drug resistance in a soil bacterium that is not a vertebrate pathogen suggests that chemotherapeutic use of antibiotics is not required for the development of multiple drug resistance. PMID:485137

  12. 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

  13. In vitro susceptibility testing of nonsporing anaerobes to ten antimicrobial agents.

    PubMed

    Rao, P S; Shivananda, P G

    2000-07-01

    Antibiotic susceptibility was performed on sixty clinical isolates of nonsporing anaerobes against ten antimicrobial agents. The test was performed on Muller Hinton Agar and Wilkins Chalgren blood agar by preparing suspension of freshly isolated colonies in BHI broth. Apart from Metronidazole and Chloramphenicol newer antibiotics like Minocycline, Secnidazole, Tinidazole, Clarithromycin, Roxithromycin were also tried. Antimicrobial agents like Metronidazole, Secnidazole, Tinidazole and Minocycline were 100% sensitive, followed by Chloramphenicol, Clarithromycin and Roxithromycin. These newer agents can be good alternatives for the treatment of non sporing anaerobes. PMID:11218673

  14. In vitro susceptibility testing of nonsporing anaerobes to ten antimicrobial agents.

    PubMed

    Rao, P S; Shivananda, P G

    2000-04-01

    Antibiotic Susceptibility was performed on sixty clinical isolates of nonsporing anaerobes against ten antimicrobial agents. The test was performed on Muller Hinton Agar and Wilkins Chalgren blood agar by preparing suspension of freshly isolated colonies in BHI broth. Apart from Metronidazole and Chloramphenicol newer antibiotics like Minocycline, Secnidazole, Tinidazole, Clarithromycin, Roxithromycin were also tried. Antimicrobial agents like metronidazole, Secnidazole, Tinidazole and Minocycline were 100% sensitive, followed by Chloramphenicol, Clarithromycin and Roxithromycin. These newer agents can be good alternatives for the treatment of non sporing anaerobes. PMID:11217270

  15. Antimicrobial resistance of Salmonella isolates in a Spanish hospital.

    PubMed Central

    Muńoz, P; Díaz, M D; Rodríguez-Créixems, M; Cercenado, E; Peláez, T; Bouza, E

    1993-01-01

    We studied 961 clinical Salmonella isolates (one per patient) seen in one Spanish hospital from 1988 to 1991. The incidence of non-Salmonella typhi Salmonella infections per 100,000 admissions increased from 3.93 to 5.98. Overall rates of resistance to ampicillin, chloramphenicol, and co-trimoxazole were 32, 11, and 2%, respectively. Resistance to chloramphenicol increased from 9 to 16% during the study period, while resistance to each of the other drugs remained stable. Variations related to serogroups were observed. PMID:8517717

  16. Periconceptional and Gestational Exposure to Antibiotics and Childhood Asthma

    PubMed Central

    Chu, Shuyuan; Yu, Hongping; Chen, Yan; Chen, Qian; Wang, Bin; Zhang, Jun

    2015-01-01

    Background Previous studies suggest that maternal antibiotics exposure during pregnancy may increase the risk of childhood asthma, but the results were inconsistent. Furthermore, most studies did not examine periconception period as an exposure window. We aim to assess the associations between maternal exposure to specific antibiotics before and during pregnancy and the risk of asthma in early childhood. Methods Data from the Collaborative Perinatal Project were used. Maternal exposure to antibiotics before and during pregnancy was recorded at each prenatal visit. A total of 39,907 singleton children were followed up to 7 years of age. Multilevel multiple logistic regression models were used to control for potential confounders and account for multiple pregnancies per woman. Results Maternal use of penicillin or chloramphenicol was associated with an increased risk of asthma in the offspring (adjusted odds ratio = 1.21, 95% confidence interval 1.08–1.36 for penicillin; 1.72 [1.14–2.59] for chloramphenicol). The risk was significantly increased if penicillin or chloramphenicol was used in the 1st trimester (1.09 [1.04–1.13] for penicillin and 1.23 [1.01–1.51] for chloramphenicol). Conclusion Maternal exposure to certain antibiotics is associated with childhood asthma by 7 years of age. Early pregnancy may be a sensitive window. PMID:26488397

  17. 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

  18. In vitro susceptibility of Bordetella parapertussis to various antimicrobial agents.

    PubMed Central

    Watanabe, M; Haraguchi, Y

    1989-01-01

    The in vitro activity of 18 antimicrobial agents against 32 strains of Bordetella parapertussis isolated from whooping cough patients was studied. The most active antimicrobial agents were piperacillin and minocycline, followed (in descending order of activity) by moxalactam, erythromycin, cefoperazone, tetracycline, ampicillin, cefotaxime, chloramphenicol, josamycin, sulfamethoxazole, and nalidixic acid. Isolates were resistant to benzylpenicillin, cephalothin, cefatrizine, cefaclor, streptomycin, and cephalexin. PMID:2764546

  19. A novel case of chronic conjunctivitis in a 58-year-old woman caused by Raoultella.

    PubMed

    Zuberbuhler, B; Abedin, A; Roudsari, A

    2014-10-01

    A 58-year-old woman presented to eye emergency with a chronic conjunctivitis which was diagnosed by laboratory microbiological testing to be due to the environmental pathogen Raoultella planticola. The organism was sensitive to Chloramphenicol and the patient made a rapid recovery on these drops. This is the first report of this organism infecting the eye.

  20. 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

  1. An in vitro study on the effects of nisin on the antibacterial activities of 18 antibiotics against Enterococcus faecalis.

    PubMed

    Tong, Zhongchun; Zhang, Yuejiao; 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.

  2. [The effect of amphoteric tensides on the pharmaceutic availability of model drugs according to formulation].

    PubMed

    Bialik, W; Kuferska, E; Jaworski, E

    1992-01-01

    Pharmaceutical availability of chloramphenicol from o/w emulsion ointment and salicylic acid from tablets as influenced by the kind and concentration of amphoteric tenside has been investigated. It has been shown that amphoterics investigated variously influenced drug release, what makes possible to obtain the form of drug showing desired rate of release. PMID:8769076

  3. Isolation of Acanthamoeba culbertsoni from a patient with meningitis.

    PubMed

    Lalitha, M K; Anandi, V; Srivastava, A; Thomas, K; Cherian, A M; Chandi, S M

    1985-04-01

    A case of amoebic meningitis, presumably primary, was encountered in the Christian Medical College Hospital, Vellore, South India, in November 1983. The patient, a 40-year-old man, had cerebrospinal fluid rhinorrhea before the meningitis developed. Acanthamoeba culbertsoni was repeatedly demonstrated in and cultured from the cerebrospinal fluid. The patient responded dramatically to a combination therapy of penicillin and chloramphenicol. PMID:3988911

  4. 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. PMID:25736080

  5. Isolation and characterization of chromosomal promoters of Streptococcus salivarius subsp. thermophilus.

    PubMed Central

    Slos, P; Bourquin, J C; Lemoine, Y; Mercenier, A

    1991-01-01

    A promoter probe vector, pTG244, was constructed with the aim of isolating transcription initiation signals from Streptococcus thermophilus (Streptococcus salivarius subsp. thermophilus). pTG244 is based on the Escherichia coli-streptococcus shuttle vector pTG222, into which the promoterless chloramphenicol acetyltransferase gene of Bacillus pumilus (cat-86) was cloned. Random Sau3A fragments from the S. thermophilus A054 chromosomal DNA were cloned upstream of the cat-86 gene by using E. coli as the host. The pool of recombinant plasmids were introduced into S. thermophilus and Lactococcus lactis subsp. lactis in order to search for promoter activity in these hosts. For S. thermophilus, it was necessary to first select erythromycin-resistant transformants and then to screen for chloramphenicol resistance among these. Direct selection of chloramphenicol-resistant clones was, however, possible in L. lactis subsp. lactis. Six fragments exhibiting promoter activity were characterized in S. thermophilus by measuring the levels of cat-86 transcription and/or chloramphenicol acetyltransferase specific activity. Three of the promoter-carrying fragments were sequenced. The 5' ends of their corresponding mRNAs were determined by S1 mapping and shown to correspond to a purine residue in all cases. Upstream from these potential transcription start points, sequences homologous to the E. coli sigma 70 and the Bacillus subtilis vegetative sigma 43 (or sigma A) consensus promoters were identified. Images PMID:1854195

  6. Evaluation of a bi-layer wound dressing for burn care. II. In vitro and in vivo bactericidal properties.

    PubMed

    Martineau, Lucie; Shek, Pang N

    2006-03-01

    We have recently designed a medicated bi-layer wound dressing to address the key requirements for treating external, contaminated war wounds. This study assessed the in vitro and in vivo bactericidal efficacies of the DRDC hydrogel/polyurethane wound dressing. Chloramphenicol- and chlorhexidine-loaded DRDC dressings produced significantly larger zones of inhibition against Pseudomonas aeruginosa than the other medicated dressings for 4 d. Chlorhexidine-loaded Allevyn and Hydrasorb remained bactericidal for 48 h only. Chloramphenicol-loaded Hydrasorb and Allevyn remained bactericidal for 1 and 3 d, respectively. Ps. aeruginosa and Staphylococcus epidermidis counts in wounds treated with chlorhexidine- and chloramphenicol-loaded DRDC dressings for 24 h were 1-3-log lower than those of control wounds. While Ps. aeruginosa counts in the wounds on day 4 were comparable following daily changes of either antiseptic-loaded dressings, chlorhexidine showed a 75% greater bactericidal efficacy against Staph. epidermidis than chloramphenicol. Though increasing the frequency of dressing changes led to a greater reduction in the wound bacterial load, the contamination levels of all antiseptic-treated wounds remained below 10(5) CFU/g of wound. Cerium nitrate-loaded dressings did not exert any bactericidal effect, irrespective of the experimental conditions. These data show that the DRDC dressing is effective in delivering medications, such as an antimicrobial agent, to the wound bed. PMID:16455202

  7. 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. PMID:9164164

  8. 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...

  9. 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.

  10. ACSSuT Multi-Drug Resistance Among Salmonella Isolates of Animal Origin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: Multi-drug resistant (MDR) Salmonella Typhimurium DT104 (DT104) emerged in the mid-1990’s in humans and animals with infection resulting in increased morbidity and mortality. DT104 was characterized by resistance to Ampicillin, Chloramphenicol, Streptomycin, Sulfa, and Tetracycline (AC...

  11. 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.

  12. Regulation of a ribonucleoside reductase during the early generative phase in Acetabularia.

    PubMed

    de Groot, E J; Schweiger, H G

    1985-02-01

    The activity of a ribonucleoside reductase was estimated during the life cycle of Acetabularia. During the early generative phase the enzyme activity was dramatically increased. Regulation of the ribonucleoside reductase was observed even in the absence of the nucleus. The increase in activity was inhibited by chloramphenicol but not by cycloheximide. These results indicate that the enzyme is translated on 70 S ribosomes.

  13. An in vitro study on the effects of nisin on the antibacterial activities of 18 antibiotics against Enterococcus faecalis.

    PubMed

    Tong, Zhongchun; Zhang, Yuejiao; 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

  14. Primary amebic meningoencephalitis.

    PubMed

    Rai, Ruchi; Singh, D K; Srivastava, A K; Bhargava, Anudita

    2008-12-01

    We report an 8 month-old infant with primary amebic meningoencephalitis (PAME) due to Naegleria fowleri. The child was treated with amphotericin B, chloramphenicol and rifampicin for 3 weeks. PAME is an almost universally fatal condition with very few survivors till date. Our patient was one of the rare survivors who recovered after treatment and was discharged without any residual neurological deficit.

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

    PubMed Central

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

    2015-01-01

    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. PMID:25552415

  16. Unique Plasmids Generated via pUC Replicon Mutagenesis in an Error-Prone Thermophile Derived from Geobacillus kaustophilus HTA426

    PubMed Central

    Kobayashi, Jyumpei; Tanabiki, Misaki; Doi, Shohei; Kondo, Akihiko; Ohshiro, Takashi

    2015-01-01

    The plasmid pGKE75-catA138T, which comprises pUC18 and the catA138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CATA138T), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-catA138T in an error-prone thermophile, generating the mutant plasmid pGKE75αβ-catA138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75αβ-catA138T contained no mutation in the catA138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75αβ-catA138T is attributable to increases in intracellular CATA138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75αβ-catA138T. The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75αβ-catA138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli. PMID:26319877

  17. Incidence of wound infection in three different departments and the antibiotic sensitivity pattern of the isolates in a Saudi Arabian hospital.

    PubMed

    Abussaud, M J

    1996-01-01

    The incidence of wound infection in three different departments: general surgery (GS), interdisciplinary intensive care (IIC) and pediatric intensive care (PIC) in King Khalid Hospital in Hail was studied during the hot months June-October. A total of 2,331 wounds were examined. One hundred and ninety-three of them were infected (8%). The overall monthly infection rates in these departments were significantly different. They were 18, 9 and 4% in the IIC, GS, and PIC respectively. The monthly infection rates varied, the highest rates occurred in July. Two hundred and eighty three strains were isolated from the infected wounds. Seventy ones were isolated during June, and 69, 60, 42 and 41 strains were isolated during July, October, August and September, respectively- Thirty five per cent of these strains were identified as Staphylococcus aureus, 31% as Escherichia coli, 25% as Pseudomonas aeruginosa, and 10% as Klebsiella. S. aureus was the predominant organism during July; while E. coli predominated during August and October. S. aureus and P. aeruginosa strains were isolated in equal numbers during June. The sensitivity of these bacteria to ampicillin, cephalotin, cefotaxime, cefoxitin, tetracycline, chloramphenicol and gentamicin was tested by the standard disk diffusion technique. The most effective antibiotic against S. aureus was chloramphenicol (78%) followed by gentamicin (63%) and tetracycline (57%); against E. coli cefotaxime (82%) followed by cefoxitin (77%), cephalitin (62%) and chloramphenicol (53%); against Klebsiella cefotaxime (95%) followed by chloramphenicol (90%), tetracycline (70%), cephalotin and gentamicin (each 66%); and cefoxitin (60%); against P. aeruginosa cefotaxime (56%), gentamicin (33%) and chloramphenicol (29%).

  18. Unique plasmids generated via pUC replicon mutagenesis in an error-prone thermophile derived from Geobacillus kaustophilus HTA426.

    PubMed

    Kobayashi, Jyumpei; Tanabiki, Misaki; Doi, Shohei; Kondo, Akihiko; Ohshiro, Takashi; Suzuki, Hirokazu

    2015-11-01

    The plasmid pGKE75-catA138T, which comprises pUC18 and the catA138T gene encoding thermostable chloramphenicol acetyltransferase with an A138T amino acid replacement (CATA138T), serves as an Escherichia coli-Geobacillus kaustophilus shuttle plasmid that confers moderate chloramphenicol resistance on G. kaustophilus HTA426. The present study examined the thermoadaptation-directed mutagenesis of pGKE75-catA138T in an error-prone thermophile, generating the mutant plasmid pGKE75(αβ)-catA138T responsible for substantial chloramphenicol resistance at 65°C. pGKE75(αβ)-catA138T contained no mutation in the catA138T gene but had two mutations in the pUC replicon, even though the replicon has no apparent role in G. kaustophilus. Biochemical characterization suggested that the efficient chloramphenicol resistance conferred by pGKE75(αβ)-catA138T is attributable to increases in intracellular CATA138T and acetyl-coenzyme A following a decrease in incomplete forms of pGKE75(αβ)-catA138T. The decrease in incomplete plasmids may be due to optimization of plasmid replication by RNA species transcribed from the mutant pUC replicon, which were actually produced in G. kaustophilus. It is noteworthy that G. kaustophilus was transformed with pGKE75(αβ)-catA138T using chloramphenicol selection at 60°C. In addition, a pUC18 derivative with the two mutations propagated in E. coli at a high copy number independently of the culture temperature and high plasmid stability. Since these properties have not been observed in known plasmids, the outcomes extend the genetic toolboxes for G. kaustophilus and E. coli.

  19. Effects of UV light disinfection on antibiotic-resistant coliforms in wastewater effluents

    SciTech Connect

    Meckes, M.C.

    1982-02-01

    Total coliforms and total coliforms resistant to streptomycin, tetracycline, or chloramphenicol were isolated from filtered activated sludge effluents before and after UV light irradiation. Although the UV irradiation effectively disinfected the wastewater effluent, the percentage of the total surviving coliform population resistant to tetracycline or chloramphenicol was significantly higher than the percentage of the total coliform population resistant to those antibiotics before UV irradiation. This finding was attributed to the mechanism of R-factor mediated resistance to tetracycline. No significant difference was noted for the percentage of the surviving total coliform population resistant to streptomycin before or after UV irradiation. Multiple drug resistant to patterns of 300 total coliform isolates revealed that 82% were resistant to two or more antibiotics. Furthermore, 46% of these isolates were capable of transferring antibiotic resistance to a sensitive strain of Escherichia coli.

  20. 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

  1. Changing trends in the antibiograms of Salmonella isolates at a tertiary care hospital in Hyderabad.

    PubMed

    Lakshmi, V; Ashok, R; Susmita, J; Shailaja, V V

    2006-01-01

    The present study was undertaken to compare the changing trends of antibiograms of Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A isolates. A total of 80 isolates of salmonella obtained from blood cultures between 2001-2004 were included in the study. Identification and antibiotic sensitivities of the isolates were performed by using mini API (bio Merieux, France). Sixty isolates were identified as Salmonella enterica serovar Typhi and 20 were identified as Salmonella enterica serovar Paratyphi A. More than 67% of S.typhi and 80% of S.paratyphi A isolates were sensitive to chloramphenicol. Sensitivity of S.typhi isolates to cephalosporins was found to have increased from 2001-2004 while that of S.paratyphi A showed a decline. With increasing resistance to ciprofloxacin and the possibility of re-emergence of sensitivity to chloramphenicol, the policy of empirical treatment of enteric fever needs to be rationalized. PMID:16505555

  2. 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.

  3. BIOSYNTHESIS OF MYCOBACILLIN, A NEW ANTIFUNGAL PEPTIDE I.

    PubMed Central

    Banerjee, Arun B.; Bose, S. K.

    1964-01-01

    Banerjee, Arun B. (University of Calcutta, Calcutta, India), and S. K. Bose. Biosynthesis of mycobacillin, a new antifungal peptide. I. Role of nucleic acid. J. Bacteriol. 87:1397–1401. 1964.—The biosynthesis of mycobacillin, a cyclic polypeptide antifungal antibiotic, was studied in relation to the effect of chloramphenicol, 6-azathymine, and 5-bromouracil on the process. It was found that chloramphenicol inhibits both mycobacillin synthesis and growth, whereas nucleic acid base analogues inhibit only growth and nucleic acid synthesis but not mycobacillin formation. A change in the concentration of labeled aspartic acid in the general metabolic pool led to a corresponding change in the specific activity of aspartic acid isolated from different peptide fragments of the mycobacillin molecule, suggesting that mycobacillin synthesis occurs by way of linear addition of amino acid to the peptide chain. PMID:14188719

  4. Changing trends in the antibiograms of Salmonella isolates at a tertiary care hospital in Hyderabad.

    PubMed

    Lakshmi, V; Ashok, R; Susmita, J; Shailaja, V V

    2006-01-01

    The present study was undertaken to compare the changing trends of antibiograms of Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A isolates. A total of 80 isolates of salmonella obtained from blood cultures between 2001-2004 were included in the study. Identification and antibiotic sensitivities of the isolates were performed by using mini API (bio Merieux, France). Sixty isolates were identified as Salmonella enterica serovar Typhi and 20 were identified as Salmonella enterica serovar Paratyphi A. More than 67% of S.typhi and 80% of S.paratyphi A isolates were sensitive to chloramphenicol. Sensitivity of S.typhi isolates to cephalosporins was found to have increased from 2001-2004 while that of S.paratyphi A showed a decline. With increasing resistance to ciprofloxacin and the possibility of re-emergence of sensitivity to chloramphenicol, the policy of empirical treatment of enteric fever needs to be rationalized.

  5. 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.

  6. 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.

  7. Fungi responsible for skin mycoses in Turin (Italy).

    PubMed

    Filipello Marchisio, V; Preve, L; Tullio, V

    1996-01-01

    There has been a recent increase in the frequency of skin mycoses, coupled with changes in the epidemiology and distribution of the species responsible. Periodic epidemiological analyses of these disease are thus required to ensure their efficacious control. Hair fragments, skin scrapings, specimens from vesicles and blisters and nail parings were seeded on Petri dishes loaded with Sabouraud or Mycosel agar, supplemented with chloramphenicol and with chloramphenicol plus cycloheximide respectively. Parts of each specimen were also mounted in 10% potassium hydroxide for examination under the microscope. Yeasts prevailed over dermatophytes. Microsporum canis was the most frequent dermatophyte, followed by Trichophyton rubrum, T. mentagrophytes and Epidermophyton floccosum. Tinea corporis was the most common mycosis, followed by tinea unguis, tinea capitis and tinea pedis. Men were chiefly bearers of tinea cruris and tinea pedis, women of tinea corporis, and children and teenagers of tinea capitis. Some examples of the transmission of infection through interhuman contact, via animals and from the soil are also presented.

  8. 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.

  9. Molecular biology of coal bio-desulfurization; Quarterly technical progress report, October 1--December 31, 1990

    SciTech Connect

    Young, K.D.; Gallagher, J.R.

    1991-01-25

    The aim of this project is to use the techniques of molecular genetics to identify, clone, sequence, and enhance the expression of proteins which remove sulfur covalently bound to coal. This includes the movement and expression of these proteins into bacterial species which may be more useful in the industrial application of a biological desulfurization process. This quarter we finalized the initial cloning and sequencing of the dibenzothiophene (DBT) metabolic (``dox``) genes from strain C18. In addition, we constructed several mutations in single dox genes and have begun to dissect the contribution of each gene product in the DBT degradation pathway. Using a probe derived from DNA adjacent to a transposon which inactivated DBT metabolism, the DBT active genes from A15 have been cloned and identified on cosmids. We have also electroporated Thiobacillus ferrooxidans with a plasmid containing a chloramphenicol resistant transposon. Colonies of T. ferrooxidans resistant to chloramphenicol were obtained.

  10. MITOCHONDRIAL BIOGENESIS IN NEUROSPORA CRASSA

    PubMed Central

    Howell, Neil; Zuiches, Carol A.; Munkres, Kenneth D.

    1971-01-01

    The isolation of a new class of mutants permitting facultative anaerobiosis in Neurospora crassa is described. Backcross analyses to the obligate aerobe prototroph (An-) indicate single nuclear gene inheritance (An-/An+). An+ and An- are indistinguishable in morphology and growth rates under aerobic conditions. Anaerobic growth requires nutritional supplements that are dispensable for aerobic growth. Conidiogenesis, conidial germination, and vegetative growth rate are suppressed by anaerobiosis. An+ mutants produce substantial quantities of ethanol under anaerobic conditions. Anaerobiosis and chloramphenicol both affect mitochondrial enzyme activity and morphology. Chloramphenicol inhibition leads to reduction in cytochrome oxidase and swollen mitochondria with few cristae. Anaerobiosis leads to reduction in both cytochrome oxidase and malate dehydrogenase activities, enlarged mitochondria with fewer cristae, enlarged nuclei, and other alterations in cellular morphology. The fine structure of anaerobically grown cells changes with the time of anaerobic growth. We conclude that either inhibition of mitochondrial membrane synthesis or inhibition of respiration might lead to the observed alterations in mitochondria. PMID:4329155

  11. [Acute epiglottitis in adults].

    PubMed

    Castillo, A

    1992-09-01

    The author presents the clinical history of 14 patients, from 21 to 48 years of age, 10 men and 4 women, with a final diagnosis of acute epiglottitis who were hospitalized at Gorgas Army Hospital or at the San Fernando Clinic. All the patients had pharyngitis and dysphagia, a few with nasal voice, stridor and difficulty breathing, as the chief complaint. All the patients were initially intubated orally for diagnostic purposes and immediately after nasotracheal intubation was done until the patient improved in 2 or 3 days (one patient remained intubated for 5 days). All patients were kept in the Intensive Care Unit and were treated with Ampicillin and Chloramphenicol IV and lately with a second generation cephalosporin (Cefamandole). The patients allergic to Penicillin were treated with Clindamycin and Chloramphenicol. Corticosteroids were not used in any of the patients. There were no sequelae and none of the patients expired. PMID:1439005

  12. 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.

  13. Effect of UV light disinfection on antibiotic-resistant coliforms in wastewater effluents.

    PubMed Central

    Meckes, M C

    1982-01-01

    Total coliforms and total coliforms resistant to streptomycin, tetracycline, or chloramphenicol were isolated from filtered activated sludge effluents before and after UV light irradiation. Although the UV irradiation effectively disinfected the wastewater effluent, the percentage of the total surviving coliform population resistant to tetracycline or chloramphenicol was significantly higher than the percentage of the total coliform population resistant to those antibiotics before UV irradiation. This finding was attributed to the mechanism of R-factor-mediated resistance to tetracycline. No significant difference was noted for the percentage of the surviving total coliform population resistant to streptomycin before or after UV irradiation. Multiple drug resistance patterns of 300 total coliform isolates revealed that 82% were resistant to two or more antibiotics. Furthermore, 46% of these isolates were capable of transferring antibiotic resistance to a sensitive strain of Escherichia coli. PMID:7059170

  14. [Sensitivity of Proteus hauseri bacteria to chemotherapeutic preparations depending on the cultivation conditions and on the composition of the nutrient medium].

    PubMed

    Shvidenko, I G

    1977-05-01

    Sensitivity of 227 Proteus strains isolated from patients was studied comparatively using the agar-diffusion method (disks) and the method of serial dilutions. Marked differences in the numbers of the strains resistant to benzylpenicillin and chloramphenicol were found with the above methods. It was shown that the ingredients of Ploskirev's medium significantly (by 2.8--13.5 times) inhibited the antibacterial activity of streptomycin, neomycin, monomycin, kanamycin, ampicillin and nalidixic acid and had practically no effect on the activity of benzylpenicillin, chloramphenicol and furazolidone. The values of the MIC of the drugs used in the experiment with liquid media correlated with those obtained with Sabouro's medium, which provided recommendation of the latter for determination of Proteus sensitivity by the method of serial dilutions in the solid medium, Cultivation of Proteus at a temperature of 40 degrees C resulted in a decrease of the resistance to most of the drugs tested by (by 3--12.4 times).

  15. 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

  16. Dissemination of Salmonella enterica serotype agona and multidrug-resistant Salmonella enterica serotype typhimurium in Cuba.

    PubMed

    Cabrera, Roberto; Ruiz, Joaquim; Ramírez, Margarita; Bravo, Laura; Fernández, Anabel; Aladueña, Ana; Echeíta, Aurora; Gascón, Joaquim; Alonso, Pedro L; Vila, Jordi

    2006-06-01

    The molecular epidemiology, antimicrobial susceptibility, and mechanisms of resistance of 34 Salmonella spp. strains causing acute gastroenteritis, isolated from different provinces in Cuba, were determined. Sixty-four percent of the strains showed multiresistance. Salmonella typhimurium was the most frequent with 15 strains (44%), 13 of which belonged to phagotype 104 and presented similar genetic profiles of pulsed field gel electrophoresis. High levels of resistance to tetracycline (53%), spectinomycin (50%), ampicillin (44%), and chloramphenicol (41%) were found. Resistance to tetracycline was associated with the tet G and tet A genes. Resistance to ampicillin was caused by the presence of beta-lactamases, mainly the CARB type. The floR gene was the main mechanism of resistance to chloramphenicol. Our results showed an antimicrobial susceptible clone of Salmonella enterica serotype Agona in two separate regions. This is the first report of the widespread dissemination of a multiresistant clone of S. enterica serotype Typhimurium definitive phage type 104 in Cuba.

  17. 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. PMID:25133348

  18. The in vitro activity of 15 antimicrobial agents against bacterial isolates from dogs.

    PubMed

    Awji, Elias Gebru; Damte, Dereje; Lee, Seung-Jin; Lee, Joong-Su; Kim, Young-Hoan; Park, Seung-Chun

    2012-08-01

    The in vitro activity of 15 antimicrobial agents against clinical isolates of Staphylococcus pseudintermedius, Staphylococcus aureus, Escherichia coli, Pasteurella spp. and Streptococcus canis from dogs was investigated. For Staphylococcus spp., the highest frequency of resistance was observed for penicillin, followed by ampicillin, tetracycline and chloramphenicol. The highest frequency of resistance in E. coli isolates was recorded for tetracycline and streptomycin. Pasteurella spp. and S. canis had the highest resistance rate for tetracycline and chloramphenicol. Most isolates showed full susceptibility to low-level resistance to colistin, florfenicol and fluoroquinolones. Further studies using larger number of isolates from both healthy and diseased dogs would provide a broader picture of antimicrobial resistance at a national level and promote prudent use of antimicrobial agents in companion animals. PMID:22516694

  19. Evaluation of certain veterinary drug residues in food. Sixty-second report of the Joint FAO/WHO Expert Committee on food additives.

    PubMed

    2004-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of residues of certain veterinary drugs in food and to recommend maximum levels for such residues in food. The first part of the report considers conclusions on specific toxicological end-points, lipid-soluble residues of veterinary drugs with MRLs in milk, statistical methods for the estimation of MRLs, and the Committee's review and comments on documents provided by Codex Committees. Summaries follow of the Committee's evaluations of toxicological and residue data on a variety of veterinary drugs: five antibacterial agents (cefuroxime, chloramphenicol, flumequine, lincomycin, pirlimycin), four insecticides (cyhalothrin, cypermethrin and alpha-cypermethrin, doramectin, phoxim), and two production aids (melengestrol acetate, ractopamine). The Committee's comments on chloramphenicol found at low levels in animal products are also summarized. Annexed to the report is a summary of the Committee's recommendations on these drugs, including acceptable daily intakes and proposed maximum residue limits. PMID:15587045

  20. Comprehensive screening of veterinary drugs in honey by ultra-high-performance liquid chromatography coupled to mass spectrometry.

    PubMed

    Staub Spörri, Aline; Jan, Philippe; Cognard, Emmanuelle; Ortelli, Didier; Edder, Patrick

    2014-01-01

    In the context of multi-residue screening in honey, a complete methodology was developed for 200 veterinary drugs comprising a sample preparation step and an ultra-high-performance liquid chromatography (UHPLC) coupled to time-of-flight (TOF) mass spectrometry analysis. In addition, specific analytical strategies were developed for two compounds, streptomycin and chloramphenicol, using UHPLC and tandem mass spectrometry (MS/MS). Methodologies were then applied to real honey samples obtained from the Swiss market. PMID:24499104

  1. 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

    2015-01-01

    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. PMID:26782556

  2. 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

    2015-01-01

    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. PMID:26404603

  3. Antimicrobial Resistance in Salmonella in the United States from 1948 to 1995

    PubMed Central

    Tadesse, Daniel A.; Singh, Aparna; Zhao, Shaohua; Bartholomew, Mary; Womack, Niketta; Ayers, Sherry; Fields, Patricia I.

    2016-01-01

    We conducted a retrospective study of 2,149 clinical Salmonella strains to help document the historical emergence of antimicrobial resistance. There were significant increases in resistance to older drugs, including ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline, which were most common in Salmonella enterica serotype Typhimurium. An increase in multidrug resistance was observed for each decade since the 1950s. These data help show how Salmonella evolved over the past 6 decades, after the introduction of new antimicrobial agents. PMID:26856840

  4. 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

  5. A new culture medium for recovering the agents of Cryptococcosis from environmental sources.

    PubMed

    Castro e Silva, Dulcilena de Matos; Santos, Dayane C S; Pukinskas, Sandra R B S; Oshida, Julia T U; Oliveira, Lidiane; Carvallho, Anderson F; Melhem, Márcia S C

    2015-06-01

    The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans. PMID:26273249

  6. 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.

  7. Induction of Nitrate-Dependent Fe(II) Oxidation by Fe(II) in Dechloromonas sp. Strain UWNR4 and Acidovorax sp. Strain 2AN

    PubMed Central

    Chakraborty, Anirban

    2013-01-01

    We evaluated the inducibility of nitrate-dependent Fe(II)-EDTA oxidation (NDFO) in non-growth, chloramphenicol-amended, resting-cell suspensions of Dechloromonas sp. strain UWNR4 and Acidovorax sp. strain 2AN. Cells previously incubated with Fe(II)-EDTA oxidized ca. 6-fold more Fe(II)-EDTA than cells previously incubated with Fe(III)-EDTA. This is the first report of induction of NDFO by Fe(II). PMID:23144134

  8. 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. PMID:23039919

  9. 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

  10. [Correlation between the synthesis of extracellular proteases and the synthesis of the red pigment prodigiosin in Serratia marcescens].

    PubMed

    Loriia, Zh K; Briukner, B; Egorov, N S

    1977-01-01

    A correlation has been established between synthesis of exocellular protease and synthesis of a red pigment prodigiosine by Serratia marcescens. Chloramphenicol, an inhibitor of protein synthesis, inhibits also synthesis of the pigment. Leucine, an inductor of synthesis of the exocellular protease by Serratia marcescens VI, induces also synthesis of the pigment. A mixture of 18 natural amino acids, asparagine and ammonium ions represses both synthesis of the enzyme and the pigment.

  11. Relative impact of clinical evidence and over-the-counter prescribing on topical antibiotic use for acute infective conjunctivitis

    PubMed Central

    Davis, Helen; Mant, David; Scott, Caroline; Lasserson, Daniel; Rose, Peter W

    2009-01-01

    Background Acute infective conjunctivitis is a common presentation in general practice. In 2005, three placebo-controlled clinical trials showed that use of topical antibiotics had a small effect on time to clinical resolution. In the same year, chloramphenicol eye drops were made available for sale over the counter. Aim To compare the relative impact of clinical trial evidence and a change to over-the-counter availability on community use of topical chloramphenicol. Design of study Observational study using mainly routinely collected data for England. Setting National prescribing data for England and local data from general practices in Oxfordshire, England. Method Data were collated from three sources: GP prescriptions from the Prescription Pricing Authority, wholesale supply to pharmacists from IMS Health, and an audit of delayed prescribing and non-prescribing from electronic consultation records for acute conjunctivitis, in four general practices. Results The number of general practice prescriptions for topical chloramphenicol fell from 2.3 million in 2004 to 1.9 million in 2007, a reduction of 15.5%. In contrast, over-the-counter sales by pharmacists have increased steadily. The net effect of these changes has been a 47.8% increase in total chloramphenicol use during 2005–2007, with 1.1 million additional packs being used in 2007 compared to 2004. Conclusion Making an antibiotic available over the counter increases its use substantially. This is in conflict with the important public health message that antibiotic use needs to be reduced to combat resistance. These findings support the views of the Chief Medical Officer that no more antibiotics should currently be made available over the counter. PMID:20875257

  12. Multiple clones within multidrug-resistant Salmonella enterica serotype Typhimurium phage type DT104. The Greek Nontyphoidal Salmonella Study Group.

    PubMed

    Markogiannakis, A; Tassios, P T; Lambiri, M; Ward, L R; Kourea-Kremastinou, J; Legakis, N J; Vatopoulos, A C

    2000-03-01

    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.

  13. 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

  14. Comparative in vitro activities of teichomycin and other antibiotics against JK diphtheroids.

    PubMed Central

    Jadeja, L; Fainstein, V; LeBlanc, B; Bodey, G P

    1983-01-01

    The in vitro susceptibilities of 98 isolates of JK diphtheroids to teichomycin, vancomycin, and eight other antibiotics was investigated. Teichomycin and vancomycin were extremely active against all the isolates tested. The 90% minimal inhibitory concentrations for teichomycin and vancomycin were 1.56 and 0.78 micrograms/ml, respectively. The majority of the isolates were resistant to ticarcillin, methicillin, erythromycin, tetracycline, chloramphenicol, clindamycin, and cephalothin. PMID:6227280

  15. Mycotic keratitis due to Engyodontium album: first case report from India.

    PubMed

    Thamke, D C; Mendiratta, D K; Dhabarde, A; Shukla, A K

    2015-01-01

    Engyodontium album is a rare and an unusual human pathogen. It is a common inhabitant of waste and moist material and frequently isolated from substrates such as paper, jute, linen and painted walls. This fungus grew within 3 days on SDA with chloramphenicol from corneal scrapping of a 70-year-old male farmer with a history of trauma by unknown vegetative matter. The fungus can be confused with Tritirachium sp and Beauveria sp. PMID:25865991

  16. 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

  17. Atypical, fljB-Negative Salmonella enterica subsp. enterica Strain of Serovar 4,5,12:i:− Appears To Be a Monophasic Variant of Serovar Typhimurium

    PubMed Central

    Echeita, M. Aurora; Herrera, Silvia; Usera, Miguel A.

    2001-01-01

    An fljB-negative, multidrug-resistant Salmonella enterica serovar 4,5,12:i:− phage type DT U302 strain (resistant to ampicillin, chloramphenicol, sulfonamide, gentamicin, streptomycin, tetracycline, and sulfamethoxazole-trimethoprim) emerged and spread in Spain in 1997. Sequences specific for Salmonella serovar Typhimurium and phage type DT 104 and U302 were present in this atypical Salmonella strain, suggesting that it is a monophasic Salmonella serovar Typhimurium variant. PMID:11474028

  18. Atypical, fljB-negative Salmonella enterica subsp. enterica strain of serovar 4,5,12:i:- appears to be a monophasic variant of serovar Typhimurium.

    PubMed

    Echeita, M A; Herrera, S; Usera, M A

    2001-08-01

    An fljB-negative, multidrug-resistant Salmonella enterica serovar 4,5,12:i:- phage type DT U302 strain (resistant to ampicillin, chloramphenicol, sulfonamide, gentamicin, streptomycin, tetracycline, and sulfamethoxazole-trimethoprim) emerged and spread in Spain in 1997. Sequences specific for Salmonella serovar Typhimurium and phage type DT 104 and U302 were present in this atypical Salmonella strain, suggesting that it is a monophasic Salmonella serovar Typhimurium variant.

  19. A new culture medium for recovering the agents of Cryptococcosis from environmental sources

    PubMed Central

    Silva, Dulcilena de Matos Castro e; Santos, Dayane C.S.; Pukinskas, Sandra R.B.S.; Oshida, Julia T.U.; Oliveira, Lidiane; Carvallho, Anderson F.; Melhem, Márcia S.C.

    2015-01-01

    The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans. PMID:26273249

  20. 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.

  1. Improved vector for promoter screening in lactococci.

    PubMed Central

    Bojovic, B; Djordjevic, G; Topisirovic, L

    1991-01-01

    Fragments of Lactococcus lactis subsp. lactis NP45 chromosomal DNA provided promoter activity in Escherichia coli when cloned into the promoter probe vector pGKV210. Only 13% of these recombinant plasmids promoted detectable cat-86 activity when transferred to L. lactis, i.e., expressed chloramphenicol resistance. In these promoter-containing versions of pGKV210, the cat-86 gene specifies chloramphenicol-inducible chloramphenicol acetyltransferase expression. This could be a limiting factor for cloning of promoters with lower activity in L. lactis. Therefore, we have constructed a new promoter probe vector, pBV5030, with the mutated version of the cat-86 gene, which is constitutively expressed when transcriptionally activated by the insertion of a promoter. We found that in L. lactis IL1403 the constitutively expressed cat-86 gene (on a pBV5030 derivative) has four times higher activity than the inducible version of the same gene (on a pGKV210 derivative) when both have the same promoter inserted upstream of the cat-86 gene. These results suggest that plasmid pBV5030 could be a more efficient vector for the cloning of promoters from lactococci. PMID:1901705

  2. Characterization of antimicrobial-resistant phenotypes and genotypes among Salmonella enterica recovered from pigs on farms, from transport trucks, and from pigs after slaughter.

    PubMed

    Gebreyes, Wondwossen A; Davies, Peter R; Turkson, Paa-Kobina; Morrow, W E; Funk, Julie A; Altier, Craig; Thakur, Siddhartha

    2004-04-01

    The main objectives of this study were to determine antimicrobial resistance patterns among Salmonella serotypes and to evaluate the role of transport trucks in dissemination of antimicrobial-resistant strains of Salmonella. Salmonella from groups of nursery and finishing pigs on farms, from trucks, and from pigs after slaughter were compared using serotyping, patterns of antimicrobial resistance, and pulsed-field gel electrophoresis patterns. The five farms included in the study yielded 858 isolates representing 27 Salmonella serovars. The most common resistance observed (80% of all isolates) was to tetracycline; resistance to ampicillin (42%), chloramphenicol (31%), amoxicillin/clavulanic acid (30%), and piperacillin (31%) also were common. We found a correlation between serovar and antimicrobial resistance. High correlation was found between Salmonella Typhimurium var. Copenhagen and chloramphenicol resistance (Spearman rank correlation, rho = 0.7). Multidrug resistance was observed primarily in Salmonella Typhimurium var. Copenhagen (94%) and Salmonella Typhimurium (93%) and was much less common in the other common serovars, including Salmonella Derby (7%) and Salmonella Heidelberg (8%). Of the 225 isolates exhibiting the most common pentaresistance pattern in this study, amoxicillin/clavulanic acid-ampicillin-chloramphenicol-piperacillin-tetracycline, 220 (98%) were Salmonella Typhimurium var. Copenhagen, and 86% of the isolates of this serovar had this pattern. Isolates from the trucks were similar, based on pulsed-field gel electrophoresis patterns, to those from the cecum and mesenteric lymph nodes of pigs on two of the farms, suggesting the probable infection of pigs during transport. Class I integrons were also common among various serovars.

  3. Characterization of multiple-antibiotic-resistant Salmonella typhimurium stains: molecular epidemiology of PER-1-producing isolates and evidence for nosocomial plasmid exchange by a clone.

    PubMed Central

    Vahaboglu, H; Dodanli, S; Eroglu, C; Oztürk, R; Soyletir, G; Yildirim, I; Avkan, V

    1996-01-01

    We characterized epidemiologic and genetic features of nosocomially originated multiple-antibiotic-resistant Salmonella typhimurium isolates from two hospitals. A total of 32 multiply resistant strains, isolated during a 28-month period, were studied. Four resistance phenotypes were distinguished on the basis of the results of disc diffusion tests. Group 1 was resistant to chloramphenicol, gentamicin, tobramycin, amikacin, and the newer cephalosporins because of the production of an extended-spectrum beta-lactamase (PER-1). Group 2 exhibited the same pattern plus resistance to sulfamethoxazole-trimethoprim (Sxt). Except for Sxt resistance, dominant phenotypes of both groups were transferred on an identical plasmid, pSTI1 (81 MDa). Group 3 was resistant to ampicillin, chloramphenicol, gentamicin, tobramycin, and Sxt. This pattern was also transferred on an 81-MDa plasmid (pSTI2) which differed from pSTI1 on the basis of EcoRI and HindIII restriction fragments. Group 4 was resistant to ampicillin, chloramphenicol, and tetracycline, and a 74-MDa nonconjugative plasmid was detected. Restriction fragment length polymorphism of RNA-encoding DNA and arbitrarily primed PCR tests revealed that bacteria from groups 1, 2, and 3 were clonally related. Epidemiologic data also supported the clonal-dissemination hypothesis. We concluded that S. typhimurium isolates acquire and exchange multiple-resistance plasmids in hospital microflora. PMID:8940427

  4. An in vitro antibacterial study of savory essential oil and geraniol in combination with standard antimicrobials.

    PubMed

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

    2014-11-01

    The chemical composition and antibacterial activity of Satureja kitaibelii Wierzb. ex Heuff. (savory) essential oil were examined, as well as the association between it and standard antimicrobials: tetracycline and chloramphenicol. The antibacterial activities of geraniol, the main constituent of S. kitaibelii oil, individually and in combination with standard antimicrobials were also determined. The interactions of the essential oil and geraniol with antimicrobials toward five selected strains were evaluated using the microdilution checkerboard assay in combination with chemometric methods. Oxygenated monoterpenes were the most abundant compound class in the oil (59.7%), with geraniol (50.4%) as the major compound. The essential oil exhibited in vitro antibacterial activity against all tested bacterial strains, but the activities were lower than those of the standard antimicrobials. The combinations savory oil-chloramphenicol, savory oil-tetracycline and geraniol-chloramphenicol produced predominantly synergistic interactions (FIC indices in the range 0.21-0.87) and substantial reductions in the MIC values of antimicrobials against Gram-negative bacteria, the pharmacological treatment of which is very difficult nowadays. In the PCA and HCA analyses these combinations form a separate group. PMID:25532298

  5. Molecular identification of Salmonella Infantis isolated from backyard chickens and detection of their resistance genesby PCR.

    PubMed

    Ghoddusi, A; Nayeri Fasaei, B; Karimi, V; Ashrafi Tamai, I; Moulana, Z; Zahraei Salehi, T

    2015-01-01

    This study aims at molecular identification of Salmonella Infantis isolated from backyard chickens and the detection of their antibiotic resistance genes. A total of 46 Salmonella-suspected samples isolated from backyard chickens of northern Iran were collected. Serotyping was done by the traditional method and then confirmed by PCR. Antimicrobial susceptibility of the isolates against 13 antimicrobial agents was determined by the standard disk diffusion method. There were 44 samples identified as Salmonella. Serotyping results showed that all 44 isolates belonged to serogroup C1 and serovar Infantis. The most resistance observed was to tetracycline and doxycycline (100%), chloramphenicol (79%) and florfenicol (72%). The floR, catI, tetA and tetG genes were used for the detection of florfenicol chloramphenicol and tetracycline resistance. In order to identify the phenotypic resistance in strains which showed resistance genes by PCR, colony PCR and culture on plates each containing antibiotic was performed simultaneously. All the Salmonella Infantis resistant to florfenicol and chloramphenicol harbored floR and catI. None of the Salmonella resistant to tetracycline carried tetA or tetG. The result of colony PCR and culture in antibiotic medium confirmed the results of PCR and indicated phenotypic resistance in these samples. PMID:27175192

  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. Context-specific inhibition of translation by ribosomal antibiotics targeting the peptidyl transferase center

    PubMed Central

    Marks, James; Kannan, Krishna; Roncase, Emily J.; Klepacki, Dorota; Kefi, Amira; Orelle, Cédric; Vázquez-Laslop, Nora; Mankin, Alexander S.

    2016-01-01

    The first broad-spectrum antibiotic chloramphenicol and one of the newest clinically important antibacterials, linezolid, inhibit protein synthesis by targeting the peptidyl transferase center of the bacterial ribosome. Because antibiotic binding should prevent the placement of aminoacyl-tRNA in the catalytic site, it is commonly assumed that these drugs are universal inhibitors of peptidyl transfer and should readily block the formation of every peptide bond. However, our in vitro experiments showed that chloramphenicol and linezolid stall ribosomes at specific mRNA locations. Treatment of bacterial cells with high concentrations of these antibiotics leads to preferential arrest of translation at defined sites, resulting in redistribution of the ribosomes on mRNA. Antibiotic-mediated inhibition of protein synthesis is most efficient when the nascent peptide in the ribosome carries an alanine residue and, to a lesser extent, serine or threonine in its penultimate position. In contrast, the inhibitory action of the drugs is counteracted by glycine when it is either at the nascent-chain C terminus or at the incoming aminoacyl-tRNA. The context-specific action of chloramphenicol illuminates the operation of the mechanism of inducible resistance that relies on programmed drug-induced translation arrest. In addition, our findings expose the functional interplay between the nascent chain and the peptidyl transferase center. PMID:27791002

  8. [Therapeutic aspects of salmonellosis in veal calves].

    PubMed

    Groothuis, D G; van Miert, A S

    1985-01-01

    The present investigation was undertaken to improve dosage regimens--using amoxycillin, chloramphenicol or trimethoprim/sulphadiazin--in Salmonella dublin or Salmonella typhimurium infected vealcalves. The pharmacokinetic of these drugs was studied after i/v, oral, and i/m administration (bio-availability, local irritation at the injection site, volume of distribution, and elimination half-life). The most important conclusion was that amoxycillin, chloramphenicol, and trimethoprim were suitable for oral administration to vealcalves, although the bioavailability of chloramphenicol and trimethoprim was significantly less when administered with a milk replacer concurrently. In vitro the antibacterial activities of these drugs were compared with each other. Addition of sulphadiazin to trimethoprim lowered its MIC for S. dublin, but it reduced the killing rate compared to that of trimethoprim alone. In the efficacy studies the activities of several serum enzymes and the plasma concentrations of Fe, Zn, Cu, and bilirubine were measured. Using optimal dose regimens based on MIC values and blood levels treatment with either of the three drugs was of equal efficacy.

  9. 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

  10. Integron presence in a multiresistant Morganella morganii isolate.

    PubMed

    Rojas, Laura; Vinuesa, Teresa; Tubau, Fe; Truchero, Consol; Benz, Roland; Viñas, Miguel

    2006-06-01

    A multiresistant strain of Morganella morganii was isolated from a patient affected by several severe pathologies. The isolate was found to be resistant to the following antimicrobials: ampicillin, nalidixic acid, cefalothin, cefoxitin, ceftriaxone, ciprofloxacin, chloramphenicol, streptomycin, erythromycin, gentamicin, novobiocin, penicillin, rifampicin, tetracycline and violet crystal. Mechanisms leading to this multiresistance were studied. Porins of M. morganii multiresistant and wild-type strains were analysed by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and were characterised by their ability to form channels in planar black lipid bilayers. The channels formed by porins from multiresistant and susceptible strains suggested that the porins of the multiresistant strain were not responsible for resistance. A 6.6 kb plasmid (pML2003) was detected, isolated and studied. pML2003 included two integrons. Direct sequencing revealed that one of the integrons contained two cassettes, aminoglycoside adenyltransferase (aadB) and chloramphenicol acetyltransferase (catB3) conferring resistance to aminoglycosides and chloramphenicol, respectively. The second integron contained carbenicillinase (blaP1b) and adenyltransferase (aadA2), which confer resistance to beta-lactamases and streptomycin, respectively.

  11. Ribulose 1,5-Diphosphate Carboxylase Synthesis in Euglena: II. Effect of Inhibitors on Enzyme Synthesis during Regreening and Subsequent Transfer to Darkness.

    PubMed

    Lord, J M; Armitage, T L; Merrett, M J

    1975-11-01

    Dark-grown Euglena gracilis Klebs strain Z Pringsheim cells, which have been partially regreened in the light, show a striking, continued synthesis of the chloroplast enzyme ribulose 1,5-diphosphate carboxylase on transfer back into darkness. This dark synthesis of the enzyme was completely prevented by the addition of 15 mug/ml of cycloheximide to the culture medium but was unaffected, for at least 8 hours, by the addition of 1 mg/ml of d-threo-chloramphenicol. The addition of either cycloheximide or d-threo-chloramphenicol to dark-grown cultures at the onset of illumination completely inhibited the light-induced synthesis of ribulose 1,5-diphosphate carboxylase. When cells which had been illuminated in the presence of d-threo-chloramphenicol, and hence were unable to synthesize ribulose 1,5-diphosphate carboxylase, were transferred to darkness in the absence of this inhibitor, synthesis of the carboxylase then occurred. Dark-grown cells which had been illuminated in the presence of cycloheximide failed to synthesize the enzyme when placed in the dark in the absence of cycloheximide. The addition of 5-fluorouracil to regreening cultures to prevent light-induced transcriptional steps completely blocked the synthesis of ribulose 1,5-diphosphate carboxylase.

  12. Prevalence, antibiotic resistance and RAPD typing of Campylobacter species isolated from ducks, their rearing and processing environments in Penang, Malaysia.

    PubMed

    Adzitey, Frederick; Rusul, Gulam; Huda, Nurul; Cogan, Tristan; Corry, Janet

    2012-03-15

    We report for the first time on the prevalence, antibiotic resistance and RAPD types of Campylobacter species in ducks and duck related environmental samples in Malaysia. Samples were examined by enrichment in Bolton Broth followed by plating onto modified Charcoal Cefoperazone Deoxycholate agar (mCCDA) and/or plating directly onto mCCDA. A total of 643 samples were screened, and the prevalence of Campylobacter spp. in samples from different sources ranged from 0% to 85%. The method of isolation had a significant (P<0.05) effect on the isolation rate. One hundred and sixteen Campylobacter isolates, comprising of 94 Campylobacter jejuni, 19 Campylobacter coli and three Campylobacter lari, were examined for their sensitivity to 13 antibiotics. Majority of the C. jejuni isolates were resistant to cephalothin (99%), tetracycline (96%), suphamethoxazole/trimethoprim (96%), and very few were resistant to gentamicin (5%), chloramphenicol (7%) and erythromycin (1%). All C. coli isolates were resistant to cephalothin, nalidixic acid, norfloxacin and tetracycline but susceptible to chloramphenicol, erythromycin and gentamicin. The three C. lari isolates were resistant to all the antibiotics tested except chloramphenicol and gentamicin (1/3 and 2/3 susceptible, respectively). Genetic diversity of Campylobacter isolates were determined using random amplification of polymorphic DNA (RAPD). C. jejuni and C. coli isolates belong to fifty-eight and twelve RAPD types, respectively.

  13. Multivariable Analysis of the Association Between Antimicrobial Use and Antimicrobial Resistance in Escherichia coli Isolated from Apparently Healthy Pigs in Japan.

    PubMed

    Makita, Kohei; Goto, Masaki; Ozawa, Manao; Kawanishi, Michiko; Koike, Ryoji; Asai, Tetsuo; Tamura, Yutaka

    2016-01-01

    The objective of this study was to investigate the association between antimicrobial agent use and antimicrobial resistance in Escherichia coli isolated from healthy pigs using data from 2004 to 2007 in the Japanese Veterinary Antimicrobial Resistance Monitoring System (JVARM). Fecal E. coli isolates from 250 pigs (one isolate each from a pig per farm) were examined for antimicrobial resistance. Information on the use of antimicrobials within preceding 6 months and types of farms recorded in JVARM was collected and statistically analyzed against the resistance patterns. In the univariate analysis, associations between both therapeutic and feed additive use of antimicrobials, and resistance to dihydrostreptomycin, gentamicin, kanamycin, ampicillin, cefazolin, ceftiofur, oxytetracycline, chloramphenicol, trimethoprim, nalidixic acid, enrofloxacin, colistin, and bicozamycin, and husbandry factors were investigated. In multivariable analysis, generalized estimating equations were used to control geographical intraclass correlation. Confounding for structurally unrelated associations was tested using generalized linear models. The results suggested direct and cross selections in the associations between use of aminoglycosides in reproduction farms and resistance to kanamycin, use of tetracyclines in larger farms and resistance to oxytetracycline, use of beta-lactams and resistance to ampicillin, use of phenicols and resistance to chloramphenicol, and use of fluoroquinolones and resistance to nalidixic acid and enrofloxacin. Coselection was suggested in the use of tetracyclines and chloramphenicol resistance. The associations between use of beta-lactams and dihydrostreptomycin resistance, use of macrolides and ampicillin and oxytetracycline resistance, and use of colistin and kanamycin resistance were significant, but were confounded by the simultaneous use of homologous antimicrobials.

  14. 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

  15. 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

  16. Antimicrobial resistance patterns and prevalence of class 1 and 2 integrons in Shigella flexneri and Shigella sonnei isolated in Uzbekistan

    PubMed Central

    2010-01-01

    Background Shigella is a frequent cause of bacterial dysentery in the developing world. Treatment with effective antibiotics is recommended for shigellosis, but options become limited due to globally emerging resistance. One of the mechanisms for the development of resistance utilizes integrons. This study described the antibiotic susceptibility and the presence of class 1 and 2 integrons in S. flexneri and S. sonnei isolated in Uzbekistan. Results We studied 31 isolates of S. flexneri and 21 isolates of S. sonnei isolated in Uzbekistan between 1992 and 2007 for the susceptibility or resistance to ampicillin (Am), chloramphenicol (Cl), tetracycline (Te), co-trimoxazole (Sxt), kanamycin (Km), streptomycin (Str), gentamicin (Gm), cefazolin (Czn), cefoperazone (Cpr), cefuroxime (Cur), ceftazidime (Ctz), nalidixic acid (NA) and ciprofloxacin (Cip). Am/Str/Cl/Te and Am/Str/Cl/Te/Sxt resistance patterns were found most frequently in S. flexneri. Single isolates were resistant to aminoglycoside, quinolones and cephalosporins. The resistance patterns were different in the two species. Integrons were detected in 93.5% of S. flexneri (29/31) and 81.0% of S. sonnei (17/21) isolates. In addition, 61.3% of S. flexneri (19/31) isolates and 19.0% of S. sonnei (4/21) isolates carried both classes of integrons. In 29.0% of S. flexneri (9/31) isolates, only class 1 integrons were identified. In S. flexneri isolates, the presence of class 1 integrons was associated with resistance to ampicillin and chloramphenicol. Only Class 2 integrons were present in 61.9% of S. sonnei (13/21) isolates. Conclusions Our study documents antibiotic resistance among Shigella spp. in Uzbekistan. Ninety percent of Shigella strains were resistant to previously used antibiotics. Differences among S. flexneri and S. sonnei isolates in patterns of antimicrobial resistance to routinely used shigellosis antibiotics were observed. The majority of S. flexneri were resistant to ampicillin, chloramphenicol

  17. 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

  18. Structural Development during Germination of Different Populations of Mitochondria from Pea Cotyledons 1

    PubMed Central

    Malhotra, S. S.; Spencer, Mary

    1973-01-01

    The crude mitochondrial fraction from pea cotyledons can, from days 1 to 7 of germination, be separated into three fractions by sucrose density gradient centrifugation. When seeds were grown in water (control) or cycloheximide (120 micrograms per milliliter of medium) for 4 days, the originally different populations of mitochondria acquired a uniform density and separated together in band 1 (density, 1.205 grams per milliliter). The oxidative and phosphorylative activities of mitochondria obtained from 4-day-old control and 4-day-old cycloheximide-treated pea seeds were the same. However, mitochondria from pea seeds that were grown in d-threo-chloramphenicol (1.5 milligrams per milliliter of medium) or erythromycin (0.5 milligram per milliliter of medium) for 4 days separate into three bands (fully developed mitochondria in the top band [band 1] and partially developed mitochondria in the lower two bands [bands 2 and 3]). Separation patterns and oxidative and phosphorylative activities were the same for mitochondria separated from 4-day-old cotyledons treated with d-threo-chloramphenicol or erythromycin and from 1-day-old cotyledons grown in water. This indicated that these inhibitors prevented the partially developed mitochondria originally in bands 2 and 3 from developing further. In contrast, cycloheximide did not seem to interfere with the mitochondrial structural development. These results along with those obtained from the experiments on the effects of d-threo-chloramphenicol, erthromycin, and cycloheximide on 14C-leucine incorporation into mitochondrial membrane proteins suggest that the increase in mitochondrial activity during germination may be a result of structural development (membrane synthesis) in pre-existing mitochondria. PMID:16658608

  19. Evaluation of Trimethoprim/Sulfamethoxazole (SXT), Minocycline, Tigecycline, Moxifloxacin, and Ceftazidime Alone and in Combinations for SXT-Susceptible and SXT-Resistant Stenotrophomonas maltophilia by In Vitro Time-Kill Experiments

    PubMed Central

    Cai, Xuejiu; Zhao, Jin; Cui, Junchang

    2016-01-01

    Background The optimal therapy for infections caused by Stenotrophomonas maltophilia (S. maltophilia) has not yet been established. The objective of our study was to evaluate the efficacy of trimethoprim/sulfamethoxazole (SXT), minocycline, tigecycline, moxifloxacin, levofloxacin, ticarcillin-clavulanate, polymyxin E, chloramphenicol, and ceftazidime against clinical isolated S. maltophilia strains by susceptibility testing and carried out time-kill experiments in potential antimicrobials. Methods The agar dilution method was used to test susceptibility of nine candidate antimicrobials, and time-killing experiments were carried out to evaluate the efficacy of SXT, minocycline, tigecycline, moxifloxacin, levofloxacin, and ceftazidime both alone and in combinations at clinically relevant antimicrobial concentrations. Results The susceptibility to SXT, minocycline, tigecycline, moxifloxacin, levofloxacin, ticarcillin-clavulanate, chloramphenicol, polymyxin E, and ceftazidime were 93.8%, 95.0%, 83.8%, 80.0%, 76.3%, 76.3%, 37.5%, 22.5%, and 20.0% against 80 clinical consecutively isolated strains, respectively. Minocycline and tigecycline showed consistent active against 22 SXT-resistant strains. However, resistance rates were high in the remaining antimicrobial agents against SXT-resistant strains. In time-kill experiments, there were no synergisms in most drug combinations in time-kill experiments. SXT plus moxifloxacin displayed synergism when strains with low moxifloxacin MICs. Moxifloxacin plus Minocycline and moxifloxacin plus tigecycline displayed synergism in few strains. No antagonisms were found in these combinations. Overall, compared with single drug, the drug combinations demonstrated lower bacterial concentrations. Some combinations showed bactericidal activity. Conclusions In S. maltophilia infections, susceptibility testing suggests that minocycline and SXT may be considered first-line therapeutic choices while tigecycline, moxifloxacin, levofloxacin

  20. Erythromycin resistance genes in group A streptococci in Finland. The Finnish Study Group for Antimicrobial Resistance.

    PubMed

    Kataja, J; Huovinen, P; Skurnik, M; Seppälä, H

    1999-01-01

    Streptococcus pyogenes isolates (group A streptococcus) of different erythromycin resistance phenotypes were collected from all over Finland in 1994 and 1995 and studied; they were evaluated for their susceptibilities to 14 antimicrobial agents (396 isolates) and the presence of different erythromycin resistance genes (45 isolates). The erythromycin-resistant isolates with the macrolide-resistant but lincosamide- and streptogramin B-susceptible phenotype (M phenotype) were further studied for their plasmid contents and the transferability of resistance genes. Resistance to antimicrobial agents other than macrolides, clindamycin, tetracycline, and chloramphenicol was not found. When compared to our previous study performed in 1990, the rate of resistance to tetracycline increased from 10 to 93% among isolates with the inducible resistance (IR) phenotype of macrolide, lincosamide, and streptogramin B (MLSB) resistance. Tetracycline resistance was also found among 75% of the MLSB-resistant isolates with the constitutive resistance (CR) phenotype. Resistance to chloramphenicol was found for the first time in S. pyogenes in Finland; 3% of the isolates with the IR phenotype were resistant. All the chloramphenicol-resistant isolates were also resistant to tetracycline. Detection of erythromycin resistance genes by PCR indicated that, with the exception of one isolate with the CR phenotype, all M-phenotype isolates had the macrolide efflux (mefA) gene and all the MLSB-resistant isolates had the erythromycin resistance methylase (ermTR) gene; the isolate with the CR phenotype contained the ermB gene. No plasmid DNA could be isolated from the M-phenotype isolates, but the mefA gene was transferred by conjugation.

  1. Presence of multidrug-resistant enteric bacteria in dairy farm topsoil.

    PubMed

    Burgos, J M; Ellington, B A; Varela, M F

    2005-04-01

    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. To help circumvent the problem of bacterial antibiotic resistance, it is first necessary to understand the scope of the problem. However, it is 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 salicylate, a component of aspirin and known model modulator of multiple antibiotic resistance (mar) genes, influence bacterial multi-drug 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 (MIC) of antibiotics for the isolates ranged from 6 to >50 microg/mL for chloramphenicol, 2 to 8 microg/mL for nalidixic acid, 25 to >300 microg/mL for penicillin G, and 1 to >80 microg/mL for tetracycline. On the other hand, many of the isolates had significantly enhanced MIC for the same antibiotics in the presence of 5 mM salicylate. Plasmid DNA extracted from 12 randomly chosen isolates ranged in size from 6 to 12.5 kb and, in several cases, conferred resistance to chloramphenicol and penicillin G. It is concluded that enteric bacteria from dairy farm topsoil are multidrug resistant and harbor antibiotic-resistance plasmids. A role for dairy topsoil in zoonoses is suggested, implicating this environment as a reservoir for development of bacterial resistance against clinically relevant

  2. 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. PMID:16998868

  3. 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.

  4. The in-vitro antibacterial effects of organic salts, chemical disinfectants and antibiotics against pathogens of black disease in fairy shrimp of Thailand.

    PubMed

    Saejung, C; Hatai, K; Sanoamuang, L

    2014-01-01

    The antibacterial effects of organic salts, chemical disinfectants and antibiotics were evaluated on cultures of Aeromonas hydrophila C03, Aeromonas sobria C26, A. sobria C29, Aeromonas caviae C24 and Acinetobacter sp. SH-94B, the pathogens that cause black disease found in fairy shrimps (Streptocephalus sirindhornae Sanoamuang et al. (2000) and Branchinella thailandensis Sanoamuang, Saengphan & Murugan) of Thailand. The minimal inhibitory concentrations (MICs) of organic salts (sodium chloride and potassium chloride) and antibiotics (oxytetracycline dihydrate, streptomycin sulphate, kanamycin monosulphate, chloramphenicol and ampicillin) were determined using the agar-dilution method. The effect of chemical disinfectants (sodium hypochlorite and chlorine dioxide) was evaluated by exposing bacteria to different concentrations of these chemicals for different periods of time. Interestingly, all strains were intrinsically resistant to 0.25-3% sodium chloride and potassium chloride. The effect of sodium hypochlorite was greater than that of chlorine dioxide, and 5-20 μg mL(-1) of sodium hypochlorite was sufficient to inhibit the growth of these bacteria, but the exposure time varied, depending on the bacterial species. Of the antibiotics tested, chloramphenicol and oxytetracycline dihydrate completely inhibited the selected strains. Chloramphenicol showed the highest antibacterial effect against all pathogenic species--the MIC and minimal bactericidal concentration (MBC) ranged from 0.8 to 3.1 μg mL(-1) from 3.1 to 6.25 μg mL(-1), respectively. To achieve control of black disease during cultivation of fairy shrimp, data derived from this study can be used as a basis for further toxicity tests in vivo.

  5. Development of UDP-glucuronosyltransferase activity toward digitoxigenin-monodigitoxoside in neonatal rats.

    PubMed

    Watkins, J B; Klaassen, C D

    1985-01-01

    Glucuronidation is low or undetectable in embryonic and early fetal tissues and changes to adult levels at rates depending on the acceptor, tissue, and species. Because other data indicate there may be a specific UDP-glucuronosyltransferase (GT) in the liver of adult rats that glucuronidates digitoxigenin-monodigitoxoside (DIG), the development of GT activity in neonatal rats toward DIG was compared with that of other acceptors. Conjugation of p-nitrophenol and 1-naphthol was higher at birth and decreased to adult levels by 20 days of age. Glucuronidation of chloramphenicol, morphine, valproic acid, and bilirubin increased from birth to adult activity by 20 days of age. Conjugation of phenolphthalein, estrone, and diethylstilbestrol was low in 1-day-old rats and higher than adult in 20-day-old animals. In contrast, glucuronidation of DIG was barely detectable (9% of adult) in 20-day-old rats. The concentration of UDP-glucuronic acid was 50% of adult levels at birth and increased to adult values by 10 days of age. Administration of 3-methylcholanthrene on days 6 to 9 after birth significantly stimulated GT activity toward 1-naphthol, p-nitrophenol, and morphine, whereas phenobarbital precociously increased conjugation of chloramphenicol, valproic acid, morphine, and diethylstilbestrol. Pregnenolone-16 alpha-carbonitrile enhanced the development of GT activity toward morphine, chloramphenicol, valproic acid, bilirubin, diethylstilbestrol, and estrone. Glucuronidation of DIG was not increased after 3-methylcholanthrene or phenobarbital, but could be induced after pregnenolone-16 alpha-carbonitrile to 7% of adult values in 10-day-old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. 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. PMID:27294335

  7. A meta-analysis of the proportion of antimicrobial resistant human Salmonella isolates in Ethiopia

    PubMed Central

    2014-01-01

    Background Antimicrobial resistant Salmonella is a global problem and recently, a strain on the verge of pan-resistance was reported. In Ethiopia, the therapeutic management of Salmonellosis is difficult because drug sensitivity tests are not routinely carried out and treatment alternatives are not available in most health care facilities. The objectives of this study were to estimate the temporal changes and proportions of drug resistant isolates in Ethiopia. Methods Published studies on drug resistant Salmonella isolates were searched in Medline, Google Scholar and the lists of references of articles. Eligible studies were selected by using inclusion and exclusion criteria. Generic, methodological and statistical information were extracted from the eligible studies. The extracted data included the proportions of ampicillin, co-trimoxazole, chloramphenicol, ceftriaxone, ciprofloxacin and multi-drug resistant isolates. Pooled proportions were estimated by a random effects model. Results The odds of multi-drug resistant isolates in the 2000s was higher than before the 1990s (OR =18.86, 95% CI = 13.08, 27.19). The pooled proportions of ampicillin, co-trimoxazole, chloramphenicol, ciprofloxacin and multi-drug resistant isolates in the 2000s were 86.01%, 68.01%, 62.08%, 3.61% and 79.56% respectively. S. Concord (>97%) was resistant to ampicillin, co-trimoxazole, chloramphenicol and ceftriaxone. Conclusion The proportion of drug resistant isolates has increased since the 1970s. All drugs currently used for the treatment of Salmonellosis but ciprofloxacin are not reliable for an empirical therapy. Alternative drugs should be included in the essential drug list and measures should be taken to re-enforce the drug use policy. PMID:25213011

  8. Development of a rapid multiplexed assay for the direct screening of antimicrobial residues in raw milk.

    PubMed

    McGrath, Terry F; McClintock, Laura; Dunn, John S; Husar, Gregory M; Lochhead, Michael J; Sarver, Ronald W; Klein, Frank E; Rice, Jennifer A; Campbell, Katrina; Elliott, Christopher T

    2015-06-01

    Antimicrobial residues found to be present in milk can have both health and economic impacts. For these reasons, the widespread routine testing of milk is required. Due to delays with sample handling and test scheduling, laboratory-based tests are not always suited for making decisions about raw material intake and product release, especially when samples require shipping to a central testing facility. Therefore, rapid on-site screening tests that can produce results within a matter of minutes are required to facilitate rapid intake and product release processes. Such tests must be simple for use by non-technical staff. There is increasing momentum towards the development and implementation of multiplexing tests that can detect a range of important antimicrobial residues simultaneously. A simple in situ multiplexed planar waveguide device that can simultaneously detect chloramphenicol, streptomycin and desfuroylceftiofur in raw dairy milk, without sample preparation, has been developed. Samples are simply mixed with antibody prior to an aliquot being passed through the detection cartridge for 5 min before reading on a field-deployable portable instrument. Multiplexed calibration curves were produced in both buffer and raw milk. Buffer curves, for chloramphenicol, streptomycin and desfuroylceftiofur, showed linear ranges (inhibitory concentration (IC)20-IC80) of 0.1-0.9, 3-129 and 12-26 ng/ml, whilst linear range in milk was 0.13-0.74, 11-376 and 2-12 ng/ml, respectively, thus meeting European legislated concentration requirements for both chloramphenicol and streptomycin, in milk, without the need for any sample preparation. Desfuroylceftiofur-contaminated samples require only simple sample dilution to bring positive samples within the range of quantification. Assay repeatability and reproducibility were lower than 12 coefficient of variation (%CV), whilst blank raw milk samples (n = 9) showed repeatability ranging between 4.2 and 8.1%CV when measured on all

  9. Antimicrobial susceptibility monitoring of bacterial pathogens isolated from respiratory tract infections in dogs and cats across Europe: ComPath results.

    PubMed

    Morrissey, Ian; Moyaert, Hilde; de Jong, Anno; El Garch, Farid; Klein, Ulrich; Ludwig, Carolin; Thiry, Julien; Youala, Myriam

    2016-08-15

    ComPath is a pan-European resistance monitoring programme collecting bacterial pathogens from dogs and cats. We present data for respiratory tract infection (RTI) isolates collected between 2008 and 2010. Antimicrobial minimal inhibitory concentrations (MICs) were determined and susceptibility calculated following Clinical Laboratory Standards Institute (CLSI) standards for veterinary medicine. The main pathogen from dogs was Staphylococcus intermedius Group (49/215, 22.8%) which was >90% susceptible to most antimicrobials (including oxacillin - 93.9%; 3 isolates confirmed mecA-positive) but only 59.2%, 73.5% and 87.8% susceptible to tetracycline, chloramphenicol and penicillin. Bordetella bronchiseptica (48/215, 22.3%), streptococci (36/215, 16.7%), Escherichia coli (24/215, 11.2%) and Pasteurella multocida (23/215, 10.7%) were also found in dog RTI. There are no breakpoints for Bordetella bronchiseptica. Most streptococci were penicillin- chloramphenicol-, ampicillin- and pradofloxacin-susceptible. None were enrofloxacin-resistant but 6 isolates (16.7%) were of intermediate susceptibility. The least active agent against streptococci was tetracycline (47.2% susceptible). For E. coli, 37.5% were ampicillin-susceptible but 83.3% were amoxicillin/clavulanic acid-susceptible. Only chloramphenicol showed susceptibility>90% against E. coli, with 66.7% tetracycline-susceptible and 79.2% to 87.5% susceptibility to enrofloxacin, trimethoprim-sulfamethoxazole or pradofloxacin. P. multocida were susceptible to pradofloxacin (no other breakpoints are available). The main pathogen from cats was P. multocida (82/186, 44.1%), where only pradofloxacin has breakpoints (100% susceptible). Streptococci were also collected from cats (25/186, 13.4%) and were >90% susceptible to all antimicrobials except tetracycline (36% susceptible). Most susceptibility was calculated with human-derived breakpoints and some antimicrobials had no breakpoints. Therefore predictions of clinical utility

  10. Multicentre surveillance of antimicrobial resistance in enterococci and staphylococci from Colombian hospitals, 2001-2002.

    PubMed

    Arias, C A; Reyes, J; Zúñiga, M; Cortés, L; Cruz, C; Rico, C L; Panesso, D

    2003-01-01

    Invasive isolates of staphylococci and enterococci were collected from 15 tertiary care centres in five Colombian cities from 2001 to 2002. A total of 597 isolates were available for analysis. Identification was confirmed by both automated methods and multiplex PCR assays in a central laboratory. Staphylococcus aureus and coagulase-negative staphylococci (CoNS) corresponded to 49.6% and 29.6% of isolates, respectively, and 20.8% were identified as enterococci. MICs of ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, linezolid, oxacillin, rifampicin, teicoplanin, tetracycline, trimethoprim/sulfamethoxazole (SXT) and vancomycin were determined using an agar dilution method as appropriate. Screening for vancomycin-resistant S. aureus was also carried out on brain-heart infusion agar plates supplemented with vancomycin. The presence of mecA and van genes was investigated in methicillin-resistant staphylococci and glycopeptide-resistant enterococci (GRE), respectively. All staphylococci were susceptible to vancomycin, teicoplanin and linezolid. No VISA isolates were found. In S. aureus and CoNS, the lowest rates of resistance were found for SXT (7.4%) and chloramphenicol (10.7%), respectively. Resistance to oxacillin in S. aureus and CoNS was 52% and 73%, respectively. The mecA gene was detected in 97.5% of methicillin-resistant S. aureus isolates. In enterococci, resistance to glycopeptides was 9.7%: vanA (58.3%) and vanB (41.7%) genes were found. Pulsed-field gel electrophoresis indicated that the GRE isolates were closely related. Rates of resistance to ampicillin, ciprofloxacin, chloramphenicol, rifampicin and high levels of gentamicin and streptomycin were 9.7%, 27.4%, 8.9%, 43%, 17% and 28.2%, respectively. All enterococci were susceptible to linezolid.

  11. Bacterial spectrum and resistance patterns in corneal infections at a Tertiary Eye Care Center in South China

    PubMed Central

    Wang, Nan; Huang, Qiang; Tan, Yi-Wei; Lin, Li-Ping; Wu, Kai-Li

    2016-01-01

    AIM To investigate the spectrum and antibiotic susceptibility of bacteria isolated from patients with suspected corneal infections in Zhongshan Ophthalmic Center in South China over the past four years retrospectively. METHODS Totally 1943 corneal scrapes from patients with corneal infections from 2010 to 2013 were cultured and processed using standard microbiological procedures to identify bacterial isolates. Simultaneously, the bacterial isolates were tested for antibiotic susceptibility to 8 antibiotics (ceftazidime, cefuroxim, cefazolin, levofloxacin, ofloxacin, neomycin, tobramycin, chloramphenicol) using the Kirby-Bauer disc diffusion technique. RESULTS Of the total 1943 scrapes, 397 (20.43%) were culture-positive, of which 294 (74.06%) were gram-positive (GP) and 103 (25.94%) were gram-negative (GN) bacteria. Of the GP organisms, the most prevalent genera were Staphylococcus spp. (56.17%, n=223), Kocuria spp. (5.29%, n=21) and Micrococcus spp. (1.26%, n=5). On the other hand, the most prevalent genera were Pseudomonas spp. (12.85%, n=51), Burkholderia spp. (2.02%, n=8) and Acinetobacter spp. (1.51%, n=6) for the GN organisms. Among five antibiotics that have eye drop products, the resistant to neomycin of GP (7.82%, 95% CI: 4.72%-10.92%) and GN isolates (9.71%, 95% CI: 4.01%-15.41%) was lowest, while the resistant to chloramphenicol was highest (GP: 34.35%, 95% CI: 28.92%-39.78%; GN: 60.19%, 95% CI: 50.74%-69.64%). CONCLUSION Staphylococcus spp. was the most common bacterial pathogens isolated from patients with corneal infections in this setting. High percentages of GP and GN bacteria were mostly susceptible to neomycin and highly resistant to chloramphenicol. PMID:27158607

  12. Phenotyping of Salmonella serotypes isolated from natural sources of water in rural areas of East Sikkim.

    PubMed

    Poonia, S; Singh, T S; Tsering, D C

    2015-01-01

    The objective of the study was to conduct bacteriological analysis of water with special reference to Salmonella spp from natural sources of rural habitations of East Sikkim. A total of 28 Salmonella serovars isolated were biotyped, phage typed and tested for their anti-microbial susceptibility. All the isolates of Salmonella enterica serovar Typhi belonged to Biotype I. Four isolates of S. typhi belonged to phage type A. All S. paratyphi A isolates belong to phage 2. All the isolates were sensitive to chloramphenicol, cefixime and amikacin. Untreated natural water sources are unsafe for human consumption. PMID:25560018

  13. 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.

  14. DNA, a possible site of action of aluminum in Rhizobium spp

    SciTech Connect

    Johnson, A.C.; Wood, M. )

    1990-12-01

    Al was found to penetrate the cell envelopes of both sensitive and tolerant Rhizobium strains and bind to DNA in vivo. Despite causing a reduction viability, Al stimulated DNA synthesis in the sensitive strain, which suggested that an excision repair mechanism was operating. The Al-stimulated DNA synthesis was reduced by the simultaneous addition of chloramphenicol. In contrast to the sensitive strain, DNA synthesis was unaffected by Al binding to DNA in the tolerant strain. It is proposed that Al enters the cell and binds to the DNA helix, increasing stabilization and preventing successful replication. Different repair mechanisms appear to operate in response to Al in tolerant and sensitive strains.

  15. Mechanisms of recombination and function of DNA in bacteria. Progress report, January 15, 1983-January 17, 1984

    SciTech Connect

    Guild, W.R.

    1984-01-01

    Studies on gene transfer in pneumococcus are described. Specifically transformation by chromosomal and plasmid DNAs, transfection by phage and plasmid DNA, and the novel kind of conjugative transfer of drug resistance elements among the chromosomes of streptococci are examined. Some of these elements appear to be transposons. There are numerous questions about their structure, functions, and relation to plasmids and other better characterized genetic elements. The experiments involve extensive use of transformation, cloning in both pneumococcal and E. coli systems, and analysis of DNAs by gel electrophoresis. For restriction mapping, we have concentrated on the insertion carrying genes for resistance to chloramphenicol and tetracycline.

  16. Serinol: small molecule - big impact.

    PubMed

    Andreeßen, Björn; Steinbüchel, Alexander

    2011-01-01

    The amino alcohol serinol (2-amino-1,3-propanediol) has become a common intermediate for several chemical processes. Since the 1940s serinol was used as precursor for synthesis of synthetic antibiotics (chloramphenicol). In the last years, new scopes of applications were discovered. Serinol is used for X-ray contrast agents, pharmaceuticals or for chemical sphingosine/ceramide synthesis. It can either be obtained by chemical processes based on 2-nitro-1,3-propanediol, dihydroxyacetone and ammonia, dihydroxyacetone oxime or 5-amino-1,3-dioxane, or biotechnological application of amino alcohol dehydrogenases (AMDH) or transaminases. This review provides a survey of synthesis, properties and applications for serinol. PMID:21906364

  17. 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. PMID:24779904

  18. Anthracyclic products from Streptomyces erythromogenes nov. sp. Biotransformation of daunomycin (Dn) by an acellular preparation and synergism between Dn and some known antibiotics.

    PubMed

    Abdallah, N M; Devys, M; Barbier, M

    1982-01-01

    The filtrate broth as well as mycelium of the new strain Streptomyces erythrochromogenes nov. sp. isolated from Saudi Arabian soil, produce the antitumor antibiotic daunomycin 1 and two anthracyclic derivatives: 7-deoxy 13-dihydrodaunomycinone 2 and 7-deoxy daunomycinone 4. The biotransformation of 1 to 2 and 4 by an acellular enzyme preparation from the strain was found to be NADPH and/or NADH dependent. Mixtures of daunomycin 1 with chloramphenicol or penicillin showed superior antimicrobial activities against Bacillus subtilis ICC strain, than the individual antibiotics. PMID:7113265

  19. 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. PMID:1852855

  20. Further electron microscopic studies on the expression of Escherichia coli group II capsules.

    PubMed Central

    Kröncke, K D; Golecki, J R; Jann, K

    1990-01-01

    The de novo expression of Escherichia coli K1, K5, and K12 capsules was analyzed with immunoelectron microscopy in temperature upshift experiments, with upshift from 18 degrees C (capsule restrictive) to 37 degrees C (capsule permissive). Newly produced capsular polysaccharides appeared at the cell surface atop membrane adhesion sites (Bayer's junctions). After plasmolysis of the bacteria at an early expression stage, the capsular polysaccharides were labeled at discrete sites in the periplasm by the immunogold technique. After temperature upshift in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP) or chloramphenicol, the polysaccharides were labeled in the cytoplasm. Images PMID:2188961

  1. Serinol: small molecule - big impact

    PubMed Central

    2011-01-01

    The amino alcohol serinol (2-amino-1,3-propanediol) has become a common intermediate for several chemical processes. Since the 1940s serinol was used as precursor for synthesis of synthetic antibiotics (chloramphenicol). In the last years, new scopes of applications were discovered. Serinol is used for X-ray contrast agents, pharmaceuticals or for chemical sphingosine/ceramide synthesis. It can either be obtained by chemical processes based on 2-nitro-1,3-propanediol, dihydroxyacetone and ammonia, dihydroxyacetone oxime or 5-amino-1,3-dioxane, or biotechnological application of amino alcohol dehydrogenases (AMDH) or transaminases. This review provides a survey of synthesis, properties and applications for serinol. PMID:21906364

  2. 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.

  3. PHYSIOLOGICAL ROLE OF 70S RIBOSOMES IN BACTERIA*

    PubMed Central

    Algranati, Israel D.; Gonzalez, Nelida S.; Bade, Ernesto G.

    1969-01-01

    Evidence is presented indicating that free 70S ribosomes are real components of Bacillus stearothermophilus and Escherichia coli in exponential and stationary phases of growth. After pulses of radioactive leucine or uracil were given, protein synthesis was instantaneously stopped by chloramphenicol, or polypeptide growth was allowed to proceed to completion in slowly cooled cells. Analysis of label distribution in sucrose gradients showed that 70S monomers, and not subunits, are final products of translation. A ribosome cycle for protein synthesis in bacteria is proposed. PMID:4894332

  4. 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.

  5. 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.

  6. 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.

  7. Effects of methionine and Cu2+ on the expression of tyrosinase activity in Streptomyces castaneoglobisporus.

    PubMed

    Ikeda, K; Masujima, T; Sugiyama, M

    1996-12-01

    Streptomyces castaneoglobisporus HUT6202 expresses an enzyme, tyrosinase, responsible for the production of melanin-like pigments. The present study revealed that the tyrosinase synthesis by the microorganism was induced about 80-fold, when young cells cultured for 6 h were incubated with methionine (Met) to the mid-log phase of growth, in comparison to without this amino acid. The Met-induced tyrosinase synthesis was inhibited by the addition of rifampicin and chloramphenicol, suggesting that transcriptional and translational events are necessary for the induction. We found that the addition of Cu2+ to the culture medium brings forward the period of expression of Met-induced tyrosinase activity. PMID:9010762

  8. 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

  9. Bottled beverages and typhoid fever: the Mexican epidemic of 1972-73.

    PubMed Central

    Gonzalez-Cortes, A; Gangarosa, E J; Parrilla, C; Martin, W T; Espinosa-Ayala, A M; Ruiz, L; Bessudo, D; Hernandez-Arreortua, H

    1982-01-01

    A chloramphenicol resistant strain of S. typhi which caused a very large epidemic of typhoid fever in Mexico in 1972-73 survived in opened bottles of one carbonated drink with a pH of 4.6 for two weeks and in another such drink with a pH of 5.1 for six months. Bottled beverages are potential sources of large outbreaks of enteric disease, and deserve the same type of standards sand monitoring as comparable fluids such as milk. PMID:7091481

  10. 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. PMID:27524545

  11. 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

  12. Neuroinfections due to Enterococcus faecalis in children.

    PubMed

    Benca, J; Ondrusova, A; Huttova, M; Rudinsky, B; Kisac, P; Bauer, F

    2007-06-01

    Enterococcal meningitis is a rare complication of neurosurgical procedure or high technology treatment of children and occurs mainly imunocompromised neonates with very low birth weight, severe prematurity and complicates sometime ventriculoperitoneal shunt insertion or perinatal trauma. E. faecalis caused 10 nosocomial meningitis and all strains were susceptible to vancomycin and chloramphenicol, and in our database 90% also to gentamicin and ampicillin. Mortality in our group of 10 children was 20% what is insignificantly higher than overall mortality in the whole cohort of meningitis within last 15 years in our database (15.1%). Early empiric therapy should include also ampicillin or vancomycin, if enterococcal etiology is suspected.

  13. Pasteurella anatipestifer as a cause of mortality in captive wild waterfowl.

    PubMed

    Karstad, L; Lusis, P; Long, J R

    1970-10-01

    An outbreak of Pasteurella anatipestifer infection in young wild waterfowl at the Niska Waterfowl Research Center resulted in losses of approximately 100 Blue and Snow Geese, one White-fronted Goose, five Mandarin Ducks, one Black Duck and one Wood Duck. Clinical signs included diarrhea, paralysis and tremors. Gross lesions were fibrin deposits on serosal surfaces, hemorrhages on the epicardium, consolidation of the lungs, cloudy or flaky deposits on the air sacs, and dark, swollen spleens. Microscopic lesions included fibrinous meningitis, pneumonitis, air saculitis and serositis. Swollen leg and foot joints were seen in some cases. Chloramphenicol treatment seemed to be of benefit in controlling the outbreak. PMID:16512147

  14. Use of experimental design and effective mobility calculations to develop a method for the determination of antimicrobials by capillary electrophoresis.

    PubMed

    Mamani, Mónica Cecília Vargas; Amaya-Farfan, Jaime; Reyes, Felix Guillermo Reyes; Silva, José Alberto Fracassi da; Rath, Susanne

    2008-09-15

    A capillary zone electrophoresis (CZE) method for the determination of chloramphenicol (CLP), danofloxacin (DANO), ciprofloxacin (CIPRO), enrofloxacin (ENRO), sulfamethazine (SMZ), sulfaquinoxaline (SQX) and sulfamethoxazole (SMX) is described. For the development, the effective mobilities were estimated and a central composite design was performed. The method was in-house validated for CLP, CIPRO, ENRO and SMX determination in pharmaceuticals. In comparison with the HPLC method recommended by the United States Pharmacopoeia, this CZE method exhibited the same performance, with the advantage that seven different antimicrobials in pharmaceutical formulations could be simultaneously determined. PMID:18761147

  15. 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.

  16. 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.

  17. Occurrence of Leuconostoc mesenteroides and leuconostoc-like organisms in Lagos, Nigeria.

    PubMed

    Idika, N; Lawal, S F; Odugbemi, T; Cocker, H A

    1991-12-01

    A total of 91 catalase--negative Gram-positive coccal isolates obtained from 245 clinical specimens in Lagos were characterized. Ten (11.0%) of the isolates were vancomycin resistant, they fermented glucose, sucrose, fructose, lactose, mannose, mannitol, ribose, salicin, sorbitol, arabinose and xylose with acid production. One of the isolates produced in addition gas inclusive and ethanol, thus identified as Leuconostoc mesenteroides. The ten vancomycin-resistant Gram-positive coccal organisms (VRGPC) showed variable sensitivity patterns to penicillin, tetracycline, erythromycin, ampicillin, streptomycin, chloramphenicol, cloxacillin and co-trimoxazole. The possible role of Leuconostoc spp. and VRGPC in clinical infections in hospital setting is still to be defined.

  18. In vitro susceptibility of Capnocytophaga strains to 18 antimicrobial agents.

    PubMed Central

    Sutter, V L; Pyeatt, D; Kwok, Y Y

    1981-01-01

    Twenty-seven strains of capnocytophaga were tested for their susceptibility to 18 antimicrobial agents by an agar dilution technique. All strains were susceptible to achievable blood levels of penicillin G, cefaclor, cefoxitin, cefoperazone, moxalactam, clindamycin, chloramphenicol, and tetracycline. Most were susceptible to achievable levels of cefamandole, erythromycin, and metronidazole, and more than 10% were resistant to achievable levels of cephalexin and cephradine. With antimicrobial agents used in selective media, all strains were resistant to colistin, kanamycin, and nalidixic acid at commonly recommended concentrations of bacitracin and vancomycin. PMID:7283424

  19. Cross-resistance to UV radiation of a cisplatin-resistant human cell line: Overexpression of cellular factors that recognize UV-modified DNA

    SciTech Connect

    Chao, C.C.; Huang, S.L.; Huang, H.M.; Lin-Chao, S. )

    1991-04-01

    A human cell line selected for cisplatin resistance (CPR) was irradiated with UV light and showed cross-resistance to UV light. Applying a modified chloramphenicol acetyltransferase assay, we observed that CPR cells acquired enhanced host cell reactivation of a transfected plasmid carrying UV damage. Gel mobility shift analysis indicated that two nuclear factors that recognize UV-modified DNA were overexpressed in CPR cells. In addition, factors that bind UV-modified DNA were independent from the factors that bind cisplatin-modified DNA. The significance of the identified binding factors, possibly DNA repair enzymes, is discussed.

  20. 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.

  1. Sustaining protein synthesis in the absence of rapid cell division: an investigation of plasmid-encoded protein expression in Escherichia coli during very slow growth.

    PubMed

    Flickinger, M C; Rouse, M P

    1993-01-01

    The minimum growth rate capable of supporting plasmid-encoded gene expression is determined using continuous cultures of Escherichia coli MZ9387 at dilution rates (D) as low as 5% of the maximum specific growth rate. Expression from a low copy number plasmid, pMPR166, encoding cyanase under the control of P(lac) is investigated in order to study plasmid-encoded gene expression under conditions approaching starvation. Plasmid copy number was stabilized by selection in the presence of 500 micrograms/mL chloramphenicol by constitutive expression of chloramphenicol acetyl transferase (CAT). Plasmid retention was determined by dot-blot hybridization and chloramphenicol resistance. The contribution of plasmid maintenance and cyanase expression to the maximum cell yield (Y'x/s) and the maintenance coefficient (ms) was determined for MZ9387 and MZ9387:pMPR166 under uninduced and IPTG-induced conditions. The values of Y'x/s and ms for non-plasmid-bearing cultures were 0.56 g of cell dry mass (DCM)/g of glucose and 0.26 g of glucose/g of DCM.h, respectively. The cell yield for plasmid-bearing cultures under uninduced conditions (Y 0'x/s) was 0.28 g of DCM/g of glucose, with m0s = 0.08 g of glucose/g of DCM.h. These values decreased following induction of cyanase expression. Glucose consumption in the presence of IPTG was linearly related to the growth rate at D < 0.28 h-1 but nonlinear at dilution rates greater than 50% of the maximum specific growth rate, indicating that cyanase expression alters metabolism and glucose consumption. The fraction of plasmid-free cells decreased with decreasing Damköhler number (Da). These data confirm the usefulness of Da for predicting the relationship between plasmid-free and plasmid-bearing cells where plasmids are stabilized by concentrations of antibiotic greater than the minimum plasmid-free host cell growth inhibitory concentration. Specific cyanase expression increased as the dilution rate decreased to D = 0.15 h-1. Between D = 0

  2. Susceptibilities of ampicillin-resistant strains of Salmonella other than S. typhi to 10 antimicrobial agents.

    PubMed Central

    Alós, J I; Gómez-Garcés, J L; Cogollos, R; Amor, E; Pérez-Rivilla, A

    1992-01-01

    Ampicillin-resistant strains of Salmonella other than S. typhi constitute a health problem. We tested the antimicrobial susceptibilities to 10 antibiotics of 57 of these strains isolated in a 30-month period. The rates of resistance were as follows: chloramphenicol, 40.3%; tetracycline, 33.3%; gentamicin, 5.3%; co-trimoxazole, 5.3%; nalidixic acid, 1.8%; and amoxicillin-clavulanic acid, cefotaxime, ceftriaxone, aztreonam, and ciprofloxacin, 0%. In our experience, there are alternative antibiotics with excellent in vitro activities. PMID:1416869

  3. 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.

  4. Characterization of a transcriptional promoter of human papillomavirus 18 and modulation of its expression by simian virus 40 and adenovirus early antigens

    SciTech Connect

    Thierry, F.; Heard, J.M.; Dartmann, K.; Yaniv, M.

    1987-01-01

    RNA present in cells derived from cervical carcinoma that contained human papillomavirus 18 genomes was initiated in the 1.053-kilobase BamHI fragment that covered the complete noncoding region of this virus. When cloned upstream of the chloramphenicol acetyltransferase gene, this viral fragment directed the expression of the bacterial enzyme only in the sense orientation. Initiation sites were mapped around the ATG of open reading frame E6. This promoter was active in some human and simian cell lines, and its expression was modulated positively by simian virus 40 large T antigen and negatively by adenovirus type 5 E1a antigen.

  5. 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.

  6. 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.

  7. 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.

  8. AcrAB-TolC Directs Efflux-Mediated Multidrug Resistance in Salmonella enterica Serovar Typhimurium DT104

    PubMed Central

    Baucheron, Sylvie; Tyler, Shaun; Boyd, David; Mulvey, Michael R.; Chaslus-Dancla, Elisabeth; Cloeckaert, Axel

    2004-01-01

    Multidrug-resistant Salmonella enterica serovar Typhimurium definitive phage type 104 (DT104) strains harbor a genomic island, called Salmonella genomic island 1 (SGI1), which contains an antibiotic resistance gene cluster conferring resistance to ampicillin, chloramphenicol, florfenicol, streptomycin, sulfonamides, and tetracyclines. They may be additionally resistant to quinolones. Among the antibiotic resistance genes there are two, i.e., floR and tet(G), which code for efflux pumps of the major facilitator superfamily with 12 transmembrane segments that confer resistance to chloramphenicol-florfenicol and the tetracyclines, respectively. In the present study we determined, by constructing acrB and tolC mutants, the role of the AcrAB-TolC multidrug efflux system in the multidrug resistance of several DT104 strains displaying additional quinolone resistance or not displaying quinolone resistance. This study shows that the quinolone resistance and the decreased fluoroquinolone susceptibilities of the strains are highly dependent on the AcrAB-TolC efflux system and that single mutations in the quinolone resistance-determining region of gyrA are of little relevance in mediating this resistance. Overproduction of the AcrAB efflux pump, as determined by Western blotting with an anti-AcrA polyclonal antibody, appeared to be the major mechanism of resistance to quinolones. Moreover, chloramphenicol-florfenicol and tetracycline resistance also appeared to be highly dependent on the presence of AcrAB-TolC, since the introduction of mutations in the respective acrB and tolC genes resulted in a susceptible or intermediate resistance phenotype, according to clinical MIC breakpoints, despite the presence of the FloR and Tet(G) efflux pumps. Resistance to other antibiotics, ampicillin, streptomycin, and sulfonamides, was not affected in the acrB and tolC mutants of DT104 strains harboring SGI1. Therefore, AcrAB-TolC appears to direct efflux-mediated resistance to quinolones

  9. R-plasmid transfer in a wastewater treatment plant.

    PubMed Central

    Mach, P A; Grimes, D J

    1982-01-01

    Enteric bacteria have been examined for their ability to transfer antibiotic resistance in a wastewater treatment plant. Resistant Salmonella enteritidis, Proteus mirabilis, and Escherichia coli were isolated from clinical specimens and primary sewage effluent. Resistance to ampicillin, chloramphenicol, streptomycin, sulfadiazine, and tetracycline was demonstrated by spread plate and tube dilution techniques. Plasmid mediation of resistance was shown by ethidium bromide curing, agarose gel electrophoresis, and direct cell transfer. Each donor was mated with susceptible E. coli and Shigella sonnei. Mating pairs (and recipient controls) were suspended in unchlorinated primary effluent that had been filtered and autoclaved. Suspensions were added to membrane diffusion chambers which were then placed in the primary and secondary setting tanks of the wastewater treatment plant. Resistant recombinants were detected by replica plating nutrient agar master plates onto xylose lysine desoxycholate agar plates that contained per milliliter of medium 10 micrograms of ampicillin, 30 micrograms of chloramphenicol, 10 micrograms of streptomycin, 100 micrograms of sulfadiazine, or 30 micrograms of tetracycline. Mean transfer frequencies for laboratory matings were 2.1 X 10(-3). In situ matings for primary and secondary settling resulted in frequencies of 4.9 X 10(-5) and 7.5 X 10(-5), respectively. These values suggest that a significant level of resistance transfer occurs in wastewater treatment plants in the absence of antibiotics as selective agents. Images PMID:6760813

  10. Occurrence of antibiotics in pharmaceutical industrial wastewater, wastewater treatment plant and sea waters in Tunisia.

    PubMed

    Tahrani, Leyla; Van Loco, Joris; Ben Mansour, Hedi; Reyns, Tim

    2016-04-01

    Antibiotics are among the most commonly used group of pharmaceuticals in human medicine. They can therefore reach surface and groundwater bodies through different routes, such as wastewater treatment plant effluents, surface runoff, or infiltration of water used for agricultural purposes. It is well known that antibiotics pose a significant risk to environmental and human health, even at low concentrations. The aim of the present study was to evaluate the presence of aminoglycosides and phenicol antibiotics in municipal wastewaters, sea water and pharmaceutical effluents in Tunisia. All analysed water samples contained detectable levels of aminoglycoside and phenicol antibiotics. The highest concentrations in wastewater influents were observed for neomycin and kanamycin B (16.4 ng mL(-1) and 7.5 ng mL(-1), respectively). Chloramphenicol was found in wastewater influents up to 3 ng mL(-1). It was observed that the waste water treatment plants were not efficient in completely removing these antibiotics. Chloramphenicol and florfenicol were found in sea water samples near aquaculture sites at levels up to, respectively, 15.6 ng mL(-1) and 18.4 ng mL(-1). Also aminoglycoside antibiotics were found near aquaculture sites with the highest concentration of 3.4 ng mL(-1) for streptomycin. In pharmaceutical effluents, only gentamycin was found at concentrations up to 19 ng mL(-1) over a sampling period of four months. PMID:27105406

  11. Ribosome hopping and translational frameshifting are inadequate alternatives to translational attenuation in cat-86 regulation.

    PubMed Central

    Rogers, E J; Ambulos, N P; Lovett, P S

    1991-01-01

    The induction of cat-86 by chloramphenicol has been proposed to follow the translational attenuation model. In the absence of inducer, the cat-86 gene is transcribed but remains phenotypically unexpressed because the transcripts sequester the ribosome binding site for the cat coding sequence in a stable stem-loop structure, preventing translation initiation. The translational attenuation model proposes that the natural inducer, chloramphenicol, stalls a ribosome in the leader region of cat transcripts, which causes localized melting of the downstream stem-loop structure, allowing initiation of translation of the cat-86 coding sequence. Although it is established that ribosome stalling in the cat-86 leader can induce translation of the coding sequence, several subsequent steps predicted by the model remain to be experimentally confirmed. As a consequence, the present evidence for cat-86 regulation can also be explained by two other potential control devices, ribosome hopping and translational frameshifting. Here we describe experiments designed to determine whether the alternatives to translational attenuation regulate cat-86. The results obtained are inconsistent with both competing models and are consistent with predictions made by the translational attenuation model. Images FIG. 1 FIG. 4 PMID:1720771

  12. 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.

  13. Simultaneous determination of loganin, morroniside, catalpol and acteoside in normal and chronic kidney disease rat plasma by UPLC-MS for investigating the pharmacokinetics of Rehmannia glutinosa and Cornus officinalis Sieb drug pair extract.

    PubMed

    Zhao, Min; Tao, Jinhua; Qian, Dawei; Liu, Pei; Shang, Er-xin; Jiang, Shu; Guo, Jianming; Su, Shu-lan; Duan, Jin-ao; Du, Leyue

    2016-01-15

    A sensitive and rapid method for determination of loganin, morroniside, catalpol and acteoside in rat plasma after oral administration of Rehmannia glutinosa Libosch and Cornus officinalis Sieb drug pair based on ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS). Chromatographic separation was achieved using an Acquity UPLC BEH C18 column (100mm×2.1mm, 1.7μm) at a flow rate of 0.4mL/min, using gradient mode containing 0.1% formic acid in water and acetonitrile were used as the mobile phase A and B. Loganin, morroniside, catalpol, acteoside and the internal standard (chloramphenicol) were detected by selected reaction monitoring in the negative ion mode with the mass transition of m/z 451.0→179.0 (morroniside), m/z 435.0→227.0 (loganin), m/z 407.1→199.1 (catalpol), m/z 623.2→161.0 (acteoside) and m/z 320.8→151.9 (chloramphenicol), respectively. All calibration curves showed good linearity (r>0.991). The precision was evaluated by intra-day and inter-day assays and the RSD% were all within 9.58%. The recovery ranged from 67.62 to 80.14%. The method was successfully applied to pharmacokinetic study of the analytes in normal and doxorubicin-induced chronic kidney disease rat plasma.

  14. Prevalence and antimicrobial resistance of Campylobacter in raw milk in the selected areas of Poland.

    PubMed

    Wysok, B; Wiszniewska-Łaszczych, A; Uradziński, J; Szteyn, J

    2011-01-01

    During the recent years, an immense increase in the number of food poisoning cases in people caused by Campylobacter (C.) species has occurred. Raw milk, next to poultry meat, is considered the most frequent cause of food poisoning in people caused by the subject bacteria, although it is not always possible to isolate Campylobacter cells from the incriminated milk. Most probably this difficulty is caused by low concentration of the pathogen in milk at the level of 2/3 cells/ml although even such low concentration represents risk to human health. The present study was aimed at determining the occurence of Campylobacter bacteria in milk originating from selected regions of Poland. The isolation method applied in this work was effective in recovering as few as 0.1 cell of Campylobacter per g of food. Among 150 bulk milk samples tested, Campylobacter spp. was isolated from 7 (4.6%) ones. The biochemical identification of the isolated strains conducted by means of conventional biochemical tests as well as by applying the API - Campy tests revealed that all the isolates belonged to the C. jejuni species. Determination of resistance to antibiotics was performed by means of the diffusion disks method for the following antibiotics: gentamicin, ciprofloxacin, ampicillin, chloramphenicol, erythromycin, doxycyclin and tetracycline. Among 7 isolates tested, all were susceptible to ampicillin, chloramphenicol, erythromycin and gentamicin, 28.5% to doxycyclin and 14.2% to tetracycline and ciprofloxacin.

  15. 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.

  16. Multidrug-resistant Commensal Escherichia coli in Children, Peru and Bolivia

    PubMed Central

    Pallecchi, Lucia; Benedetti, Marta; Fernandez, Connie; Vallejos, Yolanda; Guzman, Elisa; Villagran, Ana Liz; Mantella, Antonia; Lucchetti, Chiara; Bartalesi, Filippo; Strohmeyer, Marianne; Bechini, Angela; Gamboa, Herlan; Rodríguez, Hugo; Falkenberg, Torkel; Kronvall, Göran; Gotuzzo, Eduardo; Paradisi, Franco; Rossolini, Gian Maria

    2006-01-01

    Using a rapid screening method, we investigated the prevalence of fecal carriage of antimicrobial drug–resistant Escherichia coli in 3,174 healthy children from 4 urban settings in Peru and Bolivia. High resistance rates were observed for ampicillin (95%), trimethoprim-sulfamethoxazole (94%), tetracycline (93%), streptomycin (82%), and chloramphenicol (70%). Lower resistance rates were observed for nalidixic acid (35%), kanamycin (28%), gentamicin (21%), and ciprofloxacin (18%); resistance to ceftriaxone and amikacin was uncommon (<0.5%). In a random sample of 1,080 resistant E. coli isolates, 90% exhibited a multidrug-resistance (MDR) phenotype. The 2 most common MDR phenotypes (ampicillin/tetracycline/trimethoprim-sulfamethoxazole and ampicillin/tetracycline/trimethoprim-sulfamethoxazole/chloramphenicol) could be transferred en bloc in conjugation experiments. The most common acquired resistance genes were blaTEM, tet(A), tet(B), drfA8, sul1, sul2, and catI. These findings underscore the magnitude of the problem of antimicrobial drug resistance in low-resource settings and the urgent need for surveillance and control of this phenomenon. PMID:16707045

  17. [Determination of 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk by liquid chromatography-tandem mass spectromtery].

    PubMed

    Wang, Hao; Zhao, Li; Yang, Hongmei; Pan, Hongyan; Shi, Hailiang; Qian, Cong; Zhang, Shan

    2015-09-01

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the simultaneous determination of 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk. The samples were extracted with alkaline acetonitrile and McIlvaine buffer solution under ultrasonication. The separation of target compounds was performed on an Eclipse XDB-C, column (150 mm x 2.1 mm, 3.5 µm) with gradient elution at a flow rate of 0.25 mL/min, and with an injection volume of 10 µL. The identification and quantification of the compounds were completed by liquid chromatography-tandem mass spectrometry in multiple reaction monitoring ( MRM) mode. The limits of detection were all below 10.0 µg/kg. The average spiked recoveries of the method ranged from 70. 1% to 109. 9% with relative standard deviations (RSDs) of 2.89%-9.99%. After validation, the method was applied to the analysis of antibiotic residues in milk products in China. Fifty samples were screened under the well defined methodology, and the results showed that chloramphenicol, only in one sample, was monitored with the content of 0.48 µg/kg. A risk of contamination of milk with chloramphenicol has been determined to exist. Therefore this method is convenient, rapid, sensitive and reliable, and can be successfully applied to the simultaneous detection of the 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk. PMID:26753289

  18. Protein synthesis by synaptosomes from rat brain. Contribution by the intraterminal mitochondria

    PubMed Central

    Hernández, A. G.

    1974-01-01

    (1) The characteristics of protein synthesis in microsomal and synaptosomal fractions from rat brain were examined. A high sensitivity to ribonuclease and to cycloheximide, and the need for the presence of pH5 enzymes distinguished protein synthesis in microsomal fractions from protein synthesis in synaptosomes. (2) Under various conditions of incubation synaptosomal fractions prepared in sucrose showed limited protein synthesis compared with synaptosomal fractions prepared by using Ficoll. Such discrepancies could not be attributed to: (i) animal age, (ii) the metabolic state of the synaptosomal fraction, (iii) the absence of bivalent cations in the incubation medium or (iv) the temperature. (3) Protein synthesis in synaptosomal fractions was inhibited 50–65% by cycloheximide, 38–50% by chloramphenicol, 95% by puromycin, 70% by azide and 40% by deoxyglucose; ribonuclease had only a negligible inhibitory effect. (4) As a first approximation to the localization of the protein-synthetic machinery present in the synaptosomal fraction, the distribution of enzymes and radioactivity in subfractions of prelabelled synaptosomes was determined after osmotic shock with water. Approximately 60% of the total protein synthesis in the synaptosomal fraction occurred in the intraterminal mitochondria. (5) Protein synthesis in the intraterminal mitochondria did not show any fundamental difference from synthesis in somatic mitochondria, with respect to inhibition by cycloheximide and chloramphenicol. (6) It was concluded that if extramitochondrial protein synthesis occurs in synaptosomes, it must be very low. PMID:4441374

  19. Protein synthesis by brain-cortex mitochondria. Characterization of a 55S mitochondrial ribosome as the functional unit in protein synthesis by cortex mitochondria and its distinction from a contaminant cytoplasmic protein-synthesizing system

    PubMed Central

    Hernandez, A.; Burdett, I.; Work, T. S.

    1971-01-01

    Homogenates of rat brain cortex were fractionated by conventional methods of velocity sedimentation and separated into a microsomal and a washed mitochondrial fraction. By electron microscopy the mitochondrial fraction was shown to be rich in synaptosomes. The mitochondria–synaptosome fraction synthesized protein in vitro by a route that was partially inhibited by cycloheximide and partly by chloramphenicol. The relative effectiveness of the two inhibitors varied greatly with the medium used. In the mitochondria–synaptosome fraction active 80S cytoplasmic ribosomes and active 55S mitochondrial ribosomes were detected; these were also seen in the electron microscope. Mild osmotic shock of the mitochondria–synaptosome fraction followed by velocity sedimentation in sucrose–EDTA allowed isolation of a mitochondrial fraction free of synaptosomes. Protein synthesis in this fraction was entirely inhibited by chloramphenicol, but was completely resistant to cycloheximide both in a medium promoting oxidative phosphorylation and in ATP-generating medium. Ouabain had no inhibitory effect on protein synthesis in a purified mitochondrial preparation. It is concluded that brain-cortex mitochondria synthesize protein entirely on 55S mitochondrial ribosomes. ImagesPLATE 4PLATE 1PLATE 2PLATE 3 PMID:5158500

  20. Cap-independent translation by the 5' untranslated region of Theiler's murine encephalomyelitis virus.

    PubMed Central

    Bandyopadhyay, P K; Wang, C; Lipton, H L

    1992-01-01

    The RNA genome of Theiler's murine encephalomyelitis viruses, a picornavirus belonging to the genus Cardiovirus, is translated in infected cells to a polyprotein. Unlike cellular messages, the 5' end of the RNA is not capped, and the untranslated region (UTR) is quite long (1,064 nucleotides in size). In poliovirus and encephalomyocarditis virus, the 5'UTR is thought to mediate cap-independent translation. We report here experiments to determine the role of the Theiler's murine encephalomyelitis virus 5'UTR in translation. Recombinant DNAs were constructed that were transcribed into bicistronic mRNAs encoding 5' chloramphenicol acetyltransferase intercistronic sequences linked to luciferase and a poly(A) 3' tail. The sequences of the 5'UTR, either complete or with sequential 5' deletions, were inserted into the intercistronic region. Bicistronic RNA transcripts were translated in a rabbit reticulocyte lysate or used to transfect BHK-21 cells, and chloramphenicol acetyltransferase and luciferase synthesis was quantitated. The results strongly suggest that the Theiler's virus 5'UTR promotes cap-independent translation and that the 5' boundary of the relevant signals resides 3' to nucleotide 500. Monocistronic mRNAs were synthesized by using an expression vector in which the 5'UTR containing deletions at the 3' terminus was inserted 5' to the coding sequences for luciferase. Analysis of luciferase translation in a rabbit reticulocyte lysate suggests that the 3' end of the translation initiation signal lies between nucleotides 1043 and 1053. Images PMID:1404591

  1. Cap-independent translation by the 5' untranslated region of Theiler's murine encephalomyelitis virus.

    PubMed

    Bandyopadhyay, P K; Wang, C; Lipton, H L

    1992-11-01

    The RNA genome of Theiler's murine encephalomyelitis viruses, a picornavirus belonging to the genus Cardiovirus, is translated in infected cells to a polyprotein. Unlike cellular messages, the 5' end of the RNA is not capped, and the untranslated region (UTR) is quite long (1,064 nucleotides in size). In poliovirus and encephalomyocarditis virus, the 5'UTR is thought to mediate cap-independent translation. We report here experiments to determine the role of the Theiler's murine encephalomyelitis virus 5'UTR in translation. Recombinant DNAs were constructed that were transcribed into bicistronic mRNAs encoding 5' chloramphenicol acetyltransferase intercistronic sequences linked to luciferase and a poly(A) 3' tail. The sequences of the 5'UTR, either complete or with sequential 5' deletions, were inserted into the intercistronic region. Bicistronic RNA transcripts were translated in a rabbit reticulocyte lysate or used to transfect BHK-21 cells, and chloramphenicol acetyltransferase and luciferase synthesis was quantitated. The results strongly suggest that the Theiler's virus 5'UTR promotes cap-independent translation and that the 5' boundary of the relevant signals resides 3' to nucleotide 500. Monocistronic mRNAs were synthesized by using an expression vector in which the 5'UTR containing deletions at the 3' terminus was inserted 5' to the coding sequences for luciferase. Analysis of luciferase translation in a rabbit reticulocyte lysate suggests that the 3' end of the translation initiation signal lies between nucleotides 1043 and 1053.

  2. A yeast mitochondrial leader peptide functions in vivo as a dual targeting signal for both chloroplasts and mitochondria.

    PubMed

    Huang, J; Hack, E; Thornburg, R W; Myers, A M

    1990-12-01

    A fusion protein was expressed in transgenic tobacco and yeast cells to examine the functional conservation of mechanisms for importing precursor proteins from the cytosol into mitochondria and chloroplasts. The test protein consisted of the mitochondrial leader peptide from the yeast precursor to cytochrome oxidase subunit Va (prC5) fused to the reporter protein chloramphenicol acetyltransferase. This protein, denoted prC5/CAT, was transported into the mitochondrial interior in yeast and tobacco cells. In both organisms, the mitochondrial form of prC5/CAT was smaller than the primary translation product, suggesting that proteolytic processing occurred during the transport process. prC5/CAT also was translocated into chloroplasts in vivo, accumulating to approximately the same levels as in plant mitochondria. However, accumulation of prC5/CAT in chloroplasts relative to mitochondria varied with the conditions under which plants were grown. The chloroplast form of prC5/CAT also appeared to have been proteolytically processed, yielding a mature protein of the same apparent size as that seen in mitochondria of either tobacco or yeast. Chloramphenicol acetyltransferase lacking a mitochondrial targeting peptide did not associate with either chloroplasts or mitochondria. The results demonstrated that in plant cells a single leader peptide can interact functionally with the protein translocation systems of both chloroplasts and mitochondria, and raised the possibility that certain native proteins might be shared between these two organelles.

  3. 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.

  4. Ionic liquid-anionic surfactant based aqueous two-phase extraction for determination of antibiotics in honey by high-performance liquid chromatography.

    PubMed

    Yang, Xiao; Zhang, Shaohua; Yu, Wei; Liu, Zhongling; Lei, Lei; Li, Na; Zhang, Hanqi; Yu, Yong

    2014-06-01

    An ionic liquid-anionic surfactant based aqueous two-phase extraction was developed and applied for the extraction of tetracycline, oxytetracycline and chloramphenicol in honey. The honey sample was mixed with Na2EDTA aqueous solution. The sodium dodecyl sulfate, ionic liquid 1-octyl-3-methylimidazolium bromide and sodium chloride were added in the mixture. After the resulting mixture was ultrasonically shaken and centrifuged, the aqueous two phase system was formed and analytes were extracted into the upper phase. The parameters affecting the extraction efficiency, such as the volume of ionic liquid, the category and amount of salts, sample pH value, extraction time and temperature were investigated. The limits of detection of tetracycline, oxytetracycline and chloramphenicol were 5.8, 8.2 and 4.2 μg kg(-1), respectively. When the present method was applied to the analysis of real honey samples, the recoveries of analytes ranged from 85.5 to 110.9% and relative standard deviations were lower than 6.9%. PMID:24767438

  5. EBNA-2 transactivates a lymphoid-specific enhancer in the BamHI C promoter of Epstein-Barr virus.

    PubMed Central

    Sung, N S; Kenney, S; Gutsch, D; Pagano, J S

    1991-01-01

    Among the few Epstein-Barr virus (EBV) genes expressed during latency are the Epstein-Barr nuclear antigens (EBNAs), at least one of which contributes to the ability of the virus to transform B lymphocytes. We have analyzed a promoter located in the BamHI-C fragment of EBV which is responsible for the expression of EBNA-1 in some cell lines. Deletion analysis of a 1.4-kb region 5' of the RNA start site has identified a 700-bp fragment that is required for optimal promoter activity in latently infected B lymphocytes, as shown by promoter constructs linked to the chloramphenicol acetyltransferase reporter gene. This fragment is also able to enhance activity, in an orientation-independent manner, of the simian virus 40 early promoter linked to the chloramphenicol acetyltransferase gene. The enhancer element has some constitutive activity in EBV-negative lymphoid cells, which is increased in the presence of the EBNA-2 gene product. Further deletions have shown that the EBNA-2-responsive region requires a 98-bp region that contains a degenerate octamer-binding motif. In epithelial cells there was no enhancer activity regardless of the presence of EBNA-2. These results demonstrate that BamHI-C promoter activity may be dependent not on an enhancer contained in the ori-P, as was previously assumed, but rather on EBNA-2 transactivation of this more proximal enhancer located in the upstream region of the BamHI C promoter itself. Images PMID:1850003

  6. 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.

  7. Induction of beta-galactosidase in Streptomyces violaceus.

    PubMed

    Sanchez, J; Hardisson, C

    1979-07-01

    Synthesis of beta-galactosidase by Streptomyces violaceus was induced by D-galactose and L-arabinose, and to a lesser extent by lactose, D-arabinose, and methyl-beta-D-galactopyranoside. The synthesis of the enzyme was linear and started to increase 2--3 h after induction by galactose, reaching a maximum after 5--7 h. The highest level of specific activity was observed in 2% galactose, with an increase of 45 times over the basal level in glycerol. Isopropyl-beta-D-thiogalactopyranoside (IPTG) and methyl-beta-D-thiogalactopyranoside (TMG) inhibited induction by D-galactose, but did not influence enzymatic activity. Cellular extracts hydrolyzed O-nitrophenyl-beta-D-galactopyranoside, but did not significantly hydrolyze lactose, melibiose, p-nitrophenyl-alpha-D-galactopyranoside, p-nitrophenyl-beta-D-fucoside, or p-nitrophenyl-beta-D-glucopyranoside. Rifampicin and chloramphenicol inhibited beta-galactosidase synthesis in non-preinduced and in preinduced cells. The inhibition by chloramphenicol was reversible. PMID:113072

  8. 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. PMID:27534137

  9. 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

    2015-02-01

    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.

  10. 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

  11. Expression and organization of BP74, a cyclic AMP-regulated gene expressed during Dictyostelium discoideum development.

    PubMed Central

    Hopkinson, S B; Pollenz, R S; Drummond, I; Chisholm, R L

    1989-01-01

    We have characterized a cDNA and the corresponding gene for a cyclic AMP-inducible gene expressed during Dictyostelium development. This gene, BP74, was found to be first expressed about the time of aggregate formation, approximately 6 h after starvation. Accumulation of BP74 mRNA did not occur in Dictyostelium cells that had been starved in fast-shaken suspension cultures but was induced in similar cultures to which cyclic AMP pulses had been added. The BP74 cDNA and gene were characterized by DNA sequence analysis and transcriptional mapping. When the BP74 promoter region was fused with a chloramphenicol acetyltransferase reporter gene and reintroduced into Dictyostelium cells, the transfected chloramphenicol acetyltransferase gene displayed the same developmentally regulated pattern of expression as did the endogenous BP74 gene, suggesting that all of the cis-acting elements required for regulated expression were carried by a 2-kilobase cloned genomic fragment. On the basis of sequence analysis, the gene appeared to encode a protein containing a 20-residue hydrophobic sequence at the amino-terminal end and 26 copies of a 20-amino-acid repeat. Images PMID:2555685

  12. Denitrification in soil amended with thermophile-fermented compost suppresses nitrate accumulation in plants.

    PubMed

    Ishikawa, Kazuto; Ohmori, Takashi; Miyamoto, Hirokuni; Ito, Toshiyuki; Kumagai, Yoshifumi; Sonoda, Masatoshi; Matsumoto, Jirou; Miyamoto, Hisashi; Kodama, Hiroaki

    2013-02-01

    NO (3) (-) is a major nitrogen source for plant nutrition, and plant cells store NO (3) (-) in their vacuoles. Here, we report that a unique compost made from marine animal resources by thermophiles represses NO (3) (-) accumulation in plants. A decrease in the leaf NO (3) (-) content occurred in parallel with a decrease in the soil NO (3) (-) level, and the degree of the soil NO (3) (-) decrease was proportional to the compost concentration in the soil. The compost-induced reduction of the soil NO (3) (-) level was blocked by incubation with chloramphenicol, indicating that the soil NO (3) (-) was reduced by chloramphenicol-sensitive microbes. The compost-induced denitrification activity was assessed by the acetylene block method. To eliminate denitrification by the soil bacterial habitants, soil was sterilized with γ irradiation and then compost was amended. After the 24-h incubation, the N(2)O level in the compost soil with presence of acetylene was approximately fourfold higher than that in the compost soil with absence of acetylene. These results indicate that the low NO (3) (-) levels that are often found in the leaves of organic vegetables can be explained by compost-mediated denitrification in the soil.

  13. 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.

  14. 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. PMID:24001605

  15. Prevalence and antimicrobial resistance of Campylobacter in raw milk in the selected areas of Poland.

    PubMed

    Wysok, B; Wiszniewska-Łaszczych, A; Uradziński, J; Szteyn, J

    2011-01-01

    During the recent years, an immense increase in the number of food poisoning cases in people caused by Campylobacter (C.) species has occurred. Raw milk, next to poultry meat, is considered the most frequent cause of food poisoning in people caused by the subject bacteria, although it is not always possible to isolate Campylobacter cells from the incriminated milk. Most probably this difficulty is caused by low concentration of the pathogen in milk at the level of 2/3 cells/ml although even such low concentration represents risk to human health. The present study was aimed at determining the occurence of Campylobacter bacteria in milk originating from selected regions of Poland. The isolation method applied in this work was effective in recovering as few as 0.1 cell of Campylobacter per g of food. Among 150 bulk milk samples tested, Campylobacter spp. was isolated from 7 (4.6%) ones. The biochemical identification of the isolated strains conducted by means of conventional biochemical tests as well as by applying the API - Campy tests revealed that all the isolates belonged to the C. jejuni species. Determination of resistance to antibiotics was performed by means of the diffusion disks method for the following antibiotics: gentamicin, ciprofloxacin, ampicillin, chloramphenicol, erythromycin, doxycyclin and tetracycline. Among 7 isolates tested, all were susceptible to ampicillin, chloramphenicol, erythromycin and gentamicin, 28.5% to doxycyclin and 14.2% to tetracycline and ciprofloxacin. PMID:21957744

  16. [Determination of 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk by liquid chromatography-tandem mass spectromtery].

    PubMed

    Wang, Hao; Zhao, Li; Yang, Hongmei; Pan, Hongyan; Shi, Hailiang; Qian, Cong; Zhang, Shan

    2015-09-01

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was established for the simultaneous determination of 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk. The samples were extracted with alkaline acetonitrile and McIlvaine buffer solution under ultrasonication. The separation of target compounds was performed on an Eclipse XDB-C, column (150 mm x 2.1 mm, 3.5 µm) with gradient elution at a flow rate of 0.25 mL/min, and with an injection volume of 10 µL. The identification and quantification of the compounds were completed by liquid chromatography-tandem mass spectrometry in multiple reaction monitoring ( MRM) mode. The limits of detection were all below 10.0 µg/kg. The average spiked recoveries of the method ranged from 70. 1% to 109. 9% with relative standard deviations (RSDs) of 2.89%-9.99%. After validation, the method was applied to the analysis of antibiotic residues in milk products in China. Fifty samples were screened under the well defined methodology, and the results showed that chloramphenicol, only in one sample, was monitored with the content of 0.48 µg/kg. A risk of contamination of milk with chloramphenicol has been determined to exist. Therefore this method is convenient, rapid, sensitive and reliable, and can be successfully applied to the simultaneous detection of the 35 antibiotic residues of tetracyclines, sulfonamides, penicillins, macrolides and amphenicols in milk.

  17. The antibiotic sensitivity patterns of Bacillus anthracis isolated from the Kruger National Park.

    PubMed

    Odendaal, M W; Pieterson, P M; de Vos, V; Botha, A D

    1991-03-01

    Forty-four isolates of Bacillus anthracis made from carcasses and soil in different localities of an endemic anthrax area in the Kruger National Park, South Africa, were tested by standard disc diffusion for their susceptibility to 18 different antibiotics. These were ampicillin, penicillin G, sulphatriad, streptomycin, clindamycin, gentamicin, fusidic acid, trimethoprim, sulphamethoxazole, chloramphenicol, erythromycin, methicillin, tetracycline (2 different concentrations), novobiocin, cefotaxime, netilmicin, cefamandole and cefoxitin. All the isolates were susceptible to ampicillin, streptomycin, chloramphenicol, erythromycin, tetracycline, methicillin and netilmicin. More than 90% of the isolates were sensitive to clindamycin, gentamicin and cefoxitin, whereas only 84.1% of the isolates were sensitive to penicillin G, 86.4% to novobiocin and 68.18% to cefamandole. Complete resistance in 100% of the isolates was encountered with trimethoprim and sulphamethoxazole, with 95.45% for sulphatriad. Moderate sensitivity occurred with penicillin G (15.9% of the isolates), clindamycin (6.8%), novobiocin (13.6%), fusidic acid (84.1%), cefotaxime (100%), cefamandole (31.8%) and cefoxitin (6.8%). The relevance of the findings to the therapeutic uses of different types of antibiotic in human clinical cases referred to in the literature is discussed. PMID:1905000

  18. 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. PMID:21153729

  19. A novel in vivo model for evaluating agents that protect against ultraviolet A-induced photoaging.

    PubMed

    Takeuchi, T; Uitto, J; Bernstein, E F

    1998-04-01

    Increasing evidence demonstrates that ultraviolet A radiation (UVA) contributes to photoaging, which results in the accumulation of massive amounts of abnormal elastic material in the dermis of photoaged skin. To study UVA-induced photoaging in an in vivo system, we utilized a line of transgenic mice containing the human elastin promoter linked to a chloramphenicol acetyl transferase reporter gene. Our prior work demonstrates promoter activation in response to ultraviolet B radiation (UVB), UVA, and psoralen plus ultraviolet A radiation in the skin of these mice. The addition of psoralen (8-MOP) prior to administration of UVA results in substantial increases in promoter activation, as compared with UVA alone. To demonstrate the utility of these mice as a model of UVA-induced photodamage, we administered four lotions to the skin of our transgenic mice that included: a sunscreen containing octyl methoxycinnamate and benzophenone-3 with a sun protection factor (SPF) of 15, the UVA filter butyl methoxydibenzoylmethane, the SPF 15 sunscreen and the UVA filter together, and the lotion vehicle alone. Following sunscreen administration, mice received a single psoralen plus ultraviolet A radiation treatment. All sunscreens decreased chloramphenicol acetyl transferase activity with the SPF 15 sunscreen, the UVA filter, and the combination SPF 15 sunscreen and UVA filter, resulting in increasing degrees of protection against psoralen plus ultraviolet A radiation. These results demonstrate that this model functions as a rapid and sensitive model of UVA photodamage for the identification and comparison of compounds that protect against UVA-induced photoaging.

  20. Mycotoxigenic Fungi and Natural Co-Occurrence of Mycotoxins in Rainbow Trout (Oncorhynchus mykiss) Feeds

    PubMed Central

    Greco, Mariana; Pardo, Alejandro; Pose, Graciela

    2015-01-01

    Samples of rainbow trout feed were analyzed with the aim to determine the mycobiota composition and the co-occurrence of mycotoxins. A total of 28 samples of finished rainbow trout feed from hatcheries in the provinces of Río Negro and Neuquén, Argentina, were studied. Fungal counts were obtained on three culture media in the ranges of <10 to 4.2 × 104 CFU/g on Dichloran Rose Bengal Chloramphenicol Agar (DRBC), <10 to 5.1 × 104 CFU/g on Dichloran Chloramphenicol Peptone Agar (DCPA) and <10 to 3.6 × 104 CFU/g on Dichloran 18% Glycerol Agar (DG18). The most frequent mycotoxigenic fungi were Eurotium (frequency (Fr) 25.0%), followed by Penicillium (Fr 21.4%) and Aspergillus (Fr 3.6%). The most prevalent mycotoxigenic species were E. repens (Fr 21.4%) and E. rubrum (Fr 14.3%). All samples were contaminated with mycotoxins: 64% samples were contaminated with T-2 toxin (median 70.08 ppb), 50% samples with zearalenone (median 87.97 ppb) and aflatoxins (median 2.82 ppb), 25% with ochratoxin A (median 5.26 ppb) and 3.57% samples with deoxynivalenol (median 230 ppb). Eight samples had a fumonisins contamination level below the limit of detection. Co-occurrence of six mycotoxins was determined in 7% of the samples. PMID:26556374

  1. Occurrence of antibiotics in pharmaceutical industrial wastewater, wastewater treatment plant and sea waters in Tunisia.

    PubMed

    Tahrani, Leyla; Van Loco, Joris; Ben Mansour, Hedi; Reyns, Tim

    2016-04-01

    Antibiotics are among the most commonly used group of pharmaceuticals in human medicine. They can therefore reach surface and groundwater bodies through different routes, such as wastewater treatment plant effluents, surface runoff, or infiltration of water used for agricultural purposes. It is well known that antibiotics pose a significant risk to environmental and human health, even at low concentrations. The aim of the present study was to evaluate the presence of aminoglycosides and phenicol antibiotics in municipal wastewaters, sea water and pharmaceutical effluents in Tunisia. All analysed water samples contained detectable levels of aminoglycoside and phenicol antibiotics. The highest concentrations in wastewater influents were observed for neomycin and kanamycin B (16.4 ng mL(-1) and 7.5 ng mL(-1), respectively). Chloramphenicol was found in wastewater influents up to 3 ng mL(-1). It was observed that the waste water treatment plants were not efficient in completely removing these antibiotics. Chloramphenicol and florfenicol were found in sea water samples near aquaculture sites at levels up to, respectively, 15.6 ng mL(-1) and 18.4 ng mL(-1). Also aminoglycoside antibiotics were found near aquaculture sites with the highest concentration of 3.4 ng mL(-1) for streptomycin. In pharmaceutical effluents, only gentamycin was found at concentrations up to 19 ng mL(-1) over a sampling period of four months.

  2. Prevalence of Antibiotic-Resistant Fecal Escherichia coli Isolates from Penned Broiler and Scavenging Local Chickens in Arusha, Tanzania.

    PubMed

    Rugumisa, Bernadether T; Call, Douglas R; Mwanyika, Gaspary O; Mrutu, Rehema I; Luanda, Catherine M; Lyimo, Beatus M; Subbiah, Murugan; Buza, Joram J

    2016-08-01

    We compared the prevalence of antibiotic-resistant Escherichia coli isolates from household-level producers of broiler (commercial source breeds) and local chickens in the Arusha District of Tanzania. Households were composed of a single dwelling or residence with independent, penned broiler flocks. Free-range, scavenging chickens were mixed breed and loosely associated with individual households. A total of 1,800 E. coli isolates (1,200 from broiler and 600 from scavenging local chickens) from 75 chickens were tested for their susceptibility against 11 antibiotics by using breakpoint assays. Isolates from broiler chickens harbored a higher prevalence of antibiotic-resistant E. coli relative to scavenging local chickens, including sulfamethoxazole (80.3 versus 34%), followed by trimethoprim (69.3 versus 27.7%), tetracycline (56.8 versus 20%), streptomycin (52.7 versus 24.7%), amoxicillin (49.6 versus 17%), ampicillin (49.1 versus 16.8%), ciprofloxacin (21.9 versus 1.7%), and chloramphenicol (1.5 versus 1.2%). Except for resistance to chloramphenicol, scavenging local chickens harbored fewer resistant E. coli isolates (P < 0.05). Broiler chickens harbored more isolates that were resistant to ≥7 antibiotics (P < 0.05). The higher prevalence of antibiotic-resistant E. coli from broiler chickens correlated with the reported therapeutic and prophylactic use of antibiotics in this poultry population. We suggest that improved biosecurity measures and increased vaccination efforts would reduce reliance on antibiotics by these households. PMID:27497131

  3. [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.

  4. The in vitro effect of six antimicrobials against Mycoplasma putrefaciens, Mycoplasma mycoides subsp. mycoides LC and Mycoplasma capricolum subsp. capricolum isolated from sheep and goats in Jordan.

    PubMed

    Al-Momani, W; Nicholas, R A J; Janakat, S; Abu-Basha, E; Ayling, R D

    2006-01-01

    Respiratory disease in sheep and goats is a major problem in Jordan and is often associated with Mycoplasma species. Without effective vaccines, control is mainly by chemotherapy, but the uncontrolled use of antimicrobials has led to concerns about the potential development of antimicrobial resistance. The in vitro effect of chloramphenicol, florfenicol, enrofloxacin, tylosin, erythromycin and oxytetracycline was determined against 32 isolates of Mycoplasma species-M. mycoides subsp. mycoides LC (6), M. capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions of Jordan. The antimicrobial susceptibility showed some Mycoplasma species-specific differences, with M. capricolum subsp. capricolum being more susceptible to tylosin and erythromycin. Chloramphenicol and florfenicol were the least effective for all three Mycoplasma species. No trends or significant differences in antimicrobial susceptibilities were observed between sheep and goat isolates, between milk or nasal swab isolates, or between isolates from different regions of Jordan. Some isolates of M. capricolum subsp. capricolum and M. putrefaciens showed higher MIC levels with oxytetracycline, as did two isolates of M. mycoides subsp. mycoides LC with tylosin, possibly indicating signs of development of antimicrobial resistance. PMID:17405622

  5. 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. PMID:12688126

  6. R-plasmid transfer in a wastewater treatment plant.

    PubMed

    Mach, P A; Grimes, D J

    1982-12-01

    Enteric bacteria have been examined for their ability to transfer antibiotic resistance in a wastewater treatment plant. Resistant Salmonella enteritidis, Proteus mirabilis, and Escherichia coli were isolated from clinical specimens and primary sewage effluent. Resistance to ampicillin, chloramphenicol, streptomycin, sulfadiazine, and tetracycline was demonstrated by spread plate and tube dilution techniques. Plasmid mediation of resistance was shown by ethidium bromide curing, agarose gel electrophoresis, and direct cell transfer. Each donor was mated with susceptible E. coli and Shigella sonnei. Mating pairs (and recipient controls) were suspended in unchlorinated primary effluent that had been filtered and autoclaved. Suspensions were added to membrane diffusion chambers which were then placed in the primary and secondary setting tanks of the wastewater treatment plant. Resistant recombinants were detected by replica plating nutrient agar master plates onto xylose lysine desoxycholate agar plates that contained per milliliter of medium 10 micrograms of ampicillin, 30 micrograms of chloramphenicol, 10 micrograms of streptomycin, 100 micrograms of sulfadiazine, or 30 micrograms of tetracycline. Mean transfer frequencies for laboratory matings were 2.1 X 10(-3). In situ matings for primary and secondary settling resulted in frequencies of 4.9 X 10(-5) and 7.5 X 10(-5), respectively. These values suggest that a significant level of resistance transfer occurs in wastewater treatment plants in the absence of antibiotics as selective agents.

  7. Modified Artificial Diet for Rearing of Tobacco Budworm, Helicoverpa armigera, using the Taguchi Method and Derringer's Desirability Function

    PubMed Central

    Assemi, H.; Rezapanah, M.; Vafaei-Shoushtari, R.

    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. PMID:23425103

  8. Microbial receptor assay for rapid detection and identification of seven families of antimicrobial drugs in milk: collaborative study.

    PubMed

    Charm, S E; Chi, R

    1988-01-01

    A microbial competitive receptor assay for detecting residues of antibiotic families in milk was studied collaboratively by 13 laboratories. The drugs and levels (ppb) tested in this study include penicillin G, 4.8; cephapirin, 5.0; cloxacillin, 100; tetracycline, 2000; chlortetracycline, 2000; oxytetracycline, 2000; erythromycin, 200; lincomycin, 400; clindamycin, 400; sulfamethazine, 75; sulfamethoxazole, 50; sulfisoxazole, 50; streptomycin, 1000; novobiocin, 50; and chloramphenicol, 800. In this method, microbial cells added to a milk sample provide specific binding sites for which 14C or 3H labeled drug competes with drug residues in the sample. The 14C or 3H binding to the specific binding sites is measured in a scintillation counter and compared with a zero standard milk. If the sample is statistically different from the zero standard, it is positive. The assay takes about 15 min. The binding reaction occurs between the receptor site and the drug functional group, so all members of a drug family are detected. In this case, beta-lactams, tetracyclines, macrolides, aminoglycosides, novobiocin, chloramphenicol, and sulfonamides, including p-aminobenzoic acid (PABA) and its other analogs, are detectable. The incidence of false negative determinations among samples is about 1%; the incidence of false positives is about 3%. For negative cases, the relative standard deviations for repeatability ranged from 0 to 5% and for reproducibility from 0 to 6%. For positive cases, relative standard deviations ranged from 0 to 13% for repeatability and from 0 to 14% for reproducibility. The method has been adopted official first action. PMID:3260233

  9. Assessment of antibiotic resistance phenotype and integrons in Salmonella enterica serovar Typhimurium isolated from swine.

    PubMed

    Rayamajhi, Nabin; Kang, Sang Gyun; Kang, Mi Lan; Lee, Hee Soo; Park, Kyung Yoon; Yoo, Han Sang

    2008-10-01

    Salmonella enterica serovar Typhimurium (S. Typhimurium) isolated and identified from swine were subjected for the analysis of antibiotic resistance pattern and clinically important class 1 and 2 integrons. In addition, S. Typhimurium isolates exhibiting ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline and florfenicol (ACSSuTF) resistance pattern as described in most Salmonella enterica serotype Typhimurium definitive type 104 (DT104) were characterized by polymerase chain reaction. All the isolates were resistant to more than four antibiotics and showed the highest resistance to streptomycin (94.1%), followed by tetracycline (90.1%), ampicillin (64.7%), chloramphenicol (56.8%) and gentamicin (54.9%). MIC value for the ten isolates ranged between 0.125-2 mug/ml for ciprofloxacin. Among the beta-lactams used, only one of the isolate exhibited resistance to ceftiofur (MIC 8 microg/ml). Sixty eight percent of these multi drug resistance (MDR) S. Typhimurium isolates carried clinically important class 1 integron with 1kb (aadA) and/or 2kb (dhfrXII-orfF-aadA2) resistance gene cassettes. This study reports the increasing trend of multi drug resistance (MDR) S. Typhimurium with clinically important class 1 integron in pigs. In addition, emergence of the ACSSuTF-type resistance in S. Typhimurium PT other than DT104 may limit the use of resistance gene markers in its detection methods by PCR. PMID:18981675

  10. Expression of R-plasmid functions during anaerobic growth of an Escherichia coli K-12 host.

    PubMed Central

    Burman, L G

    1977-01-01

    The normal habitat of enteric bacteria is largely anaerobic. Expression of the three characteristic properties of R-plasmids, drug resistance, vegetative replication, and fertility, was therefore studied in Escherichia coli K-12 during anaerobiosis. Replication and drug resistance functions were not altered in the 45 R-plasmids studies, whereas the expression of fertility varied considerably among different R-plasmids during anaerobiosis. The R-plasmids could be divided into three groups, one showing a strong, the second a moderate, and the third little or no reduction of fertility by anaerobiosis. Plasmid-determined sensitivity to F-, N-, and I-specific phage, respectively, was well, although not absolutely, correlated with each of three groups mentioned. Anaerobiosis-aerobiosis appears to change the fertility of type F R-plasmids by influencing the degree of repression of their fertility functions such as the formation of sex pili. Although the minimum inhibitory concentrations of ampicillin, chloramphenicol, streptomycin, and tetracycline were unaltered by anaerobiosis, sulfonamide was found to be four- to eightfold less active under this condition in both resistant and sensitive strains. A surprisingly high frequency and uniformity of minimal inhibitory concentrations was observed for R-plasmid-mediated resistance to streptomycin and chloramphenicol. PMID:326771

  11. Prevalence of Antibiotic-Resistant Fecal Escherichia coli Isolates from Penned Broiler and Scavenging Local Chickens in Arusha, Tanzania.

    PubMed

    Rugumisa, Bernadether T; Call, Douglas R; Mwanyika, Gaspary O; Mrutu, Rehema I; Luanda, Catherine M; Lyimo, Beatus M; Subbiah, Murugan; Buza, Joram J

    2016-08-01

    We compared the prevalence of antibiotic-resistant Escherichia coli isolates from household-level producers of broiler (commercial source breeds) and local chickens in the Arusha District of Tanzania. Households were composed of a single dwelling or residence with independent, penned broiler flocks. Free-range, scavenging chickens were mixed breed and loosely associated with individual households. A total of 1,800 E. coli isolates (1,200 from broiler and 600 from scavenging local chickens) from 75 chickens were tested for their susceptibility against 11 antibiotics by using breakpoint assays. Isolates from broiler chickens harbored a higher prevalence of antibiotic-resistant E. coli relative to scavenging local chickens, including sulfamethoxazole (80.3 versus 34%), followed by trimethoprim (69.3 versus 27.7%), tetracycline (56.8 versus 20%), streptomycin (52.7 versus 24.7%), amoxicillin (49.6 versus 17%), ampicillin (49.1 versus 16.8%), ciprofloxacin (21.9 versus 1.7%), and chloramphenicol (1.5 versus 1.2%). Except for resistance to chloramphenicol, scavenging local chickens harbored fewer resistant E. coli isolates (P < 0.05). Broiler chickens harbored more isolates that were resistant to ≥7 antibiotics (P < 0.05). The higher prevalence of antibiotic-resistant E. coli from broiler chickens correlated with the reported therapeutic and prophylactic use of antibiotics in this poultry population. We suggest that improved biosecurity measures and increased vaccination efforts would reduce reliance on antibiotics by these households.

  12. Characterization and adsorption of Lactobacillus virulent phage P1.

    PubMed

    Chen, X; Xi, Y; Zhang, H; Wang, Z; Fan, M; Liu, Y; Wu, W

    2016-09-01

    Bacteriophage infection of lactic acid bacteria is considered an important problem worldwide in the food fermentation industry, as it may produce low quality or unsafe foods, cause fermentation failure, and result in economic losses. To increase current knowledge on the properties of Lactobacillus virulent phages, we evaluated the effect of divalent cations, temperature, pH, and chloramphenicol on the adsorption ability of Lactobacillus virulent phage P1. Phage P1 was isolated from the abnormal fermentation liquid of Lactobacillus plantarum IMAU10120. The results showed that this phage belonged to the Siphoviridae family. The latent period of this phage was 45min, and the burst time was 90min. Burst size was 132.88±2.37 phage counts expressed per milliliter per infective center. This phage showed good tolerance at different temperatures, but incubation at 50°C only affected its adsorption. Adsorption rate reached a maximum value between 30 and 42°C. A high adsorption value of phage infectivity was obtained from pH 6 to 8. Moreover, calcium ions promoted and increased the adsorption capacity of phage P1, but magnesium ions had negative effects. Chloramphenicol had no effect on phage adsorption. This study increased current knowledge on the characterization and biological aspects of Lactobacillus virulent phages, and may provide some basic information that can be used to design successful antiphage strategies in the food industry. PMID:27372579

  13. 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.

  14. Mycotoxigenic fungi and natural co-occurrence of mycotoxins in rainbow trout (Oncorhynchus mykiss) feeds.

    PubMed

    Greco, Mariana; Pardo, Alejandro; Pose, Graciela

    2015-11-05

    Samples of rainbow trout feed were analyzed with the aim to determine the mycobiota composition and the co-occurrence of mycotoxins. A total of 28 samples of finished rainbow trout feed from hatcheries in the provinces of Río Negro and Neuquén, Argentina, were studied. Fungal counts were obtained on three culture media in the ranges of <10 to 4.2 × 10⁴ CFU/g on Dichloran Rose Bengal Chloramphenicol Agar (DRBC), <10 to 5.1 × 10⁴ CFU/g on Dichloran Chloramphenicol Peptone Agar (DCPA) and <10 to 3.6 × 10⁴ CFU/g on Dichloran 18% Glycerol Agar (DG18). The most frequent mycotoxigenic fungi were Eurotium (frequency (Fr) 25.0%), followed by Penicillium (Fr 21.4%) and Aspergillus (Fr 3.6%). The most prevalent mycotoxigenic species were E. repens (Fr 21.4%) and E. rubrum (Fr 14.3%). All samples were contaminated with mycotoxins: 64% samples were contaminated with T-2 toxin (median 70.08 ppb), 50% samples with zearalenone (median 87.97 ppb) and aflatoxins (median 2.82 ppb), 25% with ochratoxin A (median 5.26 ppb) and 3.57% samples with deoxynivalenol (median 230 ppb). Eight samples had a fumonisins contamination level below the limit of detection. Co-occurrence of six mycotoxins was determined in 7% of the samples.

  15. 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.

  16. Antimicrobial activity of commonly used antibiotics and DNA fingerprint analysis of Pseudomonas aeruginosa obtained from clinical isolates and unchlorinated drinking water in Korea, 2010.

    PubMed

    Kim, Jung Rae; Lee, Do Kyung; An, Hyang Mi; Kim, Mi Jin; Lee, Si Won; Cha, Min Kyeong; Lee, Kang Oh; Ha, Nam Joo

    2011-08-01

    Pseudomonas aeruginosa exists in various environments, and can cause mild or serious infections resulting in a wide range of symptoms. In this study, we collected bacterial isolates from hospitalized patients and unchlorinated drinking water, in Korea, 2010. The water-borne and clinical isolates were compared using colony morphology, antimicrobial susceptibility testing, and random amplification of polymorphic DNA analysis. We first compared morphological features of the water-borne and clinical isolates. The clearest difference in colony morphology was colony shape; five water-borne isolate colonies (83%) had a smooth, circular morphology, while nine (75%) clinical isolate colonies had a rough, irregular morphology. Minimum inhibitory concentrations analyses were performed to determine antimicrobial resistant patterns; using ceftazidime, gentamicin, tigecycline, chloramphenicol, meropenem, and tobramycin according to Clinical and Laboratory Standard Institute (CLSI, 2009) methodology. All waterborne isolates were not resistant to gentamicin, tobramycin, and meropenem. The clinical isolates were resistant to every antibiotic except chloramphenicol. Genotyping was performed using the repetitive extragenic palindromic polymerase-chain-reaction. The DNA fingerprinting patterns did not reveal genetic similarity between the water-borne and clinical P. aeruginosa isolates. On the contrary, they showed that genetically distinct populations have been established in each of these environments. We have revealed significant morphological, clinical and genetic differences between water-borne and clinical isolates of the same bacterial species.

  17. 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.

  18. 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.

  19. Clinically significant anaerobic bacteria isolated from patients in a South African academic hospital: antimicrobial susceptibility testing.

    PubMed

    Naidoo, S; Perovic, O; Richards, G A; Duse, A G

    2011-09-27

    BACKGROUND. Increasing resistance to some antimicrobial agents among anaerobic bacteria has made susceptibility patterns less predictable. METHOD. This was a prospective study of the susceptibility data of anaerobic organisms isolated from clinical specimens from patients with suspected anaerobic infections from June 2005 until February 2007. Specimens were submitted to the microbiology laboratory at Charlotte Maxeke Johannesburg Academic Hospital, where microscopy, culture and susceptibility testing were performed the using E test® strip minimum inhibitory concentration method. Results were interpreted with reference to Clinical and Laboratory Standards Institute guidelines for amoxicillin-clavulanate, clindamycin, metronidazole, penicillin, ertapenem, cefoxitin, ceftriaxone, chloramphenicol and piperacillin-tazobactam. RESULTS. One hundred and eighty anaerobic isolates were submitted from 165 patients. The most active antimicrobial agents were chloramphenicol (100% susceptible), ertapenem (97.2%), piperacillin-tazobactam (99.4%) and amoxicillin-clavulanic acid (96.7%). Less active were metronidazole (89.4%), cefoxitin (85%), clindamycin (81.7%), ceftriaxone (68.3%) and penicillin (33.3%). CONCLUSION. Susceptibility testing should be performed periodically to identify emerging trends in resistance and to modify empirical treatment of anaerobic infections.

  20. Mycoflora of poultry feeds and ochratoxin-producing ability of isolated Aspergillus and Penicillium species.

    PubMed

    Rosa, C A R; Ribeiro, J M M; Fraga, M J; Gatti, M; Cavaglieri, L R; Magnoli, C E; Dalcero, A M; Lopes, C W G

    2006-03-10

    In Brazil, commercial feedstuffs are an important component in modern animal husbandry, but there is no information available about fungal contamination and ochratoxin A (OTA) production. The aims of this study were to determine the mycoflora incidence in poultry feeds and evaluate OTA production. In addition, the ability to produce OTA by several Aspergillus and Penicillium species was investigated. A total of 96 samples of poultry feeds were collected from four factories in Rio de Janeiro. Samples were examined for total moulds, for Aspergillus and Penicillium spp. occurrence and for their relative densities on dichloran rose bengal chloramphenicol and dichloran 18% glycerol media. The capacity to produce ochratoxin A by selected Aspergillus and Penicillium species was determined by HPLC. Total mould counts were generally higher than 1 x 10(5 )CFU ml(-1). Aspergillus and Penicillium species were isolated in the highest numbers. Aspergillus flovus and Penicillium citrinum were the most prevalent species. There was a high percentage of potential OTA producers (46%). The amount of OTA produced on this substrate was enough to cause adverse effects in animals. Several strains isolated from poultry feeds were able to produce high levels of OTA on chloramphenicol yeast medium. OTA in raw materials needs to be surveyed and storage practices must be investigated to determine occurrence and establish the livestock toxicological risk in poultry feed.

  1. Characterization of the structure and catalytic activity of Legionella pneumophila VipF.

    PubMed

    Young, Byron H; Caldwell, Tracy A; McKenzie, Aidan M; Kokhan, Oleksandr; Berndsen, Christopher E

    2016-10-01

    The pathogenic bacteria Legionella pneumophila is known to cause Legionnaires' Disease, a severe pneumonia that can be fatal to immunocompromised individuals and the elderly. Shohdy et al. identified the L. pneumophila vacuole sorting inhibitory protein VipF as a putative N-acetyltransferase based on sequence homology. We have characterized the basic structural and functional properties of VipF to confirm this original functional assignment. Sequence conservation analysis indicates two putative CoA-binding regions within VipF. Homology modeling and small angle X-ray scattering suggest a monomeric, dual-domain structure joined by a flexible linker. Each domain contains the characteristic beta-splay motif found in many acetyltransferases, suggesting that VipF may contain two active sites. Docking experiments suggest reasonable acetyl-CoA binding locations within each beta-splay motif. Broad substrate screening indicated that VipF is capable of acetylating chloramphenicol and both domains are catalytically active. Given that chloramphenicol is not known to be N-acetylated, this is a surprising finding suggesting that VipF is capable of O-acetyltransferase activity. Proteins 2016; 84:1422-1430. © 2016 Wiley Periodicals, Inc. PMID:27315603

  2. Photo-activated porphyrin in combination with antibiotics: therapies against Staphylococci.

    PubMed

    Dastgheyb, Sana S; Eckmann, David M; Composto, Russell J; Hickok, Noreen J

    2013-12-01

    Staphylococcal infections have become difficult to treat due to antibiotic insensitivity and resistance. Antimicrobial combination therapies may minimize acquisition of resistance and photodynamic therapy is an attractive candidate for these combinations. In this manuscript, we explore combined use of antibiotics and meso-tetra (4-aminophenyl) porphine (TAPP), a cationic porphyrin, for treatment of Staphylococcus aureus contamination. We characterize the antimicrobial activity of photoactivated TAPP and show that activity is largely lost in the presence of a radical scavenger. Importantly, TAPP can be reactivated with continued, albeit attenuated, antibacterial activity. We then show that the antimicrobial activity of illuminated TAPP is additive with chloramphenicol and tobramycin for S. aureus and Escherichia coli, and synergistic for MRSA and Staphylococcus epidermidis. Chloramphenicol+methylene blue, another photosensitizer, also show additivity against S. aureus. In contrast, ceftriaxone and vancomycin do not strongly augment the low level effects of TAPP against S. aureus. Eukaryotic cells exhibit a dose-dependent toxicity with illuminated TAPP. Our results suggest that even sub-minimum inhibitory concentrations of photo-activated TAPP could be used to boost the activity of waning antibiotics. This may play an important role in treatments reliant on antibiotic controlled release systems where augmentation with photo-active agents could extend their efficacy.

  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. Multiple antibiotic resistance in Pseudomonas aeruginosa: evidence for involvement of an efflux operon.

    PubMed Central

    Poole, K; Krebes, K; McNally, C; Neshat, S

    1993-01-01

    An outer membrane protein of 50 kDa (OprK) was overproduced in a siderophore-deficient mutant of Pseudomonas aeruginosa capable of growth on iron-deficient minimal medium containing 2,2'-dipyridyl (0.5 mM). The expression of OprK in the mutant (strain K385) was associated with enhanced resistance to a number of antimicrobial agents, including ciprofloxacin, nalidixic acid, tetracycline, chloramphenicol, and streptonigrin. OprK was inducible in the parent strain by growth under severe iron limitation, as provided, for example, by the addition of dipyridyl or ZnSO4 to the growth medium. The gene encoding OprK (previously identified as ORFC) forms part of an operon composed of three genes (ORFABC) implicated in the secretion of the siderophore pyoverdine. Mutants defective in ORFA, ORFB, or ORFC exhibited enhanced susceptibility to tetracycline, chloramphenicol, ciprofloxacin, streptonigrin, and dipyridyl, consistent with a role for the ORFABC operon in multiple antibiotic resistance in P. aeruginosa. Sequence analysis of ORFC (oprK) revealed that its product is homologous to a class of outer membrane proteins involved in export. Similarly, the products of ORFA and ORFB exhibit homology to previously described bacterial export proteins located in the cytoplasmic membrane. These data suggest that ORFA-ORFB-oprK (ORFC)-dependent drug efflux contributes to multiple antibiotic resistance in P. aeruginosa. We propose, therefore, the designation mexAB (multiple efflux) for ORFAB. Images PMID:8226684

  5. Species distribution, antibiotic resistance and virulence traits in enterococci from meat in Tunisia.

    PubMed

    Klibi, Naouel; Said, Leila Ben; Jouini, Ahlem; Slama, Karim Ben; López, Maria; Sallem, Rym Ben; Boudabous, Abdellatif; Torres, Carmen

    2013-03-01

    Antimicrobial resistance and the mechanisms implicated were studied in 119 enterococci from 105 meat samples from Tunisian markets. Almost 24.5% of recovered enterococci showed resistance against four or more antimicrobial agents and these isolates were identified to the species level. Enterococcus faecalis was the most prevalent species (41%). High percentages of erythromycin and tetracycline resistances were found among our isolates, and lower percentages were identified to aminoglycosides, ciprofloxacin and chloramphenicol. All tetracycline-resistant isolates carried the tet(M) and/or tet(L) genes. The erm(B) gene was detected in 78.5% of erythromycin-resistant isolates, ant(6)-Ia gene in 58.8% of streptomycin-resistant isolates, and cat(A) gene in one chloramphenicol-resistant isolate. Forty-eight isolates carried the gelE gene and exhibited gelatinase activity. The hyl and esp genes were detected in one and three Enterococcus faecium isolates, respectively. Streptomycin-resistant isolates showed a high genetic diversity by PFGE and MLST. Meat might play a role in the spread through the food chain of enterococci with these virulence and resistance characteristics to humans.

  6. Transcription of the procyclic acidic repetitive protein genes of Trypanosoma brucei.

    PubMed Central

    Clayton, C E; Fueri, J P; Itzhaki, J E; Bellofatto, V; Sherman, D R; Wisdom, G S; Vijayasarathy, S; Mowatt, M R

    1990-01-01

    The procyclic acidic repetitive protein (parp) genes of Trypanosoma brucei encode a small family of abundant surface proteins whose expression is restricted to the procyclic form of the parasite. They are found at two unlinked loci, parpA and parpB; transcription of both loci is developmentally regulated. The region of homology upstream of the A and B parp genes is only 640 base pairs long and may contain sequences responsible for transcriptional initiation and regulation. Transcription upstream of this putative promoter region is not developmentally regulated and is much less active than that of the parp genes; the polymerase responsible is inhibited by alpha-amanitin, whereas that transcribing the parp genes is not. Transcription of the parp genes is strongly stimulated by low levels of UV irradiation. The putative parp promoter, when placed upstream of the chloramphenicol acetyltransferase gene, is sufficient to cause production of chloramphenicol acetyltransferase in a T. brucei DNA transformation assay. Taken together, these results suggest that a promoter for an alpha-amanitin-resistant RNA polymerase lies less than 600 nucleotides upstream of the parp genes. Images PMID:2342468

  7. Characterization of Escherichia coli Isolates from an Urban Lake Receiving Water from a Wastewater Treatment Plant in Mexico City: Fecal Pollution and Antibiotic Resistance.

    PubMed

    Rosas, Irma; Salinas, Eva; Martínez, Leticia; Cruz-Córdova, Ariadnna; González-Pedrajo, Bertha; Espinosa, Norma; Amábile-Cuevas, Carlos F

    2015-10-01

    The presence of enteric bacteria in water bodies is a cause of public health concerns, either by directly causing water- and food-borne diseases, or acting as reservoirs for antibiotic resistance determinants. Water is used for crop irrigation; and sediments and aquatic plants are used as fertilizing supplements and soil conditioners. In this work, the bacterial load of several micro-environments of the urban lake of Xochimilco, in Mexico City, was characterized. We found a differential distribution of enteric bacteria between the water column, sediment, and the rhizoplane of aquatic plants, with human fecal bacteria concentrating in the sediment, pointing to the need to assess such bacterial load for each micro-environment, for regulatory agricultural purposes, instead of only the one of the water, as is currently done. Resistance to tetracycline, ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole was common among Escherichia coli isolates, but was also differentially distributed, being again higher in sediment isolates. A distinct distribution of chloramphenicol minimum inhibitory concentrations (MIC) among these isolates suggests the presence of a local selective pressure favoring lower MICs than those of isolates from treated water. Fecal bacteria of human origin, living in water bodies along with their antibiotic resistance genes, could be much more common than typically considered, and pose a higher health risk, if assessments are only made on the water column of such bodies.

  8. Variability of the tandem repeat region of the Escherichia coli tolA gene.

    PubMed

    Zhou, Kai; Vanoirbeek, Kristof; Aertsen, Abram; Michiels, Chris W

    2012-06-01

    An intragenic tandem repeat (TR) region has been previously reported in the tolA gene of Escherichia coli. In silico analysis of 123 E. coli tolA sequences from Genbank and PCR analysis of the tolA TR region from 111 additional E. coli strains revealed that this TR region is highly variable. Nine different TR sizes with 8 up to 16 repeat units were found in in silico analysis and 6 of these were also found by PCR analysis. The 13-unit TR emerged as the predominant type using both approaches (47.2% and 86.5%, respectively). Remarkably, TRs in pathogenic strains appeared to be more variable than those in non-pathogens. To demonstrate the occurrence of TR variation in a clonal population, a selection system for TR deletion events was constructed by inserting the 13-unit TR region of MG1655 in frame into a plasmid-borne chloramphenicol acetyltransferase (cat) gene. The resulting cat gene no longer conferred chloramphenicol resistance unless the insert size was reduced by TR contraction. Using this system, Cm-resistant revertants with a TR contraction were recovered at a frequency of 1.1 × 10(-7), and contraction was shown to be recA-dependent and enhanced in a DNA repair-deficient mutS background. PMID:22659144

  9. Biochemical characteristic of biofilm of uropathogenic Escherichia coli Dr(+) strains.

    PubMed

    Zalewska-Piątek, Beata; Wilkanowicz, Sabina; Bruździak, Piotr; Piątek, Rafał; Kur, Józef

    2013-07-19

    Urinary tract infections caused by Escherichia coli are very common health problem in the developed countries. The virulence of the uropathogenic E. coli Dr(+) IH11128 is determined by Dr fimbriae, which are homopolymeric structures composed of DraE subunits with the DraD protein capping the fiber. In this study, we have analyzed the structural and biochemical properties of biofilms developed by E. coli strains expressing Dr fimbriae with or without the DraD tip subunit and the surface-exposed DraD protein. We have also demonstrated that these E. coli strains form biofilms on an abiotic surface in a nutrient-dependent fashion. We present evidence that Dr fimbriae are necessary during the first stage of bacterial interaction with the abiotic surface. In addition, we reveal that the DraD alone is also sufficient for the initial surface attachment at an even higher level than Dr fimbriae and that chloramphenicol is able to reduce the normal attachment of the analyzed E. coli. The action of chloramphenicol also shows that protein synthesis is required for the early events of biofilm formation. Additionally, we have identified reduced exopolysaccharide coverage in E. coli that express only Dr fimbrial polyadhesins at the cell surface with or without the DraD capping subunit.

  10. T-2 mycotoxin inhibits mitochondrial protein synthesis

    SciTech Connect

    Pace, J.G.; Watts, M.R.; Canterbury, W.J.

    1988-01-01

    The authors investigated the effect of T-2 toxin on rat liver mitochondrial protein synthesis. Isolated rat liver mitochondria were supplemented with an S-100 supernatant from rat liver and an external ATP-generating system. An in-vitro assay employing cycloheximide, and inhibitor of cytoplasmic protein synthesis, and chloramphenicol, and inhibitor of mitochondrial protein synthesis, to distinguish mitochondrial protein synthesis from the cytoplasmic process. Amino acid incorporation into mitochondria was dependent on the concentration of mitochondria and was inhibited by chloramphenicol. The rate of uptake of tritium leucine into mitochondrial protein was unaffected by the addition of T-2 toxin and was not a rate-limiting step in incorporation. However, 0.02 micrograms/ml of T-2 toxin decreased the rate of protein synthesis inhibition correlated with the amount of T-2 toxin taken up by the mitochondria. While T-2 toxin is known to inhibit eukaryotic protein synthesis, this is the first time T-2 was shown to inhibit mitochondrial protein synthesis.

  11. 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. PMID:12647952

  12. 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

  13. A SAM-dependent methyltransferase cotranscribed with arsenate reductase alters resistance to peptidyl transferase center-binding antibiotics in Azospirillum brasilense Sp7.

    PubMed

    Singh, Sudhir; Singh, Chhaya; Tripathi, Anil Kumar

    2014-05-01

    The genome of Azospirillum brasilense harbors a gene encoding S-adenosylmethionine-dependent methyltransferase, which is located downstream of an arsenate reductase gene. Both genes are cotranscribed and translationally coupled. When they were cloned and expressed individually in an arsenate-sensitive strain of Escherichia coli, arsenate reductase conferred tolerance to arsenate; however, methyltransferase failed to do so. Sequence analysis revealed that methyltransferase was more closely related to a PrmB-type N5-glutamine methyltransferase than to the arsenate detoxifying methyltransferase ArsM. Insertional inactivation of prmB gene in A. brasilense resulted in an increased sensitivity to chloramphenicol and resistance to tiamulin and clindamycin, which are known to bind at the peptidyl transferase center (PTC) in the ribosome. These observations suggested that the inability of prmB:km mutant to methylate L3 protein might alter hydrophobicity in the antibiotic-binding pocket of the PTC, which might affect the binding of chloramphenicol, clindamycin, and tiamulin differentially. This is the first report showing the role of PrmB-type N5-glutamine methyltransferases in conferring resistance to tiamulin and clindamycin in any bacterium.

  14. One Pot Synthesis, Photophysical and X-ray Studies of Novel Highly Fluorescent Isoquinoline Derivatives with Higher Antibacterial Efficacy Based on the In-vitro and Density Functional Theory.

    PubMed

    Asiri, Abdullah M; Khan, Salman A; Al-Thaqafy, Saad H; Sharma, Kamlesh

    2015-05-01

    Series of 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 agreement with their chemical structures. Structure of the compound was further conformed by X-ray crystallographic. UV-vis and fluorescence spectroscopy measurements provided 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, extenction 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. The minimum inhibitory concentration was then 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. Furthermore, quantum chemistry calculations using DFT/6-31-G* level of theory confirm the results. Dipole moment and frontier molecular orbitals were also investigated.

  15. A complex androgen-responsive enhancer resides 2 kilobases upstream of the mouse Slp gene.

    PubMed Central

    Loreni, F; Stavenhagen, J; Kalff, M; Robins, D M

    1988-01-01

    Neighboring genes encoding the mouse sex-limited protein (Slp) and fourth component of complement (C4) show extensive homology. In contrast to C4, however, Slp is regulated by androgen. One region of the Slp gene capable of hormonal response following transfection was located about 2 kilobases upstream of the transcription start site, where the C4 and Slp sequences diverge. This region, delimited here to a 0.75-kilobase fragment, showed cryptic promoter activity as well as androgen responsiveness in either orientation in front of the bacterial chloramphenicol acetyltransferase coding region. When this fragment was placed upstream of a viral long terminal repeat, increased chloramphenicol acetyltransferase expression derived from the viral promoter. Proteins from nuclear extracts specifically bound to four sequences within the region, near sites that are DNase I hypersensitive in vivo and reflect the hormonal and developmental regulation of Slp. Like several other cellular enhancers, this androgen-responsive element seems to be modular in nature and complex in its function. Images PMID:3165490

  16. Mycotoxigenic fungi and natural co-occurrence of mycotoxins in rainbow trout (Oncorhynchus mykiss) feeds.

    PubMed

    Greco, Mariana; Pardo, Alejandro; Pose, Graciela

    2015-11-01

    Samples of rainbow trout feed were analyzed with the aim to determine the mycobiota composition and the co-occurrence of mycotoxins. A total of 28 samples of finished rainbow trout feed from hatcheries in the provinces of Río Negro and Neuquén, Argentina, were studied. Fungal counts were obtained on three culture media in the ranges of <10 to 4.2 × 10⁴ CFU/g on Dichloran Rose Bengal Chloramphenicol Agar (DRBC), <10 to 5.1 × 10⁴ CFU/g on Dichloran Chloramphenicol Peptone Agar (DCPA) and <10 to 3.6 × 10⁴ CFU/g on Dichloran 18% Glycerol Agar (DG18). The most frequent mycotoxigenic fungi were Eurotium (frequency (Fr) 25.0%), followed by Penicillium (Fr 21.4%) and Aspergillus (Fr 3.6%). The most prevalent mycotoxigenic species were E. repens (Fr 21.4%) and E. rubrum (Fr 14.3%). All samples were contaminated with mycotoxins: 64% samples were contaminated with T-2 toxin (median 70.08 ppb), 50% samples with zearalenone (median 87.97 ppb) and aflatoxins (median 2.82 ppb), 25% with ochratoxin A (median 5.26 ppb) and 3.57% samples with deoxynivalenol (median 230 ppb). Eight samples had a fumonisins contamination level below the limit of detection. Co-occurrence of six mycotoxins was determined in 7% of the samples. PMID:26556374

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. A new inducible expression system in a transformed green alga, Chlorella vulgaris.

    PubMed

    Niu, Y F; Zhang, M H; Xie, W H; Li, J N; Gao, Y F; Yang, W D; Liu, J S; Li, H Y

    2011-01-01

    Genetic transformation is useful for basic research and applied biotechnology. However, genetic transformation of microalgae is usually quite difficult due to the technical limitations of existing methods. We cloned the promoter and terminator of the nitrate reductase gene from the microalga Phaeodactylum tricornutum and used them for optimization of a transformation system of the microalga Chlorella vulgaris. This species has been used for food production and is a promising candidate as a bioreactor for large-scale production of value-added proteins. A construct was made containing the CAT (chloramphenicol acetyltransferase) reporter gene driven by the nitrate reductase promoter. This construct was transferred into the C. vulgaris genome by electroporation. Expression of CAT in transgenic Chlorella conferred resistance to the antibiotic chloramphenicol and enabled growth in selective media. Overall efficiency for the transformation was estimated to be approximately 0.03%, which is relatively high compared with other available Chlorella transformation systems. Expression of CAT was induced in the presence of nitrate and inhibited in the presence of ammonium as a sole nitrogen source. This study presented an inducible recombinant gene expression system, also providing more gene regulation elements with potential for biotechnological applications.

  2. The 5'-untranslated regions of picornavirus RNAs contain independent functional domains essential for RNA replication and translation.

    PubMed Central

    Rohll, J B; Percy, N; Ley, R; Evans, D J; Almond, J W; Barclay, W S

    1994-01-01

    The role of the 5'-untranslated region (5'UTR) in the replication of enteroviruses has been studied by using a series of poliovirus type 3 (PV3) replicons containing the chloramphenicol acetyltransferase reporter gene in which the 5'UTR was replaced by the 5'UTR of either coxsackievirus B4 or human rhinovirus 14 or composite 5'UTRs derived from sequences of PV3, human rhinovirus 14, coxsackievirus B4, or encephalomyocarditis virus. The results indicate that efficient replication of an enterovirus genome requires a compatible interaction between the 5'-terminal cloverleaf structure and the coding and/or 3'-noncoding regions of the genome. A crucial determinant of this interaction is the stem-loop formed by nucleotides 46 to 81 (stem-loop d). The independence of the cloverleaf structure formed by the 5'-terminal 88 nucleotides and the ribosome landing pad or internal ribosome entry site (IRES) was investigated by constructing a 5'UTR composed of the PV3 cloverleaf and the IRES from encephalomyocarditis virus. Chloramphenicol acetyltransferase gene-containing replicons and viruses containing this recombinant 5'UTR showed levels of replication similar to those of the corresponding genomes containing the complete PV3 5'UTR, indicating that the cloverleaf and the IRES may be regarded as functionally independent and nonoverlapping elements. Images PMID:8207812

  3. A central composite rotatable design (CCRD) approach to study the combined effect of antimicrobial agents against bacterial pathogens.

    PubMed

    Santos, Fernanda Godoy; Mendonça, Layanne Andrade; Mantovani, Hilário Cuquetto

    2015-09-01

    The combination of antimicrobial agents has been proposed as a therapeutic strategy to control bacterial diseases and to reduce the emergence of antibiotic-resistant strains in clinical environments. In this study, the interaction between the lantibiotic bovicin HC5 with chloramphenicol, gentamicin, nisin, lysostaphin and hydrogen peroxide against Staphylococcus aureus O46 was evaluated by MIC assays. The central composite rotatable design (CCRD), a robust and economic statistical design, was used to combine concentration levels of different antimicrobials agents with distinct mechanisms of action and the presence of significant interactions among the antimicrobials was determined by regression analysis. According to the adjusted model, there were no significant interactions between bovicin HC5 and gentamicin, lysostaphin, nisin or hydrogen peroxide. However, bovicin HC5 showed a significant interaction (P < 0.02) with chloramphenicol. This is the first study applying the CCRD approach to evaluate the combined effect of antimicrobials against S. aureus. Based on our results, this approach is an effective strategy to determine synergistic interactions between antimicrobial agents applied in human and veterinary medicine against bacterial pathogens. PMID:26081601

  4. Expanding the bactericidal action of the food color additive phloxine B to gram-negative bacteria.

    PubMed

    Rasooly, Reuven

    2005-08-01

    Phloxine B (D&C red no. 28) is a color additive for food, drugs, and cosmetics. It has been previously shown to have anti-Staphylococcus aureus activities. In this work, the effect of Phloxine B on various gram-negative bacteria and other gram-positive bacteria including Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus subtilis, Bacillus aureus, Salmonella, Escherichia coli and Shigella was studied, along with the mechanism of anti-microbial activity. In the presence of fluorescent light, the viable count for gram-positive bacteria, (Bacillus spp. and S. aureus) decreased in a dose and time dependent manner when incubated with Phloxine B. The viability of gram-positive bacteria was reduced by 99.99% in 40 min, while there was no effect on gram-negative bacteria (Salmonella choleraesuis, E. coli and Shigella flexneri). However, the use of ethylenediaminetetraacetic acid (EDTA) expands the spectrum of activity for Phloxine B to include gram-negative bacteria. EDTA increased membrane-permeability by releasing lipopolysaccharide. Overall, in an Agar diffusion test the light-dependent bactericidal activity of 1 microg of Phloxine B had a potency of 0.64 units of chloramphenicol and 0.5 units of tetracycline when tested on B. cereus, and had a potency of 0.7 units of chloramphenicol and 0.2 units of tetracycline when tested on S. aureus. The data suggest that the dye may have some potential anti-microbial applications.

  5. 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

  6. [Resistance of Haemophilus sp. to antibiotics].

    PubMed

    Janicka, G; Gospodarek, E; Mikucka, A; Białek, M; Wojak, I; Krawiecka, D

    1998-02-01

    The present study was undertaken to determine the in vitro drug resistance of Haemophilus influenzae (68 isolates) and H. parainfluenzae (17 isolates). The tests susceptibility to Ampicillin, Amoxicilin/Clavulanic Acid, Cefaclor, Cefuroxime, Cotrimoxazole, Aztreonam, Ceftriaxone, Tetracycline, Ciprofloxacin, Rifampicin and Chloramphenicol were performed with a standard disk-diffusion method. The NCCLS methodology and susceptibility interpretative criteria were applied as described by the disk manufacturer. Beta-lactamase production was detected with nitrocefin impregnated disk (Cefinase, BBL Microbiology System). Resistance in nosocomially acquired Haemophilus isolates to several antibiotics was observed. Of the Haemophilus isolates 28.2% were Ampicillin in resistant, all were susceptible to the combination of Amoxicillin/Clavulanic acid. The Ampicillin-resistant strains were beta-lactamase producers. We observed the high resistance (70.1%) to Tetracycline and (28.2%) to SXT (Cotrimoxazole). All isolates of Haemophilus were susceptible to Ciprofloxacin. The low resistance percentages to Rifampin (1.2%), Aztreonam (3.5%) and Chloramphenicol (3.5%) was observed.

  7. Transformation of Rhodococcus fascians by High-Voltage Electroporation and Development of R. fascians Cloning Vectors

    PubMed Central

    Desomer, Jan; Dhaese, Patrick; Montagu, Marc Van

    1990-01-01

    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 105/μg of DNA to 107/μg 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. Images PMID:16348290

  8. Automated microbiological assay of thiamin in serum and red cells.

    PubMed Central

    Icke, G; Nicol, D

    1994-01-01

    AIMS--To develop a sensitive, direct, automated method for the measurement of serum and red cell thiamin. METHODS--A microbiological assay using a chloramphenicol resistant strain of Lactobacillus fermenti as the test organism was developed. Addition of chloramphenicol and cycloheximide to the assay medium suppressed bacterial and yeast contamination and enabled tests to be automated without recourse to aseptic procedures. Evaluation of the assay included precision analysis and estimation of thiamin recovery. Results obtained on red cell extracts were compared with an established colorimetric (thiochrome) method. RESULTS--Acceptable intrabatch and interbatch precision was obtained and good recovery of thiamin added to serum was obtained. Non-parametric reference ranges based on the results from 505 healthy people were: serum thiamin 11.3-35.0 nmol/l and red cell thiamin 190-400 nmol/l. Results were not age or gender related. The method gave results for red cell thiamin which were significantly higher than those obtained with an established thiochrome method. CONCLUSIONS--This automated microbiological assay is sensitive to 2.0 nmol/l of thiamin and allows tests to be set up at the rate of 100 per hour and after 20-22 hours allows incubation results to be read at 60 per hour. The method has proved reliable, suitable for the assay of large numbers of samples, and relatively inexpensive to perform. PMID:8089221

  9. Comparative antibiogram of coagulase-negative Staphylococci (CNS) associated with subclinical and clinical mastitis in dairy cows

    PubMed Central

    Bansal, B. K.; Gupta, D. K.; Shafi, T. A.; Sharma, S.

    2015-01-01

    Aim: The present study was planned to determine the in vitro antibiotic susceptibility of coagulase-negative Staphylococci (CNS) strains isolated from clinical and subclinical cases of mastitis in dairy cows. Antibiotic sensitivity profile will be helpful to recommend early therapy at the field level prior to availability of CST results. Materials and Methods: The milk samples from cases of clinical mastitis received in Mastitis Laboratory, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana and those of subclinical mastitis collected during routine screening of state dairy farms, were subjected to microbial culture. Identification of CNS organisms was done by standard biochemical tests. Antibiotic sensitivity testing, based on 30 antibiotics belonging to 12 groups, was done on 58 randomly selected CNS isolates (clinical isolates: 41, subclinical isolates: 17). Results: Isolates were highly susceptible to chloramphenicol (98.3%), gentamicin (93.1%), streptomycin (91.4%), linezolid (91.4%), ceftixozime (87.9%), cloxacillin (86.2%), clotrimazole (86.2%), bacitracin (86.2%), enrofloxacin (84.5%) and ceftrioxone + tazobactum (70.7%), while resistance was observed against amoxicillin (77.6%), penicillin (75.9%), ampicillin (74.1%) and cefoperazone (51.7%). Overall, isolates from clinical cases of mastitis had a higher resistance than subclinical isolates. Conclusion: CNS isolates were susceptible to chloramphenicol, gentamicin and streptomycin, while higher resistance was recorded against routinely used penicillin group. PMID:27047108

  10. Dissemination of antibiotic resistance genes in representative broiler feedlots environments: identification of indicator ARGs and correlations with environmental variables.

    PubMed

    He, Liang-Ying; Liu, You-Sheng; Su, Hao-Chang; Zhao, Jian-Liang; Liu, Shuang-Shuang; Chen, Jun; Liu, Wang-Rong; Ying, Guang-Guo

    2014-11-18

    Livestock operations are known to harbor elevated levels of antibiotic resistance genes (ARGs) that may pose a threat to public health. Broiler feedlots may represent an important source of ARGs in the environment. However, the prevalence and dissemination mechanisms of various types of ARGs in the environment of broiler feedlots have not previously been identified. We examined the occurrence, abundance and variation of ARGs conferring resistance to chloramphenicols, sulfonamides and tetracyclines in the environments of two representative types of broiler feedlots (free range and indoor) by quantitative PCR, and assessed their dissemination mechanisms. The results showed the prevalence of various types of ARGs in the environmental samples of the broiler feedlots including manure/litter, soil, sediment, and water samples, with the first report of five chloramphenicol resistance genes (cmlA, floR, fexA, cfr, and fexB) in broiler feedlots. Overall, chloramphenicol resistance genes and sulfonamides sul genes were more abundant than tetracyclines tet genes. The ARG abundances in the samples from indoor boiler feedlots were generally different to the free range feedlots, suggesting the importance of feeding operations in ARG dissemination. Pearson correlation analysis showed significant correlations between ARGs and mobile genetic element genes (int1 and int2), and between the different classes of ARGs themselves, revealing the roles of horizontal gene transfer and coselection for ARG dissemination in the environment. Further regression analysis revealed that fexA, sul1 and tetW could be reliable indicator genes to surrogate anthropogenic sources of ARGs in boiler feedlots (correlations of fexA, sul1 and tetW to all ARGs: R = 0.95, 0.96 and 0.86, p < 0.01). Meanwhile, significant correlations were also identified between indicator ARGs and their corresponding antibiotics. In addition, some ARGs were significantly correlated with typical metals (e.g., Cu, Zn, and As with

  11. Antimicrobial resistance and molecular epidemiology of vancomycin-resistant enterococci from North America and Europe: a report from the SENTRY antimicrobial surveillance program.

    PubMed

    Deshpande, Lalitagauri M; Fritsche, Thomas R; Moet, Gary J; Biedenbach, Douglas J; Jones, Ronald N

    2007-06-01

    Increases in prevalence of vancomycin-resistant enterococci (VRE) have been documented globally since its emergence in the 1980s. A SENTRY Antimicrobial Surveillance Program (2003) objective monitored VRE isolates with respect to antimicrobial susceptibility trends, geographic resistance variability, and clonal dissemination. In 2003, VRE isolates from North America (United States and Canada, n = 839, 26 sites) and Europe (n = 56, 10 sites) were susceptibility tested using Clinical and Laboratory Standards Institute (CLSI) reference methodologies. Based on resistance profiles, 155 isolates displayed similar multidrug-resistant (MDR) profiles and were temporally related; these were subsequently submitted for typing by pulsed-field gel electrophoresis (PFGE). Most of the submitted isolates were Enterococcus faecium (91.0%) and Enterococcus faecalis (7.8%). Among VRE, the VanA phenotype was more prevalent in North America (76%) than Europe (40%), and all isolates had elevated resistance rates to other antimicrobial classes including the following: 1) chloramphenicol resistance among E. faecalis being greater in North America than in Europe (28.6% versus 7.1%, respectively) but reversed among E. faecium (0.5% and 15.0%, the latter due to clonal occurrences); 2) ciprofloxacin resistance in North America >99% for both species and in Europe varying from 85.7% to 87.5%; 3) rare occurrences of linezolid resistance in North America (0.8% to 1.8%) due to G2576U ribosomal mutation; 4) higher quinupristin/dalfopristin resistance observed among European E. faecium strains (10.0% versus 0.6%); and 5) higher rifampin resistance rates among European E. faecalis (21.4% versus 5.4%). Thirty-five MDR epidemic clusters were identified by PFGE in 21 North American and 2 European medical centers including the following: 1) VanA (20 sites, 27 clonal occurrences) and VanB (1 site, 2 clonal occurrences); 2) elevated quinupristin/dalfopristin MIC results (not vatD/E, 3 sites); and 3

  12. Altered myoelectric activity in the experimental blind loop syndrome.

    PubMed

    Justus, P G; Fernandez, A; Martin, J L; King, C E; Toskes, P P; Mathias, J R

    1983-09-01

    Nutrient malabsorption and diarrhea are characteristic of the blind loop syndrome. Alterations in motility have been implicated as a cause of bacterial overgrowth, but the possibility that altered motility may result from alterations in the flora has not been explored. The purpose of this study was to characterize the myoelectric activity of the small intestine in the blind loop rat model. Eight groups of rats were studied: rats with self-filling blind loops, which develop bacterial overgrowth; rats with self-emptying blind loops, which are surgical controls that do not develop overgrowth; unoperated litter mates; rats with self-filling blind loops and unoperated controls treated with chloramphenicol, 200 mg/d i.p.; rats with surgically removed self-filling blind loops; operated control rats; and gnotobiotic rats with self-filling blind loops. In the untreated rats with self-filling blind loops, there was altered myoelectric activity characterized by an increased percentage of slow waves occupied by action potentials and by organized activity similar to the migrating action potential complex. Migrating action potential complex activity and percentage of slow waves occupied by action potentials were significantly decreased with chloramphenicol therapy; that decrease correlated with a decrease in aerobes and anaerobes. Migrating action potential complex activity was abolished in rats with surgically removed self-filling blind loops; they also showed a significant decrease in percentage of slow waves occupied by action potentials. Gnotobiotic rats with self-filling blind loops showed no alteration in myoelectric activity. These data indicate: (a) bacterial overgrowth is associated with a significant increase in percentage of slow waves occupied by action potentials and migrating action potential complex activity; (b) chloramphenicol significantly reduced both percentage of slow waves occupied by action potentials and migrating action potential complex activity; and (c

  13. In Vitro Activities of Nisin and Nisin Derivatives Alone and In Combination with Antibiotics against Staphylococcus Biofilms

    PubMed Central

    Field, Des; O’ Connor, Rory; Cotter, Paul D.; Ross, R. Paul; Hill, Colin

    2016-01-01

    The development and spread of pathogenic bacteria that are resistant to the existing catalog of antibiotics is a major public health threat. Biofilms are complex, sessile communities of bacteria embedded in an organic polymer matrix which serve to further enhance antimicrobial resistance. Consequently, novel compounds and innovative methods are urgently required to arrest the proliferation of drug-resistant infections in both nosocomial and community environments. Accordingly, it has been suggested that antimicrobial peptides could be used as novel natural inhibitors that can be used in formulations with synergistically acting antibiotics. Nisin is a member of the lantibiotic family of antimicrobial peptides that exhibit potent antibacterial activity against many Gram-positive bacteria. Recently we have used bioengineering strategies to enhance the activity of nisin against several high profile targets, including multi-drug resistant clinical pathogens such as methicillin-resistant Staphylococcus aureus, vancomycin-resistant enterococci, staphylococci, and streptococci associated with bovine mastitis. We have also identified nisin derivatives with an enhanced ability to impair biofilm formation and to reduce the density of established biofilms of methicillin resistant S. pseudintermedius. The present study was aimed at evaluating the potential of nisin and nisin derivatives to increase the efficacy of conventional antibiotics and to assess the possibility of killing and/or eradicating biofilm-associated cells of a variety of staphylococcal targets. Growth curve-based comparisons established that combinations of derivatives nisin V + penicillin or nisin I4V + chloramphenicol had an enhanced inhibitory effect against S. aureus SA113 and S. pseudintermedius DSM21284, respectively, compared to the equivalent nisin A + antibiotic combinations or when each antimicrobial was administered alone. Furthermore, the metabolic activity of established biofilms treated with nisin

  14. Dissemination of antibiotic resistance genes in representative broiler feedlots environments: identification of indicator ARGs and correlations with environmental variables.

    PubMed

    He, Liang-Ying; Liu, You-Sheng; Su, Hao-Chang; Zhao, Jian-Liang; Liu, Shuang-Shuang; Chen, Jun; Liu, Wang-Rong; Ying, Guang-Guo

    2014-11-18

    Livestock operations are known to harbor elevated levels of antibiotic resistance genes (ARGs) that may pose a threat to public health. Broiler feedlots may represent an important source of ARGs in the environment. However, the prevalence and dissemination mechanisms of various types of ARGs in the environment of broiler feedlots have not previously been identified. We examined the occurrence, abundance and variation of ARGs conferring resistance to chloramphenicols, sulfonamides and tetracyclines in the environments of two representative types of broiler feedlots (free range and indoor) by quantitative PCR, and assessed their dissemination mechanisms. The results showed the prevalence of various types of ARGs in the environmental samples of the broiler feedlots including manure/litter, soil, sediment, and water samples, with the first report of five chloramphenicol resistance genes (cmlA, floR, fexA, cfr, and fexB) in broiler feedlots. Overall, chloramphenicol resistance genes and sulfonamides sul genes were more abundant than tetracyclines tet genes. The ARG abundances in the samples from indoor boiler feedlots were generally different to the free range feedlots, suggesting the importance of feeding operations in ARG dissemination. Pearson correlation analysis showed significant correlations between ARGs and mobile genetic element genes (int1 and int2), and between the different classes of ARGs themselves, revealing the roles of horizontal gene transfer and coselection for ARG dissemination in the environment. Further regression analysis revealed that fexA, sul1 and tetW could be reliable indicator genes to surrogate anthropogenic sources of ARGs in boiler feedlots (correlations of fexA, sul1 and tetW to all ARGs: R = 0.95, 0.96 and 0.86, p < 0.01). Meanwhile, significant correlations were also identified between indicator ARGs and their corresponding antibiotics. In addition, some ARGs were significantly correlated with typical metals (e.g., Cu, Zn, and As with

  15. Regulation of glutamine synthetase, aspartokinase, and total protein turnover in Klebsiella aerogenes.

    PubMed

    Fulks, R M; Stadtman, E R

    1985-12-13

    When suspensions of Klebsiella aerogenes are incubated in a nitrogen-free medium there is a gradual decrease in the levels of acid-precipitable protein and of aspartokinase III (lysine-sensitive) and aspartokinase I (threonine-sensitive) activities. In contrast, the level of glutamine synthetase increases slightly and then remains constant. Under these conditions, the glutamine synthetase and other proteins continue to be synthesized as judged by the incorporation of [14C]leucine into the acid-precipitable protein fraction and into protein precipitated by anti-glutamine synthetase antibodies, by the fact that growth-inhibiting concentrations of chloramphenicol also inhibit the incorporation of [14C]leucine into protein and into protein precipitated by anti-glutamine synthetase antibody, and by the fact that chloramphenicol leads to acceleration in the loss of aspartokinases I and III and promotes a net decrease in the level of glutamine synthetase and its cross-reactive protein. The loss of aspartokinases I and III in cell suspensions is stimulated by glucose and is inhibited by 2,4-dinitrophenol. Glucose also stimulates the loss of aspartokinases and glutamine synthetase in the presence of chloramphenicol. Cell-free extracts of K. aerogenes catalyze rapid inactivation of endogenous glutamine synthetase as well as exogenously added pure glutamine synthetase. This loss of glutamine synthetase is not associated with a loss of protein that cross-reacts with anti-glutamine synthetase antibodies. The inactivation of glutamine synthetase in extracts is not due to adenylylation. It is partially prevented by sulfhydryl reagents, Mn2+, antimycin A, 2,4-dinitrophenol, EDTA, anaerobiosis and by dialysis. Following 18 h dialysis, the capacity of extracts to catalyze inactivation of glutamine synthetase is lost but can be restored by the addition of Fe2+ (or Ni2+) together with ATP (or other nucleoside di- and triphosphates. After 40-60 h dialysis Fe3+ together with NADH (but

  16. Antimicrobial activity of essential oil of Eucalyptus globulus against fish pathogenic bacteria.

    PubMed

    Park, Joon-Woo; Wendt, Mitchell; Heo, Gang-Joon

    2016-06-01

    The antibacterial activities of the essential oil of Eucalyptus globulus (EOEG) was determined against 7 fish pathogenic bacteria (Edwardsiella tarda, Streptococcus iniae, S. parauberis, Lactococcus garviae, Vibrio harveyi, V. ichthyoenteri and Photobacterium damselae) obtained from farmed olive flounder. The inhibitory activity was evaluated by three methods: Disc diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). According to the disc diffusion test, as the concentration of EOEG (5-40 µg) rises, the inhibitory zone increases in size. Compared with amoxicillin, tetracycline and chloramphenicol, EOEG showed similar antibacterial activity. The MIC of EOEG ranged from 7.8 to 125 mg/mL and MBC values ranged from 62 to 250 mg/mL. These results show that EOEG has antimicrobial activity against all seven bacteria, but there was no marked difference between each genus. From these results, it is suggested that EOEG can be used as an antimicrobial agent against fish bacterial diseases in the fish industry.

  17. An improved method of electroporation for introducing biologically active foreign genes into cultured mammalian cells

    SciTech Connect

    Tatsuka, Masaaki; Orita, Satoshi; Yagi, Takashi; Kakunaga, Takeo )

    1988-09-01

    The authors have developed a modified, reproducible, and efficient method for introducing cloned genes into mammalian cells by using an electric field followed by treatment with sodium butyrate. Transfection frequencies with plasmid pSV2-neo, consisting of an antibiotic (G418) resistance gene and simian virus 40 (SV40) early promoter, by electroporation were higher than those by calcium phosphate DNA precipitation. Treatment with sodium butyrate following electroporation significantly increased the frequency of transfection in various types of cell lines and primary cultured cells including human skin fibroblasts. Treatment with sodium butyrate also increased the transient expression of the gene for chloramphenicol acetyltransferase when the gene was introduced into BALB/c 3T3 cells by eletroporation. Electroporation combined with sodium butyrate treatment is an improved method for stable and transient biochemical transformation of foreign genes in cultured mammalian cells.

  18. Efflux Pump Blockers in Gram-Negative Bacteria: The New Generation of Hydantoin Based-Modulators to Improve Antibiotic Activity

    PubMed Central

    Otręebska-Machaj, Ewa; Chevalier, Jacqueline; Handzlik, Jadwiga; Szymańska, Ewa; Schabikowski, Jakub; Boyer, Gérard; Bolla, Jean-Michel; Kieć-Kononowicz, Katarzyna; Pagès, Jean-Marie; Alibert, Sandrine

    2016-01-01

    Multidrug resistant (MDR) bacteria are an increasing health problem with the shortage of new active antibiotic agents. Among effective mechanisms that contribute to the spread of MDR Gram-negative bacteria are drug efflux pumps that expel clinically important antibiotic classes out of the cell. Drug pumps are attractive targets to restore the susceptibility toward the expelled antibiotics by impairing their efflux activity. Arylhydantoin derivatives were investigated for their potentiation of activities of selected antibiotics described as efflux substrates in Enterobacter aerogenes expressing or not AcrAB pump. Several compounds increased the bacterial susceptibility toward nalidixic acid, chloramphenicol and sparfloxacin and were further pharmacomodulated to obtain a better activity against the AcrAB producing bacteria. PMID:27199950

  19. Phytochrome activation of two nuclear genes requires cytoplasmic protein synthesis.

    PubMed Central

    Lam, E; Green, P J; Wong, M; Chua, N H

    1989-01-01

    We have investigated the effects of protein synthesis inhibitors on light-induced expression of two plant nuclear genes, Cab and rbcS, in wheat, pea and transgenic tobacco. Light activation of these two genes is very sensitive to cycloheximide, an inhibitor of cytoplasmic protein synthesis but not to chloramphenicol, an inhibitor of organellar protein synthesis. Studies with chimeric gene constructs in transgenic tobacco seedlings show that cycloheximide exerts its effect at the transcriptional level. As a control, we show that the expression of the cauliflower mosaic virus (CaMV) 35S promoter is enhanced by cycloheximide treatment, irrespective of the coding sequence used. Escape-time analyses with green wheat seedlings show that the cycloheximide block for Cab gene expression is after the primary signal transduction step linked to phytochrome photoconversion. Our results suggest that phytochrome activation of Cab and rbcS is mediated by a labile protein factor(s) synthesized on cytoplasmic ribosomes. Images PMID:2583082

  20. Epidemic typhus in the United States associated with flying squirrels.

    PubMed

    Duma, R J; Sonenshine, D E; Bozeman, F M; Veazey, J M; Elisberg, B L; Chadwick, D P; Stocks, N I; McGill, T M; Miller, G B; MacCormack, J N

    1981-06-12

    Between July 1977 and January 1980, seven cases of sporadic, nonepidemic "epidemic" typhus (Rickettsia prowazekii) were discovered in Virginia, West Virginia, and North Carolina. The reservoir seemed to be the southern flying squirrel (Glaucomys volans), an animal indigenous to the eastern United States; however, the vector or mode of acquisition was not evident. Diagnosis was established principally through complement fixation, indirect immunofluorescence, and toxin neutralization tests. Patients' ages were 11 to 81 years. Most were white women. Six had abrupt onset of illness. Headaches, fever, myalgias, and exanthems were among the presenting complaints. The disease seemed milder than classic louse-born epidemic typhus, but in some instances, it was life-threatening. All patients responded to tetracycline or chloramphenicol. This entity probably is more common than reported, is difficult to recognize, and is produced by an organism seemingly identical to that producing louse-born epidemic typhus. PMID:6785459

  1. A clinico-epidemiological study of epidemic typhus in Africa.

    PubMed

    Perine, P L; Chandler, B P; Krause, D K; McCardle, P; Awoke, S; Habte-Gabr, E; Wisseman, C L; McDade, J E

    1992-05-01

    Epidemic, louse-borne typhus persists in the rugged, mountainous areas of Ethiopia and much of northeastern and central Africa as well as in the rural highlands of Central and South America, where the conditions of living favor the harboring of body lice and where antibiotic treatment and effective louse-control measures are unavailable. The historical significance and current epidemiology of typhus, including the reservoir of Rickettsia prowazekii in flying squirrels in the United States, are reviewed, and the clinical presentation, laboratory findings, and hospital course in the cases of 60 patients admitted with epidemic, louse-borne typhus to the St. Paul's Hospital in Addis Ababa, Ethiopia, are described. Treatment of this disease with oral doxycycline, tetracycline, or chloramphenicol prevents complications and results in prompt resolution of symptoms. PMID:1600020

  2. Use of imported folk remedies and medications in the Wisconsin Hmong community.

    PubMed

    Werner, M A; Knobeloch, L M; Erbach, T; Anderson, H A

    2001-01-01

    The Marathon County Health Department and the Wisconsin Division of Public Health evaluated several imported drugs and folk remedies that were being used by 2 Hmong families. These included a powdered blend of folk remedies that had been purchased in California and 5 packets of medication that had been imported from Thailand. The powdered folk remedy contained arsenic sulfide as a principal ingredient. The drug packets contained acetyl aspirin, acetaminophen and chloramphenicol. The purity of these drugs was not assessed, and their dates of manufacture could not be determined. To discourage use of folk remedies and imported drugs, the county health department issued a press release that was published in a Hmong community newsletter. Despite these efforts, many Asian immigrants may continue to use these products.

  3. Antimicrobial resistance determinant microarray for analysis of multi-drug resistant isolates

    NASA Astrophysics Data System (ADS)

    Taitt, Chris Rowe; Leski, Tomasz; Stenger, David; Vora, Gary J.; House, Brent; Nicklasson, Matilda; Pimentel, Guillermo; Zurawski, Daniel V.; Kirkup, Benjamin C.; Craft, David; Waterman, Paige E.; Lesho, Emil P.; Bangurae, Umaru; Ansumana, Rashid

    2012-06-01

    The prevalence of multidrug-resistant infections in personnel wounded in Iraq and Afghanistan has made it challenging for physicians to choose effective therapeutics in a timely fashion. To address the challenge of identifying the potential for drug resistance, we have developed the Antimicrobial Resistance Determinant Microarray (ARDM) to provide DNAbased analysis for over 250 resistance genes covering 12 classes of antibiotics. Over 70 drug-resistant bacteria from different geographic regions have been analyzed on ARDM, with significant differences in patterns of resistance identified: genes for resistance to sulfonamides, trimethoprim, chloramphenicol, rifampin, and macrolide-lincosamidesulfonamide drugs were more frequently identified in isolates from sources in Iraq/Afghanistan. Of particular concern was the presence of genes responsible for resistance to many of the last-resort antibiotics used to treat war traumaassociated infections.

  4. Biochemical Studies on Development of Mitochondria in Pea Cotyledons during the Early Stage of Germination

    PubMed Central

    Nawa, Yoshihiko; Asahi, Tadashi

    1973-01-01

    l-Leucine-U-14C was incorporated into mitochondrial protein in pea (Pisum sativum var. Alaska) cotyledons during the imbibing stages. Incorporation was almost completely inhibited by cycloheximide but not by chloramphenicol. Both antibiotics did not affect increases in mitochondrial activities and components of the cotyledons during imbibition. Therefore, mitochondrial development seems to be achieved by a transfer of protein pre-existing in the cytoplasm into the mitochondria rather than by de novo synthesis of mitochondrial protein. Cycloheximide stimulated an increase in bile saltsoluble protein of mitochondria in imbibing pea cotyledons. The recovery of cytochrome oxidase activity after sucrose density gradient centrifugation was enhanced, and the morphological properties of mitochondria were altered by cycloheximide. Images PMID:16658422

  5. Appearance and Disappearance of Cyanide-Resistant Respiration in Vigna mungo Cotyledons during and following Germination of the Axis

    PubMed Central

    Morohashi, Yukio; Matsushima, Hisashi

    1983-01-01

    Mitochondrial preparations isolated from black gram (Vigna mungo L.) cotyledons exhibited cyanide-resistant respiration which was of mitochondrial origin. The appearance and the disappearance of this alternative respiration took place during and following imbibition. During the first 6 hours of imbibition, the respiration was completely inhibited by cyanide, but after this time the alternative respiration markedly developed, reaching a maximal cyanide-resistance 12 to 16 hours after the start of imbibition. Subsequently, the alternative respiration gradually disappeared. The actions of cycloheximide and chloramphenicol indicated that the appearance was dependent on cytoplasmic protein synthesis and that the disappearance depended on both cytoplasmic and mitochondrial protein synthesis. The alternative pathway contributed to state 4 respiration, but not to state 3 respiration, in mitochondria from 1-day-old cotyledons. On day 3, it contributed to neither state 3 nor state 4. PMID:16663192

  6. Novel Functions of an Iron-Sulfur Flavoprotein from Trichomonas vaginalis Hydrogenosomes

    PubMed Central

    Smutná, Tamara; Pilarová, Katerina; Tarábek, Ján; Tachezy, Jan

    2014-01-01

    Iron-sulfur flavoproteins (Isf) are flavin mononucleotide (FMN)- and FeS cluster-containing proteins commonly encountered in anaerobic prokaryotes. However, with the exception of Isf from Methanosarcina thermophila, which participates in oxidative stress management by removing oxygen and hydrogen peroxide, none of these proteins has been characterized in terms of function. Trichomonas vaginalis, a sexually transmitted eukaryotic parasite of humans, was found to express several iron-sulfur flavoprotein (TvIsf) homologs in its hydrogenosomes. We show here that in addition to having oxygen-reducing activity, the recombinant TvIsf also functions as a detoxifying reductase of metronidazole and chloramphenicol, both of which are antibiotics effective against a variety of anaerobic microbes. TvIsf can utilize both NADH and reduced ferredoxin as electron donors. Given the prevalence of Isf in anaerobic prokaryotes, we propose that these proteins are central to a novel defense mechanism against xenobiotics. PMID:24663020

  7. Genetic transformation of marine cyanobacterium Synechococcus sp. CC9311 (Cyanophyceae) by electroporation

    NASA Astrophysics Data System (ADS)

    Chen, Huaxin; Lin, Hanzhi; Jiang, Peng; Li, Fuchao; Qin, Song

    2013-03-01

    Synechococcus sp. CC9311 is a marine cyanobacterium characterized by type IV chromatic acclimation (CA). A genetic transformation system was developed as a first step to elucidate the molecular mechanism of CA. The results show that Synechococcus sp. CC9311 cells were sensitive to four commonly used antibiotics: ampicillin, kanamycin, spectinomycin, and chloramphenicol. An integrative plasmid to disrupt the putative phycoerythrin lyase gene mpeV, using a kanamycin resistance gene as selectable marker, was constructed by recombinant polymerase chain reaction. The plasmid was then transformed into Synechococcus sp. CC9311 via electroporation. High transformation efficiency was achieved at a field strength of 2 kV/cm. DNA analysis showed that mpeV was fully disrupted following challenge of the transformants with a high concentration of kanamycin. In addition, the transformants that displayed poor growth on agar SN medium could be successfully plated on agarose SN medium.

  8. The Synthesis of Ribosomes in E. coli

    PubMed Central

    Britten, R. J.; McCarthy, B. J.; Roberts, R. B.

    1962-01-01

    The incorporation of C14 leucine into the protein moiety of ribosomes has been studied as a sequel to the studies of ribosomal RNA synthesis. In contrast to the latter studies, labeled leucine is incorporated directly into 50S and 30S ribosomes without measurable delay by precursor stages. There is, however, evidence of some transfer of radioactivity from the 43S group of particles to the 50S. The inhibition of protein synthesis by chloramphenicol results in the accumulation of material similar to the eosome—the primary precursor in ribosome synthesis. There is also evidence for the synthesis of some neosome. The results of the studies of ribosomal RNA and protein synthesis are combined into a model of ribosome synthesis. Finally, consideration is made of the significance of these studies of ribosome synthesis for general problems of protein synthesis and information transfer. PMID:13873182

  9. Expression of a human placental alkaline phosphatase gene in transfected cells: Use as a reporter for studies of gene expression

    SciTech Connect

    Henthorn, P.; Zervos, P.; Raducha, M.; Harris, H.; Kadesch, T.

    1988-09-01

    The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is appropriate for use as a reporter in studies of gene regulation since its expression is dependent on the presence of exogenous transcription control elements. The overall assay to detect the expression of the gene is quantitative, very rapid, and inexpensive. Cotransfections of cells with pSV2Apap and a related plasmid carrying the bacterial chloramphenicol acetyltransferase gene (pSV2Acat) indicate that transcription of these two genes is detected with roughly the same sensitivity.

  10. Thermolabile repression of cephalosporinase synthesis in Citrobacter freundii.

    PubMed

    Sawai, T; Nakajima, S; Morohoshi, T; Yamagishi, S

    1977-11-01

    An unusual regulatory system of cephalosporinase synthesis in Citrobacter freundii has been found. When the bacteria are grown at 20 C, the cephalosporinase is synthesized as a typical inducible enzyme and benzylpenicillin acts as an effective inducer. The enzyme, however, is synthesized in the absence of the inducer at growth temperatures above 25 C. when the growth temperature is shifted from 20 C to 37 C, the induction of enzyme synthesis is observed after about one half of the organism doubling time, but it does not occur in the presence of chloramphenicol. The reverse control mutants, the enzyme constitutive synthesis of which is markedly depressed by benzylpenicillin, were isolated from the C. freundii wild strain. The possibility that the enzyme synthesis is governed by a regulatory system analogous to the its mutant of the lac operon in Escherichia coli was suggested. PMID:203825

  11. Human Cytomegalovirus Infection Upregulates the Mitochondrial Transcription and Translation Machineries

    PubMed Central

    Weekes, M. P.; Antrobus, R.; Rorbach, J.; van Haute, L.; Umrania, Y.; Smith, D. L.; Minczuk, M.; Lehner, P. J.; Sinclair, J. H.

    2016-01-01

    ABSTRACT Infection with human cytomegalovirus (HCMV) profoundly affects cellular metabolism. Like in tumor cells, HCMV infection increases glycolysis, and glucose carbon is shifted from the mitochondrial tricarboxylic acid cycle to the biosynthesis of fatty acids. However, unlike in many tumor cells, where aerobic glycolysis is accompanied by suppression of mitochondrial oxidative phosphorylation, HCMV induces mitochondrial biogenesis and respiration. Here, we affinity purified mitochondria and used quantitative mass spectrometry to determine how the mitochondrial proteome changes upon HCMV infection. We found that the mitochondrial transcription and translation systems are induced early during the viral replication cycle. Specifically, proteins involved in biogenesis of the mitochondrial ribosome were highly upregulated by HCMV infection. Inhibition of mitochondrial translation with chloramphenicol or knockdown of HCMV-induced ribosome biogenesis factor MRM3 abolished the HCMV-mediated increase in mitochondrially encoded proteins and significantly impaired viral growth under bioenergetically restricting conditions. Our findings demonstrate how HCMV manipulates mitochondrial biogenesis to support its replication. PMID:27025248

  12. Clonal Occurrence of Salmonella Weltevreden in Cultured Shrimp in the Mekong Delta, Vietnam.

    PubMed

    Noor Uddin, Gazi Md; Larsen, Marianne Halberg; Barco, Lisa; Minh Phu, Tran; Dalsgaard, Anders

    2015-01-01

    This study investigated the occurrence, serovar and antimicrobial resistance of Salmonella spp. in shrimp samples from intensive and extensive farms located in three different provinces in the Mekong Delta, Vietnam. Shrimp from 11 of the 48 farms all contained S. Weltevreden, except for one farm yielding S. Agona, with no difference in Salmonella occurrence between the two production systems. Pulsed field gel electrophoresis (PFGE) of S. Weltevreden showed closely related XbaI pulse types, suggesting a clonal relationship despite the farms and shrimp samples being epidemiologically unrelated. S. Weltevreden was susceptible to most antimicrobials tested, with a few strains being resistant to florfenicol, chloramphenicol, sulfamethoxazole or trimethoprim. Future studies of the ecology of S. Weltevreden should establish if this serovar may survive better and even multiply in warm-water shrimp farm environments compared to other Salmonella serovars. PMID:26222547

  13. Isolation of multiple drug-resistant enteric bacteria from feces of wild Western Lowland Gorilla (Gorilla gorilla gorilla) in Gabon.

    PubMed

    Mbehang Nguema, Pierre Philippe; Okubo, Torahiko; Tsuchida, Sayaka; Fujita, Shiho; Yamagiwa, Juichi; Tamura, Yutaka; Ushida, Kazunari

    2015-05-01

    Prevalence of drug-resistant bacteria in wildlife can reveal the actual level of anthropological burden on the wildlife. In this study, we isolated two multiple drug-resistant strains, GG6-2 and GG6-1-1, from 27 fresh feces of wild western lowland gorillas in Moukalaba-Doudou National Park, Gabon. Isolates were identified as Achromobacter xylosoxidans and Providencia sp., respectively. Minimum inhibitory concentrations of the following 12 drugs-ampicillin (ABPC), cefazolin (CEZ), cefotaxime (CTX), streptomycin (SM), gentamicin (GM), kanamycin (KM), tetracycline (TC), nalidixic acid (NA), ciprofloxacin (CPFX), colistin (CL), chloramphenicol (CP) and trimethoprim (TMP)-were determined. Isolate GG6-2 was resistant to all antimicrobials tested and highly resistant to CTX, SM, TC, NA and TMP. Isolate GG6-1-1 was resistant to ABPC, CEZ, TC, CL, CP and TMP.

  14. Efflux Pump Blockers in Gram-Negative Bacteria: The New Generation of Hydantoin Based-Modulators to Improve Antibiotic Activity.

    PubMed

    Otręebska-Machaj, Ewa; Chevalier, Jacqueline; Handzlik, Jadwiga; Szymańska, Ewa; Schabikowski, Jakub; Boyer, Gérard; Bolla, Jean-Michel; Kieć-Kononowicz, Katarzyna; Pagès, Jean-Marie; Alibert, Sandrine

    2016-01-01

    Multidrug resistant (MDR) bacteria are an increasing health problem with the shortage of new active antibiotic agents. Among effective mechanisms that contribute to the spread of MDR Gram-negative bacteria are drug efflux pumps that expel clinically important antibiotic classes out of the cell. Drug pumps are attractive targets to restore the susceptibility toward the expelled antibiotics by impairing their efflux activity. Arylhydantoin derivatives were investigated for their potentiation of activities of selected antibiotics described as efflux substrates in Enterobacter aerogenes expressing or not AcrAB pump. Several compounds increased the bacterial susceptibility toward nalidixic acid, chloramphenicol and sparfloxacin and were further pharmacomodulated to obtain a better activity against the AcrAB producing bacteria. PMID:27199950

  15. The Effect of Light and Inhibitors on Chloroplast and Cytoplasmic RNA Synthesis

    PubMed Central

    Ingle, J.

    1968-01-01

    Chloroplast RNA is synthesized in dark-grown radish cotyledons at about one-third the rate of that in the light. The synthesis, however, continues for longer in the dark and the percentage of chloroplast RNA can approach that in light-grown tissue. Light stimulates the synthesis and accumulation of both cytoplasmic and chloroplast RNA, but shows a 4-fold greater stimulation of the chloroplast RNA. Chloramphenicol, streptomycin and cycloheximide inhibit the synthesis of chloroplast RNA with little effect on cytoplasmic RNA. 5-Fluorouracil inhibits the synthesis of cytoplasmic more than chloroplast RNA. Synthesis of the 0.56 × 106 mol wt chloroplast RNA is inhibited much less than the other ribosomal RNA components by actinomycin D. PMID:5699149

  16. Comparative examination of probe labeling methods for microarray hybridization

    NASA Astrophysics Data System (ADS)

    Burke, David I.; Woodward, Karen; Setterquist, Robert A.; Kawasaki, Ernest S.

    2001-06-01

    For detection of differential gene expression, confocal laser based scanners are now capable of analyzing microarrays using one to five wavelengths. This allows investigators to choose among several labeling methods. Here we compare direct incorporation and indirect methods (amino-allyl and dendrimers) for labeling cDNA probes. We assessed reproducible sensitivity of each probe preparation method in two ways. First, by comparing hybridization intensities for limit of signal detection and second by measuring the lowest detectable concentration of a known ratio of mixed DNA (spikes). Limit of detection assay was done using arrays of mixed targets consisting of a serially diluted human specific gene fragment (HU1) and an undiluted DNA of chloramphenicol acetyl tranferase (CAT) gene. Then, individual single target arrays of CAT and HU1 DNA were used to determine the lowest detectable spike ratio of each labeling method. The results of this study will be presented and their significance for the analysis of microarrays will be discussed.

  17. A plasmid in Streptococcus pneumoniae.

    PubMed Central

    Smith, M D; Guild, W R

    1979-01-01

    Plasmid deoxyribonucleic acid has been detected in three related laboratory strains of Streptococcus pneumoniae. Strains D39S, R36, and R36NC each contain a minimum of two copies per cell of a 2.0-megadalton plasmid (pDP1). A plasmid twice as large as this smaller one is also present in much lower quantity in these strains, but neither plasmid is present in four strains related to these or in a drug-resistant clinical isolate from Paris. The plasmid yield was not amplified in the presence of chloramphenicol. No phenotype has been correlated with the presence of pDP1, which has existed in strains carried for many years in laboratory collections. Images PMID:33961

  18. Inactivation of antibiotics and the dissemination of resistance genes.

    PubMed

    Davies, J

    1994-04-15

    The emergence of multidrug-resistant bacteria is a phenomenon of concern to the clinician and the pharmaceutical industry, as it is the major cause of failure in the treatment of infectious diseases. The most common mechanism of resistance in pathogenic bacteria to antibiotics of the aminoglycoside, beta-lactam (penicillins and cephalosporins), and chloramphenicol types involves the enzymic inactivation of the antibiotic by hydrolysis or by formation of inactive derivatives. Such resistance determinants most probably were acquired by pathogenic bacteria from a pool of resistance genes in other microbial genera, including antibiotic-producing organisms. The resistance gene sequences were subsequently integrated by site-specific recombination into several classes of naturally occurring gene expression cassettes (typically "integrons") and disseminated within the microbial population by a variety of gene transfer mechanisms. Although bacterial conjugation once was believed to be restricted in host range, it now appears that this mechanism of transfer permits genetic exchange between many different bacterial genera in nature.

  19. Nasopharyngeal carriage of Streptococcus pneumoniae in adults infected with human immunodeficiency virus in Jakarta, Indonesia.

    PubMed

    Harimurti, Kuntjoro; Saldi, Siti R F; Dewiasty, Esthika; Khoeri, Miftahuddin M; Yunihastuti, Evi; Putri, Tiara; Tafroji, Wisnu; Safari, Dodi

    2016-01-01

    This study investigated the distribution of serotype and antimicrobial susceptibility of Streptococcus pneumoniae carried by adults infected with human immunodeficiency virus (HIV) in Jakarta, Indonesia. Specimens of nasopharyngeal swab were collected from 200 HIV infected adults aged 21 to 63 years. Identification of S. pneumoniae was done by optochin susceptibility test and PCR for the presence of psaA and lytA genes. Serotyping was performed with sequential multiplex PCR and antibiotic susceptibility with the disk diffusion method. S. pneumoniae strains were carried by 10% adults with serotype 6A/B 20% was common serotype among cultured strains in 20 adults. Most of isolates were susceptible to chloramphenicol (80%) followed by clindamycin (75%), erythromycin (75%), penicillin (55%), and tetracycline (50%). This study found resistance to sulphamethoxazole/trimethoprim was most common with only 15% of strains being susceptible. High non-susceptibility to sulphamethoxazole/trimethoprim was observed in S. pneumoniae strains carried by HIV infected adults in Jakarta, Indonesia.

  20. A plant scaffold attached region detected close to a T-DNA integration site is active in mammalian cells.

    PubMed Central

    Dietz, A; Kay, V; Schlake, T; Landsmann, J; Bode, J

    1994-01-01

    Integration of foreign genes into plant genomes by the Agrobacterium T-DNA transfer system has been considered to occur at random. It has been speculated that the chromosomal structure of the integration site might affect the expression pattern of the introduced genes. To gain insight into the molecular structure of T-DNA integration sites and its possible impact on gene expression, we have examined plant DNA sequences in the vicinity of T-DNA borders. Analysis of a transgenic petunia plant containing a chloramphenicol acetyltransferase (CAT) gene regulated by the hemoglobin promoter (PAR) from Parasponia andersonii revealed a scaffold attachment region (SAR) close to one T-DNA end. In addition to having strong binding affinities for both animal and plant nuclear scaffolds this petunia SAR element is as active in mammalian cells as the authentic elements from mammalian sources. Images PMID:8052530

  1. Multidetection of antibiotics in liver tissue by ultra-high-pressure-liquid-chromatography-tandem mass spectrometry.

    PubMed

    Freitas, Andreia; Barbosa, Jorge; Ramos, Fernando

    2015-01-22

    A multiresidue quantitative screening method covering 39 antibiotics from 7 different families by ultra-high-pressure-liquid-chromatography-tandem mass spectrometry (UHPLC-MS/MS) is described. Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol are simultaneously detected in liver tissue. A simple sample treatment method consisting of extraction with a mixture of acetonitrile and ethylenediaminetetraacetic acid (EDTA) followed by solid-phase extraction (SPE) with a hydrophilic-lipophilic balanced (HLB) cartridge was developed. The methodology was validated, in accordance with Decision 2002/657/EC, by evaluating the following required parameters: decision limit (CCα), detection capability (CCβ), specificity, repeatability and reproducibility. The precision, in terms of the relative standard deviation, was under 22% for all of the compounds, and the recoveries were between 80% and 110%. The CCα and CCβ were determined according to the maximum residue limit (MRL) or the minimum required performance limit (MRPL), when established.

  2. Tests in vitro and in pots with certain chemicals for inhibition of Pseudomonas solanacearum.

    PubMed

    el-Goorani, M A; Abo-el-Dahab, M K; Wagih, E E

    1978-01-01

    Twenty one isolates of Pseudomonas solanacearum E. F. Smith (Race 3) from various parts of Egypt were inhibited in vitro by Ampicillin, Chloramphenicol, Kanamycin, Oxytetracycline. Tetracycline, Penicillin G, Streptomycin, Nabam (Dithane A-40), Maneb (Dithane M-22), Zinc-ion maneb complex (Dithane M-45), and the insecticide Chlorthion. On the basis of in vitro-sensitivity to the selected 11 chemicals the 21 isolates could not be separated into different groups. Carbendazin (Bavistin), Benomyl, Drazoxolon (Mil-Col), and Temik proved ineffective in inhibiting the in vitro growth of all isolates at all tested concentrations. Preliminary investigations indicate that drenching the soil with solutions of Dithane M-22 (0.25% w/v, Dithane M-45 (0.25%) w/v, or insecticide Chlorthion (50 microgram/ml active ingredient) decreased the incidence of potato wilt disease that developed in soil, artificially infested with P. solanacearum. PMID:696045

  3. Antimicrobial susceptibility of Staphylococcus intermedius and Staphylococcus schleiferi isolated from dogs.

    PubMed

    Vanni, M; Tognetti, R; Pretti, C; Crema, F; Soldani, G; Meucci, V; Intorre, L

    2009-10-01

    The susceptibility to 23 antimicrobial agents was determined in 114 isolates of Staphylococcus intermedius and eight isolates of Staphylococcus schleiferi of canine origin. Overall, 73% of S. intermedius isolates and 37.5% of S. schleiferi isolates were susceptible to all the 23 antimicrobials tested. The large majority of S. intermedius strains retained susceptibility to antimicrobials currently employed in treatment of pyoderma (cephalosporins, cotrimoxazole and association amoxicillin-clavulanic acid) as well as to those effective against staphylococci (fusidic acid, rifampicin and fluoroquinolones). Resistance in S. intermedius was observed mainly against macrolides, chloramphenicol and lincosamides, while S. schleiferi isolates retained susceptibility to all antimicrobials except three of six fluoroquinolones. Although, our results confirm susceptibility to antimicrobials currently employed in pyoderma treatment, the several different resistance patterns observed for S. intermedius emphasize the importance of antimicrobial susceptibility testing of canine staphylococci to choose the most appropriate treatment of infections and to allow the prudent use of antimicrobial drugs in companion animals.

  4. Physiological characteristics of the biocontrol yeast Pichia anomala J121.

    PubMed

    Fredlund, Elisabeth; Druvefors, Ulrika; Boysen, Marianne E; Lingsten, Karl-Johan; Schnürer, Johan

    2002-08-01

    The yeast Pichia anomala J121 prevents mold spoilage and enhances preservation of moist grain in malfunctioning storage systems. Development of P. anomala J121 as a biocontrol agent requires in-depth knowledge about its physiology. P. anomala J121 grew under strictly anaerobic conditions, at temperatures between 3 degrees C and 37 degrees C, at pH values between 2.0 and 12.4, and at a water activity of 0.92 (NaCl) and 0.85 (glycerol). It could assimilate a wide range of C- and N-sources and produce killer toxin. A selective medium containing starch, nitrate, acetic acid, and chloramphenicol was developed for P. anomala. P. anomala was equally sensitive as Candida albicans to common antifungal compounds. Growth ability at a range of environmental conditions contributes to the competitive ability of the biocontrol yeast P. anomala J121. PMID:12702290

  5. An Outbreak of Bartonella bacilliformis in an Endemic Andean Community

    PubMed Central

    Sanchez Clemente, Nuria; Ugarte-Gil, Cesar; Solorzano, Nelson; Maguiña, Ciro; Moore, David

    2016-01-01

    Background Bartonellosis affects small Andean communities in Peru, Colombia and Ecuador. Research in this area has been limited. Methods Retrospective review of 191 cases of bartonellosis managed in Caraz District Hospital, Peru, during the last outbreak (2003). Results The majority of cases (65%) were 14 years old and younger. There was a peak in acute cases after the rainy season; chronic cases presented more constantly throughout the year. The sensitivity of blood smear against blood culture in acute disease was 25%. The most commonly used treatment for chronic disease was rifampicin; chloramphenicol was used to treat most acute cases. Complications arose in 6.8% and there were no deaths. Conclusions Diagnostic and treatment algorithms for acute and chronic bartonellosis have been developed without a strong evidence base. Preparation of ready-to-go operational research protocols for future outbreaks would strengthen the evidence base for diagnostic and treatment strategies and enhance opportunities for control. PMID:26991495

  6. Repression of In Vivo Synthesis of the Mitochondrial Elongation Factors T and G in Saccharomyces fragilis

    PubMed Central

    Richter, D.

    1973-01-01

    In vivo synthesis of the mitochondrial elongation factors T and G in the yeast Saccharomyces fragilis can be repressed. Enzymatic activity assays and immunochemical titration methods reveal that cells grown in the presence of 8% glucose or in the absence of oxygen contain relatively lower amounts of mitochondrial elongation factors than cells grown in the presence of lactate. In contrast, in vivo production of the cytoplasmic elongation factors 1 and 2 does not respond to such a change of extracellular conditions. The rate of growth does not affect the level of the mitochondrial elongation factors. Production of both enzymes is almost constant during logarithmic growth, but decreases when the stationary phase is reached. Chloramphenicol, an inhibitor of mitochondrial protein synthesis, does not block but, rather, seems to enhance the in vivo synthesis of mitochondrial T or G. PMID:4717524

  7. Differential antimicrobial susceptibility between human and chicken isolates of vancomycin-resistant and sensitive Enterococcus faecium.

    PubMed

    Chen, Han Yuan; Hill, Robert L R; Kirk, Monica; Casewell, Mark W; Beighton, David

    2002-01-01

    To compare the differential antimicrobial susceptibilities of Enterococcus faecium from humans and whole chicken carcasses, MICs of 12 antimicrobial agents were determined for 54 clinical-isolates (31 vancomycin-resistant [VREF]) and 60 chicken-isolates (29 VREF). Chicken VREF were slightly but consistently more resistant to vancomycin, teicoplanin and avoparcin, compared with human VREF (P<0.01). MICs of LY333328 were chloramphenicol than human VREF (P<0.001). PMID:11814766

  8. Multi-residue and multi-class determination of antibiotics in gilthead sea bream (Sparus aurata) by ultra high-performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Freitas, Andreia; Leston, Sara; Rosa, João; Castilho, Maria da Conceição; Barbosa, Jorge; Rema, Paulo; Pardal, Miguel Ângelo; Ramos, Fernando

    2014-01-01

    This paper describes a method for the determination of 41 antibiotics from seven different classes in gilthead sea bream (Sparus aurata) by ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). Sulfonamides, trimethoprim, tetracyclines, macrolides, quinolones, penicillins and chloramphenicol were simultaneously determined. Fourteen procedures for sample treatment were tested and an extraction with acetonitrile and ethylenediaminetetra acetic acid was found to be the best option. The methodology was validated in accordance with Decision 2002/657/EC. Precision in terms of relative standard deviation (RSD) was under 17% for all compounds, and the recoveries ranged from 92% to 111%. CCα and CCβ were determined according to the maximum residue limit or the minimum required performance limit, when necessary. The validation provided evidence that the method was suitable for application in routine analysis for the detection and confirmation of antibiotics in muscle of gilthead sea bream, an important and intensively produced fish in aquaculture.

  9. Comparative study of novel versus conventional two-wavelength spectrophotometric methods for analysis of spectrally overlapping binary mixture

    NASA Astrophysics Data System (ADS)

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

    2015-09-01

    Smart spectrophotometric methods have been applied and validated for the simultaneous determination of a binary mixture of chloramphenicol (CPL) and prednisolone acetate (PA) without preliminary separation. Two novel methods have been developed; the first method depends upon advanced absorbance subtraction (AAS), while the other method relies on advanced amplitude modulation (AAM); in addition to the well established dual wavelength (DW), ratio difference (RD) and constant center coupled with spectrum subtraction (CC-SS) methods. Accuracy, precision and linearity ranges of these methods were determined. Moreover, selectivity was assessed by analyzing synthetic mixtures of both drugs. The proposed methods were successfully applied to the assay of drugs in their pharmaceutical formulations. No interference was observed from common additives and the validity of the methods was tested. The obtained results have been statistically compared to that of official spectrophotometric methods to give a conclusion that there is no significant difference between the proposed methods and the official ones with respect to accuracy and precision.

  10. Functionalized chitosan/NIPAM (HEMA) hybrid polymer networks as inserts for ocular drug delivery: synthesis, in vitro assessment, and in vivo evaluation.

    PubMed

    Verestiuc, Liliana; Nastasescu, Oana; Barbu, Eugen; Sarvaiya, Indrajeetsinh; Green, Keith L; Tsibouklis, John

    2006-06-15

    A series of hybrid polymeric hydrogels, prepared by the reaction of acrylic acid-functionalized chitosan with either N-isopropylacrylamide or 2-hydroxyethyl methacrylate monomers, were synthesized, pressed into minitablets, and investigated for their ability to act as controlled release vehicles for ophthalmic drug delivery. For comparison, interpolymeric complex analogues synthesized using the same monomers and pure, unfunctionalized chitosan were examined by means of an identical characterization protocol. The effects of network structure and composition upon the swelling properties, adhesion behavior, and drug release characteristics were investigated. Comparative in vitro studies employing chloramphenicol, atropine, norfloxacin, or pilocarpine informed the selection of drug-specific carrier compositions for the controlled delivery of these compounds. In addition, in vivo (rabbit model) experiments involving the delivery of pilocarpine indicated that chitosan-based hybrid polymer networks containing 2-hydroxyethyl methacrylate are useful carriers for the delivery of this therapeutic agent. PMID:16555266

  11. Isolation and characterization of a variant of human papillomavirus type 11 from a nasal inverting (Schneiderian) papilloma.

    PubMed

    Pater, A; Gardner, H; Respler, D S; Jahn, A; Pater, M M

    1988-06-01

    We have previously reported the presence of a variant of human papillomavirus (HPV) type 11 in a nasal inverting papilloma [Respler et al., 1987]. In the present study, we have cloned molecularly the DNA of this variant at its unique restriction enzyme Bam HI site into lambda BF101 phage. Restriction enzyme mapping and DNA sequencing revealed that the genome of this virus contained an extra 531 base pair (bp) which was the repeat of most of the noncoding region (ncr) of HPV 11. Insertion of transcriptional control elements, including the repeated sequence, in front of the chloramphenicol acetyltransferase (CAT) gene resulted in a 5- to 30-fold increase in expression in transfected cells, as compared to the constructs containing a single ncr of HPV 11. This increased expression was due to enhanced levels of CAT RNA the synthesis of which is initiated by the viral promoter element. PMID:2839608

  12. Antimicrobial susceptibility patterns of enterobacteriaceae isolated from HIV-infected patients in Kinshasa

    PubMed Central

    Iyamba, Jean-Marie Liesse; Wambale, José Mulwahali; Takaisi-Kikuni, Ntondo za Balega

    2014-01-01

    Introduction People infected by Human Immunodeficiency Virus (HIV) are susceptible to develop severe bacterial infections. We set out to determine the frequency and the sensitivity to antibiotics of enterobaceriaceae isolated from urine and feces of HIV-infected persons. Methods Urine and feces samples were collected from HIV-infected patients of the Centre de Traitement Ambulatoire de Kabinda (CTA/Kabinda, Kinshasa) and analyzed at the Reference National Laboratory for HIV/AIDS and Sexually Transmitted Infections. The isolated enterobacteriaceae strains were identified by conventional microbiological methods. Antibiotic sensitivity pattern was carried out by disc diffusion method. Results The following bacteria pathogens were isolated: Escherichia coli, Klebsiella, Enterobacter, Proteus, and Providencia. Most species were sensitive to cefotaxim, ceftriaxon, and gentamicin and resistant to chloramphenicol, cotrimoxazole, tetracycline, and norfloxacin. Conclusion The results of the present study show that the most frequently bacteria isolated were Esherichia coli and cefotaxim, ceftriaxon, and gentamicin were the most active antibiotics. PMID:25392725

  13. Quinolone and Cephalosporin Resistance in Enteric Fever

    PubMed Central

    Capoor, Malini Rajinder; Nair, Deepthi

    2010-01-01

    Enteric fever is a major public health problem in developing countries. Ciprofloxacin resistance has now become a norm in the Indian subcontinent. Novel molecular substitutions may become frequent in future owing to selective pressures exerted by the irrational use of ciprofloxacin in human and veterinary therapeutics, in a population endemic with nalidixic acid-resistant strains. The therapeutics of ciprofloxacin-resistant enteric fever narrows down to third- and fourth-generation cephalosporins, azithromycin, tigecycline and penems. The first-line antimicrobials ampicillin, chloramphenicol and co-trimoxazole need to be rolled back. Antimicrobial surveillance coupled with molecular analysis of fluoroquinolone resistance is warranted for reconfirming novel and established molecular patterns for therapeutic reappraisal and for novel-drug targets. This review explores the antimicrobial resistance and its molecular mechanisms, as well as novel drugs in the therapy of enteric fever. PMID:20927288

  14. Genetic transformation and gene expression in white pine (pinus strobus)

    SciTech Connect

    Minocha, R.

    1987-10-01

    The objectives of the study were: (1) to develop protocols for transformation of white pine (Pinus strobus) embryonic tissue; and (2) to analyze the regulation of foreign gene expression in Pinus strobus. A number of Agrobacterium tumefaciens strains containing chimeric genes for neomycin phosphotransferase (NPTII for kanamycin resistance) and chloramphenicol acetyl transferase (CAT) under the control of either a constitutive promoter (NOS-nopaline synthase) or light-inducible promoters (RuBisCO small subunit and chlorophyll a/b binding protein) were used. A variety of tissues from white pine seedlings and mature trees was used. The techniques for transformation were modified from those used for tobacco transformation. The results show that white pine tissue from young seedlings is high suitable for transformation by A. tumefaciens. Whereas the normal tissues are very sensitive to kanamycin, transformed callus was quite resistant to this antibiotic.

  15. Synthesis, evaluation and molecular docking studies of amino acid derived N-glycoconjugates as antibacterial agents.

    PubMed

    Baig, Noorullah; Singh, Rajnish Prakash; Chander, Subhash; Jha, Prabhat Nath; Murugesan, Sankaranarayanan; Sah, Ajay K

    2015-12-01

    Six amino acid derived N-glycoconjugates of d-glucose were synthesized, characterized and tested for antibacterial activity against G(+)ve (Bacillus cereus) as well as G(-)ve (Escherichia coli and Klebsiella pneumoniae) bacterial strains. All the tested compounds exhibited moderate to good antibacterial activity against these bacterial strains. The results were compared with the antibacterial activity of standard drug Chloramphenicol, where results of A5 (Tryptophan derived glycoconjugates) against E. coli and A4 (Isoleucine derived glycoconjugates) against K. pneumoniae bacterial strains are comparable with the standard drug molecule. In silico docking studies were also performed in order to understand the mode of action and binding interactions of these molecules. The docking studies revealed that, occupation of compound A5 at the ATP binding site of subunit GyrB (DNA gyrase, PDB ID: 3TTZ) via hydrophobic and hydrogen bonding interactions may be the reason for its significant in vitro antibacterial activity.

  16. Identification of the promoter sequences involved in the cell specific expression of the rat somatostatin gene.

    PubMed Central

    Andrisani, O M; Hayes, T E; Roos, B; Dixon, J E

    1987-01-01

    DNA sequences containing the 5' flanking region of the rat somatostatin gene were linked to the coding sequence of the bacterial chloramphenicol acetyl transferase gene. This recombinant plasmid is active in expressing CAT activity in the neuronally derived, somatostatin producing CA-77 cell line. Deletion analyses of the somatostatin promoter show that the sequences proximal to position -60, relative to the cap site are required for expression of this promoter. A 4 base pair deletion of residues -46 through -43 within the somatostatin promoter results in a down mutation in vivo suggesting the existence of an element critical for the expression of the promoter in CA-77 cells. In addition, the somatostatin recombinant and its 5' deletion constructs preferentially express CAT activity in CA-77 cells, whereas only basal level of expression is observed in HeLa, BSC40, and RIN-5F cell lines, pointing to the cell specific nature of this promoter. Images PMID:2886975

  17. Spectrum and treatment of anaerobic infections.

    PubMed

    Brook, Itzhak

    2016-01-01

    Anaerobes are the most predominant components of the normal human skin and mucous membranes bacterial flora, and are a frequent cause of endogenous bacterial infections. Anaerobic infections can occur in all body locations: the central nervous system, oral cavity, head and neck, chest, abdomen, pelvis, skin, and soft tissues. Treatment of anaerobic infection is complicated by their slow growth in culture, by their polymicrobial nature and by their growing resistance to antimicrobials. Antimicrobial therapy is frequently the only form of therapy needed, whereas in others it is an important adjunct to drainage and surgery. Because anaerobes generally are isolated mixed with aerobes, the antimicrobial chosen should provide for adequate coverage of both. The most effective antimicrobials against anaerobes are: metronidazole, the carbapenems (imipenem, meropenem, doripenem, ertapenem), chloramphenicol, the combinations of a penicillin and a beta-lactamase inhibitors (ampicillin or ticarcillin plus clavulanate, amoxicillin plus sulbactam, piperacillin plus tazobactam), tigecycline, cefoxitin and clindamycin. PMID:26620376

  18. Prevalence and characteristics of Aeromonas species isolated from processed channel catfish.

    PubMed

    Wang, C; Silva, J L

    1999-01-01

    From August 1994 to May 1995, 238 channel catfish fillets collected from three processing plants in the Mississippi Delta at four time periods were tested for the presence of Aeromonas species. Identification of Aeromonas spp. was accomplished using an automated Vitek bioassay system with gram-negative and nonfermenter cards. Approximately 36.1% were positive for A. hydrophila, 35.7% for A. sobria, and 10.9% for A. caviae. All three Aeromonas spp. were found in all three processing plants, and the incidence of A. hydrophila contamination appeared to be higher in summer than other seasons. Eighty-six percent of the Aeromonas isolates were hemolytic on 5% sheep blood agar plates. Most isolates were susceptible to chloramphenicol, neomycin, streptomycin, and trimethoprim-sulfamethoxazole and resistant to ampicillin and bacitracin. Results suggest that Aeromonas spp. are prevalent in processed channel catfish, and most isolates are hemolytic and resistant to ampicillin and bacitracin. PMID:9921825

  19. Effect of foliar application of antibiotics and gibberellic acid on the rhizosphre microflora of pea, infected with Verticillium dahliae.

    PubMed

    Ramarao, P; Isaac, I

    1980-01-01

    A study was made of the effects of foliar spray of bacitracin, chloramphenicol and gibberellic acid on the rhizosphere microflora of pea seedlings (Pisum sativum L.) infected with Verticillium dahliae. The antibiotics increased fungus and actinomycete counts and reduced the bacterial populations in the rhizosphere. Gibberellic acid at 10 ppm concentration reduced all three groups of microorganisms while at 100 ppm fungi and actinomycetes increased slightly. Invariably the rhizosphere effect was as follows: bacteria leads to fungi leads to actinomycetes. Foliar sprays also affected percentage occurrence of particular genera of fungi in the rhizosphere; for example, Trichoderma spp. were stimulated by all the treatments, the maximum being with 10 ppm gibberellic acid, even though the total fungus count was reduced. The disease severity was markedly reduced by foliar sprays. PMID:7191387

  20. Conjugal transfer of broad-host-range incompatibility group P and Q plasmids from Escherichia coli to Actinobacillus actinomycetemcomitans.

    PubMed Central

    Goncharoff, P; Yip, J K; Wang, H; Schreiner, H C; Pai, J A; Furgang, D; Stevens, R H; Figurski, D H; Fine, D H

    1993-01-01

    The first example of conjugal transfer of DNA from Escherichia coli to the periodontal pathogen Actinobacillus actinomycetemcomitans is presented. Derivatives of the incompatibility group P (IncP) plasmid RK2 successfully transferred from an E. coli donor to an A. actinomycetemcomitans recipient. The resulting A. actinomycetemcomitans transconjugants transferred the plasmids back to E. coli recipients. The IncP transfer functions were also used in trans to mobilize the IncQ plasmid pBK1 from E. coli to A. actinomycetemcomitans. The IncP and IncQ plasmids both transferred into A. actinomycetemcomitans at high frequencies (0.3 to 0.5 transconjugants per donor) and showed no gross deletions, insertions, or rearrangements. Determinations of MICs of various antibiotics for the A. actinomycetemcomitans transconjugant strains demonstrated the expression of ampicillin, chloramphenicol, and kanamycin resistance determinants. Images PMID:8335386