Sample records for carbenicillin

  1. Influences of Cefotaxime and Carbenicillin on Shoot Regeneration from Soybean Cotyledonary-node%头孢青霉素和羧苄青霉素对大豆子叶节茎再生的影响

    Institute of Scientific and Technical Information of China (English)

    可庆波; 王丽艳; 王艳; 张春涛; 殷奎德


    试验通过向茎诱导培养基和茎伸长培养基中添加不同浓度的头孢青霉素(Cef)和羧苄青霉素(Cb)来研究二者对大豆子叶节茎再生潜在的影响。一定浓度的Cef或Cb对大豆子叶节部位茎的再生有促进或抑制作用。不同基因型大豆的最适Cef或Cb添加浓度不同。培养基中添加100 mg.L-1 Cef或120 mg.L-1 Cb能显著提高垦农18号(K18)子叶节部位丛生芽的诱导及可伸长茎的生成和伸长;150 mg.L-1 Cef对垦农4号(K4)子叶节部位茎的再生有显著的促进作用1,20 mg.L-1 Cb和200 mg.L-1 Cb分别显著提高K4丛生芽的再生和随后可伸长茎的生成及伸长。经121℃高压灭菌处理的Cef和Cb对大豆子叶节部位丛生芽的诱导和可伸长茎的生成及伸长有显著的抑制作用。%The influences of cefotaxime and carbenicillin on shoot regeneration potential of soybean cotyledonary-node were investigated by supplying with various concentrations of cefotaxime and carbenicillin to shoot induction medium and shoot elongation medium.The capability of shoot regeneration from soybean cotyledonary-node could be promoted or suppressed by supplying with some certain concentration of cefotaxime and carbenicillin.However,the most appropriate treatment is genotype-dependent.The buds regeneration,shoot multiplication and elongation of cotyledonary-node of K18 can be enhanced significantly by supplying with 100 mg·L-1 cefotaxime or 120 mg·L-1 carbenicillin in culture medium;the capability of shoot regeneration of K4 could be promoted with 150 mg·L-1 cefotaxime or 120 mg·L-1 carbenicillin on buds regeneration and 200 mg·L-1 on shoot multiplication and elongation.Autoclaved cefotaxime and carbenicillin showed detrimental effects on buds regeneration and shoot multiplication and elongation respectively.

  2. Drug: D07614 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available biotic, penicillin, broad-spectrum Same as: C06869 ATC code: J01CA03 Semisynthetic penicillin: moderate spectrum...Penicillins with extended spectrum J01CA03 Carbenicillin D07614 Carbenicillin (INN) Antiinfectives [BR:br083

  3. Effects of antibiotics on plantlet regeneration via organogenesis in Populus euphratica

    Institute of Scientific and Technical Information of China (English)

    Ding Xia; Chen Xiao-yang; Li Wei; Du Zhi-yan


    Four antibiotics, kanamycin, geneticin, carbenicillin and cefotaxime, were evaluated for their effect on the regeneration of adventitious buds, shoot differentiation, rooting from regenerated shoots of Populus euphratica as well as on their control of Agrobacterium-mediated transformations. Results showed that the optimum concentration ranges of kanamycin and geneticin were 15-20 and 10-15 mg·L-1 at the stage of transgenic plantlet selection. The inhibitory effects of cefotaxime and carbenicillin varied among different genotypes of Agrobacterium. The inhibition of cefotaxime on Agrobacterium C58 was stronger than that of carbenicillin.LBA4404 and their optimum concentrations were both 150 mg·L-1.

  4. Heterologous viral expression systems in fosmid vectors increase the functional analysis potential of metagenomic libraries. (United States)

    Terrón-González, L; Medina, C; Limón-Mortés, M C; Santero, E


    The extraordinary potential of metagenomic functional analyses to identify activities of interest present in uncultured microorganisms has been limited by reduced gene expression in surrogate hosts. We have developed vectors and specialized E. coli strains as improved metagenomic DNA heterologous expression systems, taking advantage of viral components that prevent transcription termination at metagenomic terminators. One of the systems uses the phage T7 RNA-polymerase to drive metagenomic gene expression, while the other approach uses the lambda phage transcription anti-termination protein N to limit transcription termination. A metagenomic library was constructed and functionally screened to identify genes conferring carbenicillin resistance to E. coli. The use of these enhanced expression systems resulted in a 6-fold increase in the frequency of carbenicillin resistant clones. Subcloning and sequence analysis showed that, besides β-lactamases, efflux pumps are not only able contribute to carbenicillin resistance but may in fact be sufficient by themselves to convey carbenicillin resistance.

  5. Drug: D02190 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available IALS, PENICILLINS J01CA Penicillins with extended spectrum J01CA03 Carbenicillin D02190 Carbenicillin sodium...17H16N2O6S. 2Na 422.0524 422.3633 D02190.gif Antibacterial Same as: C13967 ATC code: J01CA03 Semisynthetic penicillin: moderate spect...rum penicillin penicillin binding proteins inhibitor ko00550 Peptidoglycan biosynth

  6. In-vitro activity of 21 antimicrobial agents against Neisseria gonorrhoeae in Brussels.


    Gordts, B; Vanhoof, R; Hubrechts, J M; Dierickx, R; Coignau, H; Butzler, J P


    The minimum inhibitory concentrations (MIC) of 21 antimicrobial agents was measured for 80 strains of Neisseria gonorrhoeae isolated in Brussels in 1978. Bimodal distributions were found for penicillin G, ampicillin, amoxycillin, carbenicillin, and cephalexin. Of the strains, 17.5% were relatively resistant to penicillin G (MIC greater than 0.08 microgram/ml) 27.5% to ampicillin (MIC greater than 0.16 microgram/ml), 23.8% to amoxycillin, and 43.3% to carbenicillin. Cefotaxime was the most act...

  7. Plasmid-determined resistance to fosfomycin in Serratia marcescens. (United States)

    Mendoza, C; Garcia, J M; Llaneza, J; Mendez, F J; Hardisson, C; Ortiz, J M


    Multiple-antibiotic-resistant strains of Serratia marcescens isolated from hospitalized patients were examined for their ability to transfer antibiotic resistance to Escherichia coli by conjugation. Two different patterns of linked transferable resistance were found among the transconjugants. The first comprised resistance to carbenicillin, streptomycin, and fosfomycin; the second, and more common, pattern included resistance to carbenicillin, streptomycin, kanamycin, gentamicin, tetracycline, chloramphenicol, sulfonamide, and fosfomycin. The two types of transconjugant strains carried a single plasmid of either 57 or 97 megadaltons in size. Both of these plasmids are present in parental S. marcescens strains resistant to fosfomycin. The 57-megadalton plasmid was transformed into E. coli.

  8. Stability of antibiotics and amino acids in two synthetic L-amino acid solutions commonly used for total parenteral nutrition in children

    DEFF Research Database (Denmark)

    Colding, H; Andersen, G E


    The stability and interaction at 29 degrees C of ampicillin, carbenicillin, gentamicin, and polymyxin B were examined in a common electrolyte solution, invertose darrow, and in two synthetic l-amino acid solutions, one commercial (vamin with fructose; Vitrum) and the other a neonatal preparation ...

  9. Effects of Antibiotics on Callus Growth and Shoot Regeneration of Sisal%抗生素对剑麻愈伤组织生长和植株再生的影响

    Institute of Scientific and Technical Information of China (English)

    张燕梅; 李俊峰; 周文钊; 林映雪


    The dose response assay was performed to insight into the effects of kanamycin, hygromycin, cefotaxime, and carbenicillin on the callus growth, adventitious buds regeneration and root induction of sisal. The optimum concentrations of cefotaxime and carbenicillin inhibiting Agrobacterium was determined. The results showed that the callus growth and adventitious buds regeneration were greatly inhibited by hygromycin, followed by kanamycin, cefotaxime and carbenicillin. Root induction was inhibited by each of the four antibiotics, with the order of cefotaxime > kanamycin > carbenicillin > hygromycin. The growth of Agrobacterium was completely inhibited by 200 mg/L cefotaxime or carbenicillin. In conclusion, 100 mg/L kanamycin, 50 mg/L hygromycin, 200 mg/L cefotaxime or carbenicillin was recommended to screen transformed tissue in Agrobacterium-mediated transformation of sisal.%研究卡那霉素、潮霉素、头孢霉素和羧苄青霉素对剑麻愈伤组织生长、不定芽分化及生根的影响,并确定抑制农杆菌生长的头孢霉素、羧苄青霉素的质量浓度.结果表明:潮霉素明显抑制剑麻愈伤组织生长和不定芽分化,卡那霉素次之,头孢霉素和羧苄青霉素相对较小;4种抗生素均能抑制剑麻生根,其抑制效果依次为头孢霉素>卡那霉素>羧苄青霉素>潮霉素.头孢霉素和羧苄青霉素的浓度大于200 mg/L时,农杆菌生长完全被抑制.在筛选剑麻转化体时,卡那霉素和潮霉素的使用浓度分别以100、50 mg/L为宜,羧苄青霉素和头孢霉素的使用浓度均为200 mg/L.

  10. Antimicrobial activity of beta-lactams against multiresistant micro-organisms from the family Enterobacteriaceae, and genus Pseudomonas. (United States)

    Niebla, A; González, I; Vallín, C


    The antimicrobial activity of twenty beta-lactams was determined against multiresistant micro-organisms from the Enterobacteriaceae family (450) and the genus Pseudomonas (90). The antimicrobial susceptibility was assessed by the disk diffusion method. The most effective antibiotics were cephalosporins of the second and third generation, and non-classical beta-lactams (imipenem and moxalactam). A pronounced resistance was found to carbenicillin, ampicillin, cephalotin and cefazolin. These resistance patterns corresponded to a high consumption of these antibiotics.




    Microbial contribution increasing rapidly due to invasion by the pathogenic organisms like bacteria, fungi and virus in the present disease burden of human health. To treat these diseases many potent and broad spectrum antibiotics were discovered e.g., ampicillin, amoxicillin, carbenicillin, ofloxacin and tetracycline etc., Even though antibiotics are life saving drugs in therapeutics but they are potentially harmful. These harmful effects include allergic and anaphylactic reaction, developme...

  12. Optimizing a Synthetic Signaling System, Using Mathematical Modeling to Direct Experimental Work (United States)


    control mechanism operating in diverse areas of biology, e.g., phage immunity, tryptophan synthesis, and lactose metabolism 2. Jacob and Monod described...signaling system and reporter plasmids and grown on Luria Broth (LB) plates with appropriate antibiotics (50 µg/ml carbenicillin (Teknova, Hollister, CA... antibiotics (20mg/L Gentamicin for the Agrobact.), at 30oC / 2 days. 2. Inoculate this starter culture into 250 mL of LB media (containing the same

  13. Bacteriostatic and bactericidal activities of 24 antimicrobial agents against Campylobacter fetus subsp. jejuni.


    Vanhoof, R.; Gordts, B; Dierickx, R; Coignau, H; Butzler, J P


    The bacteriostatic and bactericidal activities of 24 antimicrobial agents were tested with the Dynatech MIC 2000 system against 86 strains of Campylobacter fetus subsp. jejuni from human sources. The penicillins (penicillin G, ampicillin, amoxycillin, carbenicillin) had poor activity. Ampicillin and amoxycillin were equally active. Cefotaxime revealed a rather good activity. Erythromycin, gentamicin, tobramycin, amikacin, and furazolidone were the most active compounds. Two strains (2.3%) wer...

  14. Influences of antibiotics on plantlet regeneration via organogenesis in loblolly pine (Pinus taeda L.)

    Institute of Scientific and Technical Information of China (English)

    TANG Wei; Latoya Harris; Ronald J. Newton


    Three antibiotics ampicillin, carbenicillin, and cefotaxime were evaluated for their effects on induction, growth, and differentiation of organogenic calli, as well as rooting of regenerated shoots of three loblolly pine (Pinus taeda L.) genotypes. Of the antibiotics administered, cefotaxime maximally increased the frequency of callus formation and growth rate of organogenic calli, carbenicillin maximally increased the frequency of shoot regeneration and the average number of adventitious shoots per piece of organogenic callus, ampicillin maximally decreased the rooting frequency of regenerated shoots and mean number of roots per regenerated shoot, in comparison with antibiotic-free media. Compared with the control, ampicillin minimally increased the frequency of callus formation, cefotaxime minimally increased the frequency of shoot regeneration, and carbenicillin minimally decreased the rooting frequency of regenerated shoots in three loblolly pine genotypes tested. All three antibiotics increased the frequencies of callus formation and shoot regeneration, and reduced the rooting frequency of regenerated shoots suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into loblolly pine need to select a suitable antibiotic. This investigation could be useful for optimizing genetic transformation of conifers.

  15. Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes. (United States)

    Guo, M; Zhang, Y L; Meng, Z J; Jiang, J


    Micro-Tom is the smallest known variety of tomatoes. An orthogonal experimental design L(16) (4(5)) was used to optimize Agrobacterium-mediated transformation of cotyledon explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were investigated to determine their effect on transformation frequency: the concentration of bacterial suspension, time of dip in bacterial suspension, co-cultivation time, and concentration of carbenicillin. We also examined the effect of these parameters on contamination rate, necrosis rate, mortality, cut-surface browning rate, and undamaged explant rate. Both the bacterial and carbenicillin concentrations had a significant influence on the rate of infected explants. The time of co-cultivation also had a significant influence on the transformation parameters. The optimal transformation protocol consisted of an Agrobacterium suspension of 0.5 × 10(8) cells/mL (OD(600) = 0.5) and an infection time of 5 min, one day of co-cultivation and 500 mg/L carbenicillin. Under these conditions, the transformation efficiency of the shoots reached 5.1%; the mean transformation frequency was 3.9% (N = 838).

  16. Interaction of zwitterionic penicillins with the OmpF channel facilitates their translocation. (United States)

    Danelon, Christophe; Nestorovich, Ekaterina M; Winterhalter, Mathias; Ceccarelli, Matteo; Bezrukov, Sergey M


    To study translocation of beta-lactam antibiotics of different size and charge across the outer bacterial membrane, we combine an analysis of ion currents through single trimeric outer membrane protein F (OmpF) porins in planar lipid bilayers with molecular dynamics simulations. Because the size of penicillin molecules is close to the size of the narrowest part of the OmpF pore, penicillins occlude the pore during their translocation. Favorably interacting penicillins cause time-resolvable transient blockages of the small-ion current through the channel and thereby provide information about their dynamics within the pore. Analyzing these random fluctuations, we find that ampicillin and amoxicillin have a relatively high affinity for OmpF. In contrast, no or only a weak interaction is detected for carbenicillin, azlocillin, and piperacillin. Molecular dynamics simulations suggest a possible pathway of these drugs through the OmpF channel and rationalize our experimental findings. For zwitterionic ampicillin and amoxicillin, we identify a region of binding sites near the narrowest part of the channel pore. Interactions with these sites partially compensate for the entropic cost of drug confinement by the channel. Whereas azlocillin and piperacillin are clearly too big to pass through the channel constriction, dianionic carbenicillin does not find an efficient binding region in the constriction zone. Carbenicillin's favorable interactions are limited to the extracellular vestibule. These observations confirm our earlier suggestion that a set of high-affinity sites at the narrowest part of the OmpF channel improves a drug's ability to cross the membrane via the pore.

  17. Agrobacterium tumefaciens-mediated transformation of Vigna mungo (L.) Hepper. (United States)

    Karthikeyan, A S; Sarma, K S; Veluthambi, K


    Transformed Vigna mungo (blackgram) calli were obtained by cocultivating segments of primary leaves with Agrobacterium tumefaciens vir helper strains harbouring the binary vector pGA472 having kanamycin resistance gene as plant transformation marker. Transformed calli were selected on Murashige and Skoog medium supplemented with 50 mg/l kanamycin and 500 mg/l carbenicillin. Transformed calli were found to be resistant to kanamycin up to 900 mg/l concentration. Expression of kanamycin resistance gene in transformed calli was demonstrated by neomycin phosphotransferase assay. Stable integration of transferred DNA into V. mungo genome was confirmed by Southern blot analysis.

  18. [Evaluation of the influence of deep geomagnetic impacting on growth rate and sensitivity to antibiotics in Escherichia coli]. (United States)

    Kolmakov, V M; Kulikov, V Iu; Evstropov, A N; Voronin, A Iu


    The short-term (1-7 days) and prolonged (30-90 days) adaptation of bacteria to hypomagnetic conditions was studied by observing changes in growth rate and resistance to antibiotics in E. coli used as an experimental model. The decrease of the growth rate of bacterial cells in the exponential phase was found to occur. The effect disappeared when the screened culture was placed under normal geomagnetic conditions for 1 hour daily. After prolonged screening an increase in the resistance of these bacteria to kanamycin and carbenicillin was registered. These changes were not hereditary: after the cessation of screening the sensitivity of the bacteria to antibiotics was restored.

  19. [Production of hydrolases by lactic acid bacteria and bifidobacteria and their antibiotic resistance]. (United States)

    Novik, G I; Astanovich, N I; Riabaia, N E


    It was demonstrated that bifidobacteria and lactic acid bacteria B. adolescentis and Lactobacillus sp. synthesized extracellular enzymes cleaving glycoside bonds in the molecules of dextran, pectic acid, and soluble starch. The maximal production of extracellular beta-galactosidase by B. adolescentis 91-BIM and 94-BIM at a rate of 0.08 and 0.03 U/mg h was observed during the exponential growth phase at 5 and 12 h of cultivation, respectively. The cultures of bifidobacteria retained 60-70% of beta-galactosidase and alpha-amylase activities after six months of storage. The bifidobacterium strains studied were resistant to amphotericin and aminoglycosides (gentamicin, kanamycin, and netromycin). The lactam antibiotics (ampicillin, benzylpenicillin, bicillin 3, bicillin 5, and carbenicillin), the preparations inhibiting protein synthesis at the level of ribosomes (lincomycin), RNA polymerase inhibitors (rifampin), cephalosporin, and Maxipime inhibited the growth of bifidobacteria. Rifampin, erythromycin, amphotericin, Maxipime, Fortum, doxycycline, levomycetin, streptomycin, and the aminoglycosides netromycin, gentamicin, and kanamycin did not have an effect on the growth of Lactobacillus sp., whereas semisynthetic derivatives of penicillin, carbenicillin and ampicillin, inhibited its growth as well as Oxamp and lincomycin. The lactam antibiotics benzylpenicillin, bicillin 3, and bicillin 5 inhibited the growth of lactic acid bacilli by 30-90%.

  20. In-vitro activity of 21 antimicrobial agents against Neisseria gonorrhoeae in Brussels. (United States)

    Gordts, B; Vanhoof, R; Hubrechts, J M; Dierickx, R; Coignau, H; Butzler, J P


    The minimum inhibitory concentrations (MIC) of 21 antimicrobial agents was measured for 80 strains of Neisseria gonorrhoeae isolated in Brussels in 1978. Bimodal distributions were found for penicillin G, ampicillin, amoxycillin, carbenicillin, and cephalexin. Of the strains, 17.5% were relatively resistant to penicillin G (MIC greater than 0.08 microgram/ml) 27.5% to ampicillin (MIC greater than 0.16 microgram/ml), 23.8% to amoxycillin, and 43.3% to carbenicillin. Cefotaxime was the most active antibiotic, with MICs in the nanogram range; 3.8% and 5% of the strains were relatively resistant to cephaloridine and cephalexin respectively, but no strains were resistant to cefazolin, cefuroxime, or cefotaxime. Resistance to tetracycline, doxycycline, minocycline, erythromycin, and spiramycin (MIC greater than 1 microgram/ml) was found in 6.3%, 2.5%, 5%, and 51.3% of the strains respectively. A very good correlation was present between chloramphenicol and thiamphenicol, with 16.3% and 10% of relatively resistant strains respectively. Only two isolates showed an MIC greater than 1.25 microgram/ml for rifampicin, and 10% of the strains needed greater than or equal to 12 microgram/ml of spectinomycin for complete inhibition of growth. A very high energy was found for the 20 : 1 combination of sulphamethoxazole and trimethoprim, with only one isolate resistant to this combination. None of the strains tested produced beta-lactamase.

  1. Influence of antibiotics on indirect organogenesis of Teak

    Directory of Open Access Journals (Sweden)

    Evandro Vagner Tambarussi


    Full Text Available Agrobacterium is the largest method employed to transform woody plants. The bacterium is required to introduce the transgene into the plant nuclear genome. After transferring T-DNA to the plant cell, the bacteria affect plant growth negatively and have to be eliminated from plant tissue culture medium through the use of antibiotics. The effect of different antibiotics (timentin, cefotaxime and carbenicillin on in vitro shoot regeneration of teak (Tectona grandis L. f. was compared in hypocotyl, mature cotyledon and cotyledonary segments explants. Timentin and cefotaxime (100-300 mg l-1 did not affect shoot regeneration and the number of shoots per explant. Moreover, at these concentrations, the two antibiotics seem to stimulate shoot regeneration. Carbenicillin at a dosage of 300 mg l-1 as well as cefotaxime and timentin at a dosage of 500 mg l-1 induced abundant calli formation and inhibited regeneration. Our data show that cefotaxime and timentin (300 mg l-1 can be harmless to teak regeneration and can be used as bactericide agents during Agrobacterium-mediated transformation of Tectona grandis. Furthermore, we discuss the effect of antibiotic degradation on plant morphogenesis and its effect on regeneration from different explants.

  2. Effects of Combined Treatment with Sansanmycin and Macrolides on Pseudomonas aeruginosa and Formation of Biofilm

    Institute of Scientific and Technical Information of China (English)



    Objective To observe the effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm. Methods Micro-dilution method was used to determine the minimal inhibitory concentrations (MICs) of sansanmycin, gentamycin, carbenicillin, polymyxin B, roxithromycin, piperacillin, and tazobactam. PA1 and PA27853 biofilms were observed under optical microscope after staining and under SEM after treatment with sansanmycin at different dosages and combined treatment with sansanmycin and roxithromycin. Viable bacteria in PA1 and PA27853 biofilms were counted after treatment with sansanmycin at different dosages or combined treatment with sansanmycin and roxithromycin. Results The MIC of sansanmycin was lower than that of gentamycin and polymyxin B, but was higher than that of carbenicillin. Roxithromycin enhanced the penetration of sansanmycin to PA1 and PA27853 strains through biofilms. PA1 and PA27853 biofilms were gradually cleared with the increased dosages of sansanmycin or with the combined sansanmycin and roxithromycin. Conclusion Sub-MIC levels of roxithromycin and sansanmycin substantially inhibit the generation of biofilms and proliferation of bacteria. Therefore, combined antibiotics can be used in treatment of intractable bacterial infection.

  3. Sinorhizobium morelense sp. nov., a Leucaena leucocephala-associated bacterium that is highly resistant to multiple antibiotics. (United States)

    Wang, En Tao; Tan, Zhi Yuan; Willems, Anne; Fernández-López, Manuel; Reinhold-Hurek, Barbara; Martínez-Romero, Esperanza


    Sinorhizobium morelense sp. nov. is described to designate a group of bacteria isolated from root nodules of Leucaena leucocephala. S. morelense shows 98% 16S rRNA gene sequence similarity to some Sinorhizobium species and to Ensifer adhaerens. This novel species is distinguished from other Sinorhizobium species and from E. adhaerens by DNA-DNA hybridization, 165 rRNA gene restriction fragments and sequence and some distinctive phenotypic features. Strains of this species are highly resistant to some antibiotics, such as carbenicillin (1 mg ml(-1)), kanamycin (500 microg ml(-1)) and erythromycin (300 microg ml(-1)). They do not form nodules, but a nodulating strain, Lc57, is closely related to the novel species. Strain Lc04T (= LMG 21331T = CFN E1007T) is designated as the type strain of this novel species.

  4. Study of Pseudomonas Aeroginosa resistance to Penicillines, Cephalosporins and Aminoglycosides

    Directory of Open Access Journals (Sweden)

    Maleknezhad P


    Full Text Available Drug therapy and prophylaxy in infectious diseases, from hygienic and economical point of view, are very important. Infections caused by pseudomonas aeroginosa were particularly severe, with high mortality rates. In the recent years pseudomonas aeroginosa continued to cause the most severe, life-thereating infections in burned patients, in spite of the introduction of a wide variety of antibiotics advised specifically for their anti pseudomonal activity. The aim of this study, in which many cases of ps.aeroginosa infections are assessed is to identify the drug resistance of this bacteria to penicillines, cephalosporins and aminoglycosides by antibiotic sensitivity test (disk ager diffusion. Results as percent of resistance to each antibiotic were 89% to carbenicillin, 55% to piperacillin, 89% to mezlocillin, 89.5% to ticarcillin+clavulonic acid, 85% to ceftriaxone, 95% to tobramycin, 5% of all isolates were not sensitive to any antibiotics.

  5. Suppression of damping-off in maize seedlings by Pseudomonas corrugata. (United States)

    Pandey, A; Palni, L M; Hebbar, K P


    Two strains of Pseudomonas corrugata, (1 and 7), isolated from subtropical and temperate soils in Sikkim Himalaya, respectively, were subjected to Petri-dish as well as plant-based bioassay to examine their potential for disease suppression against three major pathogens of maize. A mixture of Pythium ultimum, P. arrhenomanes and Fusarium graminearum was introduced in the soil; maize seed inoculated with one of the two strains of Pseudomonas corrugata (1 or 7) were sown in pots containing such soil. The bacterial inoculation resulted in significant disease suppression as well as growth promotion of seedlings. The bacterial strains were also evaluated for their intrinsic antibiotic resistance against a range of concentrations of ten antibiotics. While the bacteria were found to be sensitive to gentamycin and rifampicin, they exhibited resistance against ampicillin, carbenicillin and penicillin, even at high concentrations.

  6. [Evaluation of an automated procedure determining the minimum inhibitory concentrations (MIC). ]. (United States)

    Thabaut, A; Durosoir, J L; Meyran, M


    The ABAC system allows to distribute simultaneously and automatically a standardized inoculum into microtube-cuvettes containing in lyophilized broth medium twofold serial dilutions of the antibiotics. After an 18 hours incubation time, The system prints automatically the MIC. We have compared the MIC of beta-lactam antibiotics (ampicillin, carbenicillin, cephalothin, cefoxitin, cefamandole, cefuroxime and cefotaxime) and 6 aminoglycoside (gentamicin, tobramycin, netilmycin, amikacin, kanamycin, lividomycin) obtained by the ABAC system and by the Agar dilution method for 302 gram negative bacilli. We also made a comparison of the MIC of 8 antibiotics (oxacillin, oleandomycin, spiramycin, erythromycin, clindamycin, pristinamycin, doxycycline, vancomycin) obtained by the 2 methods for 117 Staphylococcus aureus strains. The evaluation shows that the reproducibility of the results obtained by the ABAC system is good. The statistical analysis shows that the correlation between the MIC obtained with the 2 methods is excellent and that there is no significant discrepancy.

  7. Bacteriostatic and bactericidal activities of 24 antimicrobial agents against Campylobacter fetus subsp. jejuni. (United States)

    Vanhoof, R; Gordts, B; Dierickx, R; Coignau, H; Butzler, J P


    The bacteriostatic and bactericidal activities of 24 antimicrobial agents were tested with the Dynatech MIC 2000 system against 86 strains of Campylobacter fetus subsp. jejuni from human sources. The penicillins (penicillin G, ampicillin, amoxycillin, carbenicillin) had poor activity. Ampicillin and amoxycillin were equally active. Cefotaxime revealed a rather good activity. Erythromycin, gentamicin, tobramycin, amikacin, and furazolidone were the most active compounds. Two strains (2.3%) were resistant to erythromycin. One strain (1.2%) was completely resistant to tobramycin. The tetracyclines (tetracyline, doxycycline, minocycline) were generally effective, but 8% of the strains were totally resistant to them. Minocycline was the most active. Chloramphenicol, thiamphenicol, and clindamycin had good activity. The bacteriostatic and bactericidal distributions for colistin, nalidixic acid, and metronidazole were broad.

  8. Antibacterial Activities of Neolignans Isolated from the Seed Endotheliums of Trewia nudiflora

    Institute of Scientific and Technical Information of China (English)

    LIGuo-Hong; ZHAOPei-Ji; SHENYue-Mao; ZHANGKe-Qin


    Five neolignans including four new ones were obtained from the seed endotheliums of Trewia nudiflora L. Their structures were determined to be 9'-methyl americanol A(1), 9'-methyl isoamericanol A(2), 9'-ethyl americanol A(3), 9'-butyl americanol A(4), and americanin (5). Two acetylated products 3,4-diacetyl americanin (5a) and 3,4,9-triacetyl americanin (5b) had been prepared from compound 5. All of these compounds were investigated on antibacterial assays when carbenicillin sodium, streptomycin sulfate and rifampicin were used as positive controls. Compounds 4 and 5 exhibited antibacterial activities against gram-positive bacterium Staphylococcus aureus and gram-negative bacterium Mycobacterium tuberculosis at the minimum inhibitory concentrations (MIC) of 50μg/mL and 100μg/mE respectively, but 5a and 5b did not exhibit antibacterial activity at 200μg/disk.

  9. Investigation of Burkholderia cepacia nosocomial outbreak with high fatality in patients suffering from diseases other than cystic fibrosis. (United States)

    Shehabi, Asem A; Abu-Al-Soud, Waleed; Mahafzah, Azmi; Khuri-Bulos, Najwa; Abu Khader, Ilham; Ouis, Ibn-Sina; Wadström, Torkel


    Over a 1-y period, 26 inpatients at the Jordan University Hospital in Amman were detected with bacteraemia (23 cases) or respiratory tract colonized with B. cepacia (3 cases). A combination of genetic identification and molecular typing has proved that all cases were caused by a single epidemic strain of B. cepacia genomovar IIIa. Nosocomial infections could be documented in 21/26 (81%) patients, mostly with severe underlying or malignant diseases other than cystic fibrosis, but the source of infection was undetected. The overall mortality related to infection with B. cepacia was 42%. All B. cepacia isolates were resistant to ampicillin, amikacin, carbenicillin and gentamicin; and mostly susceptible to piperacillin, chloramphenicol, cotri-moxazole, tetracycline, ceftazidime, and tazocin (62-88%). This study demonstrates the nosocomial and high fatality of B. cepacia genomovar IIIa in Jordanian patients suffering from diseases other than cystic fibrosis.

  10. Inhibition of Four Kinds of Antibiotics on Endopyhtic Bacteria in ‘Zoji’ Strawberry and Their Effects on the Growth of Its in Vitro Shoots%4种抗生素对草莓‘章姬’内生菌的抑制及对试管苗生长的影响

    Institute of Scientific and Technical Information of China (English)

    王梅; 汤浩茹; 刘淑芳; 刘祥林; 李靖


    4种常用抗生素对‘章姬'草莓内生菌的抑制结果表明,卡那霉素(Kanamycin)不仅对内生菌无抑制效果,还对试管苗的生长有毒害作用.头孢霉素(Cefotaxime)和羧苄青霉素(Carbenicillin)的抑菌效果不明显,但对试管苗生长有促进作用.青霉素G钠(Penicinllin G sodium)对内生菌抑制效果很好,尤其是当其浓度为400mg/L时抑菌率可达100%,同时还对植株的生长有明显促进作用.

  11. The bactericidal activity of β-lactam antibiotics is increased by metabolizable sugar species

    DEFF Research Database (Denmark)

    Thorsing, Mette; Bentin, Thomas; Givskov, Michael;


    Here, the influence of metabolizable sugars on the susceptibility of Escherichia coli to β-lactam antibiotics was investigated. Notably, monitoring growth and survival of mono- and combination-treated planktonic cultures showed a 1000- to 10 000-fold higher antibacterial efficacy of carbenicillin...... and cefuroxime in the presence of certain sugars, whereas other metabolites had no effect on β-lactam sensitivity. This effect was unrelated to changes in growth rate. Light microscopy and flow cytometry profiling revealed that bacterial filaments, formed due to β-lactam-mediated inhibition of cell division......, rapidly appeared upon β-lactam mono-treatment and remained stable for up to 18 h. The presence of metabolizable sugars in the medium did not change the rate of filamentation, but led to lysis of the filaments within a few hours. No lysis occurred in E. coli mutants unable to metabolize the sugars, thus...

  12. In-vitro activity of antimicrobial agents against Neisseria gonorrhoeae in Brussels. (United States)

    Vanhoof, R; Vanderlinden, M P; Hubrechts, J M; Butzler, J P; Yourassowsky, E


    The minimum inhibitory concentrations (MICs) of 18 antimicrobial agents against 104 strains of Neisseria gonorrhoeae isolated in the Brussels area between January and October 1976 have been measured. The MICs for penicillin G, ampicillin, amoxycillin, carbenicillin, and cephalexin showed a bimodal distribution. The second modus strains of cephalexin (MIC = 6.25 microgram/ml) were relatively resistant to penicillin G (MIC greater than or equal to 0.08 microgram/ml). About 51% of all strains were relatively resistant to penicillin G, 40.5% to ampicillin (MIC greater than or equal to 0.16 microgram/ml), 46% to amoxycillin, and 47.5% to carbenicillin. For cephalexin and cephaloridine, 25% and 8.5% respectively of all strains were relatively resistant (MIC greater than 3.12 microgram/ml). For cefazolin all MICs fell into a range of 0.097--3.12 microgram/ml. Resistance to tetracycline, doxycycline, minocycline, erythromycin, and spiramycin (MIC greater than or equal to 1 microgram/ml) was found in 9.5%, 7%, 6%, 36.5%, and 71% respectively of all isolates. No strains were resistant to rifampicin. For chloramphenicol and thiamphenicol the MICs ranged from 0.39 to 12.5 microgram/ml and from 0.195 to 3.12 microgram/ml respectively. The results for sulphamethoxazole, trimethoprim, and the combination of sulphamethoxazole and trimethoprim in a 20:1 ratio are given and discussed. The fractional inhibitory concentration (FIC) indices have also been calculated. No beta-lactamase-producing strains were found, and a contingency coefficient C has been determined for all the pairs of antibiotics investigated.

  13. Chromosomal integration of a cephalosporinase gene from Acinetobacter baumannii into Oligella urethralis as a source of acquired resistance to beta-lactams. (United States)

    Mammeri, Hedi; Poirel, Laurent; Mangeney, Nicole; Nordmann, Patrice


    Clinical Oligella urethralis isolate COH-1, which was uncommonly resistant to penicillins and narrow-spectrum cephalosporins, was recovered from a 55-year-old patient with a urinary tract infection. Shotgun cloning into Escherichia coli and expression experiments gave recombinant clones expressing either an AmpC beta-lactamase-type phenotype of resistance or a carbenicillin-hydrolyzing beta-lactamase-type phenotype of resistance. The AmpC beta-lactamase identified (ABA-1), which had a pI value of 8.2, had 98% amino acid identity with a chromosomally encoded cephalosporinase of Acinetobacter baumannii. A 820-bp insertion sequence element, ISOur1, belonging to the IS6 family of insertion sequence elements, was identified immediately upstream of bla(ABA-1), providing a -35 promoter sequence and likely giving rise to a hybrid promoter region. The carbenicillin-hydrolyzing beta-lactamase identified (CARB-8), which had a pI value of 6.4, differed from CARB-5 by two amino acid substitutions. Hybridization of CeuI fragment I-restricted DNA fragments of O. urethralis COH-1 with bla(ABA-1)-, bla(CARB-8)-, and 16S rRNA-specific probes indicated the chromosomal integration of the beta-lactamase genes. PCR and hybridization experiments failed to detect bla(CARB-8)- and bla(ABA-1)-like genes in three O. urethralis reference strains, indicating that the beta-lactamase genes identified were the source of acquired resistance in O. urethralis COH-1. This is one of the few examples of the interspecies transfer and the chromosomal integration of a gene encoding a naturally occurring beta-lactamase.

  14. Quantifying Selective Pressures Driving Bacterial Evolution Using Lineage Analysis (United States)

    Lambert, Guillaume; Kussell, Edo


    Organisms use a variety of strategies to adapt to their environments and maximize long-term growth potential, but quantitative characterization of the benefits conferred by the use of such strategies, as well as their impact on the whole population's rate of growth, remains challenging. Here, we use a path-integral framework that describes how selection acts on lineages—i.e., the life histories of individuals and their ancestors—to demonstrate that lineage-based measurements can be used to quantify the selective pressures acting on a population. We apply this analysis to Escherichia coli bacteria exposed to cyclical treatments of carbenicillin, an antibiotic that interferes with cell-wall synthesis and affects cells in an age-dependent manner. While the extensive characterization of the life history of thousands of cells is necessary to accurately extract the age-dependent selective pressures caused by carbenicillin, the same measurement can be recapitulated using lineage-based statistics of a single surviving cell. Population-wide evolutionary pressures can be extracted from the properties of the surviving lineages within a population, providing an alternative and efficient procedure to quantify the evolutionary forces acting on a population. Importantly, this approach is not limited to age-dependent selection, and the framework can be generalized to detect signatures of other trait-specific selection using lineage-based measurements. Our results establish a powerful way to study the evolutionary dynamics of life under selection and may be broadly useful in elucidating selective pressures driving the emergence of antibiotic resistance and the evolution of survival strategies in biological systems.

  15. Transferring cucumber mosaic virus-white leaf strain coat protein gene into Cucumis melo L. and evaluating transgenic plants for protection against infections

    Energy Technology Data Exchange (ETDEWEB)

    Gonsalves, C.; Xue, B.; Yepes, M.; Fuchs, M.; Ling, K.; Namba, S. (Cornell Univ., Geneva, NY (United States). Dept. of Plant Pathology)


    A single regeneration procedure using cotyledon examples effectively regenerated five commercially grown muskmelon cultivars. This regeneration scheme was used to facilitate gene transfers using either Agrobacterium tumefaciens or microprojectile bombardment methods. In both cases, the transferred genes were from the T-DNA region of the binary vector plasmid pGA482GG/cp cucumber mosaic virus-white leaf strain (CMV-WL), which contains genes that encode neomycin phosphotransferase II (NPT II), [beta]-glucuronidase (GUS), and the CMV-WL coat protein (CP). Explants treated with pGA482GG/cpCMV-WL regenerated shoots on Murashige and Skoog medium containing 4.4 [mu]m 6-benzylaminopurine (BA), kanamycin (Km) at 150 mg[center dot]liter[sup [minus]1] and carbenicillin (Cb) at 500 mg[center dot]liter[sup [minus]1]. The authors' comparison of A. tumefaciens- and microprojectile-mediated gene transfer procedures shows that both methods effectively produce nearly the same percentage of transgenic plants. R[sub 0] plants were first tested for GUS or NPT II expression, then the polymerase chain reaction (PCR) and other tests were used to verify the transfer of the NPT II, GUS, and CMV-WL CP genes.

  16. Contribution of midgut bacteria to blood digestion and egg production in aedes aegypti (diptera: culicidae (L.

    Directory of Open Access Journals (Sweden)

    Pimenta Paulo FP


    Full Text Available Abstract Background The insect gut harbors a variety of microorganisms that probably exceed the number of cells in insects themselves. These microorganisms can live and multiply in the insect, contributing to digestion, nutrition, and development of their host. Recent studies have shown that midgut bacteria appear to strengthen the mosquito's immune system and indirectly enhance protection from invading pathogens. Nevertheless, the physiological significance of these bacteria for mosquitoes has not been established to date. In this study, oral administration of antibiotics was employed in order to examine the contribution of gut bacteria to blood digestion and fecundity in Aedes aegypti. Results The antibiotics carbenicillin, tetracycline, spectinomycin, gentamycin and kanamycin, were individually offered to female mosquitoes. Treatment of female mosquitoes with antibiotics affected the lysis of red blood cells (RBCs, retarded the digestion of blood proteins and reduced egg production. In addition, antibiotics did not affect the survival of mosquitoes. Mosquito fertility was restored in the second gonotrophic cycle after suspension of the antibiotic treatment, showing that the negative effects of antibiotics in blood digestion and egg production in the first gonotrophic cycle were reversible. Conclusions The reduction of bacteria affected RBC lysis, subsequently retarded protein digestion, deprived mosquito from essential nutrients and, finally, oocyte maturation was affected, resulting in the production of fewer viable eggs. These results indicate that Ae. aegypti and its midgut bacteria work in synergism to digest a blood meal. Our findings open new possibilities to investigate Ae. aegypti-associated bacteria as targets for mosquito control strategies.


    Directory of Open Access Journals (Sweden)



    Full Text Available Introduction. It is very important to know the resistant bacteria to common used antibiotics in our community. Staphylococcus coagulase positive was the main cause of infection in infectious disease. This study was done to demonstrate the pattern of resistance to batalactamase antibiotics among staphiococci. Methods. During a period of five month, 38 coagulase positive staphylococcus isolates were identified from various clinical specimens from 600 patients at the AI-Zahra university hospital (affiliated to IUMSHS. Results. Betalactamase production assays using rapid acidometric and iodometric tests showed that 78.9 percent and 73.6 percent of isolates were positive, respectively. The differnce in determination rate between acidometric and iodometric tests was not statistically significant. Moreover the acidometric test was cheaper and more easy to perform than iodometric test. In vitro sensitivity testing using the disc diffusion method showed that all of isolates were resistant to carbenicillin, ampicillin and amoxicillin, while 7.9 percent and 13.2 percent were resistant to cefazolin and cephalexin, respectively. Discussion. We recommend use of cefazolin, cephalexin and oxacillin for treatment of patients with staphylococcus infections.

  18. A Highly Sensitive Resonance Rayleigh Scattering Method for the Determination of Penicillin Antibiotics with Potassium Ferricyanide

    Institute of Scientific and Technical Information of China (English)

    DUAN Hui; LIU Zhong-Fang; LIU Shao-Pu; KONG Ling


    Heated in a boiling water bath, penicillin antibiotics such as amoxicillin, ampicillin, sodium cloxacillin, sodium carbenicillin and sodium benzylpenicillin could react with K3[Fe(CN)6] to form combined products in a dilute HCl medium.As a result, resonance Rayleigh scattering (RRS) intensity was enhanced greatly and new RRS spectra appeared.The maximum scattering wavelengths of the five combined products are all located at 330 nm.The scattered intensity increments (△I) of the combined products are directly proportional to the concentrations of the antibiotics in a certain range.The methods exhibit high sensitivity, and the detection limits for the five penicillin antibiotics are between 4.61 and 5.62 ng·mL-1.The spectral characteristics of RRS and the optimum reaction conditions were investigated.The mechanism of reaction and the reasons for the enhancement of resonance light scattering were discussed.The effects of coexisting substances have been examined, and the results indicated that the method had a good selectivity.It can be applied to the determination of penicillin antibiotics in capsule, tablet, human serum and urine samples.

  19. An Efficient Agrobacterium-Mediated Transformation of Strawberry cv. Camarosa by a Dual Plasmid System

    Directory of Open Access Journals (Sweden)

    Fatemeh Haddadi


    Full Text Available An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100% of shoot formation and the highest mean number of shoots (24 produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86% in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet.

  20. Recycling antibiotics into GUMBOS: a new combination strategy to combat multi-drug-resistant bacteria. (United States)

    Cole, Marsha R; Hobden, Jeffery A; Warner, Isiah M


    The emergence of multi-drug-resistant bacteria, coupled with the lack of new antibiotics in development, is fast evolving into a global crisis. New strategies utilizing existing antibacterial agents are urgently needed. We propose one such strategy in which four outmoded β-lactam antibiotics (ampicillin, carbenicillin, cephalothin and oxacillin) and a well-known antiseptic (chlorhexidine di-acetate) were fashioned into a group of uniform materials based on organic salts (GUMBOS) as an alternative to conventional combination drug dosing strategies. The antibacterial activity of precursor ions (e.g., chlorhexidine diacetate and β-lactam antibiotics), GUMBOS and their unreacted mixtures were studied with 25 clinical isolates with varying antibiotic resistance using a micro-broth dilution method. Acute cytotoxicity and therapeutic indices were determined using fibroblasts, endothelial and cervical cell lines. Intestinal permeability was predicted using a parallel artificial membrane permeability assay. GUMBOS formed from ineffective β-lactam antibiotics and cytotoxic chlorhexidine diacetate exhibited unique pharmacological properties and profound antibacterial activity at lower concentrations than the unreacted mixture of precursor ions at equivalent stoichiometry. Reduced cytotoxicity to invasive cell types commonly found in superficial and chronic wounds was also observed using GUMBOS. GUMBOS show promise as an alternative combination drug strategy for treating wound infections caused by drug-resistant bacteria.


    Directory of Open Access Journals (Sweden)

    Pavan Kumar Padarthi


    Full Text Available Microbial contribution increasing rapidly due to invasion by the pathogenic organisms like bacteria, fungi and virus in the present disease burden of human health. To treat these diseases many potent and broad spectrum antibiotics were discovered e.g., ampicillin, amoxicillin, carbenicillin, ofloxacin and tetracycline etc., Even though antibiotics are life saving drugs in therapeutics but they are potentially harmful. These harmful effects include allergic and anaphylactic reaction, development of resistance, destruction of normal non-pathogenic bacterial flora and selective toxicity like aplastic anemia, kidney damage. As the microbial resistance make anti-microbial therapy very complex, there is a definite need of novel anti-microbials or drugs for combination therapy with standard antibiotics. Our aim was to synthesize and explore the anti-microbial activity of chalcones and its derived pyrimidines against various pathological micro organisms. Novel imidazole derived chalcones were synthesized and characterization was carried out by analyzing melting point, IR, 1H NMR data. The synthesized chalcones and pyrimidines are tested for their antimicrobial activity against various bacteria as well as fungi. Further synthesis of novel heterocyclic chalcones, structural elucidation, spectral analysis, biological activity of synthesized chalcones and its derived pyrimidines gives a hope for enhanced biological action using QSAR Studies.

  2. Prevalence of some mastitis causes in dromedary camels in Abu Dhabi, United Arab Emirates

    Directory of Open Access Journals (Sweden)

    A.A. Al-Juboori1


    Full Text Available The present study was designed to determine the prevalence of different types of mastitis in camels in U.A.E. and to identify the causative microorganisms and their sensitivity to different antimicrobial agents. From 162 lactating she-camels, 630 milk samples were collected from different cities in Abu Dhabi Emirate/UAE. The overall prevalence of mastitis was 18.52% (7.94% on quarter basis, the prevalence of clinical and sub clinical mastitis was found to be 24.70% and 11.67% on animal basis, respectively; it being 9.70% and 5.86% on quarter basis, respectively. The hind quarters were more frequently affected than the fore quarters. Bacteriological examination of milk samples revealed that Staphylococcus was the chief etiological agents both in clinical and sub clinical mastitis (41.67% in camels, followed by Streptococcus spp. (21.67%, Enterobacter spp. (15.00%, C. pyogenes (10.00%, Micrococcus spp. (5.00%, Pasteurells spp. (5.00% and Pseudomonas aeruginosa (1.66%. Most of the Staphylococcus spp., Streptococcus spp. and C. pyogenes strains were sensitive to carbenicillin, gentamycin, kanamycin, and erythromycin, but resistant to colistin and sulphamethoxazole. Other pathogens like Enterobacter, Micrococcus, Pasteurella spp. and Ps. aeuroginosa isolates showed variable sensitivities to the antimicrobials.

  3. Biodegradation and bioremediation of endosulfan contaminated soil. (United States)

    Kumar, Mohit; Lakshmi, C Vidya; Khanna, Sunil


    Among the three mixed bacterial culture AE, BE, and CE, developed by enrichment technique with endosulfan as sole carbon source, consortium CE was found to be the most efficient with 72% and 87% degradation of alpha-endosulfan and beta-endosulfan, respectively, in 20 days. In soil microcosm, consortium AE, BE and CE degraded alpha-endosulfan by 57%, 88% and 91%, respectively, whereas beta-endosulfan was degraded by 4%, 60% and 67% after 30 days. Ochrobacterum sp., Arthrobacter sp., and Burkholderia sp., isolated and identified on the basis of 16s rDNA gene sequence, individually showed in situ biodegradation of alpha-endosulfan in contaminated soil microcosm by 61, 73, and 74, respectively, whereas degradation of beta-endosulfan was 63, 75, and 62, respectively, after 6 weeks of incubation over the control which showed 26% and 23 % degradation of alpha-endosulfan and beta-endosulfan, respectively. Population survival of Ochrobacterum sp., Arthrobacter sp., and Burkholderia sp., by plate count on Luria Broth with carbenicillin showed 75-88% survival of these isolates as compared to 36-48% of survival obtained from PCR fingerprinting. Arthrobacter sp. oxidized endosulfan to endosulfan sulfate which was further metabolized but no known metabolite of endosulfan sulfate was detected.

  4. Initial identification and sensitivity to antimicrobial agents of Salmonella sp.isolated from poultry products in the state of Ceara, Brazil

    Directory of Open Access Journals (Sweden)

    WF Oliveira


    Full Text Available The objective of this research was to isolate and to verify the sensitivity to antimicrobial agents of strains of Salmonella sp. isolated from poultry products in the state of Ceara, Brazil. A total number of 114 samples was collected from 63 broiler carcasses derived from two processing plants and two supermarkets, and 51 excreta samples were collected in broiler farms located in the state of Ceara, which used three live production stages. Each excreta sample consisted of a fresh excreta pool from 100 birds. Samples were submitted to microbiological analyses, and the isolated Salmonella strains were tested for antimicrobial sensitivity. No Salmonella was isolated from excreta samples, while broiler carcass samples showed a high contamination rate of11.8%. Three serotypes were identified: Salmonella enterica serovar Enteritidis, 50%; Salmonella enterica serovar Panama 33%, and Salmonella enterica serovar Newport, 17%. As to the susceptibility tests to antimicrobial agents, 100% of the isolated Salmonella strains showed resistance to Ampicillin and Tetracycline, and sensitivity to Gentamycin, Netilmycin, Carbenicillin, Chloramphenicol.

  5. Prevalence and distribution of Vibrio parahaemolyticus in finfish from Cochin (south India). (United States)

    Sudha, Santha; Divya, Puthenkandathil S; Francis, Bini; Hatha, Ammanamveetil A M


    Finfish samples obtained from four retail outlets in Cochin between June 2009 and June 2010 were investigated for the occurrence of Vibrio parahaemolyticus. A total of 182 samples were collected and suspect isolates were identified using standard biochemical tests and were further confirmed by a species-specific tlh gene. V. parahaemolyticus was detected in 45.1% of samples, with demersal fish being more affected than pelagic species. The bacterium was isolated more frequently from the skin and gills of pelagic fish, while the intestine yielded greater numbers of V. parahaemolyticus in demersal fish. The highest incidence of antibiotic resistance was recorded against ampicillin and streptomycin, followed by carbenicillin, cefpodoxime, cephalothin, colistin and amoxycillin; the lowest was against nalidixic acid, tetracycline, chloramphenicol and ciprofloxacin. Multiple drug resistance was prevalent among isolates. Although only a fraction of strains are pathogenic for humans, the time-temperature abuse in markets provides ample scope for these strains to multiply to dangerous levels. The multidrug resistant nature of the strains adds to the gravity of the problem. High V. parahaemolyticus incidence rates in market finfish samples from areas in and around Cochin clearly indicates that control measures should be adopted to reduce post-harvest contamination in seafood and time-temperature abuse in markets to diminish the risk of V. parahaemolyticus infection associated with seafood destined for human consumption.

  6. Prevalence and distribution of Vibrio parahaemolyticus in finfish from Cochin (south India

    Directory of Open Access Journals (Sweden)

    Ammanamveetil A.M. Hatha


    Full Text Available Finfish samples obtained from four retail outlets in Cochin between June 2009 and June 2010 were investigated for the occurrence of Vibrio parahaemolyticus. A total of 182 samples were collected and suspect isolates were identified using standard biochemical tests and were further confirmed by a species-specific tlh gene. V. parahaemolyticus was detected in 45.1% of samples, with demersal fish being more affected than pelagic species. The bacterium was isolated more frequently from the skin and gills of pelagic fish, while the intestine yielded greater numbers of V. parahaemolyticus in demersal fish. The highest incidence of antibiotic resistance was recorded against ampicillin and streptomycin, followed by carbenicillin, cefpodoxime, cephalothin, colistin and amoxycillin; the lowest was against nalidixic acid, tetracycline, chloramphenicol and ciprofloxacin. Multiple drug resistance was prevalent among isolates. Although only a fraction of strains are pathogenic for humans, the time-temperature abuse in markets provides ample scope for these strains to multiply to dangerous levels. The multidrug resistant nature of the strains adds to the gravity of the problem. High V. parahaemolyticus incidence rates in market finfish samples from areas in and around Cochin clearly indicates that control measures should be adopted to reduce post-harvest contamination in seafood and time-temperature abuse in markets to diminish the risk of V. parahaemolyticus infection associated with seafood destined for human consumption.

  7. [Infections in the child with acute leukemia]. (United States)

    Carrillo, J M; Jiménez, E; Jiménez, R


    One hundred and twenty-five febrile episodes in 82 children with acute leukemia were studied; 46% of the patients were from urban and 54% from rural areas. The origin of the fever was identified in 91% of the episodes, prevailing pneumonia, septicemia, chickenpox and herpes zoster. The etiological agent was identified in 46% of the cases. A viral predominance was evident, and among them varicela-zoster, following in importance gram-negative bacteria. Histoplasma capsulatum and Pneumocystis carinii were isolated in two occassions each. Sepsis was found more frequently in children with active leukemia than in those in remission (p less than 0.001). Forty-four febrile episodes occurred in patients with less than 1,000 neutrophils/ul. The daily-risk rate of infection was higher in children fom rural than in those from urban areas (p less than 0.001). After clinical and laboratory studies, methicillin and gentamicin were used, in addition to carbenicillin or trimethoprim-sulfamethoxazole is selected cases. This treatment was effective in 86% of the cases. Twelve (15%) children died, 6 of whom were in remission at that moment.

  8. Commonly prescribed β-lactam antibiotics induce C. trachomatis persistence/stress in culture at physiologically relevant concentrations.

    Directory of Open Access Journals (Sweden)

    Jennifer eKintner


    Full Text Available Chlamydia trachomatis, the most common bacterial sexually transmitted disease agent worldwide, enters a viable, non-dividing and non-infectious state (historically termed persistence and more recently referred to as the chlamydial stress response when exposed to penicillin G in culture. Notably, penicillin G-exposed chlamydiae can reenter the normal developmental cycle upon drug removal and are resistant to azithromycin-mediated killing. Because penicillin G is less frequently prescribed than other β-lactams, the clinical relevance of penicillin G-induced chlamydial persistence/stress has been questioned. The goal of this study was to determine whether more commonly used penicillins also induce C. trachomatis serovar E persistence/stress. All penicillins tested, as well as clavulanic acid, induced formation of aberrant, enlarged reticulate bodies (called aberrant bodies or AB characteristic of persistent/stressed chlamydiae. Exposure to the penicillins and clavulanic acid also reduced chlamydial infectivity by >95%. None of the drugs tested significantly reduced chlamydial unprocessed 16S rRNA or genomic DNA accumulation, indicating that the organisms were viable, though non-infectious. Finally, recovery assays demonstrated that chlamydiae rendered essentially non-infectious by exposure to ampicillin, amoxicillin, carbenicillin, piperacillin, penicillin V and clavulanic acid recovered infectivity after antibiotic removal. These data definitively demonstrate that several commonly used penicillins induce C. trachomatis persistence/stress at clinically relevant concentrations.

  9. Identification of Pseudomonas aeruginosa genes associated with antibiotic susceptibility

    Institute of Scientific and Technical Information of China (English)


    Pseudomonas aeruginosa causes acute and chronic infections in humans and these infections are difficult to treat due to the bacteria’s high-level of intrinsic and acquired resistance to antibiotics. To address this problem, it is crucial to investigate the molecular mechanisms of antibiotic resistance in this organism. In this study, a P. aeruginosa transposon insertion library of 17000 clones was constructed and screened for altered susceptibility to seven antibiotics. Colonies grown on agar plates con- taining antibiotics at minimum inhibitory concentrations (MICs) and those unable to grow at ? MIC were collected. The transposon-disrupted genes in 43 confirmed mutants that showed at least a three-fold increase or a two-fold decrease in suscep- tibility to at least one antibiotic were determined by semi-random PCR and subsequent sequencing analysis. In addition to nine genes known to be associated with antibiotic resistance, including mexI, mexB and mexR, 24 new antibiotic resis- tance-associated genes were identified, including a fimbrial biogenesis gene pilY1 whose disruption resulted in a 128-fold in- crease in the MIC of carbenicillin. Twelve of the 43 genes identified were of unknown function. These genes could serve as targets to control or reverse antibiotic resistance in this important human pathogen.

  10. [Incidence of alginate-coding gene in carbapenem-resistant Pseudomonas aeruginosa strains]. (United States)

    Bogiel, Tomasz; Kwiecińska-Piróg, Joanna; Kozuszko, Sylwia; Gospodarek, Eugenia


    Pseudomonas aeruginosa rods are one of the most common isolated opportunistic nosocomial pathogens. Strains usually are capable to secret a capsule-like polysaccharide called alginate important for evasion of host defenses, especially during chronic pulmonary disease of patients with cystic fibrosis. Most genes for alginate biosynthesis and lysis are encoded by the operon. The aim of our study was to evaluate the incidence of algD sequence, generally use for alginate-coding gene detection, in 120 P. aeruginosa strains resistant to carbapenems. All isolates were obtained in the Department of Clinical Microbiology University Hospital no. 1 of dr A. Jurasz Collegium Medicum of L. Rydygier in Bydgoszcz Nicolaus Copernicus University in Toruń. Examined strains demonstrated resistance to carbenicillin (90,0%), ticarcillin (89,2%) and ticarcillin clavulanate (86,7%). All strains were susceptible to colistin. The majority of examined strains was susceptible to ceftazidime and cefepime (40,8% each) and norfloxacin (37,5%). Presence of algD gene - noted in 112 (93,3%) strains proves that not every strain is capable to produce alginate. It was also found out that differences in algD genes incidence in case of different clinical material that strains were isolated from were not statistically important.

  11. [Occurrence and susceptibility to antibiotics of carbapenem-resistant Pseudomonas aeruginosa strains between 1998 and 2009]. (United States)

    Bogiel, Tomasz; Mikucka, Agnieszka; Skalski, Tomasz; Gospodarek, Eugenia


    P. aeruginosa rods are dangerous pathogens mainly responsible for nosocomial infections of different localization. Resistance to carbapenems, observed among them, is a serious threat due to ability to be transmitted between bacterial species. The aim of our study was to retrospectively evaluate the frequency of isolation and susceptibility to antibiotics of imipenem- and meropenem-resistant P. aeruginosa strains isolated between 1998 and 2009 from patients of University Hospital No 1 of dr A. Jurasz in Bydgoszcz. Study shows increasing number of isolation that type of strains from 19 in 1998 to 144 in 2009. Among all isolated P. aeruginosa strains majority was obtained from patients of the Intensive Care Units, Rehabilitation and Surgery Clinics. Examined strains of P. aeruginosa rods were mainly isolated from urine (20.5%), bronchoalveolar lavage (17.7%) and wound swabs (14.5%) samples. The isolates demonstrated frequently resistance to carbenicillin (> or 66.7%), ticarcillin (> or = 72.7%) and cefotaxime (> or = 75.6%). The lowest rate of resistant strains was observed in case of ceftazidime (< or = 68.8%), aztreonam (< or = 47.4%) and colistin (< or = 1.7%) suggesting the highest activity of that antimicrobials against infections caused by examined strains.

  12. Meropenem as an Alternative Antibiotic Agent for Suppression of Agrobacterium in Genetic Transformation of Orchid

    Institute of Scientific and Technical Information of China (English)

    CAO Ying; Niimi Yoshiyuki; HU Shang-lian


    A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies (PLBs) has been reported in this article. The in vitro activities of meropenem and four comparator antibacterial agents against three Agrobacterium tumefaciens strains, LBA4404, EHA101, and GV3101, were assessed. In addition, the effect of meropenem on the growth of Dendrobium phalaenopsis PLBs was determined. Compared with other commonly used antibiotics (including ampicillin,carbenicillin, cefotaxime, and cefoperazone), meropenem showed the highest activity in suppressing all tested A.tumefaciens strains (minimum inhibitory concentration [MIC] < 0.5 mg L-1, which is equal to minimum bactericidal concentration [MBC]). Meropenem, at all tested concentrations, except for 10 mg L-1 concentration, had little negative effect on the growth of orchid tissues. The A. tumefaciens strain EHA101 in genetic transformation with vector pIG121Hm in infected PLBs of the orchid was visually undetectable after a two-month subculture in 1/2 MS medium with 50 mg L-1 meropenem and 25 mg L-1 hygromacin. The expression and incorporation of the transgenes were confirmed by GUS histochemical assay and PCR analysis. Meropenem may be an alternative antibiotic for the effective suppression of A. tumefaciens in genetic transformation.

  13. ISPpu22, a novel insertion sequence in the oprD porin gene of a carbapen- em-resistant Pseudomonas aeruginosa isolate from a burn patient in Tehran, Iran

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    Davood Kalantar-Neyestanaki


    Full Text Available Background and Objectives: The oprD mutation and AmpC overproduction are the main mechanisms of intrinsic resistance to carbapenems such as imipenem and meropenem in Pseudomonas aeruginosa.Materials and Methods: In this study, we investigated intrinsic resistance to carbapenems including mutation of oprD and AmpC overproduction in a carbapenem-resistant P. aeruginosa isolated from a burn patient by phenotypic and molecular methods.Results: In our study, the carbapenem-resistant P. aeruginosa isolate was resistant to imipenem, meropenem, cefepime, gentamicin, ceftriaxone, carbenicillin, aztreonam and ciprofloxacin but was susceptible to ceftazidime and polymyxin B. The minimum inhibitory concentrations (MICs against imipenem, meropenem and ceftazidime were 64 μg/ml, 16 μg/ml and 2μg/ml, respectively. The isolate was ESBLs and AmpC overproducer. No carbapenemase activity was detected by Modified Hodge test (MHT. This isolate was carrying only blaOXA-10. PCR amplification and sequencing of oprD performed on isolate resulted in PCR product of 2647bp. Sequence analysis of the 2647bp product revealed insertion of a sequence of 1232 bp at position 8 in coding region of oprD.Conclusion: According to the results of this study, oprD mutation and AmpC overproduction can cause the main mechanism of resistance of P. aeruginosa to carbapenems.Keywords: ISPpu22, oprD, AmpC, Carbapenem-resistant P. aeruginosa

  14. The Prevalence and Antimicrobial Susceptibility of Bacterial Uropathogens Isolated from Pediatric Patients

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    R Ranjbar


    Full Text Available "nBackground: Urinary tract infection (UTI is considered as the most common bacterial infectious disease seen among the pediatric patients. The aim of this study was to investigate the prevalence and antimicrobial susceptibility of bacterial uropathogens isolated from the pediatric patients with urinary tract infections."nMethods: This descriptive study was conducted in Children Medial Center, Tehran, Iran from March 2006 to Feb 2007. Clean-catch midstream urine specimens were obtained from the patients and cultured on the appropriate bacteriological media. Bacterial isolates were identified by standard biochemical and serological tests. Antimicrobial susceptibility testing was performed according to CLSI guidelines."nResults: From 14199 urine specimens, 16.2% had positive results for bacterial cultures. Nine hundred twenty one strains were identified as Escherichia coli; 412 as Klebsiella spp., 285 as Coagulase negative Staphylocococci, 202 as Enterococcus spp., 158 as Pseudomonas spp., and 83 as Staphylococcus aureus. E. coli isolates showed high resistance to carbenicillin (68%, ampicillin (96%, trimethoprim-sulfomethoxazol (70% and kanamycin (65%. More than 30% of isolates of Klebsiella spp., Pseudomonas spp. and Enterobacter spp. have shown high degree of resistance to commonly used antibiotics."nConclusion: Our findings reinforce the need for ongoing investigation to show trends in antibiotic resistance, which can help to prescribing of antibiotics in clinics.

  15. Cephalosporin resistance in Pseudomonas aeruginosa, with special reference to the proposed trapping of antibiotics by beta-lactamase. (United States)

    Livermore, D M; Williams, J D; Davy, K W


    Resistance of Pseudomonas aeruginosa strains to newer cephalosporins is often associated with stable derepression of synthesis of the chromosomal beta-lactamase. Similar resistance is developed by enzyme inducible (i.e. normal) strains in response to beta-lactamase inducers. By comparing the responses of otherwise isogenic P. aeruginosa beta-lactamase inducibility mutants to antipseudomonal cephalosporins alone or in combination with potent beta-lactamase inducers we confirmed that resistance to cefotaxime, ceftriaxone, cefoperazone, and ceftazidime and latamoxef was caused by beta-lactamase action. The low-level resistance to carbenicillin and cefsulodin which was exhibited by some fully beta-lactamase derepressed strains was not confirmed to be beta-lactamase determined and may have reflected concurrent target or permeability changes. The mechanism whereby the enzyme protected the cell against cefotaxime and ceftriaxone was also investigated. These agents are reportedly stable to the enzyme and some workers have suggested that resistance entails their being trapped rather than hydrolysed. However, the use of a novel model of cellular beta-lactamase function indicated that a hydrolytic resistance mechanism remained likely.

  16. [Genetic transformation of buckwheat ( Fagopyrum esculentum Moench ) with AtNHX1 gene and regeneration of salt-tolerant transgenic plants]. (United States)

    Cheng, Li-Hong; Zhang, Bo; Xu, Zi-Qin


    The Arabidopsis thaliana tonoplast Na+ /H+ antiporter gene, AtNHX1, was transferred into buckwheat by Agrobacterium-mediated method. Transgenic buckwheat plants were regenerated and selected on MS basal medium supplemented with 2.0mg/L 6-BA, 1.0mg/L KT, 0.lmg/L IAA, 50mg/L kanamycin and 500mg/L carbenicillin. 426 seedlings from 36 resistant calli originated from 864 explants (transformed about at 4.17 percentage) exhibited resistance to kanamycin. The transformants were confirmed by PCR, Southern blotting, RT-PCR and Northern blotting analysis. After stress treatment for 6 weeks with 200mmol/L NaCl, transgenic plants survived, while wild-type plants did not. After 3 days of stress treatment through different concentrations of NaCl, transgenic plants accumulated higher concentration of Na+ and proline than the control plants. However, the K+ concentration of transgenic plants declined in comparison with the control plants. Moreover, the rutin content of the roots, stems and leaves of transgenic buckwheat increased than those of the control plants. These results showed that it could be possible to improve the salt-tolerance of crops with genetic technology.

  17. An efficient Agrobacterium-mediated transformation of strawberry cv. Camarosa by a dual plasmid system. (United States)

    Haddadi, Fatemeh; Aziz, Maheran Abd; Abdullah, Siti Nor Akmar; Tan, Soon Guan; Kamaladini, Hossein


    An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet.

  18. Effects of antibiotics on explant regeneration of Actinidia chinensis%抗生素对中华猕猴桃外植体再生的影响

    Institute of Scientific and Technical Information of China (English)

    尚霄丽; 谭彬; 李继东; 张建鹏; 冯建灿


    【目的】为确定中华猕猴桃遗传转化中适宜的抗生素种类和浓度,【方法】以中华猕猴桃‘伏牛95—2’叶片为外植体,研究了抗生素对外植体再生的影响。【结果】抗生素敏感性测定发现,当卡那霉素质量浓度为20mg·^(-1)时,愈伤组织及不定芽分化率均为0,因此,20mg·L^(-1)卡那霉素为适宜的筛选浓度;头孢霉素和羧苄青霉素对中华猕猴桃‘伏牛95—2’叶片分化均有抑制作用,但头孢霉素对叶片分化的抑制作用比羧苄青霉素小,结合抑菌试验,将浓度400mg.L^(-1)头孢霉素确定为适宜于中华猕猴桃‘伏牛95—2’叶片遗传转化的抑菌抗生素。I结论】卡那霉素20mg·L。和头孢霉素400mg·L^(-1)为中华猕猴桃‘伏牛95—2’遗传转化适宜的抗生素及质量浓度。%[Objective] The study is to screen the suitable type and concentration of antibiotics for the transformation of A ctinidia chine ns is. [ Method ] Effects of antibiotics on explant regeneration were studied using leaves of A ctinidia chinensis ‘Funiu 95-2' as explants. [Result] The results showed that differentiation rate of callus and adventitious shoots was zero when 20 rag. L^(-1) kanamycin was added into the medium, so the optimal kanamycin concentration for the selection of transformants was 20 mg·L-1 based on the antibiotic sensitivity measurement. Differentiation of adventitious shoots was inhibited by adding Cefotaxime and Carbenicillin into the medium, while inhibition effect of Carbenicillin was stronger than that of Cefotaxime. 400 rag. L^(-1) Cefotaxime was optimal for genetic transformation by Agrobacteriumin in Actinidia chinensis ‘Funiu 95-2' [Conclusion] The studies suggested that 20 mg.L^(-1) Kanamycin and 400 rag.L^(-1) Cefotaxime were suitable for the transformation of Actinidia chinensis ‘Funiu 95-2'.

  19. Influence of antibiotics on embryogenic tissue and Agrobacterium tumefaciens suppression in soybean genetic transformation Influência de antibióticos sobre o tecido embriogênico e a supressão de Agrobacterium tumefaciens na transformação genética de soja

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    Beatriz Wiebke


    Full Text Available The influence of different antibiotic treatments in soybean genetic transformation was evaluated. First, an assay was performed to verify how different antibiotic treatments affect soybean embryogenic tissues. The effect of carbenicillin at 500 mg L-1 was genotype-dependent. This antibiotic did not affect embryo survival of cv. IAS5, but a three-fold increase of embryo proliferation was observed for cv. Bragg, when compared to the control. On the other hand, cefotaxime at 350 and 500 mg L-1 caused death of embryogenic tissues of both cultivars. Finally, the association of cefotaxime (250 mg L-1 + vancomycin (250 mg L-1 did not affect negatively the somatic embryos of tested cultivars until 63 days of treatment. Thereafter, a second experiment was carried out to determine the efficacy of different antibiotic treatments in suppressing LBA4404 Agrobacterium tumefaciens strain in genetic transformation. On tissue culture conditions, carbenicillin at 500 and 1000 mg L-1 was not active against Agrobacterium. On the other hand, treatments with cefotaxime at 350 and 500 mg L-1, and cefotaxime + vancomycin efficiently suppressed Agrobacterium during 49 days. Data of both experiments suggested cefotaxime + vancomycin for 49-63 days as the most appropriate treatment. This is the first work reporting the effect of antibiotics on soybean tissues. By identifying an antibiotic combination that suppressed A. tumefaciens with minimal phytotoxic effects, we are able to recommend it for improvement of soybean Agrobacterium-mediated transformation procedure.Foi avaliada a influência de diferentes tratamentos com antibióticos durante a transformação genética de soja. Inicialmente, desenvolveu-se um estudo para identificar como diferentes tratamentos com antibióticos afetam o tecido embriogênico da soja. O efeito da carbenicilina a 500 mg L-1 foi genótipo-dependente. Esse antibiótico não afetou a sobrevivência dos embriões da cv. IAS5, enquanto a

  20. Effective antibiotics against 'Candidatus Liberibacter asiaticus' in HLB-affected citrus plants identified via the graft-based evaluation. (United States)

    Zhang, Muqing; Guo, Ying; Powell, Charles A; Doud, Melissa S; Yang, Chuanyu; Duan, Yongping


    Citrus huanglongbing (HLB), caused by three species of fastidious, phloem-limited 'Candidatus Liberibacter', is one of the most destructive diseases of citrus worldwide. To date, there is no established cure for this century-old and yet, newly emerging disease. As a potential control strategy for citrus HLB, 31 antibiotics were screened for effectiveness and phytotoxicity using the optimized graft-based screening system with 'Candidatus Liberibacter asiaticus' (Las)-infected citrus scions. Actidione and Oxytetracycline were the most phytotoxic to citrus with less than 10% of scions surviving and growing; therefore, this data was not used in additional analyses. Results of principal component (PCA) and hierarchical clustering analyses (HCA) demonstrated that 29 antibiotics were clustered into 3 groups: highly effective, partly effective, and not effective. In spite of different modes of actions, a number of antibiotics such as, Ampicillin, Carbenicillin, Penicillin, Cefalexin, Rifampicin and Sulfadimethoxine were all highly effective in eliminating or suppressing Candidatus Liberibacter asiaticus indicated by both the lowest Las infection rate and titers of the treated scions and inoculated rootstock. The non-effective group, including 11 antibiotics alone with three controls, such as Amikacin, Cinoxacin, Gentamicin, Kasugamycin, Lincomycin, Neomycin, Polymixin B and Tobramycin, did not eliminate or suppress Las in the tested concentrations, resulting in plants with increased titers of Las. The other 12 antibiotics partly eliminated or suppressed Las in the treated and graft-inoculated plants. The effective and non-phytotoxic antibiotics could be potential candidates for control of citrus HLB, either for the rescue of infected citrus germplasm or for restricted field application.

  1. Mutation of a novel virulence-related gene mltD in Vibrio anguillarum enhances lethality in zebra fish. (United States)

    Xu, Zinan; Wang, Ying; Han, Yin; Chen, Jixiang; Zhang, Xiao-Hua


    Vibrio anguillarum, a halophilic Gram-negative bacterium, is the causative agent of vibriosis, which is a major problem for the aquaculture industry worldwide. Previously, a virulence-related gene fragment of V. anguillarum was obtained from a suppression subtractive hybridization (SSH) library. In this study, the complete gene sequence was obtained by long and accurate PCR (LA-PCR). After sequence analysis and homologous comparison, this new virulence-related gene was revealed to encode a putative membrane-bound lytic murein transglycosylase D (MltD), which consisted of 547 amino acids, and showed 34% identity to the MltD in Escherichia coli. An mltD mutant of pathogenic V. anguillarum CW-1 was constructed by homologous recombination. Production of extracellular gelatinase and protease of the mltD mutant decreased markedly compared with those of the wild-type strain, and the hemolytic activity was totally lost. Sodium chloride challenge and antibiotic sensitivity assay showed that the resistance of the mltD mutant to high concentrations of sodium chloride, and rocephin, fortun, cefobid, gentamicin, kanamycin and carbenicillin was enhanced. Most importantly, virulence of the mltD mutant was enhanced compared with that of the wild type when it was inoculated intraperitoneally into zebra fish; the LD₅₀ of the wild type and the mutant was 3.92 × 10³ CFU and 1.01 × 10² CFU fish⁻¹, respectively. The mltD was cloned and overexpressed in E. coli, and the recombinant MltD protein showed hemolytic, phospholipase, gelatinase and diastase activities. This is the first report that MltD possibly has a virulence-related function.

  2. Prevalence and Antibiotics Susceptibility of Uropathogens in Patients from a Rural Environment, Tamilnadu

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    Maripandi Arjunan


    Full Text Available Problem statement: Urinary Tract Infections (UTIs in human, rural environment cause significant morbidity due to insanitary condition, lack of knowledge of personal hygiene, lack of patient’s compliance and economic burden. In our study, the bacterial etiologies and the resistance patterns found in human with serious UTIs and selecting optimal antimicrobial therapy. Approach: A total of 105 patients first morning mid stream urine samples, culture was done by the calibrated loop technique delivering 0.001 mL of urine plated on Cystine-Lactose-Electrolyte Deficient (CLED agar, MacConkey agar and Blood agar medium (Hi Media, India for isolation of uropathogens. All pathogens were analyzed for drug susceptibility by disk diffusion method. Results: More than 50% of them were confirmed to have UTI bacteriological positive. Women and man especially in the age group of 20-29 had higher incidence of bacterial infection. A total of 58 isolates seven different uropathogens were identified among the Escherichia coli (31% was dominant pathogens followed by Citrobacter spp., (20% and Pseudomonas aeruginosa (17.24%. In all uropathogens antibiotics susceptibility the more active agents were norfloxacin (87.93%, ciprofloxacin (70.68%, gentamicin (70.68%, nalidixic acid (68.96%, nitrofurantoine (60.34% and tetracycline (60.34% and less active in ampicillin (8.62%, rifampicin (37.93% and carbenicillin (41.37%. Conclusion: Among commonly used antimicrobial agents for the treatment of UTI, there is a trend towards increasing resistance to ampicillin and a persistently low resistance rate to norfloxacin, ciprofloxacin and gentamicin should be consider for treatment.

  3. Antibiotic resistance of Vibrio parahaemolyticus isolated from coastal seawater and sediment in Malaysia (United States)

    Drais, Ashraf Abbas; Usup, Gires; Ahmad, Asmat


    Vibrio parahaemolyticus is widely recognized pathogenic Vibrio species due to numerous outbreaks and its' wide occurrence in the marine environment. A total of 50 Vibrio parahaemolyticus isolates were isolated from seawater and sediments in Malaysia were tested for sensitivity to 19 antibiotics using disc diffusion method. It was found that all isolates were resistant towards ampicillin (10 μg), penicillin (10 μg), methicillin (5 μg), and novobiocin (5 μg); but exhibit sensitivity to chloramphenicol (30 μg) and gentamicin (100 μg). The low percentage of sensitivity towards antibiotics was observed with the following antibiotics; amoxicillin 10μg (98%), fluconazole 25μg (98%), erythromycin 15 μg (92%), vancomycin 30 μg (92%), bacitracin 10 μg (84%), carbenicillin 100 μg (84%), cephalothin 30 μg (52%), nitrofurantion 200 μg (52%), ciprofloxacin 5 μg (40%), tetracycline 30 μg (20%), kanamycin 30 μg (10%), nalidixic acid 30 μg (10%) and streptomycin 20 μg (6%). Multiple antibiotic resistance (MAR) index was found to be 0.42-0.78. All the isolates were multi-resistant to these antibiotics. This indicates that the isolates originate from high-risk source of contamination where antibiotics are often used. Thus, there is a need for supervised use of antibiotics and frequent surveillance of V. parahaemolyticus strains for antimicrobial resistance. The presence of V. parahaemolyticus in coastal water with a high value of multiple antibiotic resistance indexes (MARI) can increase the risk of exposure to human and regular monitoring program for this potential human pathogenic bacterium is important.

  4. Association of plasmid typing to biotyping and antibiotyping in the characterization of outbreaks by Acinetobacter baumannii

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    Maria Cristina Bronharo Tognim


    Full Text Available During an outbreak at an University Hospital, from April to September, in 1994, sixteen strains of Acinetobacter baumannii were isolated from patients and one strain from an enteral solution. We afterwards analyzed the outbreak by means of plasmid typing, antibiotic resistance typing and biotyping. Two main plasmid profiles were identified. Twelve strains belonged to biotype 2, and five to biotype 19. Susceptibility to amikacin and to carbenicillin allowed classification of the strains into two groups. The results show that association of those three typing methods allowed the differentiation of what was at first considered as a single outbreak into two apparently unrelated outbreaks.Durante um surto ocorrido de abril a setembro de 1994 em um Hospital Universitário, dezesseis cepas de Acinetobacter baumannii foram isoladas de pacientes e uma de solução enteral. Nós posteriormente analizamos as cepas isoladas durante o surto pelos seguintes métodos de tipagem : perfil de DNA plasmidial, perfil de antibiograma e biotipagem. Dois padrões de tipagem foram identificados pela análise do perfil plasmidial. Doze cepas foram caracterizadas como sendo do biotipo 2, e cinco do biotipo 19. O padrão de sensibilidade a amicacina e a carbenicilina possibilitou a classificação das cepas em dois grupos. Os resultados demonstraram que estes três métodos de tipagem associados possibilitaram a diferenciação do que primeiramente foi considerado como um único surto, em dois surtos aparentemente não relacionados.

  5. Molecular Characterisation of Salmonella enterica Serovar Typhi Isolated from Typhoidial Humans

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    Arunava Das


    Full Text Available Aims: Salmonella enterica serovar Typhi is the major causative agent for typhoidial fever around the globe among human population reported till date. Present research work was carried out for detection and molecular characterisation of Salmonella enterica serovar Typhi isolated from humans with Typhoidial fever by biochemical, phenotypical and virulence gene based polymerase chain reaction (PCR techniques. The isolated strains were also investigated for antibiotic susceptibility patterns as a control measure. Methodology and Results: A total of 16 clinical samples were collected from the same numbers of patients (7 males and 9 females from Coimbatore, Erode and Salem districts of Tamil Nadu and were processed via broth enrichment methods for isolation and identification of the causative agent S. enterica serovar Typhi. Microbiological and biochemical investigations revealed the presence of S. Typhi from 16 samples. The biotyping of the isolates showed that all the isolates belonged to biotype IV. The PCR analysis confirmed the presence of invA (Invasion gene, 244bp, tyv (Tyveloseepimerase gene, 615 bp, fliC-d (Phage-1 flagellin gene for d-antigen, 750 bp and viaB (Vi antigen gene, 439bp in all 16 clinical samples. The antibiotic susceptibility test that was carried out among the isolates against 12 antimicrobial agents, showed 100 % resistance to only ampicillin and 100 % sensitivity to carbenicillin, chloramphenicol, clindamycin, gentamycin, kanamycin and tetracycline.Conclusion, significance and impact of study: This study confirmed the association of virulent strains of S. enterica serovar Typhi from Typhoidial fever among human population and suggested that PCR based diagnostic could be very useful for the rapid detection of S. Typhi isolates. Present study emphasized the use of antibiotic like chloramphenicol or in combination with other antibiotics for the effective control of S. Typhi.

  6. Kluyvera, a new (redefined) genus in the family Enterobacteriaceae: identification of Kluyvera ascorbata sp. nov. and Kluyvera cryocrescens sp. nov. in clinical specimens. (United States)

    Farmer, J J; Fanning, G R; Huntley-Carter, G P; Holmes, B; Hickman, F W; Richard, C; Brenner, D J


    Kluyvera is proposed as a new genus for the group of organisms formerly known as Enteric Group 8 (synonym = API group 1). Strains of Kluyvera share the properties of most members of the family Enterobacteriaceae: they are gram-negative rods, motile with peritrichous flagella, catalase positive, and oxidase negative; they grow on MacConkey agar, ferment D-glucose with the production of acid and gas, and are susceptible to many antibiotics. Strains are usually indole positive, methyl red positive, Voges-Proskauer negative, citrate positive, H2S (triple sugar iron) negative, urea negative, phenylalanine deaminase negative, lysine decarboxylase positive, arginine dihydrolase negative, and ornithine decarboxylase positive. Kluyvera strains ferment many of the sugars and polyhydroxyl alcohols used in identification. By deoxyribonucleic acid-deoxyribonucleic acid hybridization, strains of Kluyvera were divided into three groups. Kluyvera ascorbata is proposed as the type species for the genus. Most strains of K. ascorbata have been isolated from clinical specimens. K. cryocrescens is proposed as the second species. It was occasionally isolated from clinical specimens, but it was isolated more commonly from the environment. Kluyvera species group 3 was heterogeneous, but was distinct from the two named species by deoxyribonucleic acid hybridization. This group was rare, so no species name will be proposed at this time. K. ascorbata can be differentiated from K. cryocrescens by its positive ascorbate test, inability to grow at 5 degrees C in a refrigerator, and smaller zones of inhibition around carbenicillin and cephalothin disks. The test normally used for identification does not clearly differentiate these two species. Kluyvera species are probably infrequent opportunistic pathogens. The most common source is sputum, where they are probably not clinically significant. Five strains have been from blood cultures. More information is needed about the incidence and clinical

  7. Identification of Additional Anti-Persister Activity against Borrelia burgdorferi from an FDA Drug Library

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    Jie Feng


    Full Text Available Lyme disease is a leading vector-borne disease in the United States. Although the majority of Lyme patients can be cured with standard 2–4 week antibiotic treatment, 10%–20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS. While the cause for this is unclear, persisting organisms not killed by current Lyme antibiotics may be involved. In our previous study, we screened an FDA drug library and reported 27 top hits that showed high activity against Borrelia persisters. In this study, we present the results of an additional 113 active hits that have higher activity against the stationary phase B. burgdorferi than the currently used Lyme antibiotics. Many antimicrobial agents (antibiotics, antivirals, antifungals, anthelmintics or antiparasitics used for treating other infections were found to have better activity than the current Lyme antibiotics. These include antibacterials such as rifamycins (3-formal-rifamycin, rifaximin, rifamycin SV, thiostrepton, quinolone drugs (sarafloxacin, clinafloxacin, tosufloxacin, and cell wall inhibitors carbenicillin, tazobactam, aztreonam; antifungal agents such as fluconazole, mepartricin, bifonazole, climbazole, oxiconazole, nystatin; antiviral agents zanamivir, nevirapine, tilorone; antimalarial agents artemisinin, methylene blue, and quidaldine blue; antihelmintic and antiparasitic agents toltrazuril, tartar emetic, potassium antimonyl tartrate trihydrate, oxantel, closantel, hycanthone, pyrimethamine, and tetramisole. Interestingly, drugs used for treating other non-infectious conditions including verteporfin, oltipraz, pyroglutamic acid, pidolic acid, and dextrorphan tartrate, that act on the glutathione/γ-glutamyl pathway involved in protection against free radical damage, and also the antidepressant drug indatraline, were found to have high activity against stationary phase B. burgdorferi. Among the active hits, agents that affect cell membranes, energy

  8. Transformation of pickling cucumber with chitinase-encoding genes using Agrobacterium tumefaciens. (United States)

    Raharjo, S H; Hernandez, M O; Zhang, Y Y; Punja, Z K


    Transformation of cucumber cv. Endeavor was attempted using three Agrobacterium tumefaciens strains (a supervirulent leucinopine type, an octopine type and a nopaline type), each harbouring one of three binary vectors which contained an acidic chitinase gene from petunia, and basic chitinase genes from tobacco and bean, respectively, driven by the CaMV 35S promoter. Petiole explants were inoculated with a bacterial suspension (10(8) cells·ml(-1)), cocultivated for 48-96 h and placed on Murashige and Skoog (MS) medium with 5.0 μM each of 2,4-D and BA, 50 mg·l(-1) kanamycin and 500 mg·l(-1) carbenicillin. The frequency of embryogenic callus formation ranged from 0 to 12%, depending on strains/vectors used and length of cocultivation, with the highest being obtained using the leucinopine strain with petunia acidic chitinase gene. The kanamycin-resistant embryogenic calli were used to initiate suspension cultures (in liquid MS medium with 1.0/1.0 μM 2,4-D/BA, 50 mg·l(-1) kanamycin) for multiplication of embryogenic cell aggregates. Upon plating of cell aggregates onto solid MS medium with 1.0/1.0 μM NAA/BA and 50 mg·l(-1) kanamycin, calli continued to grow and later differentiated into plantlets. Transformation by the leucinopine strain and all three vectors was confirmed by PCR amplification of the NPT II gene in transgenic calli and plants, in addition to Southern analysis. Expression of the acidic chitinase gene (from petunia) and both basic chitinase genes (from tobacco and bean) in different transgenic cucumber lines was confirmed by Western analyses.

  9. Multidrug-Resistant Vibrio cholerae O1 was Responsible for a Cholera Outbreak in 2013 in Bagalkot, North Karnataka. (United States)

    Bhattacharya, Debdutta; Dey, Shuchismita; Roy, Subarna; Parande, Mahantesh V; Telsang, M; Seema, M H; Parande, Aisha V; Mantur, Basappa G


    Cholera is a major cause of illness in the developing world. During the monsoon season, small sporadic clusters of cholera cases are reported on an annual basis in Karnataka, India. During the monsoons of 2013, there was a cholera outbreak in Badami, a remote area of Bagalkot district in Karnataka. The multi-drug-resistant Vibrio cholerae O1 serotype Ogawa was found to be responsible for this outbreak. On 5 August 2013, a 30-year-old woman presented with severe dehydration and watery diarrhea at the Aganwadi Health Centre in Badami. A total of 49 suspected cholera cases were reported, with an attack rate of 3.5%. The V. cholerae isolates exhibited resistance to a wide range of drugs, including ampicillin, co-trimoxazole, nitrofurantoin, carbenicillin, and third generation cephalosporins, and showed reduced susceptibility to third generation fluoroquinolones. All of the cephalosporin-resistant V. cholerae strains produced extended-spectrum beta-lactamase. All V. cholerae O1 isolates harbored virulent genes (ctxA, ctxB, tcpA El Tor, Tox S, VPI, ToxT, ToxR, ToxRS, ace, zot, and tcpP) and were found to be genetically similar as determined by randomly amplified polymorphic DNA fingerprinting assay. To the best of our knowledge, this is the first report of a cholera outbreak in the district of Bagalkot. The resistance of V. cholerae to commonly used antimicrobial drugs is becoming a major public health concern in the region as clinicians are left with a limited choice of antibiotics for the treatment of cholera.

  10. Promoter deletions of Klebsiella pneumoniae carbapenemase (KPC)-encoding genes (blaKPC -2) and efflux pump (AcrAB) on β-lactam susceptibility in KPC-producing Enterobacteriaceae. (United States)

    Seecoomar, Gomattie D; Marmol, Brenda C; Kwon, Dong H


    Klebsiella pneumoniae carbapenemase (KPC)-encoding genes containing promoter-deletions (bla(KPC-2a), bla(KPC-2b), and bla(KPC-2c) have disseminated in Enterobacteriaceae. The minimal inhibitory concentrations (MICs) to β-lactams in clinical KPC-producing Enterobacteriaceae range from susceptible to high-level resistant, resulting in diagnostic problems. To better understand the variability in β-lactam MICs among KPC-producing Enterobacteriaceae, three isoforms of bla(KPC-2) gene were used to transform Escherichia coli W4573 and its deletion mutant of an efflux pump (AcrAB) to examine the effects on β-lactam susceptibility. MICs to β-lactams in E. coli W4573 and its acrAB mutant strain increased 1- to 500-fold (MIC from 0.125 to 64 μg mL(-1) of aztreonam) in the bla(KPC-2a), bla(KPC-2b), and bla(KPC-2c) transformants compared with the cloning vector alone. However, transformants of the acrAB mutant strain remained susceptible to all β-lactams tested except for aztreonam and carbenicillin. Levels of the three promoters' length and carbapenemase activities in the transformants harboring the bla(KPC-2a), bla(KPC-2b), and bla(KPC-2c) were correlated to the levels of β-lactam MICs in both E. coli W4573 and its mutant of an efflux pump (AcrAB). Overall, these results suggest that promoter-deletions of bla(KPC-2) gene and AcrAB may be associated with the variability in β-lactam MICs in KPC-producing Enterobacteriaceae.

  11. Biochemical and molecular characterization of three new variants of AmpC beta-lactamases from Morganella morganii. (United States)

    Power, Pablo; Galleni, Moreno; Ayala, Juan A; Gutkind, Gabriel


    Morganella morganii produces an inducible, chromosomally encoded AmpC beta-lactamase. We describe in this study three new variants of AmpC within this species with apparent pIs of 6.6 (M19 from M. morganii strain PP19), 7.4 (M29 from M. morganii strain PP29), and 7.8 (M37 from M. morganii strain PP37). After gene sequencing, deduced amino acid sequences displayed one to six substitutions when compared to the available Morganella AmpC sequences. An AmpR-encoding gene was also found upstream of ampC, including the LysR regulators' helix-turn-helix DNA-binding domain and the putative T-N11-A-protected region in the ampR-ampC intercistronic sequence. All three AmpC variants were purified from in vitro-generated derepressed mutants and showed overall similar kinetic parameters. None of the observed amino acid changes, occurring at the surface of the protein, appear to have a major influence in their catalytic properties. Morganella AmpCs exhibit the highest catalytic efficiencies (k(cat)/K(m)) on classical penicillins, cefoxitin, narrow-spectrum cephalosporins, and cefotaxime. Cefotaxime was more effectively hydrolyzed than other oxyimino-cephalosporins, whereas cefepime was 3 log-fold less efficiently hydrolyzed than other cephalosporins such as cephalothin. Several differences with other AmpC beta-lactamases were found. Ampicillin was more efficiently hydrolyzed than benzylpenicillin. High k(cat)/K(m) values were observed for oxacillin and piperacillin, which are usually poor substrates for AmpC. A fairly efficient hydrolysis of imipenem was detected as well. Aztreonam, carbenicillin, and tazobactam were effective transient inactivators of these variants.

  12. Antibiotic Resistance Pattern of Pseudomonas Aeruginosa, Isolated from Patient with Burn Wound Infection in Guilan,Iran

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    Iraj Nikokar


    Full Text Available Background and Objectives: Antibiotic resistance of Pseudomonas aeruginosa remains a major problem in burn patients. The main objective of this study was to determine the antibiotic resistance pattern and frequency of class 1 integrons among P. aeruginosa strains isolated from patients with burn wound infections in a new Burn Centre in Guilan, Iran.Materials and Methods: The bacterial isolates were collected from 182 patients with burn wound infections and P. aeruginosa species were identified by standard bacteriological methods. The drug susceptibility test, using 11 antimicrobial agents, was performed for all the isolates via agar disk diffusion method. PCR was carried out for the detection of integrons.Results: Out of a total of 182 hospitalized patients in the burn center assessed, 86 (47% found to have P. aeruginosa in their isolates. Resistance rates to various antibiotics were as follows: cloxacillin (91.8%, cotrimoxazole (86%, cephazolin (83.7%, carbenicillin (74.4%, piperacillin (69.9%, ceftazidime (68.8%, ciprofloxacin (66.3%, tobramycin (58.2%, amikacin (48.8% and gentamicin (37.2%, while the most effective antibiotic was imipenem with a resistance rate of 23.3%. Thirty nine (45.3% isolates were detected as multi-drug resistant. The PCR results showed that 37 (43% P. aeruginosa isolates and 27 (69.2% multi-drug resistant strains harbored class 1 integrons. A significant correlation was obtained between the presence of integrons and resistance against imipenem, ceftazidime, piperacillin and ciprofloxacin (P < 0.001.Conclusion: Optimization of using antimicrobial agents and control of infection is recommended to prevent the increasing population of drug resistant organisms in the new burn centre setting in this study. Furthermore, the high frequency of class 1 integrons among multi-drug resistant strains might be responsible for dissemination of antibiotic resistance gene.

  13. Biochemical synthesis of silver nanoprticles using filamentous fungi Penicillium decumbens (MTCC-2494) and its efficacy against A-549 lung cancer cell line. (United States)

    Majeed, Shahnaz; Abdullah, Mohd Syafiq Bin; Dash, Gouri Kumar; Ansari, Mohammed Tahir; Nanda, Anima


    Biosynthesis of silver and other metallic nanoparticles is one of the emerging research area in the field of science and technology due to their potentiality, especially in the field of nano-biotechnology and biomedical sciences in order to develop nanomedicine. In our present study, Penicillium decumbens (MTCC-2494) was brought from Institute of Microbial Technology (IMTECH) Chandigarh and employed for extracellular biological synthesis of silver nanoparticles. Ag-NPs formation was appeared with a dark brown color inside the conical flask. Characterization of Ag-NPs were done by UV-Spectrophotometric analysis which showed absorption peak at 430 nm determines the presence of nanoparticles, Fourier transform infrared (FT-IR) spectroscopic analysis, showed amines and amides are the possible proteins involved in the stabilization of nanoparticles as capping agent. Atomic force Microscopy (AFM) confirmed the particle are spherical, size was around 30 to 60 nm and also the roughness of nanoparticles. Field emission scanning electron microscopy (FE-SEM) showed the topology of the nanoparticles and were spherical in shape. The biosynthesis process was found fast, ecofriendly and cost effective. Nano-silver particle was found to have a broad antimicrobial activity and also it showed good enhancement of antimicrobial activity of Carbenicillin, Piperacillin, Cefixime, Amoxicillin, Ofloxacin and Sparfloxacin in a synergistic mode. These Ag-NPs showed good anti-cancer activity at 80 μg·mL(-1)upon 24 hours of incubation and toxicity increases upon 48 hours of incubation against A-549 human lung cancer cell line and the synergistic formulation of the antibiotic with the synthesized nanoparticles was found more effective against the pathogenic bacteria studied.

  14. A selective medium for the rapid detection by an impedance technique of Pseudomonas spp. associated with poultry meat. (United States)

    Salvat, G; Rudelle, S; Humbert, F; Colin, P; Lahellec, C


    A new medium for detecting and enumerating Pseudomonas spp. associated with poultry meat spoilage by a rapid impedance technique was developed, after testing potential growth promoters for eight Pseudomonas strains and inhibitors against eight competing strains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977). Four basal media (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB)) and a synthetic medium were evaluated. Whitley impedance broth was the best basal medium for detecting variations in impedance in relation to Pseudomonas growth. The efficiency of WIB was improved by adding compounds which enhanced the growth of Pseudomonas on the synthetic medium. Among the incubation temperatures tested, 22 degrees C proved to be the best compromise between growth of Pseudomonas associated with poultry meat spoilage and inhibition of competitors. Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium: metronidazole, carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium). Preliminary results obtained from experiments with beef and pork meat showed that this medium could also be used without diamide and at an incubation temperature of 25 degrees C. The impedance technique using MCCCD medium was then compared with an official method which uses the medium of Mead and Adams (1977) on 106 samples of poultry neck skin. The linear regression coefficient between the two techniques was approximately r = 0.85. Impedance was able to detect 10(3) Pseudomonas g-1 within less than 19 h making it a promising technique for predicting poultry meat spoilage.

  15. Transformation of indica rice (Oryza sativa L. cv. RD6 mediated by Agrobacterium tumefaciens

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    Manit Kosittrakul


    Full Text Available High percentage of callus induction at 97% was obtained when seeds of rice (Oryza sativa L. cv. RD6 were cultured on modified N6 medium supplemented with 3% (w/v sucrose, 22.5 μM 2,4-D and 0.8% agar under light condition. The suitable regeneration medium was N6 medium supplemented with 3% (w/v sucrose, 2.5 μM IAA, 18 μM BA and 0.8% agar. A test had been performed to determine the effect of antibiotics on the regeneration of rice cv. RD6. It was found that kanamycin concentration up to 150 mg l-1 and hygromycin concentration at 10 mg l-1 were effective for selection of transformants. Cefotaxime and carbenicillin concentration up to 250 mg l-1 had the highest phytotoxicity to plant regeneration. Agrobacterium-mediated gene transfer protocols for rice cv. RD6 were performed using A. tumefaciens strain LBA4404, which harbored the plasmid pBI121 containing genes for β- glucuronidase (GUS and kanamycin resistance (nptII, and strain EHA105, which harbored plasmid pCAMBIA1301 containing genes for β-glucuronidase (GUS and hygromycin resistance (hptII. GUS activities were found in rice calli after co-cultivation. A number of morphologically normal fertile transgenic rice plants were obtained. Stable integration, expression and inheritance of transgenes were demonstrated by molecular and genetic analysis of transgenic plants in T0 and T1 generation. Mendelian segregation was observed in T1 progeny.

  16. MARCADORES DE PATOGENICIDADE EM Yersinia enterocolitica O: 3 ISOLADAS DE SUÍNOS DO RIO DE JANEIRO Genetic markers of pathogenicity in Yersinia enterocolitica O: 3 isolated from healthy pigs from Rio de Janeiro

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    Tereza C. A. Leal


    Full Text Available Foi realizada a caracterização genotípica e fenotípica de fatores de patogenicidade em 16 amostras de Yersinia enterocolitica O:3 isoladas de suínos sadios do Rio de Janeiro. Foi observado que apenas 6 cepas possuíam o plasmídio de virulência, pYV (+ 70 kb e apresentavam dependência ao cálcio no meio MOX a 37C. Um plasmídio críptico de cerca de 8,6 kb foi encontrado em uma cepa. Doze cepas revelaram sensibilidade à pesticina enquanto que apenas três se revelaram capazes de hidrolisar a esculina. Através de PCR com "primers" específicos, foi constatada a presença dos genes ail em 14 cepas, irp2, em 1 cepa e a ausência de psaA em todas as cepas analisadas. Quanto aos quimioterápicos, a quase totalidade das cepas mostrou-se ao mesmo tempo resistente à ampicilina e carbenicilina e sensível ao sulfazotrin e à cefoxitina. As respostas foram variadas frente ao cloranfenicol, tetraciclina, kanamicina, gentamicina e ácido nalidíxo.Sixteen Yersinia enterocolitica serotype O:3 strains, isolated from pigs from Rio de Janeiro, have been analyzed for genetic and phenotypic markers of pathogenicity. It was observed that only 6 strains harbored the pYV (+70 kb plasmid and one strain harbored a small cryptic plasmid of about 8.6 kb. Accordingly only strains harboring pYV were calcium dependent in the MOX medium at 370C. Twelve strains showed pesticin sensitivity and the esculin reaction was negative in 13 strains. PCR analysis of pathogenicity genes using specific primers showed the presence of the ail gene in 14 strains, the irp2 gene in one and the psaA in none. Most of the strains were resistant to ampicillin and carbenicillin, although they were susceptible to sulfazotrin and cefoxitin. For chloramphenicol, tetracycline, kanamycin, gentamicin and nalidixic acid the results varied among the strains.


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    Ahmed M.A. Mansour


    Full Text Available Aeromonas species are increasingly recognized as enteric pathogens. Faecal samples from 20 cow, 45 sheep; 60 goat and 60 camels were examined for the presence of Aeromonas species, which was also sought in the available drinking water (55 well water and 52 drinking chlorinated tap water were also examined. Aeromonas species was isolated more frequently from goats (21.7% than from other animal groups sampled and isolated more frequently from well water (38.2% than chlorinated supplies (23.0%. A. hydrophilia was the most dominant species isolated from different kinds of samples (13.4%. Whereas A. sobria and A. caviae were isolated in much lower rates 4.7 and 2.1% respectively. There was significant association between the isolation of Aeromonas species from all animal faeces and its presence in drinking water. All isolated strains were examined for the characteristics that are reputed to have roles in pathogenicity. The data reported in this study indicates that the distributions of virulence factors, that regulate the pathogenicity of Aeromonads, are different in clinical and enviromental samples. Aeromonas isolates exhibited multi-drug resistanc amoxicillin, carbenicillin and ampicillin. The most potent antibiotics against Aeromonas species isolated in this study were ceftriaxone, ceftazidime, cefotaxime, cefepime. Essential oils have been tested for in vitro and in vivo antimicrobial activity. Clove, Olive and Peppermint oil exhibited a wide spectrum of antimicrobial activity against all strains used in this study, showed a zone of inhibition ranging from 10.00±0.8 to 14.82±0.41 mm in diameter. Minimum Inhibitory Concentration (MIC for selected oils ranged from 12.8 to 25.6 mg mL-1. Treatment of mice with essential oil for 15 days led to enhance antibody levels in all treated groups and significant clearance of A. hydrophilia from animals. The treated animals had

  18. Isolation and Identification of Aeromonas veronii from the Obscure Puffer%暗纹东方鲀致病性维氏气单胞菌的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    常燕; 王兆慧; 刘思园; 徐磊; 曹军


    A bacteria strain was isolated from the obscure puffer (Takifugu obscurus),morphology,physiology and biochemistry,16 S rDNA polymerase chain reaction (PCR) amplification and artificially infection of health obscure puffer and ICR mice were determined.The results showed that the medianlethal dose of this bacteria to mice and obscure puffer were 2.26×106 and 5.96×104 CFU/mL,respectively.The 16 S rDNA sequence showed high similarity with Aeromonas veronii,plus the morphological,physiology and biochemistry data,this strian was determined as A.veronii,and named as NT-HV.Then,the sensitivity of this strian was tested using nine common aquatic drugs.The results showed that NT-HV was sensitive to florfenicol,trimethoprim-sulfamethoxazole,neomycin,oxygen piperacilliny,and resisted to tetracyclics,doxycycline,carbenicillin,ampicillin,suggestting the reasonable drug concentration and alternative using was necessary.%从濒死的暗纹东方鲀(Takifugu obscurus)体内分离到一株优势菌株,经形态观察、生化、16 S rDNA聚合酶链式反应(PCR)以及人工感染健康暗纹东方鲀和ICR小鼠等试验可知,该菌对小鼠和暗纹东方鲀半数致死量(LD卯)分别为2.26×106 CFU/mL、5.96×104 CFU/mL;其16 S rDNA序列与维氏气单胞菌具有高度的同源性,根据该菌的形态特征、生理生化特性,初步鉴定该菌为维氏气单胞菌(Aeromonas veronii),命名为NT-HV.选择9种水产常用药物进行药敏试验,结果显示该菌对氟苯尼考、复方新诺明、新霉素、氧哌嗪青霉素敏感,对养殖场常用的四环素、强力霉素、羧苄青霉素、氨苄青霉素不敏感,提示在药物的使用过程中应注意药物的浓度和合理轮换.

  19. 猪大肠杆菌和葡萄球菌的分离鉴定及耐药性分析%Isolation, Identification and Analysis of Drug Resistance on E.coli and Staphylococcus in Pigs

    Institute of Scientific and Technical Information of China (English)

    李淑红; 王京仁; 成钢; 曾文虎


    对湘西北地区某猪场患病的仔猪的粪便及病变的脾脏、肝脏、淋巴结、肺脏、肾水肿液进行病原菌分离鉴定,并用Kirby-Bauer法测定分离菌株对15种抗菌药的敏感性.结果表明,患病仔猪分离到大肠杆菌和猪葡萄球菌两种细菌.其中大肠杆菌对庆大霉素、头孢呋肟、丁胺卡那霉素、四环素、先锋必素敏感,对氯霉素、氧哌嗪青霉素、先锋五号、头孢曲松钠、新霉素中介,对羧苄青霉素、强力霉素、青霉素、氨苄青霉素、红霉素有较高的耐药性.葡萄球菌对先锋霉素V、丁胺卡那霉素敏感,对头孢曲松钠中介,对其他抗生素耐药.其中大肠杆菌对药物敏感率、中介率、耐药率均为33.33%;葡萄球菌的敏感率为13.33%,中介率为6.67%,耐药率为80.00%.%he pathogen were isolated and identified on feces of sick piglets and disease of the spleen, liver, lymph nodes, lung, kidney edema fluidin a pig farm in Hunan northwestern region. Susceptibility of 15 kinds antimicrobial drug was determined with the Kirby-Bauer for isolated strains. The results showed piglets were infected with E. coli and Staphylococcus. Escherichia coli was sensitive to gentamicin, cefuroxime, Amikacin, tetracycline, cefoperazone sodium, to chloramphenicol, piperacillin, pioneer V, ceftriaxone, neomycin was intermediate, to carbenicillin, doxycycline, penicillin, ampicillin, erythro-mycin had a higher resistance. Staphylococcus was sensitive to pioneer V, Amikacin, to ceftriaxone was intermediate, on the other antibiotic was resistance. Which was 33.33% the rate of drug sensitivity, intermediate and resistance to E. coli. Which was 13. 33% sensitive rate, intermediate rate was 6. 67%, resistance rate was 80. 00% to Staphylococcus.

  20. 农杆菌介导的黄瓜未受精子房培养遗传转化体系的建立%Transformation System Through Agrobacterium tumefaciens Mediated Unfertilized Ovary Culture in Cucumber

    Institute of Scientific and Technical Information of China (English)

    魏爱民; 杜胜利; 韩毅科; 刘楠


    为了建立一种直接获得转基因纯系的转化方法,首次开展了农杆菌介导的黄瓜未受精子房培养遗传转化体系研究。试验以除草剂草胺膦为选择标记,胚胎发生阶段,最适除草剂质量浓度为0.5~1.0 mg·L-1,幼苗生根阶段除草剂质量浓度为2.0 mg·L-1;以羧苄青霉素为抑菌剂,最适抑菌质量为450 mg·L-1;最佳浸染方法为OD600=0.3农杆菌工程菌,浸染10 min;共培养时间为3 d;在浸染菌液和共培养基中加入0.01%的silwet-77,有利于获得较多抗性胚胎及抗性植株。该遗传转化体系的建立,为黄瓜转基因方法研究开辟了新途径。%A new transformation technology through Agrobacterium tumefaciens mediated cucumber unfertilized ovary cul-ture was studied. Herbicide glufosinate-ammonium was used as the selection marker,the optimum herbicide selection pres-sure during embryogenesis was 0.5-1.0 mg·L-1,and optimum selection pressure during plantlet rooting was 2.0 mg·L-1;the optimum carbenicillin concentration for bacteria control was 450 mg · L-1;the optimum infection method was soaking ovary slices into A. tumefaciens solution with OD600 at 0.3 for 10 min,and then co-cultured for 3 days;0.01%surfactant silwet-77 in bacteria solution and co-culture medium was beneficial for transformation. The establishment of this new technology pro-vided a new way for cucumber transformation research.

  1. Aeromonas associated diarrhoeal disease in south Brazil: prevalence, virulence factors and antimicrobial resistance Aeromonas associadas a diarréias no sul do Brasil: prevalência, fatores de virulência, e resistência a antibiótico

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    Ivani M.F. Guerra


    Full Text Available Aeromonas were isolated from 27 (6.6% of 408 patients admitted with acute gastroenteritis in two hospitals at Rio Grande do Sul, Brazil. Isolates were classified as A. hydrophila (51.8%, A. caviae (40.8%, and A. veronii biotype sobria (7.4%. The highest prevalence of Aeromonas associated infections occurred in lactants and children. Virulence genes (aerA -aerolysin/hemolysin, ahpA -serine-protease, satA - glycerophospholipid-cholesterol acyltransferase, lipA -lipase, and ahyB -elastase and virulence factors (hemolytic, proteolitic, lipolitic activities, and biofilm formation were identified in most A. hydrophila and A. veronii biotype sobria isolates, with lower frequencies on A. caviae. All Aeromonas isolates were resistant to ampicillin, ticarcillin/clavulanic acid, cephalotin, and cephazolin, and most of them (>70% exhibited resistance to imipenem, carbenicillin, amoxillin/sulbactan, and piperacillin. Multiple-resistance, more than four antibiotics, was evidenced in 29.6% of the isolates. The most efficient antibiotics were the quinolones (ciprofloxacin and norfloxacin, and the aminoglycosides (amikacin and netilmicin.Aeromonas foram isoladas de 27 (6.6% dos 408 pacientes admitidos com gastroenterite aguda em dois hospitais do Rio Grande do Sul, Brasil. Os isolados foram classificados com A. hydrophila (51.8%, A. caviae (40.8%, e A. veronii biotype sobria (7.4%. A maior prevalência de Aeromonas ocorreu em lactantes e crianças. Genes (aerA -aerolisina/hemolisina, ahpA -serina-protease, satA - glicerofosfolipidio-colesterol aciltransferase, lipA -lipase, e ahyB -elastase e factores (atividade hemolítica, proteolítica, lipolítica, e formação de biofilme de virulência foram identificados na maioria dos isolados de A. hydrophila e A. veronii biotype sobria, com freqüências menores em A. caviae. Todos os isolados de Aeromonas apresentaram resistência a ampicilina, ticarcilina/ácido clavulânico, cefalotina e cefazolina, e a maior parte

  2. Virulence and antimicrobial resistance of Escherichia coli isolated from Tigris River and children diarrhea

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    Ibrahim IA


    Full Text Available Israa AJ Ibrahim, Rana M Al-Shwaikh, Mahmoud I IsmaeilDepartment of Biology, College of Education for Pure Science, Ibn Al-Haitham, University of Baghdad, Baghdad, IraqObjective: To investigate the virulence factors including hemolysin production, β-lactamase production, and biofilm formation. Antimicrobial resistance and plasmid content of 20 Escherichia coli isolates obtained from feces and Tigris water were screened.Methods: Ten clinical and ten environmental E. coli isolates were collected from children diarrhea and swim areas on Tigris River in Baghdad city, Iraq, respectively. The bacterial isolates were identified by cultural characteristics, Gram stain, biochemical tests, and screened for the presence of E. coli O157:H7 serotype. Bacterial E. coli isolates were investigated for hemolysin production, biofilm formation, and β-lactamase production. Antibiotics susceptibility and plasmid content were determined.Results: A total of ten clinical and ten water E. coli isolates were studied. Results showed that all E. coli isolates give negative results for latex O157:H7. Virulence factors analysis showed that 6/10 water isolates and 2/10 clinical isolates were hemolytic, 5/10 water isolates and 3/10 clinical isolates were biofilm formation, and 7/10 water isolates and 4/10 clinical isolates were β-lactamase producer. Antibiotics profile showed that all bacterial isolates were multidrug resistant. All E. coli isolates (100% were resistant to carbenicillin, cefodizime, imipenem, and piperacillin. The plasmid DNA analysis showed that all E. coli isolates contained plasmid with molecular weight range between 4.507 kbp and 5.07 kbp, but clinical isolates contained multiple small and mega plasmids.Conclusion: Our study revealed that E. coli isolates from river water exhibit a higher level of hemolysin production, β-lactamase production, and biofilm formation than feces isolates may be due to long adaptation. On the other hand, clinical E. coli

  3. Generation of a Pseudomonas aeruginosa elastase gene targeted deletion mutant by Red recombination system%采用Red重组系统敲除铜绿假单胞菌弹性蛋白酶基因

    Institute of Scientific and Technical Information of China (English)

    余华; 熊浚智; 何晓梅; 盛哈蕾; 蔡文强; 谢玮; 张克斌


    The aim of this study is to obtain the elastase activity negative strain by knocking out the elastase gene in Pseudomonas aeruginosa PAO1. Three genes of Red recombination system from λ phage were amplified and cloned into Esche-richia-Pseudomonas shuttle vector pUCP, and the pUCP-Red vector was transformed into PAO1 competent cells by electropo-ration. Then the recombinant DNA fragment which contains gentamycin antibiotic cassette flanked by two 80-bp homology sequences of elastase gene upstream and downstream locuses respectively was obtained by conventional cloning methods. And the fragment was electroporated into PAOl/pUCP-Red competent cells and screened on LB plate containing gentamycin and carben-icillin. The elastase gene knocked-out strain was verified by the methods of PCR, RT-PCR and enzyme activity assays. The elastase activity negative strain was successfully obtained in this study by using the Red recombination system. The elastase gene knocked-out strain obtained in this study provides the basis and materials for systemic study of pathogenicity mechanism of elastase in Pseudomonas aeruginosa.%目的 敲除铜绿假单胞菌弹性蛋白酶基因,获得无弹性蛋白酶活性的铜绿假单胞菌菌株.方法 采用PCR从pKD46质粒上扩增λ噬菌体的Red重组酶基因,并将其克隆到大肠杆菌和铜绿假单胞菌穿梭质粒pUCP多克隆位点上,电击转化铜绿假单胞菌PAO1感受态细胞,构建PAO1/pUCP-Red基因敲除体系.常规基因操作构建两端与弹性蛋白酶基因上、下游同源,中间为庆大霉素抗性基因的线性打靶片段;并将其电击转化pUCP-Red/PAO1感受态;采用庆大霉素和羧苄青霉素抗性平板初步筛选阳性重组菌;通过PCR、RT-PCR及弹性蛋白酶活性检测方法,鉴定菌株弹性蛋白酶基因的敲除情况.结果 本研究通过构建Red重组系统,获得了无弹性蛋白酶活性的铜绿假单胞菌菌株.结论 本研究成功敲除了铜绿假单胞菌弹性蛋

  4. 氨苄青霉素单克隆抗体及其与青霉素类抗生素交叉反应性%Anti-ampicillin Monoclonal Antibodies and Their Cross-reactivity to Penicillin Antibiotics

    Institute of Scientific and Technical Information of China (English)

    李青梅; 刘庆堂; 王寅彪; 柴书军; 王自良; 郭军庆; 职爱民; 孟红丽; 张改平


    Ampicillin ( Amp) was coupled to carrier proteins of bovine serum albumin ( BSA ) and ovalbumin (OVA) to synthetize immunogen BSA-Amp and detection antigen OVA-Amp. Four hybridoma cell lines secreting monoclonal antibodies (mAbs) specific for Amp were established by immunizing BALB/c mice with BSA-Amp u-sing hybridoma technology. The antibody titers of mAb ascites (1G7, 2A11, 2D10 and 3F6) were determined as 1:2.56×107, 1:1.28×107, 1 :5.12 × 106 and 1:5.12 × 106 in ELISA, and their half maximal inhibitory concentration (IC50) of Amp were 10.22, 3.58, 4.03,4.95 ng/mL in the inhibitive ELISA, respectively. The Amp mAbs showed significant cross-reactivity to the penicillin antibiotics of carbenicillin (1G7, 2A11, 2D10 and 3F6 ) , penicillin G (1G7 and 3F6) , and amoxicillin (1G7) , whereas no obvious cross-reactivity to dicloxacillin or clox-acillin was found.%以戊二醛法将氨苄青霉素(Ampicillin,Amp)偶联于牛血清白蛋白(BSA)和卵清白蛋白(OVA),合成免疫抗原BSA-Amp和检测抗原0VA-Amp,并进行了鉴定;用BSA-Amp免疫BALB/c小鼠,应用杂交瘤技术建立了4株分泌抗Amp单克隆抗体的杂交瘤细胞株.ELISA和竞争ELISA结果显示,单克隆抗体1G7、2A11、2D10和3F6的腹水抗体效价分别为1:2.56×107、1:1.28×107、1:5.12 ×106和1:5.12×106,Amp的半数抑制浓度(IC50)分别为10.22,3.58,4.03,4.95 ng/mL.Amp单克隆抗体与羧苄青霉素(1G7、2A11、2D10和3F6)、青霉素G(1G7和3F6)、阿莫西林(1G7)等青霉素类抗生素存在显著的交叉反应,而与双氯霉素和邻氯霉素未见明显交叉反应.


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    pneumoniae, Staphylococcus hominis, Candida spp. (5,4±3,0%. 2. The most effective antibacterial drug is meropenem. 78,7% of all isolates have sensitivity to it. The sensitivity to gatifloxacin have 73,7%; to tigecycline 71,1%; to amikacin – 67,3%, to moxifloxacin – 59,6% of cultures, to two agents (levofloxacin and pefloxacin – 55,8% of cultures respectively. 3. 100% of the family Micrococcaceae isolates were resistant to benzylpenicillin, 95,8% - to ampicillin, 87,5% - to amoxicillin and cefuroxime, 83,3% - to azithromycin, erythromycin and lincomycin. Members of the Streptococcaceae family were resistant to cefuroxime. 100% of the family Enterobacteriaceae isolates had no sensitivity to carbenicillin, 87,5% of isolates - to cefuroxim and gentamicin, 81,2% - to amoxicillin and ampicillin, 75,0% - to cefazolin, 50,0% - to ceftriaxonum and aztreonam. 4. Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter haemolyticus isolates were multi-drug resistant.