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Sample records for aeromonas veronii plasmid

  1. Small Protein B upregulates sensor kinase bvgS expression in Aeromonas veronii

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    Zhu eLiu

    2015-06-01

    Full Text Available Earlier studies reveal that Small Protein B (SmpB, a class of well-conserved tmRNA-binding proteins, is essential for the trans-translation process, which functions as a system for translation surveillance and ribosome rescue. Here, we report a previously unrecognized mechanism by which SmpB alone positively regulates the expression of a sensor kinase, BvgS, in Aeromonas veronii. A reporter plasmid was constructed in which the promoter of bvgS was used to control the expression of the enhanced green fluorescent protein (eGFP gene. When the reporter plasmid was co-transformed with a SmpB expression construct into E. coli, the relative fluorescence intensity increased about 3-fold. Transformation with a truncated form of smpB gene showed that the C-terminus had little effect, while N-terminus unexpectedly increased eGFP production. Next, a series of SmpB mutants were generated by site-directed mutagenesis. When the mutants SmpB (G11S or SmpB (E32AG was used in the experiment, eGFP expression dropped significantly compared with that of wild type SmpB. Further,purified SmpB was shown to bind the promoter regions of bvgS in the agarose gel retardation assay. Quantitative RT-PCR analysis showed that eGFP transcript levels increased approximately 25-fold in the presence of SmpB. Likewise, bvgS transcripts decreased significantly in smpB knockout A. veronii. Similar to BvgS inhibition, smpB knockout in A. veronii displayed a reduced capability in salt tolerance. Collectively, the data presented here will facilitate a deeper understanding of SmpB-mediated regulatory circuits as a transcriptional factor in A. veronii.

  2. Pan-genome analysis of Aeromonas hydrophila, Aeromonas veronii and Aeromonas caviae indicates phylogenomic diversity and greater pathogenic potential for Aeromonas hydrophila.

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    Ghatak, Sandeep; Blom, Jochen; Das, Samir; Sanjukta, Rajkumari; Puro, Kekungu; Mawlong, Michael; Shakuntala, Ingudam; Sen, Arnab; Goesmann, Alexander; Kumar, Ashok; Ngachan, S V

    2016-07-01

    Aeromonas species are important pathogens of fishes and aquatic animals capable of infecting humans and other animals via food. Due to the paucity of pan-genomic studies on aeromonads, the present study was undertaken to analyse the pan-genome of three clinically important Aeromonas species (A. hydrophila, A. veronii, A. caviae). Results of pan-genome analysis revealed an open pan-genome for all three species with pan-genome sizes of 9181, 7214 and 6884 genes for A. hydrophila, A. veronii and A. caviae, respectively. Core-genome: pan-genome ratio (RCP) indicated greater genomic diversity for A. hydrophila and interestingly RCP emerged as an effective indicator to gauge genomic diversity which could possibly be extended to other organisms too. Phylogenomic network analysis highlighted the influence of homologous recombination and lateral gene transfer in the evolution of Aeromonas spp. Prediction of virulence factors indicated no significant difference among the three species though analysis of pathogenic potential and acquired antimicrobial resistance genes revealed greater hazards from A. hydrophila. In conclusion, the present study highlighted the usefulness of whole genome analyses to infer evolutionary cues for Aeromonas species which indicated considerable phylogenomic diversity for A. hydrophila and hitherto unknown genomic evidence for pathogenic potential of A. hydrophila compared to A. veronii and A. caviae. PMID:27075453

  3. Characterization of Virulence Properties of Aeromonas veronii Isolated from Diseased Gibel Carp (Carassius gibelio

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    Jingjing Sun

    2016-04-01

    Full Text Available Aeromonas veronii is a kind of opportunistic pathogen to fish and humans, significantly impending aquaculture production. Recently, we isolated two A. veronii strains, named GYC1 and GYC2, from diseased Gibel carp (Carassius gibelio in China. Based on gyrB (DNA gyrase B subunit genes of GYC1 and GYC2, the constructed phylogenetic tree showed that the two strains were clustered with A. veronii. Sixteen virulence genes related to the pathogenicity of Aeromonas spp. were subjected to PCR assay. The genes of ompAI, ompAII, lafA, act, aer, fla, gcaT and acg were detected in the two strains, while genes of hly, ahp, lip, ast and alt were not detected. Additionally, genes eprCAI, ela and exu were only detected in the strain GYC1. Furthermore, the results of extracellular enzyme analysis revealed that the two isolates can produce hemolysin, caseinase, esterase, amylase and lecithinase, which were closely related to the pathogenicity of the two strains. However, the results showed that there was no gelatinase activity in either strain. According to the antibiotic resistant assay, the two strains were sensitive to cephalosporins and aminoglycosides, while they were resistant to penicillins and quinolones. Through this study, the virulence characteristics, including virulence genes and extracellular enzymes, the pathogenicity of A. veronii was clarified, enhancing the understanding about this pathogenic bacterium and providing the theoretical basis in disease control.

  4. Characterization of Virulence Properties of Aeromonas veronii Isolated from Diseased Gibel Carp (Carassius gibelio).

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    Sun, Jingjing; Zhang, Xiaojun; Gao, Xiaojian; Jiang, Qun; Wen, Yi; Lin, Li

    2016-01-01

    Aeromonas veronii is a kind of opportunistic pathogen to fish and humans, significantly impending aquaculture production. Recently, we isolated two A. veronii strains, named GYC1 and GYC2, from diseased Gibel carp (Carassius gibelio) in China. Based on gyrB (DNA gyrase B subunit) genes of GYC1 and GYC2, the constructed phylogenetic tree showed that the two strains were clustered with A. veronii. Sixteen virulence genes related to the pathogenicity of Aeromonas spp. were subjected to PCR assay. The genes of ompAI, ompAII, lafA, act, aer, fla, gcaT and acg were detected in the two strains, while genes of hly, ahp, lip, ast and alt were not detected. Additionally, genes eprCAI, ela and exu were only detected in the strain GYC1. Furthermore, the results of extracellular enzyme analysis revealed that the two isolates can produce hemolysin, caseinase, esterase, amylase and lecithinase, which were closely related to the pathogenicity of the two strains. However, the results showed that there was no gelatinase activity in either strain. According to the antibiotic resistant assay, the two strains were sensitive to cephalosporins and aminoglycosides, while they were resistant to penicillins and quinolones. Through this study, the virulence characteristics, including virulence genes and extracellular enzymes, the pathogenicity of A. veronii was clarified, enhancing the understanding about this pathogenic bacterium and providing the theoretical basis in disease control. PMID:27043558

  5. Characterization of Virulence Properties of Aeromonas veronii Isolated from Diseased Gibel Carp (Carassius gibelio)

    Science.gov (United States)

    Sun, Jingjing; Zhang, Xiaojun; Gao, Xiaojian; Jiang, Qun; Wen, Yi; Lin, Li

    2016-01-01

    Aeromonas veronii is a kind of opportunistic pathogen to fish and humans, significantly impending aquaculture production. Recently, we isolated two A. veronii strains, named GYC1 and GYC2, from diseased Gibel carp (Carassius gibelio) in China. Based on gyrB (DNA gyrase B subunit) genes of GYC1 and GYC2, the constructed phylogenetic tree showed that the two strains were clustered with A. veronii. Sixteen virulence genes related to the pathogenicity of Aeromonas spp. were subjected to PCR assay. The genes of ompAI, ompAII, lafA, act, aer, fla, gcaT and acg were detected in the two strains, while genes of hly, ahp, lip, ast and alt were not detected. Additionally, genes eprCAI, ela and exu were only detected in the strain GYC1. Furthermore, the results of extracellular enzyme analysis revealed that the two isolates can produce hemolysin, caseinase, esterase, amylase and lecithinase, which were closely related to the pathogenicity of the two strains. However, the results showed that there was no gelatinase activity in either strain. According to the antibiotic resistant assay, the two strains were sensitive to cephalosporins and aminoglycosides, while they were resistant to penicillins and quinolones. Through this study, the virulence characteristics, including virulence genes and extracellular enzymes, the pathogenicity of A. veronii was clarified, enhancing the understanding about this pathogenic bacterium and providing the theoretical basis in disease control. PMID:27043558

  6. Complete genome sequence of the fish pathogen Aeromonas veronii TH0426 with potential application in biosynthesis of pullulanase and chitinase.

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    Kang, Yuanhuan; Pan, Xiaoyi; Xu, Yang; Siddiqui, Shahrood A; Wang, Chunfeng; Shan, Xiaofeng; Qian, Aidong

    2016-06-10

    Aeromonas veronii TH0426 is a pathogen of the farmed yellow catfish Pelteobagrus fulvidraco but shows high-level expression of pullulanase and chitinase. Here, we present its genome sequence, which is the first reported complete genome of fish pathogen in A. veronii to date. Strain TH0426 harbors a single circular 4,923,009bp chromosome with a GC content of 58.25%. There are 4525 genes identified on its genome, including 4244 protein-coding genes, 32 rRNA genes, 120 tRNA genes, a noncoding RNA and 128 pseudo genes. We believe that the genomic information of A. veronii TH0426 would facilitate to reveal its pathogenic mechanism associated with yellow catfish, develop vaccine to decrease economic losses for fish farming, meanwhile explore the potential application in producing pullulanase and chitinase. PMID:27080448

  7. Humoral immune response and TLR9 gene expression in Pacific red snapper (Lutjanus peru) experimentally exposed to Aeromonas veronii.

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    Reyes-Becerril, Martha; Angulo, Carlos; Ascencio, Felipe

    2015-02-01

    Aquaculture production of Pacific red snapper Lutjanus peru is growing rapidly in Mexico, especially in Gulf of California. As it is a relatively new aquaculture species there are few reports evaluating its immune response to pathogens. The Gram-negative bacteria Aeromonas veronii is a heterogeneous organism that causes the disease known as motile aeromonad septicemia, which is responsible for serious economic loss in seabream culture due to bacterial infections. For the purpose of this study, juvenile Pacific red snapper specimens were intraperitoneally injected with low doses of A. veronii (1 × 10(6) CFU ml(-1)). Changes in humoral immune parameters (total protein, myeloperoxidase, lisozyme and antiprotease activities and IgM levels), as well as superoxide dismutase and catalase activities, and TLR9 gene expression were evaluated 24 and 48 h after injection. Overall, the results showed an enhanced in humoral immune parameters and SOD and CAT activities in fish infected with A. veronii compared with control group at 24 or 48 h. By real time PCR assays, the basal mRNA transcripts of TLR9 showed that were highly expressed in intestine and leucocytes compared to skin, head kidney, liver and gill. Then, the mRNA expression levels of TLR9 in head kidney, skin, liver and intestine were analyzed in non-infected and experimentally infected fish 24 and 48 h after injection. A. veronii up-regulated the expression of TLR9 at 24 or 48 h of exposure in all samples analyzed except in liver. Interestingly, intestine produced the greatest increase in transcript levels upon exposure (48 h) to A. veronii. Taken together, our results suggest that low doses of A. veronii infection inducing humoral immune system and TLR9 immune gene in Pacific red snapper that can be useful in the health control of this species. PMID:25462554

  8. Structural Studies of the Lipopolysaccharide from the Fish Pathogen Aeromonas veronii Strain Bs19, Serotype O16

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    Anna Turska-Szewczuk

    2014-03-01

    Full Text Available Chemical analyses, mass spectrometry, and NMR spectroscopy were applied to study the structure of the lipopolysaccharide (LPS isolated from Aeromonas veronii strain Bs19, serotype O16. ESI-MS revealed that the most abundant LPS glycoforms have tetra-acylated or hexa-acylated lipid A species, consisting of a bisphosphorylated GlcN disaccharide with an AraN residue as a non-stoichiometric substituent, and a core oligosaccharide composed of Hep5Hex3HexN1Kdo1P1. Sugar and methylation analysis together with 1D and 2D 1H and 13C NMR spectroscopy were the main methods used, and revealed that the O-specific polysaccharide (OPS of A. veronii Bs19 was built up of tetrasaccharide repeating units with the structure: →4-α-d-Quip3NAc-(1→3-α-l-Rhap-(1→4-β-d-Galp-(1→3-α-d-GalpNAc-(1→. This composition was confirmed by mass spectrometry. The charge-deconvoluted ESI FT-ICR MS recorded for the LPS preparations identified mass peaks of SR- and R-form LPS species, that differed by Δm = 698.27 u, a value corresponding to the calculated molecular mass of one OPS repeating unit (6dHexNAc6dHexHexHexNAc-H2O. Moreover, unspecific fragmentation spectra confirmed the sequence of the sugar residues in the OPS and allowed to assume that the elucidated structure also represented the biological repeating unit.

  9. Improvement of methyl orange dye biotreatment by a novel isolated strain, Aeromonas veronii GRI, by SPB1 biosurfactant addition.

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    Mnif, Inès; Maktouf, Sameh; Fendri, Raouia; Kriaa, Mouna; Ellouze, Semia; Ghribi, Dhouha

    2016-01-01

    Aeromonas veronii GRI (KF964486), isolated from acclimated textile effluent after selective enrichment on azo dye, was assessed for methyl orange biodegradation potency. Results suggested the potential of this bacterium for use in effective treatment of azo-dye-contaminated wastewaters under static conditions at neutral and alkaline pH value, characteristic of typical textile effluents. The strain could tolerate higher doses of dyes as it was able to decolorize up to 1000 mg/l. When used as microbial surfactant to enhance methyl orange biodecolorization, Bacillus subtilis SPB1-derived lipopeptide accelerated the decolorization rate and maximized slightly the decolorization efficiency at an optimal concentration of about 0.025%. In order to enhance the process efficiency, a Taguchi design was conducted. Phytotoxicity bioassay using sesame and radish seeds were carried out to assess the biotreatment effectiveness. The bacterium was able to effectively decolorize the azo dye when inoculated with an initial optical density of about 0.5 with 0.25% sucrose, 0.125% yeast extract, 0.01% SPB1 biosurfactant, and when conducting an agitation phase of about 24 h after static incubation. Germination potency showed an increase toward the nonoptimized conditions indicating an improvement of the biotreatment. When comparing with synthetic surfactants, a drastic decrease and an inhibition of orange methyl decolorization were observed in the presence of CTAB and SDS. The nonionic surfactant Tween 80 had a positive effect on methyl orange biodecolorization. Also, studies ensured that methyl orange removal by this strain could be due to endocellular enzymatic activities. To conclude, the addition of SPB1 bioemulsifier reduced energy costs by reducing effective decolorization period, biosurfactant stimulated bacterial decolorization method may provide highly efficient, inexpensive, and time-saving procedure in treatment of textile effluents.

  10. A Tale of Transmission: Aeromonas veronii Activity within Leech-Exuded Mucus.

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    Ott, Brittany M; Dacks, Andrew M; Ryan, Kenneth J; Rio, Rita V M

    2016-05-01

    Transmission, critical to the establishment and persistence of host-associated microbiotas, also exposes symbionts to new environmental conditions. With horizontal transmission, these different conditions represent major lifestyle shifts. Yet genome-wide analyses of how microbes adjust their transcriptomes toward these dramatic shifts remain understudied. Here, we provide a comprehensive and comparative analysis of the global transcriptional profiles of a symbiont as it shifts between lifestyles during transmission. The gammaproteobacteriumAeromonas veroniiis transmitted from the gut of the medicinal leech to other hosts via host mucosal castings, yetA. veroniican also transition from mucosal habitancy to a free-living lifestyle. These three lifestyles are characterized by distinct physiological constraints and consequently lifestyle-specific changes in the expression of stress-response genes. Mucus-boundA. veroniihad the greatest expression in terms of both the number of loci and levels of transcription of stress-response mechanisms. However, these bacteria are still capable of proliferating within the mucus, suggesting the availability of nutrients within this environment. We found thatA. veroniialters transcription of loci in a synthetic pathway that obtains and incorporatesN-acetylglucosamine (NAG; a major component of mucus) into the bacterial cell wall, enabling proliferation. Our results demonstrate that symbionts undergo dramatic local adaptation, demonstrated by widespread transcriptional changes, throughout the process of transmission that allows them to thrive while they encounter new environments which further shape their ecology and evolution. PMID:26896136

  11. Identification and Characterization of Pathogenic Aeromonas veronii Biovar Sobria Associated with Epizootic Ulcerative Syndrome in Fish in Bangladesh

    OpenAIRE

    Rahman, Mokhlasur; Colque-Navarro, Patricia; Kühn, Inger; Huys, Geert; Swings, Jean; Möllby, Roland

    2002-01-01

    Sparse information is available on the virulence factors of Aeromonas strains isolated from diseased fish, from the environment, and from humans. In the present study, 52 Aeromonas isolates obtained from epizootic ulcerative syndrome (EUS) lesions in fish, from the aquatic environment, and from children with diarrhea in Bangladesh were identified by biochemical phenotyping (i.e., PhenePlate [PhP] typing) and DNA fingerprinting and then characterized with respect to certain putative virulence ...

  12. Susceptibility to antimicrobial agents and plasmid carrying in Aeromonas hydrophila isolated from two estuarine systems.

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    Montoya, R; Dominguez, M; Gonzalez, C; Mondaca, M A; Zemelman, R

    1992-01-01

    Susceptibility to various antimicrobial agents and the presence of plasmids was investigated in eleven strains of Aeromonas hydrophila isolated from samples of sea water and these strains isolated from Aulacomya ater. Transference of resistance to Escherichia coli was attempted by conjugation and transformation experiments. The strains showed multiple resistance toward beta-lactam antibiotics and susceptibility to other antimicrobial agents. Five strains harboured plasmids with molecular weights below 5.7 MD. It was not possible to relate the resistance of the strains with the presence of their plasmids. PMID:1593967

  13. Class D tetracycline resistance determinants of R plasmids from the fish pathogens Aeromonas hydrophila, Edwardsiella tarda, and Pasteurella piscicida.

    OpenAIRE

    Aoki, T.; Takahashi, A.

    1987-01-01

    Tetracycline resistance determinants of R plasmids from the fish pathogens Aeromonas hydrophila, Edwardsiella tarda, and Pasteurella piscicida were classified as class D by their expression of resistance to tetracycline and minocycline and through their DNA structure.

  14. Draft genome sequences of two Aeromonas salmonicida subsp. salmonicida isolates harboring plasmids conferring antibiotic resistance.

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    Vincent, Antony T; Tanaka, Katherine H; Trudel, Melanie V; Frenette, Michel; Derome, Nicolas; Charette, Steve J

    2015-02-01

    The bacterium Aeromonas salmonicida is the etiological agent of furunculosis, a widespread fish disease causing important economic losses to the fish farming industry. Antibiotic treatments in fish farms may be challenging given the existence of multidrug-resistant isolates of this bacterium. Here, we report the draft genome sequences of the 2004-05MF26 and 2009-144K3 isolates, which harbor plasmids conferring antibiotic resistance. Both isolates also carry the large plasmid pAsa5, which is known to encode a type three secretion system (TTSS) and the pAsal1 plasmid which has the aopP gene producing a TTSS effector. These two isolates are good representatives of the plasmid diversity in A. salmonicida subsp. salmonicida. PMID:25724776

  15. 狐狸源致病性维氏气单胞菌的分离鉴定及耐药性分析%lsolation and identification of pathogenic Aeromonas veronii from fox

    Institute of Scientific and Technical Information of China (English)

    李伟杰; 赵耘; 刘燕; 岂晓鑫; 康凯; 陈敏

    2012-01-01

    A Aeromonas-like bacteria isolated from fox in Beijing zoo was identified with a polyphasic strategy, including the morphology, physical and chemical characteristics, 16S rRNA sequence analysis and pathogenicity. The results showed that the bacteria isolate was gram-negtive, and the culture character, physical and chemical characteristics were identical with A. Veronii, except sucrose and acetate reaction, In additionally, phylogenetic analysis based on 16S rRNA also indicated that the isolate was clustered in the genus Aeromonas, with the highest similarity to A. Veronii. Artificial infection test showed that the isolate was pathogenicity to CD-1 mice with a LD50 of 2.8ici5 CFU. The drug sensitivity test revealed that the isolate was sensitive to 14 kinds of drugs such as Ofloxaxin, Ceftazodime, and resistant to 9 kinds of drugs such as Penicillin G, Clindamycin.%本研究通过生物学特性、16S rRNA基因和4种等看家基因的序列测定与分析,对分离自北京动物园死亡狐狸病变组织的1株细菌进行鉴定,并对其进行小鼠致病力试验及药物敏感性试验.结果显示:该细菌为革兰氏阴性杆菌,培养特性、理化反应与维氏气单胞菌温和生物型基本相同;根据16S rRNA基因和dnaJ、rpoD、gyrB、cpn60等看家基因的系统进化及其同源性分析,确定所分离的细菌与维氏气单胞菌属于同一系统发育分支,各基因同源性最高分别为97.6%~99.9%.分离菌株对CD-1小鼠有较强的致病作用,其半数致死量为2.8×105 cfu/只;对氧氟沙星、头孢他啶等14种抗菌药物敏感,对青霉素G、克林霉素等9种抗菌药物耐药.

  16. Optimization of a plasmid electroporation protocol for Aeromonas salmonicida subsp. salmonicida.

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    Dallaire-Dufresne, Stéphanie; Emond-Rheault, Jean-Guillaume; Attéré, Sabrina A; Tanaka, Katherine H; Trudel, Mélanie V; Frenette, Michel; Charette, Steve J

    2014-03-01

    Aeromonas salmonicida subsp. salmonicida is a major fish pathogen. Molecular tools are required to study the virulence and genomic stability of this bacterium. An efficient electroporation-mediated transformation protocol for A. salmonicida subsp. salmonicida would make genetic studies faster and easier. In the present study, we designed the 4.1-kb pSDD1 plasmid as a tool for optimizing an electroporation protocol for A. salmonicida subsp. salmonicida. We systematically tested the electroporation conditions to develop a protocol that generates the maximum number of transformants. Under these optimal conditions (25 kV/cm, 200 Ω, 25 μF), we achieved an electroporation efficiency of up to 1×10(5) CFU/μg DNA. The electroporation protocol was also tested using another plasmid of 10.6-kb and three different strains of A. salmonicida subsp. salmonicida. The strains displayed significant differences in their electro-transformation competencies. Strain 01-B526 was the easiest to electroporate, especially with the pSDD1 plasmid. This plasmid was stably maintained in the 01-B526 transformants, as were the native plasmids, but could be easily cured by removing the selection conditions. This is the first efficient electroporation protocol reported for A. salmonicida subsp. salmonicida, and offers new possibilities for studying this bacterium.

  17. Diversity and homogeneity among small plasmids of Aeromonas salmonicida subsp. salmonicida linked with geographical origin

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    Sabrina A Attéré

    2015-11-01

    Full Text Available Furunculosis, which is caused by Aeromonas salmonicida subsp. salmonicida, is a major salmonid disease in fish farms worldwide. Several plasmids found in this bacterium confer phenotypes such drug resistance and virulence. Small plasmids (pAsa1, pAsa2, pAsa3, and pAsal1 related to ColE1- and ColE2-type replicons are usually present in its normal plasmidome. In the present study, with the objective to investigate if these plasmids display particularities related to the origin of the isolates bearing them, a total of 153 isolates, including 78 new and 75 previously described, were analyzed for the presence of small plasmids by PCR and DNA restriction fragment profiling. A geographical dichotomy between Canadian and European isolates for their propensity to do not have pAsa3 or pAsal1 was found. In addition, the genotyping analysis led to the identification of two European isolates harboring an unusual pAsal1. An investigation by next-generation sequencing (NGS of these two isolates shed light on two pAsal1 variants (pAsal1C and pAsal1D. As with pAsal1B, another pAsal1 variant previously described, these two new variants bore a second insertion sequence (ISAS5 in addition to the usual ISAS11. The characterization of these variants suggested that they could predominate over the wild-type pAsal1 in stressful conditions such as growth at temperatures of 25°C and above. To obtain a comprehensive portrait of the mutational pressure on small plasmids, 26 isolates whose DNA had been sequenced by NGS were investigated. pAsa3 and pAsal1 were more prone to mutations than pAsa1 and pAsa2, especially in the mobA gene, which encodes a relaxase and a primase. Lastly, the average copy number of each plasmid per cell was assessed using raw sequencing data. A clear trend with respect to the relative proportion per cell of each plasmid was identified. Our large-scale study revealed a geographical dichotomy in small plasmid repertoire in addition to a clear trend

  18. Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia.

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    Radu, Son; Ahmad, Noorlis; Ling, Foo Hooi; Reezal, Abdul

    2003-03-25

    A total of 87 market fish samples representing five types of fish were evaluated for the presence of Aeromonas spp. Of the samples examined, 69%, 55%, 11.5% and 2.3% harbored Aeromonas spp., A. veronii biovar sobria, A. hydrophila and A. caviae, respectively. The 60 isolated Aeromonas spp. strains were further examined for hemolytic activity, resistance to antimicrobial agents and presence of plasmids. Hemolytic activity varied widely among the isolated strains. Though all the isolates demonstrated resistance to three or more of the antibiotics tested, all were susceptible to ceptazidime. Thirty-four (56.7%) of the sixty isolates harbored plasmids, with sizes ranging from 2.3 to 15.7 kb. These results indicate that hemolytic, multiple antibiotic resistant and genetically diverse aeromonads are easily recovered from fish in this region. PMID:12485753

  19. Characterization of class 1 integrons associated with R-plasmids in clinical Aeromonas salmonicida isolates from various geographical areas

    DEFF Research Database (Denmark)

    Schmidt, A.S.; Bruun, Morten Sichlau; Larsen, J.L.;

    2001-01-01

    inserts. The gene cassettes tended to be conserved among isolates from a particular geographical area. Nineteen isolates transferred R- plasmids carrying different tet determinants to Escherichia coli in filter mating assays, and in 15 cases, the class 1 integrons were co-transferred. Transferable......Class 1 integrons were found in 26 of 40 antibiotic-resistant isolates of the fish pathogen Aeromonas salmonicida from Northern Europe and North America. Three different dhfr genes, conferring trimethoprim resistance, and one ant(3 " )1a aminoglycoside resistance gene were identified as gene...

  20. Complete Sequences of IncU Plasmids Harboring Quinolone Resistance Genes qnrS2 and aac(6')-Ib-cr in Aeromonas spp. from Ornamental Fish.

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    Dobiasova, Hana; Videnska, Petra; Dolejska, Monika

    2016-01-01

    The nucleotide sequences of three IncU plasmids from Aeromonas spp. isolated from ornamental fish are described. They had a typical IncU backbone for plasmid replication and maintenance functions, but conjugative transfer modules were disrupted. The gene qnrS2 was inserted into mpR as a mobile insertion cassette. Novel Tn3 family transposons carrying putative toxin-antitoxin and plasmid stability genes were identified. The study demonstrates high plasticity of IncU plasmids from aquatic environments. PMID:26525788

  1. Characterization of atypical Aeromonas salmonicida isolates by ribotyping and plasmid profiling

    DEFF Research Database (Denmark)

    Pedersen, Karl; Dalsgaard, Inger; Larsen, J.L.

    1996-01-01

    ) and plasmid profiles. Most epidemiologically unrelated strains had different ribotypes, whereas isolates from the same outbreak were identical. All strains, except one, carried one or more large plasmids (>55 kbp) and all strains, except two, additionally carried one or more smaller plasmids. Many strains...... isolated from the same outbreak showed different plasmid profiles although some plasmids were identical. The results suggest the existence of several atypical Aer, salmonicida. It also seems that ribotypes are stable properties for these bacteria while the plasmids are more labile....

  2. The mosaic architecture of Aeromonas salmonicida subsp. salmonicida pAsa4 plasmid and its consequences on antibiotic resistance

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    Tanaka, Katherine H.; Vincent, Antony T.; Trudel, Mélanie V.; Paquet, Valérie E.; Frenette, Michel

    2016-01-01

    Aeromonas salmonicida subsp. salmonicida, the causative agent of furunculosis in salmonids, is an issue especially because many isolates of this bacterium display antibiotic resistances, which limit treatments against the disease. Recent results suggested the possible existence of alternative forms of pAsa4, a large plasmid found in A. salmonicida subsp. salmonicida and bearing multiple antibiotic resistance genes. The present study reveals the existence of two newly detected pAsa4 variants, pAsa4b and pAsa4c. We present the extensive characterization of the genomic architecture, the mobile genetic elements and the antimicrobial resistance genes of these plasmids in addition to the reference pAsa4 from the strain A449. The analysis showed differences between the three architectures with consequences on the content of resistance genes. The genomic plasticity of the three pAsa4 variants could be partially explained by the action of mobile genetic elements like insertion sequences. Eight additional isolates from Canada and Europe that bore similar antibiotic resistance patterns as pAsa4-bearing strains were genotyped and specific pAsa4 variants could be attributed to phenotypic profiles. pAsa4 and pAsa4c were found in Europe, while pAsa4b was found in Canada. In accordance with their content in conjugative transfer genes, only pAsa4b and pAsa4c can be transferred by conjugation in Escherichia coli. The plasticity of pAsa4 variants related to the acquisition of antibiotic resistance indicates that these plasmids may pose a threat in terms of the dissemination of antimicrobial-resistant A. salmonicida subsp. salmonicida bacteria. PMID:27812409

  3. CHARACTERIZATION OF AEROMONAS VIRULENCE USING AN IMMUNOCOMPROMISED MOUSE MODEL

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    An immunocompromised mouse model was used to characterize Aeromonas strains for their ability to cause opportunistic, extraintestinal infections. A total of 34 isolates of Aeromonas (A. hydrophila [n = 12]), A. veronii biotype sobria [n = 7], A. caviae [n = 4], A. enchelia [n = 4...

  4. Dynamics of a Class 1 Integron Located on Plasmid or Chromosome in Two Aeromonas spp. Strains

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    Pérez-Valdespino, Abigail; Lazarini-Martínez, Alfredo; Rivera-González, Alejandro X.; García-Hernández, Normand; Curiel-Quesada, Everardo

    2016-01-01

    Integrons are non-mobile bacterial genetic elements that carry different cassettes conferring antibiotic resistance. Cassettes can excise or integrate by action of an integron-encoded integrase, enabling bacteria to face environmental challenges. In this work, the functionality and dynamics of two integrons carrying the same cassette arrangement (dfrA12–orfF–aadA2), but located on plasmid or chromosome in two different strains were studied. In order to demonstrate the functionality of the Class 1 integrase, circular cassette integration intermediaries were PCR amplified by PCR using extrachromosomal DNA extracted from bacteria grown in the presence or absence of cassette-encoded antibiotics. Circular aadA2 and dfrA12–orfF–aadA2 cassettes were detected in cultures grown either in the presence or absence of antibiotics in both strains. No dfrA12–orfF circular intermediates could be detected under any culture conditions. These results show that both integrons are functional. However, these elements show different dynamics and functionality since the presence of streptomycin led to detectable gene rearrangements in the variable region only in the strain with the plasmid-born integron. In addition, complete integration products were demonstrated using a receptor molecule carrying an empty integron. In this case, integration products were observed in both strains even in the absence of antibiotics, but they were more evident in the strain with the plasmid-located integron when streptomycin was present in the culture medium. This suggests that integrons in the two strains respond differently to streptomycin even though DNA sequences upstream the intI1 gene, including the lexA boxes of both integrons are identical. PMID:27733851

  5. Clinical implications of species identification in monomicrobial Aeromonas bacteremia.

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    Chi-Jung Wu

    Full Text Available Advances in Aeromonas taxonomy have led to the reclassification of aeromonads. Hereon, we aimed to re-evaluate the characteristics of Aeromonas bacteremia, including those of a novel species, Aeromonas dhakensis.A retrospective study of monomicrobial Aeromonas bacteremia at a medical center in southern Taiwan from 2004-2011 was conducted. Species identification was based on rpoB sequencing. Of bacteremia of 153 eligible patients, A. veronii (50 isolates, 32.7%, A. dhakensis (48, 31.4%, A. caviae (43, 28.1%, and A. hydrophila (10, 6.5% were the principal causative species. A. dhakensis and A. veronii bacteremia were mainly community-acquired and presented as primary bacteremia, spontaneous bacterial peritonitis, or skin and soft-tissue infection, whereas A. caviae was associated with hospital-onset bacteremia. The distribution of the AmpC β-lactamase and metallo-β-lactamase genes was species-specific: bla(AQU-1, bla(MOX, or bla(CepH was present in A. dhakensis, A. caviae, or A. hydrophila, respectively, and bla(CphA was present in A. veronii, A. dhakensis, and A. hydrophila. The cefotaxime resistance rates of the A. caviae, A. dhakensis, and A. hydrophila isolates were higher than that of A. veronii (39.5%%, 25.0%, and 30% vs. 2%, respectively. A. dhakensis bacteremia was linked to the highest 14-day sepsis-related mortality rate, followed by A. hydrophila, A. veronii, and A. caviae bacteremia (25.5%, 22.2%, 14.0%, and 4.7%, respectively; P = 0.048. Multivariate analysis revealed that A. dhakensis bacteremia, active malignancies, and a Pitt bacteremia score ≥ 4 was an independent mortality risk factor.Characteristics of Aeromonas bacteremia vary between species. A. dhakensis prevalence and its associated poor outcomes suggest it an important human pathogen.

  6. Characterization of integrons and tetracycline resistance determinants in Aeromonas spp. isolated from South African aquaculture systems.

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    Jacobs, Liezl; Chenia, Hafizah Y

    2007-03-20

    An increasing incidence of multidrug resistance amongst Aeromonas spp. isolates, which are both fish pathogens and emerging opportunistic human pathogens, has been observed worldwide. This can be attributed to the horizontal transfer of mobile genetic elements, viz.: plasmids and class 1 integrons. The antimicrobial susceptibilities of 37 Aeromonas spp. isolates, from tilapia, trout and koi aquaculture systems, were determined by disc-diffusion testing. The plasmid content of each isolate was examined using the alkaline lysis protocol. Tet determinant type was determined by amplification using two degenerate primer sets and subsequent HaeIII restriction. The presence of integrons was determined by PCR amplification of three integrase genes, as well as gene cassettes, and the qacEDelta1-sulI region. Thirty-seven Aeromonas spp. isolates were differentiated into six species by aroA PCR-RFLP, i.e., A. veronii biovar sobria, A. hydrophila, A. encheleia, A. ichtiosoma, A. salmonicida, and A. media. High levels of resistance to tetracycline (78.3%), amoxicillin (89.2%), and augmentin (86.5%) were observed. Decreased susceptibility to erythromycin was observed for 67.6% of isolates. Although 45.9% of isolates displayed nalidixic acid resistance, majority of isolates were susceptible to the fluoroquinolones. The MAR index ranged from 0.12 to 0.59, with majority of isolates indicating high-risk contamination originating from humans or animals where antibiotics are often used. Plasmids were detected in 21 isolates, with 14 of the isolates displaying multiple plasmid profiles. Single and multiple class A family Tet determinants were observed in 27% and 48.7% of isolates, respectively, with Tet A being the most prevalent Tet determinant type. Class 1 integron and related structures were amplified and carried different combinations of the antibiotic resistance gene cassettes ant(3'')Ia, aac(6')Ia, dhfr1, oxa2a and/or pse1. Class 2 integrons were also amplified, but the

  7. Envolvimento de Aeromonas em surto de doença diarréica aguda em São Bento do Una, Pernambuco Aeromonas associated with an acute diarrhea outbreak in São Bento do Una, Pernambuco

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    Ernesto Hofer

    2006-04-01

    Full Text Available No primeiro semestre de 2004, ocorreu um surto de diarréia em São Bento do Una, Pernambuco, registrando-se 2.170 casos. Nas 582 coproculturas realizadas, 145 (25% revelaram um enteropatógeno bacteriano, destacando 114 casos (19,5% com a participação de Aeromonas, representadas por Aeromonas caviae (57/9,8%, Aeromonas veronii biovar sobria (23/3,9%, Aeromonas veronii biovar veronii (15/2,6% e outras espécies (19/3,2%. Nos 31 episódios restantes (5,3%, foram detectados: V. cholerae O1 Ogawa toxigênico (18/3,1%, Salmonella spp (8/1,4%, Shigella spp (3/0,5% e Vibrio cholerae não O1/não O139 (2/0,3%.An acute diarrhea outbreak, with 2170 cases, was described during January to July, 2004, in São Bento do Una, Pernambuco. 582 stools were examined and an enteric pathogen was recovered in 25% (145 patients. Aeromonas species were the most frequent (114-19.5% and the main isolates were Aeromonas caviae (57-9.8%, Aeromonas veronii biovar sobria (23-3.9%, Aeromonas veronii biovar veronii (15-2.6% and other species (19-3.2%. The other isolated enteropathogens were Vibrio cholerae O1-Ogawa toxigenic (18-3.1%, Salmonella spp (8-1.4%, Shigella spp (3-0.5% and Vibrio cholerae non-O1/non-O139 (2-0.3%.

  8. Adhesion of Aeromonas sp. to cell lines used as models for intestinal adhesion.

    OpenAIRE

    Kirov, S M; Hayward, L. J.; Nerrie, M. A.

    1995-01-01

    Adhesion to HEp-2 cells has been shown to correlate with enteropathogenicity for Aeromonas species. Such adhesion is thought to reflect the ability of strains to adhere to human intestinal enterocytes, although HEp-2 cells are not of intestinal origin. In this study strains of Aeromonas veronii biotype sobria isolated from various sources were investigated in parallel assays for their ability to adhere to HEp-2 cells and to an intestinal cell line (Caco-2). Quantitative assays showed identica...

  9. Impact of antibiotic treatments on the expression of the R plasmid tra genes and on the host innate immune activity during pRAS1 bearing Aeromonas hydrophila infection in zebrafish (Danio rerio

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    Cantas Leon

    2012-03-01

    Full Text Available Abstract Background The transfer of R plasmids between bacteria has been well studied under laboratory conditions and the transfer frequency has been found to vary between plasmids and under various physical conditions. For the first time, we here study the expression of the selected plasmid mobility genes traD, virB11 and virD4 in the 45 kb IncU plasmid, pRAS1, conferring resistance to tetracycline, trimethoprim and sulphonamide, using an in vivo zebrafish infection- treatment model. Results Three days after oral infection of adult zebrafish with Aeromonas hydrophila harboring pRAS1, elevated expression of pro-inflammatory cytokine (TNF α, IL-1β and IL-8 and complement C3 genes in the intestine coincided with disease symptoms. Tetracycline, trimethoprim and an ineffective concentration of flumequine given 48 h prior to sampling, strongly increased expression of plasmid mobility genes, whereas an effective dosage of flumequine resulted in lower levels of mRNA copies of these genes relative to placebo treatment. Following effective treatment with flumequine, and ineffective treatments with a low concentration of flumequine, with trimethoprim or with sulphonamide, the intestinal expression of immune genes was strongly induced compared to placebo treated control fish. Conclusions Treatment of zebrafish infected with an antibiotic resistant (TcR, TmR, SuR A. hydrophila with ineffective concentrations of flumequine or the ineffective antimicrobials tetracycline and trimethoprim strongly induced expression of genes mediating conjugative transfer of the R-plasmid pRAS1. Simultaneously, there was a strong induction of selected inflammatory and immune response genes, which was again evident in fish subjected to ineffective treatment protocols. Our findings point to the essential role of therapeutic practices in escalation or control of antibiotic resistance transfer, and suggest that antibiotic substances, even in sub-inhibitory concentrations, may

  10. Characterisation of Aeromonas spp. isolated from frozen fish intended for human consumption in Mexico.

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    Castro-Escarpulli, G; Figueras, M J; Aguilera-Arreola, G; Soler, L; Fernández-Rendón, E; Aparicio, G O; Guarro, J; Chacón, M R

    2003-07-15

    A total of 82 strains of presumptive Aeromonas spp. were identified biochemically and genetically (16S rDNA-RFLP). The strains were isolated from 250 samples of frozen fish (Tilapia, Oreochromis niloticus niloticus) purchased in local markets in Mexico City. In the present study, we detected the presence of several genes encoding for putative virulence factors and phenotypic activities that may play an important role in bacterial infection. In addition, we studied the antimicrobial patterns of those strains. Molecular identification demonstrated that the prevalent species in frozen fish were Aeromonas salmonicida (67.5%) and Aeromonas bestiarum (20.9%), accounting for 88.3% of the isolates, while the other strains belonged to the species Aeromonas veronii (5.2%), Aeromonas encheleia (3.9%) and Aeromonas hydrophila (2.6%). Detection by polymerase chain reaction (PCR) of genes encoding putative virulence factors common in Aeromonas, such as aerolysin/hemolysin, lipases including the glycerophospholipid-cholesterol acyltransferase (GCAT), serine protease and DNases, revealed that they were all common in these strains. Our results showed that first generation quinolones and second and third generation cephalosporins were the drugs with the best antimicrobial effect against Aeromonas spp. In Mexico, there have been few studies on Aeromonas and its putative virulence factors. The present work therefore highlights an important incidence of Aeromonas spp., with virulence potential and antimicrobial resistance, isolated from frozen fish intended for human consumption in Mexico City. PMID:12781953

  11. Reassessment of the Enteropathogenicity of Mesophilic Aeromonas Species

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    Teunis, Peter; Figueras, Maria J.

    2016-01-01

    Cases of Aeromonas diarrhea have been described all over the world. The genus Aeromonas includes ca. 30 species, of which 10 have been isolated in association with gastroenteritis. The dominating species that account for ca. 96% of the identified strains are Aeromonas caviae, A. veronii, A. dhakensis, and A. hydrophila. However, the role of Aeromonas as a true enteropathogen has been questioned on the basis of the lack of outbreaks, the non-fulfillment of Koch’s postulates and the low numbers of acute illnesses in the only existing human challenge study. In the present study we reassess the enteropathogenicity of Aeromonas using dose response models for microbial infection and acute illness. The analysis uses the data from the human challenge study and additional data from selected outbreak investigations where the numbers exposed and the dose were reported, allowing their inclusion as “natural experiments”. In the challenge study several cases of asymptomatic shedding were found (26.3%, 15/57), however, only 3.5% (2/57) of those challenged with Aeromonas developed acute enteric symptoms (i.e., diarrhea). The “natural experiments” showed a much higher risk of illness associated with exposure to Aeromonas, even at moderate to low doses. The median dose required for 1% illness risk, was ~1.4 × 104 times higher in the challenge study (1.24 × 104 cfu) compared to natural exposure events (0.9 cfu). The dose response assessment presented in this study shows that the combined challenge and outbreak data are consistent with high infectivity of Aeromonas, and a wide range of susceptibility to acute enteric illness. To illustrate the outcomes, we simulate the risk associated with concentrations of Aeromonas found in different water and food matrices, indicating the disease burden potentially associated with these bacteria. In conclusion this study showed that Aeromonas is highly infectious, and that human susceptibility to illness may be high, similar to

  12. Effectiveness of radiation processing in elimination of Aeromonas from food

    Energy Technology Data Exchange (ETDEWEB)

    Nagar, Vandan [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India); Bandekar, Jayant R., E-mail: jrb@barc.gov.i [Food Technology Division, Bhabha Atomic Research Centre, Mumbai 400085 (India)

    2011-08-15

    Genus Aeromonas has emerged as an important human pathogen because it causes a variety of diseases including gastroenteritis and extra-intestinal infections. Contaminated water, sprouts, vegetables, seafood and food of animal origin have been considered to be the important sources of Aeromonas infection. In the present study, radiation sensitivity of indigenous strains of Aeromonas spp. from different food samples was evaluated. The decimal reduction dose (D{sub 10}) values of different Aeromonas isolates in saline at 0-4 {sup o}C were in the range of 0.031-0.046 kGy. The mixed sprouts, chicken and fish samples were inoculated with a cocktail of five most resistant isolates (A. salmonicida Y567, A. caviae A85, A. jandaei A514A, A. hydrophila CECT 839{sup T} and A. veronii Y47) and exposed to {gamma} radiation to study the effectiveness of radiation treatment in elimination of Aeromonas. D{sub 10} values of Aeromonas cocktail in mixed sprouts, chicken and fish samples were found to be 0.081{+-}0.001, 0.089{+-}0.003 and 0.091{+-}0.003 kGy, respectively. Radiation treatment with a 1.5 kGy dose resulted in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples. No recovery of Aeromonas was observed in the 1.5 kGy treated samples stored at 4 {sup o}C up to 12 (mixed sprouts) and 7 days (chicken and fish samples), even after enrichment and selective plating. This study demonstrates that a 1.5 kGy dose of irradiation treatment could result in complete elimination of 10{sup 5} CFU/g of Aeromonas spp. from mixed sprouts, chicken and fish samples.

  13. Clinical manifestations of bacteremia caused by Aeromonas species in southern Taiwan.

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    Hung-Jen Tang

    Full Text Available This study is conducted to investigate the clinical characteristics of patients with bacteremia caused by Aeromonas species.Patients with bacteremia caused by Aeromonas species during the period 2009 to 2013 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed.A total of 91 patients with bacteremia due to Aeromonas species were identified. In addition to 16 (17.6% primary bacteremia, the most common source of secondary infection is peritonitis (n = 27, 29.7%, followed by biliary tract infection (n = 18, 19.8%, and SSTI (n = 12, 13.2%, pneumonia (n = 9, 9.9%, catheter-related bloodstream infection (n =  5, 5.5%, and genitourinary tract infection (n = 4, 4.4%. A. hydrophila (n = 35, 38.5% was the most common pathogen, followed by A. veronii biovar sobria (n = 31, 34.1%, A. caviae (n = 14, 15.4%, and A. veronii biovar veronii (n = 9, 9.9%. Forty-three (47.3% patients were classified as healthcare-associated infections (HCAI causes by Aeromonas species, and patients with HCAI were more likely to have cancer, and receive immunosuppressant than patients with community-acquired bacteremia. The overall outcomes, including rate of ICU admission, acute respiratory failure, and mortality were 33.3%, 28.6%, and 23.1%, respectively. Multivariate analysis showed that the in-hospital day mortality was significantly associated only with underlying cancer (P <.001, and initial shock (P <.001.Aeromonas species should be considered one of the causative pathogens of healthcare-associated bacteremia, especially in immunocompromised patients. In addition, it can be associated with high fatality. Cancer and initial shock were the poor prognostic factors.

  14. Aeromonas dhakensis, an Increasingly Recognized Human Pathogen.

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    Chen, Po-Lin; Lamy, Brigitte; Ko, Wen-Chien

    2016-01-01

    Aeromonas dhakensis was first isolated from children with diarrhea in Dhaka, Bangladesh and described in 2002. In the past decade, increasing evidence indicate this species is widely distributed in the environment and can cause a variety of infections both in human and animals, especially in coastal areas. A. dhakensis is often misidentified as A. hydrophila, A. veronii, or A. caviae by commercial phenotypic tests in the clinical laboratory. Correct identification relies on molecular methods. Increasingly used matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) may be able to identify Aeromonas specie rapidly and accurately. A. dhakensis has shown its potent virulence in different animal models and clinical infections. Although several virulence factors had been reported, no single mechanism is conclusive. Characteristically A. dhakensis is the principal species causing soft tissue infection and bacteremia, especially among patients with liver cirrhosis or malignancy. Of note, A. dhakensis bacteremia is more lethal than bacteremia due to other Aeromonas species. The role of this species in gastroenteritis remains controversial. Third generation cephalosporins and carbapenems should be used cautiously in the treatment of severe A. dhakensis infection due to the presence of AmpC ββ-lactamase and metallo-β-lactamase genes, and optimal regimens may be cefepime or fluoroquinolones. Studies of bacterial virulence factors and associated host responses may provide the chance to understand the heterogeneous virulence between species. The hypothesis A. dhakensis with varied geographic prevalence and enhanced virulence that compared to other Aeromonas species warrants more investigations. PMID:27303382

  15. Aeromonas dhakensis, an increasingly recognized human pathogen

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    Po Lin eChen

    2016-05-01

    Full Text Available Aeromonas dhakensis was first isolated from children with diarrhea in Dhaka, Bangladesh and described in 2002. In the past decade, increasing evidence indicate this species is widely distributed in the environment and can cause a variety of infections both in human and animals, especially in coastal areas. A. dhakensis is often misidentified as A. hydrophila, A. veronii or A. caviae by commercial phenotypic tests in the clinical laboratory. Correct identification relies on molecular methods. Increasingly used matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS may be able to identify Aeromonas specie rapidly and accurately. A. dhakensis has shown its potent virulence in different animal models and clinical infections. Although several virulence factors had been reported, no single mechanism is conclusive. Characteristically A. dhakensis is the principal species causing soft tissue infection and bacteremia, especially among patients with liver cirrhosis or malignancy. Of note, A. dhakensis bacteremia is more lethal than bacteremia due to other Aeromonas species. The role of this species in gastroenteritis remains controversial. Third generation cephalosporins and carbapenems should be used cautiously in the treatment of severe A. dhakensis infection due to the presence of AmpC β-lactamase and metallo-β-lactamase genes, and optimal regimens may be cefepime or fluoroquinolones. Studies of bacterial virulence factors and associated host responses may provide the chance to understand the heterogeneous virulence between species. The hypothesis A. dhakensis with varied geographic prevalence and enhanced virulence that compared to other Aeromonas species warrants more investigations.

  16. Medicinal leech therapy and Aeromonas spp. infection.

    Science.gov (United States)

    Verriere, B; Sabatier, B; Carbonnelle, E; Mainardi, J L; Prognon, P; Whitaker, I; Lantieri, L; Hivelin, M

    2016-06-01

    While the use of medicinal leech therapy (MLT) in reconstructive and orthopaedic surgery is widely described, post-operative complications related to leeches remain a major concern. Aeromonas spp. strains are involved in the majority of reported cases. As surgical success rate is directly impacted, an adapted antibiotic prophylaxis should be instituted in order to minimize these complications. We assessed pharmaceutical process, microbiological control and related infections in order to provide data and choose the appropriate antibiotherapy for patients requiring MLT. We report a clinical and microbiological study over a 24-month period. Clinical data were collected from patients' database, and microbiological analysis both on leeches' tank water and crushed leeches were performed to characterize isolated strains and their susceptibility to antibiotics. A total of 595 leeches were used to treat 28 patients (12 in plastic surgery and 16 in orthopaedic surgery), and three documented cases of post-operative infections were reported. Aeromonas spp. isolates yielded from 62 % of analyzed batches (75 % of Aeromonas veronii). Eighteen Aeromonas spp. isolates yielded from 23 water samples and three crushed leeches. Isolates were similar in tank and crushed leeches. Strains were susceptible to fluoroquinolones, sulfamethoxazole/trimethoprim, aminosides, and third-generation cephalosporins but resistant to amoxicillin/clavulanic acid and second-generation cephalosporins. According to collected data, routine tank water microbiological analyses are mandatory in order to identify leeches' batches containing resistant strains and to discard them. In this context, the surgeon is able to select an appropriated antibiotic prophylaxis in order to avoid MLT associated serious post-operative complications. PMID:27039338

  17. A molecular study on the prevalence and virulence potential of Aeromonas spp. recovered from patients suffering from diarrhea in Israel.

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    Yigal Senderovich

    Full Text Available BACKGROUND: Species of the genus Aeromonas are native inhabitants of aquatic environments and have recently been considered emerging human pathogens. Although the gastrointestinal tract is by far the most common anatomic site from which aeromonads are recovered, their role as etiologic agents of bacterial diarrhea is still disputed. Aeromonas-associated diarrhea is a phenomenon occurring worldwide; however, the exact prevalence of Aeromonas infections on a global scale is unknown. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence and virulence potential of Aeromonas in patients suffering from diarrhea in Israel was studied using molecular methods. 1,033 diarrheal stools were sampled between April and September 2010 and Aeromonas species were identified in 17 (∼2% patients by sequencing the rpoD gene. Aeromonas species identity and abundance was: A. caviae (65%, A. veronii (29% and Aeromonas taiwanensis (6%. This is the first clinical record of A. taiwanensis as a diarrheal causative since its recent discovery from a wound infection in a patient in Taiwan. Most of the patients (77% from which Aeromonas species were isolated were negative for any other pathogens. The patients ranged from 1 to 92 years in age. Aeromonas isolates were found to possess different virulence-associated genes: ahpB (88%, pla/lip/lipH3/apl-1 (71%, act/hlyA/aerA (35%, alt (18%, ast (6%, fla (65%, lafA (41%, TTSS ascV (12%, TTSS ascF-ascG (12%, TTSS-dependent ADP-ribosylating toxins aexU (41% and aexT (6% in various combinations. Most of the identified strains were resistant to beta-lactam antibiotics but susceptible to third-generation cephalosporin antibiotics. CONCLUSIONS: Aeromonas may be a causative agent of diarrhea in patients in Israel and therefore should be included in routine bacteriological screenings.

  18. Genospecies and virulence factors of Aeromonas species in different sources in a North African country

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    Khalifa Sifaw Ghenghesh

    2014-09-01

    Full Text Available Introduction: Aeromonads of medical importance have been reported from numerous clinical, food, and water sources, but identification of genospecies and virulence factors of Aeromonas species from countries in North Africa and the Middle East are few. Methods: In total 99 Aeromonas species isolates from different sources (diarrheal children [n=23], non-diarrheal children [n=16], untreated drinking water from wells [n=32], and chicken carcasses [n=28] in Tripoli, Libya, were included in the present investigation. Genus identification was confirmed by biochemical analysis, and genospecies were determined using a combination of 16S rDNA variable region and gyrB sequence analysis. Polymerase chain reaction (PCR was used to detect genes encoding toxins from 52 of the isolates. Results: We identified 44 isolates (44% as A. hydrophila (3 [3.0%] subspecies anaerogenes, 23 [23%] subspecies dhakensis, and 18 [18%] subspecies ranae; 27 isolates (27% as A. veronii; 23 isolates (23% as A. caviae; and 5 isolates (5.0% as other genospecies. The genes encoding aerolysin (aer, cytolytic enterotoxin (act, and A. hydrophila isolate SSU enterotoxin (ast were detected in 45 (87%, 4 (7.7%, and 9 (17% of the 52 isolates tested, respectively. The gene encoding an extracellular lipase (alt was not detected. Conclusion: The majority of aeromonads from Libya fall within three genospecies (i.e. A. hydrophila, A. veronii, and A. caviae, and genes coding for toxin production are common among them.

  19. Aeromonas spp. e Plesiomonas shigelloides isoladas a partir de mexilhões (Perna perna in natura e pré-cozidos no Rio de Janeiro, RJ Aeromonas spp. and Plesiomonas shigelloides isolated from in natura and precooked mussels (Perna perna in Rio de Janeiro, RJ, Brazil

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    Christiane Soares Pereira

    2004-12-01

    Full Text Available O ecossistema aquático é o habitat de mexilhões (Perna perna, animais filtradores que refletem a qualidade ambiental através de análise microbiológica de sua carne. No presente trabalho avaliou-se a presença de patógenos emergentes (Aeromonas hydrophila e Plesiomonas shigelloides, em mexilhões in natura e pré-cozidos coletados por pescadores da Estação Experimental de Cultivo de Mexilhões situada em Jurujuba, Niterói, Rio de Janeiro. Foram analisadas 86 amostras de mexilhões (43 in natura e 43 pré-cozidos as quais foram submetidas a enriquecimento em Água Peptonada Alcalina (APA acrescida de 1 e 3% de Cloreto de Sódio (NaCl e em solução Salina de Butterfield, incubadas a 37ºC por 24 horas. Em seguida, foram semeadas em Ágar Seletivo para Pseudomonas-Aeromonas (GSP, Ágar Tiossulfato Citrato Bile Sacarose (TCBS e Ágar Inositol Bile Verde Brilhante (IBB. A análise geral dos resultados permitiu a identificação de Areomonas spp e Plesiomonas shigelloides em 86% das amostras de mexilhões in natura e pré-cozidas avaliadas. A posterior caracterização bioquímica permitiu a identificação das espécies Aeromonas media (37,10%, A. hydrophila (15,50%, A. caviae (14,80%, A. veronii biogrupo veronii (11,60%, Aeromonas sp. (7,36%, A. sobria (4,20%, A. trota (4,20%, A. schubertii (1,31%, A. jandaei (1,31%, A. veronii biogrupo sobria (0,52% e Plesiomonas shigelloides (2,10%. A relevância epidemiológica desses microrganismos em casos de gastrenterite humana, após consumo de mexilhões crus ou parcialmente cozidos, revela a importância de alertar as autoridades de Saúde Pública no Brasil, sobre a presença desses patógenos na cadeia alimentar e seus riscos para a saúde humana.The aquatic ecosystem is the habitat of mussels, filtrating animals that reflect the ambient quality through microbiological analysis. In the present investigation, we evaluated the presence of emergent pathogens (Aeromonas hydrophila and Plesiomonas

  20. Diversity and antibiotic resistance of Aeromonas spp. in drinking and waste water treatment plants.

    Science.gov (United States)

    Figueira, Vânia; Vaz-Moreira, Ivone; Silva, Márcia; Manaia, Célia M

    2011-11-01

    The taxonomic diversity and antibiotic resistance phenotypes of aeromonads were examined in samples from drinking and waste water treatment plants (surface, ground and disinfected water in a drinking water treatment plant, and raw and treated waste water) and tap water. Bacteria identification and intra-species variation were determined based on the analysis of the 16S rRNA, gyrB and cpn60 gene sequences. Resistance phenotypes were determined using the disc diffusion method. Aeromonas veronii prevailed in raw surface water, Aeromonas hydrophyla in ozonated water, and Aeromonas media and Aeromonas puntacta in waste water. No aeromonads were detected in ground water, after the chlorination tank or in tap water. Resistance to ceftazidime or meropenem was detected in isolates from the drinking water treatment plant and waste water isolates were intrinsically resistant to nalidixic acid. Most of the times, quinolone resistance was associated with the gyrA mutation in serine 83. The gene qnrS, but not the genes qnrA, B, C, D or qepA, was detected in both surface and waste water isolates. The gene aac(6')-ib-cr was detected in different waste water strains isolated in the presence of ciprofloxacin. Both quinolone resistance genes were detected only in the species A. media. This is the first study tracking antimicrobial resistance in aeromonads in drinking, tap and waste water and the importance of these bacteria as vectors of resistance in aquatic environments is discussed.

  1. Virulence diversity among bacteremic Aeromonas isolates: ex vivo, animal, and clinical evidences.

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    Po-Lin Chen

    Full Text Available BACKGROUND: The objective of this study was to compare virulence among different Aeromonas species causing bloodstream infections. METHODOLOGY/PRINCIPAL FINDINGS: Nine of four species of Aeromonas blood isolates, including A. dhakensis, A. hydrophila, A. veronii and A. caviae were randomly selected for analysis. The species was identified by the DNA sequence matching of rpoD. Clinically, the patients with A. dhakensis bacteremia had a higher sepsis-related mortality rate than those with other species (37.5% vs. 0%, P = 0.028. Virulence of different Aeromonas species were tested in C. elegans, mouse fibroblast C2C12 cell line and BALB/c mice models. C. elegans fed with A. dhakensis and A. caviae had the lowest and highest survival rates compared with other species, respectively (all P values <0.0001. A. dhakensis isolates also exhibited more cytotoxicity in C2C12 cell line (all P values <0.0001. Fourteen-day survival rate of mice intramuscularly inoculated with A. dhakensis was lower than that of other species (all P values <0.0001. Hemolytic activity and several virulence factor genes were rarely detected in the A. caviae isolates. CONCLUSIONS/SIGNIFICANCE: Clinical data, ex vivo experiments, and animal studies suggest there is virulence variation among clinically important Aeromonas species.

  2. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.

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    Wei Ching Khor

    Full Text Available Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT and RNA polymerase sigma-factor (rpoD genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

  3. Chironomids' Relationship with Aeromonas Species.

    Science.gov (United States)

    Laviad, Sivan; Halpern, Malka

    2016-01-01

    Chironomids (Diptera: Chironomidae), also known as non-biting midges, are one of the most abundant groups of insects in aquatic habitats. They undergo a complete metamorphosis of four life stages of which three are aquatic (egg, larva, and pupa), and the adult emerges into the air. Chironomids serve as a natural reservoir of Aeromonas and Vibrio cholerae species. Here, we review existing knowledge about the mutual relations between Aeromonas species and chironomids. Using 454-pyrosequencing of the 16S rRNA gene, we found that the prevalence of Aeromonas species in the insects' egg masses and larvae was 1.6 and 3.3% of the insects' endogenous microbiota, respectively. Aeromonas abundance per egg mass remained stable during a 6-month period of bacterial monitoring. Different Aeromonas species were isolated and some demonstrated the ability to degrade the insect's egg masses and to prevent eggs hatching. Chitinase was identified as the enzyme responsible for the egg mass degradation. Different Aeromonas species isolated from chironomids demonstrated the potential to protect their host from toxic metals. Aeromonas is a causative agent of fish infections. Fish are frequently recorded as feeding on chironomids. Thus, fish might be infected with Aeromonas species via chironomid consumption. Aeromonas strains are also responsible for causing gastroenteritis and wound infections in humans. Different virulence genes were identified in Aeromonas species isolated from chironomids. Chironomids may infest drinking water reservoirs, hence be the source of pathogenic Aeromonas strains in drinking water. Chironomids and Aeromonas species have a complicated mutual relationship. PMID:27242751

  4. Aeromonas hydrophila subsp. dhakensis isolated from feces, water and fish in Mediterranean Spain.

    Science.gov (United States)

    Esteve, Consuelo; Alcaide, Elena; Blasco, María Dolores

    2012-01-01

    Eight Aeromonas hydrophila-like arabinose-negative isolates from diverse sources (i.e., river freshwater, cooling-system water pond, diseased wild European eels, and human stools) sampled in Valencia (Spain) during 2004-2005, were characterized by 16S rRNA gene sequencing and extensive biochemical testing along with reference strains of most Aeromonas species. These isolates and all reference strains of A. hydrophila subsp. dhakensis and A. aquariorum showed a 16S rRNA sequence similarity of 99.8-100%, and they all shared an identical phenotype. This matched exactly with that of A. hydrophila subsp. dhakensis since all strains displayed positive responses to the Voges-Prokauer test and to the use of dl-lactate. This is the first report of A. hydrophila subsp. dhakensis recovered from environmental samples, and further, from its original isolation in India during 1993-1994. This was accurately identified and segregated from other clinical aeromonads (A. hydrophila subsp. hydrophila, A. caviae, A. veronii biovars veronii and sobria, A. trota, A. schubertii and A. jandaei) by using biochemical key tests. The API 20 E profile for all strains included in A. hydrophila subsp. dhakensis was 7047125. The prevalence of this species in Spanish sources was higher for water (9.4%) than for feces (6%) or eels (1.3%). Isolates recovered as pure cultures from diseased eels were moderately virulent (LD(50) of 3.3×10(6) CFU fish(-1)) to challenged eels in experimental trials. They were all resistant to ticarcillin, amoxicillin-clavuranic acid, cefoxitin, and imipenem, regardless of its source. Our data point to A. hydrophila subsp. dhakensis as an emerging pathogen for humans and fish in temperate countries. PMID:22472298

  5. Resistensi Bakteri Aeromonas sp. Isolat Ikan Piranha (Pygosentrus nattereri Bali Safari And Marine Park terhadap Antibiotik

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    Ni Made Herawati

    2013-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE Piranha (Pygosentrus nattereri is a fish that has been conserved at Conservation Center of Bali Safari and Marine Park. In May 2012 there was a case that twelve Piranhas suffered from illness and two of them died without any specific clinical signs. The dead Piranhas were necropsied and the tissues or parts of organs were collected for microbial examination interests. Spesies specific bacterial examination was also included using Microgen GNA TM + B-ID system test The bacterial examination found that the fishes were infected by Aeromonas hydropila and Aeromonas veronii. Further examination about antibiotic resistance tests showed that those Aeromonas spp were resistant to chloramphenicol and a combination of trimethoprim and sulphamethoxazole. It can be suggested that, next, the usage of other antibiotics is strongly recommended for the case of Piranha infected by Aeromonas spp at Bali Safari and Marine Park. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

  6. Antibiotic resistance of vibrio cholerae: special considerations of R-plasmids.

    Science.gov (United States)

    Kuwahara, S

    1978-09-01

    Studies on the transmission of R plasmid by conjugation between enterobacteria and vibrio or related bacteria were reviewed. The majority of the reports confirmed successful transmission from enterobacteria to Vibrio cholerae and related species, although the transmission frequencies were extremely low and the transmitted R plasmid was very unstable except for thermosensitive kanamycin plasmid and usual R plasmid coexisting with P plasmid. Strains of V. cholerae and Aeromonas liquefaciens as well as A. salmonicida bearing R plasmid were detected in nature. R plasmid was relatively unstable in V. cholerae strains with which transmission of R plasmid to enterobacteria was confirmed. At present, only 3 R plasmids have been obtained from naturally occurring strains of V. cholerae. Although the 2 European plasmids belong to the C incompatibility group with 98 megadalton closed covalent circular DNA molecule, one plasmid belongs to the J group with more than 25 megadalton molecular weight, and no CCC of satelite DNA was detected in bacteria harboring this plasmid.

  7. Impact of an Urban Effluent on Antibiotic Resistance of Riverine Enterobacteriaceae and Aeromonas spp.

    Science.gov (United States)

    Goñi-Urriza, Marisol; Capdepuy, Michèle; Arpin, Corinne; Raymond, Nathalie; Caumette, Pierre; Quentin, Claudine

    2000-01-01

    In order to evaluate the impact of an urban effluent on antibiotic resistance of freshwater bacterial populations, water samples were collected from the Arga river (Spain), upstream and downstream from the wastewater discharge of the city of Pamplona. Strains of Enterobacteriaceae (representative of the human and animal commensal flora) (110 isolates) and Aeromonas (typically waterborne bacteria) (118 isolates) were selected for antibiotic susceptibility testing. Most of the Aeromonas strains (72%) and many of the Enterobacteriaceae (20%) were resistant to nalidixic acid. Singly nalidixic acid-resistant strains were frequent regardless of the sampling site for Aeromonas, whereas they were more common upstream from the discharge for enterobacteria. The most common resistances to antibiotics other than quinolones were to tetracycline (24.3%) and beta-lactams (20.5%) for Enterobacteriaceae and to tetracycline (27.5%) and co-trimoxazole (26.6%) for Aeromonas. The rates of these antibiotic resistances increased downstream from the discharge at similar degrees for the two bacterial groups; it remained at high levels for enterobacteria but decreased along the 30-km study zone for Aeromonas. Genetic analysis of representative strains demonstrated that these resistances were mostly (enterobacteria) or exclusively (Aeromonas) chromosomally mediated. Moreover, a reference strain of Aeromonas caviae (CIP 7616) could not be transformed with conjugative R plasmids of enterobacteria. Thus, the urban effluent resulted in an increase of the rates of resistance to antibiotics other than quinolones in the riverine bacterial populations, despite limited genetic exchanges between enterobacteria and Aeromonas. Quinolone resistance probably was selected by heavy antibiotic discharges of unknown origin upstream from the urban effluent. PMID:10618213

  8. Plasmid Biopharmaceuticals.

    Science.gov (United States)

    Prazeres, Duarte Miguel F; Monteiro, Gabriel A

    2014-12-01

    Plasmids are currently an indispensable molecular tool in life science research and a central asset for the modern biotechnology industry, supporting its mission to produce pharmaceutical proteins, antibodies, vaccines, industrial enzymes, and molecular diagnostics, to name a few key products. Furthermore, plasmids have gradually stepped up in the past 20 years as useful biopharmaceuticals in the context of gene therapy and DNA vaccination interventions. This review provides a concise coverage of the scientific progress that has been made since the emergence of what are called today plasmid biopharmaceuticals. The most relevant topics are discussed to provide researchers with an updated overview of the field. A brief outline of the initial breakthroughs and innovations is followed by a discussion of the motivation behind the medical uses of plasmids in the context of therapeutic and prophylactic interventions. The molecular characteristics and rationale underlying the design of plasmid vectors as gene transfer agents are described and a description of the most important methods used to deliver plasmid biopharmaceuticals in vivo (gene gun, electroporation, cationic lipids and polymers, and micro- and nanoparticles) is provided. The major safety issues (integration and autoimmunity) surrounding the use of plasmid biopharmaceuticals is discussed next. Aspects related to the large-scale manufacturing are also covered, and reference is made to the plasmid products that have received marketing authorization as of today.

  9. CONTAMINATION RISK ASSESSMENT OF SOURCES OF WATER INTENDED FOR HUMAN CONSUMPTION WITH AEROMONAS SPP.

    Directory of Open Access Journals (Sweden)

    Luminita Minea

    2012-03-01

    Full Text Available mportance of Aeromonas spp as emerging pathogens in recent years has been a considerable increase, beingincluded by the World Health Organization (WHO and Environmental Protection Agency (EPA in the etiology ofwaterborne epidemics. We aimed to investigate the distribution of Aeromonas spp. in various aquatic ecosystems whichcan become potential sources of gastrointestinal infections. Simultaneously, the presence of this pathogen was evaluatedin relation to microbiological indicators set out in EU Directive 98/83/EC. Between March 2009 and May 2011 wereanalyzed by membrane filter technique untreated water samples, collected from surface sources (lakes, ponds andgroundwater (wells drilled, captured springs, wells and chlorinated water that coming from local distribution network,located in rural areas of Moldova (North-East Romania. The information was processed and interpreted statisticallyusing EpiInfo 3.5.1 software. The presence of Aeromonas spp. was detected in half of the 220 samples tested, especially,in surface waters (84% and captured springs (80.8% (χ 2=51.39; GL=4; p<0.001, with concentrations that ranged from1 to 3.8 x104CFU/100ml (median=4.9x103CFU/100ml, respectively, from 1 to 6.9x102CFU/100 ml (median =58CFU/100 ml . The main species identified were A. hydrophila (22%, A. caviae (10.2%, A. veronii (6% and A. sobria(6% (using API 20NE galeries. Although the etiology was reported in 15.9%, chlorinated water quality was much betterthan that of untreated sources (χ 2=26.56; GL=1; p<0.001, with Odd.Ratio =0.12 (0.04 to 0.31; CI=95%. Comparativeevaluation of contamination of studied ecosystems showed a higher presence of these organisms in surface water than ingroundwater (χ 2=8.71; GL=1; p<0.003, Odd.Ratio=37 (1.4 to 8.9; CI=95%, with an increased risk of infectiontransmission through ingestion or contact. Pearson coefficient for the presence in groundwater of fecal pollutionindicators (coliform bacteria, Escherichia coli, enterococci

  10. Two novel temperate bacteriophages co-existing in Aeromonas sp. ARM81 - characterization of their genomes, proteomes and DNA methyltransferases.

    Science.gov (United States)

    Dziewit, Lukasz; Radlinska, Monika

    2016-08-01

    Aeromonas species are causative agents of a wide spectrum of diseases in animals and humans. Although these bacteria are commonly found in various environments, little is known about their phages. Thus far, only one temperate Aeromonas phage has been characterized. Whole-genome sequencing of an Aeromonas sp. strain ARM81 revealed the presence of two prophage clusters. One of them is integrated into the chromosome and the other was maintained as an extrachromosomal, linear plasmid-like prophage encoding a protelomerase. Both prophages were artificially and spontaneously inducible. We separately isolated both phages and compared their genomes with other known viruses. The novel phages show no similarity to the previously characterized Aeromonas phages and might represent new evolutionary lineages of viruses infecting Aeromonadaceae. Apart from the comparative genomic analyses of these phages, complemented with their structural and molecular characterization, a functional analysis of four DNA methyltransferases encoded by these viruses was conducted. One of the investigated N6-adenine-modifying enzymes shares sequence specificity with a Dam-like methyltransferase of its bacterial host, while another one is non-specific, as it catalyzes adenine methylation in various sequence contexts. The presented results shed new light on the diversity of Aeromonas temperate phages.

  11. Presumptive and Definitive Identification of Aeromonas from infected Ornamental Gold fish (Carassius auratus auratus

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    M. Phani Kumar

    2013-06-01

    Full Text Available The paper deals with the presumptive identification of Aeromonas, definitive identification of two species of Aeromonas i.e. Aeromonas hydrophila, Aeromonas salmonicida in various organs of Gold fish infected with red disease.

  12. Correction: Structural Studies of the Lipopolysaccharide from the Fish Pathogen Aeromonas veronii Strain Bs19, Serotype O16. Mar. Drugs 2014, 12, 1298–1316

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    Anna Turska-Szewczuk

    2014-09-01

    Full Text Available We found one editorial mistake in our published paper [1]. In Line 2 of Table 4, the same composition of sugars is given for the C4 and C5 species (in the C5 species, one residue: 6dHexNAc has been missed. [...

  13. Biochemical characteristics, serogroups, and virulence factors of aeromonas species isolated from cases of diarrhoea and domestic water samples in Chennai

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    Alavandi S

    2003-01-01

    Full Text Available PURPOSE: The objective of the present study was to delineate the differences between the clinical and environmental Aeromonas species with respect to their biochemical characteristics, serogrouping and virulence factors, in order to find a phenotypic marker of enteropathogenicity. METHODS: A total of 55 Aeromonas spp. inclusive of 19 isolates from cases of diarrhoea, and 36 from water samples comprising, 10 isolates of A. hydrophila, 21 isolates each of A. sobria, and A. caviae, two isolates of A. jandaei and one isolate of A. veronii were subjected to analysis of their biochemical characteristics, serogrouping, and virulence factors. RESULTS: Among the differences recorded in the biochemical characteristics in the three major species, the most striking characteristic was fermentation of lactose, which was observed in all the 11 A. caviae isolates recovered from water samples. None of the 10 clinical isolates of A. caviae tested fermented lactose. The clinical Aeromonas isolates belonged to seven typable serogroups, O:13, O:14, O:16, O:21, O:27, O:32 and O:35. The environmental isolates belonged to eight different serogroups, such as, O:3, O:11, O:14, O:16, O:18, O:28, O:64 and O:78 and were predominated by serotypes O:18 and O:64. Among the virulence factors tested, 89% of the environmental isolates produced b haemolysin, while only 62.3% of clinical isolates were able to do so. There was no significant difference between the clinical and environmental aeromonads with respect to their enterotoxigenicity in suckling mice in vivo, cytotoxicity in vitro in Vero cell monolayers, and ability to produce siderophores. CONCLUSION: Efforts to delineate the differences between the clinical and environmental Aeromonas spp. did not reveal significant difference between them. However, difference was observed with respect to their ability to produce b haemolysin, wherein, higher percentage of environmental isolates was haemolytic. The results also suggest

  14. Identificação e resistência a antimicrobianos de espécies de Aeromonas móveis isoladas de peixes e ambientes aquáticos Identification and antimicrobial resistance of motile Aeromonas isolated from fish and aquatic environment

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    Daniela Hirsch

    2006-12-01

    Full Text Available Com o objetivo de se verificar a diversidade de espécies de Aeromonas móveis e seu perfil de susceptibilidade a antimicrobianos em pisciculturas comerciais, foram selecionadas oito tilapiculturas localizadas na região do Alto Rio Grande, Minas Gerais. De cada propriedade foram coletadas três amostras de peixes em estádio de pré-abate (vivos e saudáveis, uma amostra de água do tanque e uma amostra da água de abastecimento do sistema. De cada peixe foram coletadas amostras de lavado superficial e do parênquima renal. Diluições seriadas adequadas de cada amostra foram plaqueadas em TSA-ampicilina (10 mg/l e as amostras de rim em Ágar Sangue de cavalo a 5%. A partir de colônias isoladas positivas para o teste da oxidase foram realizados testes para identificação do gênero (testes presuntivos e das espécies de Aeromonas (testes bioquímicos. O perfil de antibiograma foi obtido pelo teste de difusão de discos de antibióticos em Ágar Mueller Hinton. Foram obtidos 75 isolados diferenciados em nove espécies de Aeromonas: A. jandaei, A. hydrophila, A. trota, A. caviae, A. sobria, A. eucrenophila, A. veronii bt veronii, A. schubertii, A. media, além de amostras classificadas como Aeromonas atípicas. Do total isolado, oito amostras foram provenientes da superfície corpórea de peixes, 14 da água de abastecimento e 53 da água do tanque. Não houve isolamento a partir dos espécimes de parênquima renal. Em relação ao perfil de resistência, 93% dos isolados foram resistentes à eritromicina, 36% à tetraciclina, 13% ao ác. nalidíxico, 9% à gentamicina, 8% à nitrofurantoína, 8% à canamicina, 5% à norfloxacina, 4% ao cloranfenicol e 3% às sulfonamidas. Dentre os isolados analisados, 43% apresentaram índice de múltipla resistência a antimicrobianos (MAR igual ou superior a 22%, ou seja, resistência a dois ou mais drogas das nove testadas. Os dados apontam para um risco iminente, tanto pelo isolamento de amostras

  15. Aeromonas caviae septicemia in immunocompetent gastrointestinal carriers

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    M. Dwivedi

    2008-12-01

    Full Text Available Aeromonas caviae strains have been isolated from blood and stool cultures of three immunocompetent patients, residents of Northern India, who presented with community acquired septicemia without any recent history of diarrhea. Cell culture infectivity test performed on Hep-2 cells have shown substantial degree of invasiveness in the isolated strains. This case unleashes a possibility of asymptomatic gastrointestinal carriage of such strains of A. caviae in a very large population of India, as several areas of India have very high rates of Aeromonas induced acute diarrhea/gastroenteritis (up to 13%. It needs to be appraised further in India as well as other countries having high rates of Aeromonas induced acute diarrhea/gastroenteritis.

  16. Extracellular synthesis of silver nanoparticles by novel Pseudomonas veronii AS41G inhabiting Annona squamosa L. and their bactericidal activity

    Science.gov (United States)

    Baker, Syed; Mohan Kumar, K.; Santosh, P.; Rakshith, D.; Satish, S.

    2015-02-01

    In present investigation extracellular synthesis of silver nanoparticles were synthesized using cell free supernatant of Pseudomonas veronii AS41G isolated from Annona squamosa L. The bacterium significantly reduced silver nitrate to generate silver nanoparticles which was characterized with hyphenated techniques. Synthesis of silver nanoparticles preliminary confirmed by UV-Visible spectrophotometry with the intense peak at 410 nm, Further FTIR analysis revealed the possible role of biomolecules in the supernatant responsible for mediating the nanoparticles formation. The XRD spectra exhibited the characteristic Bragg peaks of 1 0 0, 1 1 1, 2 0 0, and 2 2 0 facets of the face centred cubic symmetry of nanoparticles suggesting that these nanoparticles were crystalline in nature. TEM microgram showed polydispersity of nanoparticles with size ranging from 5 to 50 nm. Synthesized silver nanoparticles showed antibacterial activity against human and environmental pathogens including MRSA. The study enlightens the role of biosynthesized silver nanoparticles as an emerging alternative for drug resistant microorganisms. The obtained results are promising enough to pave the environmentally benign nanoparticle synthesis processes without use of any toxic chemicals and also envision the emerging role of endophytes towards synthesis of nanoparticles. With scanty reports available on P.veronii species, a new role has been reported in this study which will be very valuable for future researchers working on it.

  17. Resistance of Salmonds Against Aeromonas Salmonicida

    DEFF Research Database (Denmark)

    Holten-Andersen, Lars; Dalsgaard, Inger; Buchmann, Kurt

    2011-01-01

    susceptibility to this disease might exist between Baltic salmon and the widely used rainbow trout. Hence, a cohabitation challenge model was applied to investigate the relative susceptibility to infection with Aeromonas salmonicida in rainbow trout and Baltic salmon. The course of infection was monitored daily...

  18. Aeromonas spp.: An Emerging Nosocomial Pathogen.

    Science.gov (United States)

    Batra, Priyam; Mathur, Purva; Misra, Mahesh C

    2016-01-01

    Aeromonads are hallophillic, nonacid fast, nonspore forming, Gram-negative rods which are widely distributed in the soil, foodstuffs, and aquatic environment. Since times immemorial, they are important zoonotic pathogens of poikilotherms but are now emerging as important human pathogens. These emerging enteric pathogens flourish in the water distribution system by forming biofilms. They possess large number of virulence factors including inherent resistance to various antibiotics and ability to form biofilms using quorum sensing. These properties make them easy pathogens for human infections. Aeromonads are important enteric pathogens, but, with the growing level of immunosuppression in the population, they have been associated with various extraintestinal infections, such as skin and soft-tissue infections, traumatic wound infections, and lower respiratory tract/urinary tract infections. The average annual incidence of bacteremia in Southern Taiwan due to Aeromonas spp. was 76 cases/million inhabitants between 2008 and 2010. However, the incidence reported from Western countries is much lower. The case fatality rate among patients with Aeromonas bacteremia ranges from 27.5 to 46%. Aeromonads are universally resistant to the narrow-spectrum penicillin group of antibiotics such as penicillin, ampicillin, carbenicillin, and ticarcillin. They are however susceptible to piperacillin, azlocillin, second and third generation cephalosporins, and carbapenems. Most of the Aeromonas species are susceptible to aminoglycosides, tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole, quinolones, and monobactams. This manuscript is a comprehensive systematic review of the literature available on Aeromonas spp. PMID:27013806

  19. Aeromonas spp.: An emerging nosocomial pathogen

    Directory of Open Access Journals (Sweden)

    Priyam Batra

    2016-01-01

    Full Text Available Aeromonads are hallophillic, nonacid fast, nonspore forming, Gram-negative rods which are widely distributed in the soil, foodstuffs, and aquatic environment. Since times immemorial, they are important zoonotic pathogens of poikilotherms but are now emerging as important human pathogens. These emerging enteric pathogens flourish in the water distribution system by forming biofilms. They possess large number of virulence factors including inherent resistance to various antibiotics and ability to form biofilms using quorum sensing. These properties make them easy pathogens for human infections. Aeromonads are important enteric pathogens, but, with the growing level of immunosuppression in the population, they have been associated with various extraintestinal infections, such as skin and soft-tissue infections, traumatic wound infections, and lower respiratory tract/urinary tract infections. The average annual incidence of bacteremia in Southern Taiwan due to Aeromonas spp. was 76 cases/million inhabitants between 2008 and 2010. However, the incidence reported from Western countries is much lower. The case fatality rate among patients with Aeromonas bacteremia ranges from 27.5 to 46%. Aeromonads are universally resistant to the narrow-spectrum penicillin group of antibiotics such as penicillin, ampicillin, carbenicillin, and ticarcillin. They are however susceptible to piperacillin, azlocillin, second and third generation cephalosporins, and carbapenems. Most of the Aeromonas species are susceptible to aminoglycosides, tetracycline, chloramphenicol, trimethoprim-sulfamethoxazole, quinolones, and monobactams. This manuscript is a comprehensive systematic review of the literature available on Aeromonas spp.

  20. Aeromonas chitinase degrades chironomid egg masses.

    Science.gov (United States)

    Laviad, Sivan; Golan, Amnon; Shaked, Tamar; Vaizel-Ohayon, Dalit; Halpern, Malka; Pick, Elah

    2016-02-01

    Chironomids are freshwater insects that undergo a complete metamorphosis of four life stages. Chironomid egg masses can be degraded by Vibrio cholerae and some Aeromonas species. Egg mass degradation by V. cholerae requires haemagglutinin protease activity. Our aim was to identify the egg mass degrading (EMD) factor secreted by Aeromonas dhkanesis 3K1C15. Following the hypothesis that the EMD factor of A. dhkanesis is also a protease, secreted proteases were screened, but none of them proved to have the same properties as the EMD factor. Using conventional protein purification methods, we found that the active fraction included chitinases. We further confirmed chitin as a building block of the egg masses. Interestingly, by supplementing bacterial growth media with chitin, we observed unexpected EMD factor activity in Aeromonas isolates that initially were not able to degrade egg masses. Accordingly, we concluded that although strain 3K1C15 secretes chitinases constitutively, most Aeromonas strains secrete chitinases inductively. Induction of chitinases in nature presumably occurs when bacteria are attached to the egg mass habitat, in which chitin is abundant. Considering that chitinases are highly conserved across bacteria phyla, we assume that the role of this enzyme in the bacteria-insect interplay could be wider than is currently thought. PMID:26472256

  1. Aeromonas aquariorum sp. nov., isolated from aquaria of ornamental fish.

    Science.gov (United States)

    Martínez-Murcia, A J; Saavedra, M J; Mota, V R; Maier, T; Stackebrandt, E; Cousin, S

    2008-05-01

    During a survey to determine the prevalence of Aeromonas strains in water and skin of imported ornamental fish, 48 strains presumptively identified as Aeromonas were isolated but they could not be identified as members of any previously described Aeromonas species. These strains were subjected to a polyphasic approach including phylogenetic analysis derived from gyrB, rpoD and 16S rRNA gene sequencing, DNA-DNA hybridization, MALDI-TOF MS analysis, genotyping by RAPD and extensive biochemical and antibiotic susceptibility tests in order to determine their taxonomic position. Based on the results of the phylogenetic analyses and DNA-DNA hybridization data, we describe a novel species of the genus Aeromonas, for which the name Aeromonas aquariorum sp. nov. is proposed, with strain MDC47T (=DSM 18362T =CECT 7289T) as the type strain. This is the first Aeromonas species description based on isolations from ornamental fish. PMID:18450708

  2. Antimicrobial resistance profile of Aeromonas species isolated from Libya

    OpenAIRE

    Ghenghesh, Khalifa Sifaw; El-Mohammady, Hanan; Levin, Samuel Y.; Zorgani, Abdulazziz; Tawil, Khaled

    2013-01-01

    Aeromonas species are Gram-negative, oxidasepositive rods in the family Aeromonadaceae. These organisms have been isolated from untreated drinking water and different types of foods particularly seafood and frozen chicken. Aeromonas species are recognized as agents of serious infections in immunocompromised individuals including septicemia in patients with hepatic cirrhosis with a rapidly fatal outcome. Significant association of Aeromonas species with diarrhoea, particularly in children, and...

  3. Quorum Sensing Regulation in Aeromonas hydrophila

    DEFF Research Database (Denmark)

    Garde, Christian; Bjarnsholt, Thomas; Givskov, Michael;

    2010-01-01

    We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i.e., the Escheri......We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i.......e., the Escherichia coli monitor strain MH205), the C4-HSL production of A. hydrophila is interrupted by fusion of gfp(ASV). In the present in vitro study, we measure the response of the QS regulatory ahyRI locus in the monitor strain to predetermined concentrations of C4-HSL signal molecules. A minimal kinetic model...

  4. Plasmid segregation mechanisms

    DEFF Research Database (Denmark)

    Ebersbach, Gitte; Gerdes, Kenn; Charbon, Gitte Ebersbach

    2005-01-01

    Bacterial plasmids encode partitioning (par) loci that ensure ordered plasmid segregation prior to cell division. par loci come in two types: those that encode actin-like ATPases and those that encode deviant Walker-type ATPases. ParM, the actin-like ATPase of plasmid R1, forms dynamic filaments...... that segregate plasmids paired at mid-cell to daughter cells. Like microtubules, ParM filaments exhibit dynamic instability (i.e., catastrophic decay) whose regulation is an important component of the DNA segregation process. The Walker box ParA ATPases are related to MinD and form highly dynamic, oscillating...... filaments that are required for the subcellular movement and positioning of plasmids. The role of the observed ATPase oscillation is not yet understood. However, we propose a simple model that couples plasmid segregation to ParA oscillation. The model is consistent with the observed movement...

  5. High Prevalence of Plasmid-Mediated Quinolone Resistance and IncQ Plasmids Carrying qnrS2 Gene in Bacteria from Rivers near Hospitals and Aquaculture in China.

    Directory of Open Access Journals (Sweden)

    Yanping Wen

    Full Text Available Effluents from hospital and aquaculture are considered important sources of quinolone resistance. However, little information is available on the impact of this effluent on nearby rivers. In this study, 188 ciprofloxacin-resistant bacterial isolates obtained from rivers near hospitals and aquaculture were screened for plasmid-mediated quinolone resistance (PMQR genes. Species identification, antibiotic susceptibility testing, and PMQR gene transferability assessment were conducted for PMQR-positive bacteria. Representative qnrS2-encoding plasmids were subsequently sequenced using a primer-walking approach. In total, 44 isolates (23.4% were positive for qnr genes (16 qnrB2, 3 qnrS1, and 25 qnrS2 and 32 isolates (17.0% were positive for aac(6'-Ib-cr. Other PMQR genes were not detected. The qnrB2 and aac(6'-Ib-cr genes had a higher prevalence in aquaculture samples than in hospital samples, and were significantly associated with Enterobacteriaceae (p < 0.05. In contrast, the prevalence of qnrS2 was not site-related, but was significantly associated with Aeromonas spp. (p < 0.05. All PMQR isolates were resistant to three or more classes of antibiotics. Eleven qnrS2-harboring plasmids from Aeromonas spp., including a novel conjugative plasmid pHP18, were selected for sequencing. These plasmids were small in size (6,388-16,197 bp and belonged to the IncQ or IncU plasmid family, with qnrS2 being part of a mobile insertion cassette. Taken together, our findings suggest that aquaculture is a possible source for aac(6'-Ib-cr and qnrB2 dissemination, and demonstrate the ubiquity of qnrS2 in aquatic environments. Finally, Aeromonas spp. served as vectors for qnrS2 with the help of IncQ-type plasmids.

  6. High Prevalence of Plasmid-Mediated Quinolone Resistance and IncQ Plasmids Carrying qnrS2 Gene in Bacteria from Rivers near Hospitals and Aquaculture in China.

    Science.gov (United States)

    Wen, Yanping; Pu, Xiaoying; Zheng, Wei; Hu, Guang

    2016-01-01

    Effluents from hospital and aquaculture are considered important sources of quinolone resistance. However, little information is available on the impact of this effluent on nearby rivers. In this study, 188 ciprofloxacin-resistant bacterial isolates obtained from rivers near hospitals and aquaculture were screened for plasmid-mediated quinolone resistance (PMQR) genes. Species identification, antibiotic susceptibility testing, and PMQR gene transferability assessment were conducted for PMQR-positive bacteria. Representative qnrS2-encoding plasmids were subsequently sequenced using a primer-walking approach. In total, 44 isolates (23.4%) were positive for qnr genes (16 qnrB2, 3 qnrS1, and 25 qnrS2) and 32 isolates (17.0%) were positive for aac(6')-Ib-cr. Other PMQR genes were not detected. The qnrB2 and aac(6')-Ib-cr genes had a higher prevalence in aquaculture samples than in hospital samples, and were significantly associated with Enterobacteriaceae (p < 0.05). In contrast, the prevalence of qnrS2 was not site-related, but was significantly associated with Aeromonas spp. (p < 0.05). All PMQR isolates were resistant to three or more classes of antibiotics. Eleven qnrS2-harboring plasmids from Aeromonas spp., including a novel conjugative plasmid pHP18, were selected for sequencing. These plasmids were small in size (6,388-16,197 bp) and belonged to the IncQ or IncU plasmid family, with qnrS2 being part of a mobile insertion cassette. Taken together, our findings suggest that aquaculture is a possible source for aac(6')-Ib-cr and qnrB2 dissemination, and demonstrate the ubiquity of qnrS2 in aquatic environments. Finally, Aeromonas spp. served as vectors for qnrS2 with the help of IncQ-type plasmids. PMID:27427763

  7. High Prevalence of Plasmid-Mediated Quinolone Resistance and IncQ Plasmids Carrying qnrS2 Gene in Bacteria from Rivers near Hospitals and Aquaculture in China

    Science.gov (United States)

    Wen, Yanping; Pu, Xiaoying; Zheng, Wei

    2016-01-01

    Effluents from hospital and aquaculture are considered important sources of quinolone resistance. However, little information is available on the impact of this effluent on nearby rivers. In this study, 188 ciprofloxacin-resistant bacterial isolates obtained from rivers near hospitals and aquaculture were screened for plasmid-mediated quinolone resistance (PMQR) genes. Species identification, antibiotic susceptibility testing, and PMQR gene transferability assessment were conducted for PMQR-positive bacteria. Representative qnrS2-encoding plasmids were subsequently sequenced using a primer-walking approach. In total, 44 isolates (23.4%) were positive for qnr genes (16 qnrB2, 3 qnrS1, and 25 qnrS2) and 32 isolates (17.0%) were positive for aac(6′)-Ib-cr. Other PMQR genes were not detected. The qnrB2 and aac(6′)-Ib-cr genes had a higher prevalence in aquaculture samples than in hospital samples, and were significantly associated with Enterobacteriaceae (p < 0.05). In contrast, the prevalence of qnrS2 was not site-related, but was significantly associated with Aeromonas spp. (p < 0.05). All PMQR isolates were resistant to three or more classes of antibiotics. Eleven qnrS2-harboring plasmids from Aeromonas spp., including a novel conjugative plasmid pHP18, were selected for sequencing. These plasmids were small in size (6,388–16,197 bp) and belonged to the IncQ or IncU plasmid family, with qnrS2 being part of a mobile insertion cassette. Taken together, our findings suggest that aquaculture is a possible source for aac(6′)-Ib-cr and qnrB2 dissemination, and demonstrate the ubiquity of qnrS2 in aquatic environments. Finally, Aeromonas spp. served as vectors for qnrS2 with the help of IncQ-type plasmids. PMID:27427763

  8. SPP1-mediated plasmid transduction.

    OpenAIRE

    Canosi, U; Lüder, G; Trautner, T A

    1982-01-01

    The virulent Bacillus subtilis phage SPP1 transduces plasmid DNA. Plasmid-transducing phages contain only plasmid DNA. Such DNA represents a concatemer of monomeric plasmid molecules with the molecular weight of mature SPP1 DNA. Biological parameters of plasmid transduction are described.

  9. OVERVIEW: DISINFECTION OF HELICOBACTER PYLORI AND AEROMONAS SPECIES

    Science.gov (United States)

    Helicobacter pylori and Aeromonas hydrophila are contaminants listed on the USEPA's 1998 Contaminant Candidate List (CCL).The sensitivity of H. pylori to chlorine and of Aeromonas spp. to inactivation by free chlorine, chloramine and ultraviolet (UV) was examined. Selective and...

  10. Antimicrobial resistance of Aeromonas hydrophila isolated from different food sources: A mini-review.

    Science.gov (United States)

    Stratev, Deyan; Odeyemi, Olumide A

    2016-01-01

    Aeromonas hydrophila is a Gram-negative, oxidase-positive, facultative, anaerobic, opportunistic aquatic pathogen. A. hydrophila produces virulence factors, such as hemolysins, aerolysins, adhesins, enterotoxins, phospholipase and lipase. In addition to isolation from aquatic sources, A. hydrophila has been isolated from meat and meat products, milk and dairy products, and vegetables. However, various studies showed that this opportunistic pathogen is resistant to commercial antibiotics. This is attributed to factors such as the indiscriminate use of antibiotics in aquaculture, plasmids or horizontal gene transfer. In this report, we highlight the occurrence, prevalence and antimicrobial resistance of A. hydrophila isolated from different food samples. The presence of antimicrobial-resistant A. hydrophila in food poses threats to public and aquatic animal health. PMID:26588876

  11. The genome of Aeromonas salmonicida subsp. salmonicida A449: insights into the evolution of a fish pathogen

    Directory of Open Access Journals (Sweden)

    Murphy Colleen

    2008-09-01

    Full Text Available Abstract Background Aeromonas salmonicida subsp. salmonicida is a Gram-negative bacterium that is the causative agent of furunculosis, a bacterial septicaemia of salmonid fish. While other species of Aeromonas are opportunistic pathogens or are found in commensal or symbiotic relationships with animal hosts, A. salmonicida subsp. salmonicida causes disease in healthy fish. The genome sequence of A. salmonicida was determined to provide a better understanding of the virulence factors used by this pathogen to infect fish. Results The nucleotide sequences of the A. salmonicida subsp. salmonicida A449 chromosome and two large plasmids are characterized. The chromosome is 4,702,402 bp and encodes 4388 genes, while the two large plasmids are 166,749 and 155,098 bp with 178 and 164 genes, respectively. Notable features are a large inversion in the chromosome and, in one of the large plasmids, the presence of a Tn21 composite transposon containing mercury resistance genes and an In2 integron encoding genes for resistance to streptomycin/spectinomycin, quaternary ammonia compounds, sulphonamides and chloramphenicol. A large number of genes encoding potential virulence factors were identified; however, many appear to be pseudogenes since they contain insertion sequences, frameshifts or in-frame stop codons. A total of 170 pseudogenes and 88 insertion sequences (of ten different types are found in the A. salmonicida genome. Comparison with the A. hydrophila ATCC 7966T genome reveals multiple large inversions in the chromosome as well as an approximately 9% difference in gene content indicating instances of single gene or operon loss or gain. A limited number of the pseudogenes found in A. salmonicida A449 were investigated in other Aeromonas strains and species. While nearly all the pseudogenes tested are present in A. salmonicida subsp. salmonicida strains, only about 25% were found in other A. salmonicida subspecies and none were detected in other

  12. Aeromonas in Arab countries: 1995-2014.

    Science.gov (United States)

    Ghenghesh, Khalifa Sifaw; Rahouma, Amal; Zorgani, Abdulaziz; Tawil, Khaled; Al Tomi, Abdurazzaq; Franka, Ezzadin

    2015-10-01

    The aim of this review is to provide information on the prevalence, clinical syndromes, and antimicrobial resistance and therapy of Aeromonas spp. infections in Arab countries. The data were obtained by an English language literature search from 1995 to 2014 of Medline and PubMed for papers using the search terms "Aeromonas+name of Arab country (i.e. Algeria, Egypt, etc.)". Additional data were obtained from a Google search using the aforementioned terms. The organisms have been reported from diarrheal children, patients with cholera-like diarrhea, an outbreak of acute gastroenteritis and from different types of animals, foods and water source in several Arab countries in the Middle East and North Africa with predominance of A. hydrophila, A. caviae and A. sobria. Using molecular techniques few studies reported genes encoding several toxins from aeromonads isolated from different sources. Among the antimicrobials examined in the present review third generation cephalosporins, fluoroquinolones and aminoglycosides showed excellent activity and can be employed in the treatment of Aeromonas-associated human infections in Arabic countries. Whenever possible, treatment should be guided by the susceptibility testing results of the isolated organism. In the future, studies employing molecular testing methods are required to provide data on circulating genospecies and their modes of transmission in the community, and on their mechanisms of resistance to antimicrobials. Microbiology laboratories and research centers are encouraged to look for these organisms in clinical, food and water sources to attain a better understanding of the public health risks from these organisms in Arab countries. PMID:26577192

  13. Transfer in Marine Sediments of the Naturally Occurring Plasmid pRAS1 Encoding Multiple Antibiotic Resistance

    OpenAIRE

    Sandaa, Ruth-Anne; Enger, Øivind

    1994-01-01

    The results of microcosm experiments performed with the fish-pathogenic bacterium Aeromonas salmonicida acting as a donor showed that promiscuous plasmid pRAS1, which encodes tetracycline resistance, is transferred at a high frequency in marine sediments even in the absence of a selective factor. The presence of oxytetracycline resulted in an increase in the transfer frequency compared with that of a microcosm to which no selective factor was added. Transfer frequencies of 3.4 × 10-1 transcon...

  14. Chlamydial plasmids and bacteriophages.

    Science.gov (United States)

    Pawlikowska-Warych, Małgorzata; Śliwa-Dominiak, Joanna; Deptuła, Wiesław

    2015-01-01

    Chlamydia are absolute pathogens of humans and animals; despite being rather well recognised, they are still open for discovery. One such discovery is the occurrence of extrachromosomal carriers of genetic information. In prokaryotes, such carriers include plasmids and bacteriophages, which are present only among some Chlamydia species. Plasmids were found exclusively in Chlamydia (C.) trachomatis, C. psittaci, C. pneumoniae, C. suis, C. felis, C. muridarum and C. caviae. In prokaryotic organisms, plasmids usually code for genes that facilitate survival of the bacteria in the environment (although they are not essential). In chlamydia, their role has not been definitely recognised, apart from the fact that they participate in the synthesis of glycogen and encode proteins responsible for their virulence. Furthermore, in C. suis it was evidenced that the plasmid is integrated in a genomic island and contains the tetracycline-resistance gene. Bacteriophages specific for chlamydia (chlamydiaphages) were detected only in six species: C. psittaci, C. abortus, C. felis, C. caviae C. pecorum and C. pneumoniae. These chlamydiaphages cause inhibition of the developmental cycle, and delay transformation of reticulate bodies (RBs) into elementary bodies (EBs), thus reducing the possibility of infecting other cells in time. Plasmids and bacteriophages can be used in the diagnostics of chlamydioses; although especially in the case of plasmids, they are already used for detection of chlamydial infections. In addition, bacteriophages could be used as therapeutic agents to replace antibiotics, potentially addressing the problem of increasing antibiotic-resistance among chlamydia.

  15. Dynamics of Aeromonas species isolated from wastewater treatment system.

    Science.gov (United States)

    Martone-Rocha, S; Piveli, R P; Matté, G R; Dória, M C; Dropa, M; Morita, M; Peternella, F A; Matté, M H

    2010-12-01

    Aeromonas are widely distributed in the aquatic environment, and are considered to be emerging organisms that can produce a series of virulence factors. The present study was carried out in a sanitary sewage stabilization pond treatment system, located in Lins, State of São Paulo, Brazil. Most probable number was applied for estimation of the genus Aeromonas. Colony isolation was carried out on blood agar ampicillin and confirmed by biochemical characterization. Aeromonas species were isolated in 72.4% of influent samples, and in 55.2 and 48.3% of effluent from anaerobic and facultative lagoons, respectively. Thirteen Aeromonas species were isolated, representing most of the recognized species of these organisms. Even though it was possible to observe a tendency of decrease, total elimination of these organisms from the studied system was not achieved. Understanding of the pathogenic organism's dynamics in wastewater treatment systems with a reuse potential is especially important because of the risk it represents. PMID:20705981

  16. Hydrocarbon utilization by Brevibacterium, Azotomonas, Protaminobacterium, Mycococcus and Aeromonas spp

    Energy Technology Data Exchange (ETDEWEB)

    Lonsane, B.K.; Vadalkar, K.; Singh, H.D.; Baruah, J.N.

    1976-11-01

    Morphological, cultural and biochemical characteristics of 7 bacterial isolates, capable of utilizing hydrocarbons as sole source of carbon, reveal that 3 isolates belong to genus Aeromonas and one each to genera Brevibacterium, Protaminobacter, Mycococcus and Azotomonas. The isolates are studied for biomass formation on gas oil, substrate specificities for petroleum hydrocarbons and fermentation of gas oil by Brevibacterium sp. The hydrocarbon utilizing abilities of the strains of Protaminobacter, Azotomonas and Aeromonas are not known previously.

  17. Plasmids encoding therapeutic agents

    Science.gov (United States)

    Keener, William K.

    2007-08-07

    Plasmids encoding anti-HIV and anti-anthrax therapeutic agents are disclosed. Plasmid pWKK-500 encodes a fusion protein containing DP178 as a targeting moiety, the ricin A chain, an HIV protease cleavable linker, and a truncated ricin B chain. N-terminal extensions of the fusion protein include the maltose binding protein and a Factor Xa protease site. C-terminal extensions include a hydrophobic linker, an L domain motif peptide, a KDEL ER retention signal, another Factor Xa protease site, an out-of-frame buforin II coding sequence, the lacZ.alpha. peptide, and a polyhistidine tag. More than twenty derivatives of plasmid pWKK-500 are described. Plasmids pWKK-700 and pWKK-800 are similar to pWKK-500 wherein the DP178-encoding sequence is substituted by RANTES- and SDF-1-encoding sequences, respectively. Plasmid pWKK-900 is similar to pWKK-500 wherein the HIV protease cleavable linker is substituted by a lethal factor (LF) peptide-cleavable linker.

  18. AEROMONAS SALMONICIDA INFECTION IN VACCINATED RAINBOW TROUT

    DEFF Research Database (Denmark)

    Chettri, Jiwan Kumar; Skov, Jakob; Mohammad, Rezkar Jaafar;

    to that of the commercial vaccine with lower side effects as observed by the Speilberg scoring system. Gene expression analysis did not show a clear trend for Th1 or Th2 response in the vaccinated fish. Exposure of fish to saltwater increased the IgT production. Overall, the immune response in vaccinated fish, the side......In vivo testing of any candidate vaccine is influenced by the choice of challenge method and the external environmental conditions. In the present study, a comparative challenge study was performed to evaluate the efficacy of different vaccines against the bacterial pathogen Aeromonas salmonicida...... causing furunculosis. Rainbow trout (Oncorhynchus mykiss) were vaccinated with two trivalent adjuvanted experimental vaccines containing formalin-killed A. salmonicida, Vibrio anguillarum O1 and O2a and a commercial corresponding vaccine (Alpha Ject 3000). Fish were challenged by i.p. injection...

  19. Mechanisms of plasmid segregation: have multicopy plasmids been overlooked?

    Science.gov (United States)

    Million-Weaver, Samuel; Camps, Manel

    2014-09-01

    Plasmids are self-replicating pieces of DNA typically bearing non-essential genes. Given that plasmids represent a metabolic burden to the host, mechanisms ensuring plasmid transmission to daughter cells are critical for their stable maintenance in the population. Here we review these mechanisms, focusing on two active partition strategies common to low-copy plasmids: par systems type I and type II. Both involve three components: an adaptor protein, a motor protein, and a centromere, which is a sequence area in the plasmid that is recognized by the adaptor protein. The centromere-bound adaptor nucleates polymerization of the motor, leading to filament formation, which can pull plasmids apart (par I) or push them towards opposite poles of the cell (par II). No such active partition mechanisms are known to occur in high copy number plasmids. In this case, vertical transmission is generally considered stochastic, due to the random distribution of plasmids in the cytoplasm. We discuss conceptual and experimental lines of evidence questioning the random distribution model and posit the existence of a mechanism for segregation in high copy number plasmids that moves plasmids to cell poles to facilitate transmission to daughter cells. This mechanism would involve chromosomally-encoded proteins and the plasmid origin of replication. Modulation of this proposed mechanism of segregation could provide new ways to enhance plasmid stability in the context of recombinant gene expression, which is limiting for large-scale protein production and for bioremediation.

  20. Plasmid-to-plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1986-01-01

    No recombination between plasmids was observed after conjugal transfer of a plasmid into a cell carrying another plasmid. Two types of such recombination took place after transformation, one type being Rec/sup +/ dependent and suggesting a preferred site of recombination. The other much rarer type was at least partially Rec/sup +/ independent.

  1. Enhancement of protective immunity in European eel(Anguilla anguilla)against Aeromonas hydrophila and Aeromonas sobria by a recombinant Aeromonas outer membrane protein

    Institute of Scientific and Technical Information of China (English)

    Ruizhang Guan; Jing Xiong; Wenshu Huang; Songlin Guo

    2011-01-01

    To develop a vaccine,which can simultaneously prevent the diseases caused by various pathogenic bacteria in fish,we try to find a conserved outer membrane protein(OMP)antigen from different bacterial pathogens.In this study,an OMP fragment of 747 by(named as Omp-G),whichwas highly conserved in seven Aeromonas OMP sequences from the NCBI database,was amplified by PCR from one Aeromonas sobria strain(1110)and two Aeromonas hydrophila strains(1327 and B33)with the designed specific primers.The sequence was cloned into pGEX-2T(6 × His-tag)vector,expressed in Escherichia coli system,and then the recombinant protein(named as rOmp-G)was purified with nickel chelating affinity chromatography.The purified romp-G showed a good immunogenicity in rabbits and well-conserved characteristics in these three pathogens by enzyme-linked immunosorbed assay.Furthermore,the rOmp-G also showed good immunogenicity in eels(Anguilla anguilla)for eliciting significantly increased specific antibodies(P<0.01),and providing higher protection efficiencies(P<0.05)after the pathogens challenge.The values of the relative percent survival in eels were 70% and 50% for two A.hydrophila strain challenge,and 75% for A.sobria strain challenge.This is the first report of a potential vaccination in eels that simultaneously provide protectiveness against different Aeromonas pathogens with a conserved partial OMP.

  2. Traveller's diarrohea caused by Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    M. Martín Delgado

    2001-06-01

    Full Text Available Aeromonas hydrophila is a bacteria frequently found in fresh water and saline water. Certain strains of the species have been known to cause disease in fish and amphibians for several years, although some controversy exists about its ability to cause illness in humans. Nevertheless, either through open wounds or contaminated food or water, it can give rise to infection. However, almost all articles refer to isolated cases, rather than to outbreaks.In most cases it produces gastroenteritis in healthy people, whilst possibly causing septicemia in people with low inmmunity.In 1998, there was concern in the hotel sector due to a considerable rise in the number of cases of gastroenteritis among tourists holidaying in certain areas of the coast of the island of Tenerife.The Public Health Services of the Canarian Community iniciated an epidemiological study in order to establish the magnitude of the problem and the associated factors.The inicial analysis showed a large number of tourists affected but not seriously, and only in one particular area of the coast. This suggested traveller’s diarrohea probably associated with contaminated drinking water.

  3. Quorum sensing regulation in Aeromonas hydrophila.

    Science.gov (United States)

    Garde, Christian; Bjarnsholt, Thomas; Givskov, Michael; Jakobsen, Tim Holm; Hentzer, Morten; Claussen, Anetta; Sneppen, Kim; Ferkinghoff-Borg, Jesper; Sams, Thomas

    2010-03-01

    We present detailed results on the C4-HSL-mediated quorum sensing (QS) regulatory system of the opportunistic Gram-negative bacterium Aeromonas hydrophila. This bacterium contains a particularly simple QS system that allows for a detailed modeling of kinetics. In a model system (i.e., the Escherichia coli monitor strain MH205), the C4-HSL production of A. hydrophila is interrupted by fusion of gfp(ASV). In the present in vitro study, we measure the response of the QS regulatory ahyRI locus in the monitor strain to predetermined concentrations of C4-HSL signal molecules. A minimal kinetic model describes the data well. It can be solved analytically, providing substantial insight into the QS mechanism: at high concentrations of signal molecules, a slow decay of the activated regulator sets the timescale for the QS regulation loop. Slow saturation ensures that, in an A. hydrophila cell, the QS system is activated only by signal molecules produced by other A. hydrophila cells. Separate information on the ahyR and ahyI loci can be extracted, thus allowing the probe to be used in identifying the target when testing QS inhibitors. PMID:20064524

  4. Aeromonas lusitana sp. nov., Isolated from Untreated Water and Vegetables.

    Science.gov (United States)

    Martínez-Murcia, Antonio; Beaz-Hidalgo, Roxana; Navarro, Aaron; Carvalho, M João; Aravena-Román, Max; Correia, Antonio; Figueras, M José; Saavedra, M José

    2016-06-01

    During previous studies to evaluate the phylogenetic diversity of Aeromonas from untreated waters and vegetables intended for human consumption, a group of isolates formed a unique gyrB phylogenetic cluster, separated from those of all other species described so far. A subsequent extensive phenotypic characterization, DNA-DNA hybridization, 16S rRNA gene sequencing, multi-locus phylogenetic analysis of the concatenated sequence of seven housekeeping genes (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD; 4705 bp), and ERIC-PCR, were performed in an attempt to ascertain the taxonomy position of these isolates. This polyphasic approach confirmed that they belonged to a novel species of the genus Aeromonas, for which the name Aeromonas lusitana sp. nov. is proposed, with strain A.11/6(T) (=DSMZ 24095(T), =CECT 7828(T)) as the type strain. PMID:26868258

  5. Susceptibility of Aeromonas Hydophila Isolates to Antimicrobial Drugs

    Directory of Open Access Journals (Sweden)

    Igor Stojanov

    2010-05-01

    Full Text Available Aeromonas hydrophila is a microorganism widely distributed in nature: in water, soil, food. It is also part of the normal bacterial flora of many animals. As an opportune microorganism it is a secondary biological agent that contributes to the occurrence of a fish disease and its deterioration. Frequently, its presence is an indication of bad zoohygiene and zootechnical conditions in fish ponds. Reduced quality and quantity of feed, mechanical injuries, parasitosis, seasonal oscillation in temperature present some of the factors that produce favorable conditions for bacterial proliferation of aeromonas in fish ponds, so clinical symptoms of the disease occur. Aeromonas is almost always present in clinical isolates and may be unjustly accused for bad health of fish. Antibiotic therapy is applied even when the clinical findings are clear, what certainly effects the susceptibility to chemotherapeutics. The subject of our work was bacteriological examination of the material obtained from the carps with the observed skin changes and the carps without these changes. Also, antimicrobial susceptibility of Aeromonas hydrophila was tested. The aim of this research was to determined the presence of Aeromonas hydrophilia in the carp ponds and to test antibiotic susceptibility. The material consisted of the samples from the fish ponds where the carps were with and without changed skin. The method the isolation of Aeromonas hydrophila was used. The diffusion disk technique was used for testing antibiotic susceptibility. The isolates were tested for their susceptibility to Florephenikol, Flumequine, Olaqindox and Oxitetracycline. The obtained results point that antimicrobial susceptibility was the same regardless of the origin of the samples, i.e. the resistance was the same for both groups of samples (the strains isolated from the fish with skin changes and the strains from fish without changes on skin. The strains were highly resistant: 35% were resistant to

  6. [Bacteria of the genus Aeromonas and their role in aquaculture].

    Science.gov (United States)

    Kompanets, E V; Isaeva, N M; Balakhnin, I A

    1992-01-01

    Bacteria of genus Aeromonas are constant components of microbiota of fresh reservoirs where they, together with other microorganisms, play the part of natural biofilter and promote water self-purification. They are necessarily present in normal microflora of hydrobionts inhabiting fresh reservoirs. The greatest attention is paid by the researchers to Aeromonas and biotrophs in connection with epizootics in aquaculture which have become more frequent, in particular, under fish breeding. That is why the review is, to more extent, concerned in the works of this trend made by the foreign and home researchers for the last decade. PMID:1406386

  7. Persistence Mechanisms of Conjugative Plasmids

    DEFF Research Database (Denmark)

    Bahl, Martin Iain; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    maintenance in the host cell. These importantly include the ability to self-mobilize in a process termed conjugative transfer, which may occur across species barriers. Other plasmid stabilizing mechanisms include the multimer resolution system, active partitioning, and post-segregational-killing of plasmid......Are plasmids selfish parasitic DNA molecules or an integrated part of the bacterial genome? This chapter reviews the current understanding of the persistence mechanisms of conjugative plasmids harbored by bacterial cells and populations. The diversity and intricacy of mechanisms affecting the...... successful propagation and long-term continued existence of these extra-chromosomal elements is extensive. Apart from the accessory genetic elements that may provide plasmid-harboring cells a selective advantage, special focus is placed on the mechanisms conjugative plasmids employ to ensure their stable...

  8. 库拉索芦荟提取物对维氏气单胞菌抑菌作用的研究%Study on the Bacteriostasis of Aloe Vera Extracts against Aeromonas veronii

    Institute of Scientific and Technical Information of China (English)

    杨永立; 王利; 吴满林

    2013-01-01

    采用榨汁法、水煮法和醇提法制备库拉索芦荟提取物,通过K-B法和二倍稀释法测定3种方法芦荟提取物对维氏气单胞菌的抑菌作用.结果显示,用3种方法制备的库拉索芦荟提取物对维氏气单胞菌均有不同程度的抑制作用,榨汁法、水煮法和醇提法得到的库拉索芦荟提取物对3株维氏气单胞菌的平均最小抑菌浓度(minimal inhibitory concentration,MIC)分别为l、0.5和0.125 g/mL,最小杀菌浓度(minimal bactericidal concentration,MBC)分别为1、1和0.25 g/mL.库拉索芦荟提取物对3株维氏气单胞菌均有抑制作用,芦荟提取物浓度越高抑菌效果越明显,其中醇提法的抑菌效率高,水煮法的抑菌作用其次,榨汁法的抑菌效果较弱.

  9. Aeromonas hydrophila septicaemia and muscle abscesses associated with immunosuppression.

    OpenAIRE

    Slevin, N J; Oppenheim, B A; Deakin, D. P.

    1988-01-01

    We report a case of septicaemia and muscle abscesses due to Aeromonas hydrophila. The patient was immunosuppressed due to Hodgkin's disease and treatment with cytotoxic chemotherapy. The abscesses resolved after prolonged antibiotic therapy. Organisms that were once considered rarely pathogenic are emerging as increasingly important in causing potentially lethal infections in immunosuppressed patients.

  10. Aeromonas hydrophila in 2010: Characteristics of Alabama outbreaks

    Science.gov (United States)

    For a second year, epidemics associated with a virulent strain of Aeromonas hydrophila resulted in losses of hundreds of thousands of pounds of market size Alabama (AL) catfish. During this period, the Alabama Fish Farming Center diagnosed outbreaks of this strain of A. hydrophila on 25% (28/113) o...

  11. Ciprofloxacin-Resistant Aeromonas hydrophila Cellulitis following Leech Therapy

    OpenAIRE

    Giltner, Carmen L.; Bobenchik, April M; Uslan, Daniel Z.; Deville, Jaime G.; Humphries, Romney M.

    2013-01-01

    We report a case of surgical site infection with ciprofloxacin-resistant Aeromonas hydrophila following leech therapy. Antimicrobial and genetic analyses of leech and patient isolates demonstrated that the resistant isolates originated from the leech gut microbiota. These data suggest that ciprofloxacin monotherapy as a prophylaxis regimen prior to leech therapy may not be effective in preventing infection.

  12. Conjugative plasmids of Neisseria gonorrhoeae.

    Directory of Open Access Journals (Sweden)

    Emilia Pachulec

    Full Text Available Many clinical isolates of the human pathogen Neisseria gonorrhoeae contain conjugative plasmids. The host range of these plasmids is limited to Neisseria species, but presence of a tetracycline (tetM determinant inserted in several of these plasmids is an important cause of the rapid spread of tetracycline resistance. Previously plasmids with different backbones (Dutch and American type backbones and with and without different tetM determinants (Dutch and American type tetM determinants have been identified. Within the isolates tested, all plasmids with American or Dutch type tetM determinants contained a Dutch type plasmid backbone. This demonstrated that tetM determinants should not be used to differentiate between conjugal plasmid backbones. The nucleotide sequences of conjugative plasmids with Dutch type plasmid backbones either not containing the tetM determinant (pEP5233 or containing Dutch (pEP5289 or American (pEP5050 type tetM determinants were determined. Analysis of the backbone sequences showed that they belong to a novel IncP1 subfamily divergent from the IncP1alpha, beta, gamma, delta and epsilon subfamilies. The tetM determinants were inserted in a genetic load region found in all these plasmids. Insertion was accompanied by the insertion of a gene with an unknown function, and rearrangement of a toxin/antitoxin gene cluster. The genetic load region contains two toxin/antitoxins of the Zeta/Epsilon toxin/antitoxin family previously only found in Gram positive organisms and the virulence associated protein D of the VapD/VapX toxin/antitoxin family. Remarkably, presence of VapX of pJD1, a small cryptic neisserial plasmid, in the acceptor strain strongly increased the conjugation efficiency, suggesting that it functions as an antitoxin for the conjugative plasmid. The presence of the toxin and antitoxin on different plasmids might explain why the host range of this IncP1 plasmid is limited to Neisseria species. The isolated plasmids

  13. Phenotypic plasticity in bacterial plasmids.

    Science.gov (United States)

    Turner, Paul E

    2004-01-01

    Plasmid pB15 was previously shown to evolve increased horizontal (infectious) transfer at the expense of reduced vertical (intergenerational) transfer and vice versa, a key trade-off assumed in theories of parasite virulence. Whereas the models predict that susceptible host abundance should determine which mode of transfer is selectively favored, host density failed to mediate the trade-off in pB15. One possibility is that the plasmid's transfer deviates from the assumption that horizontal spread (conjugation) occurs in direct proportion to cell density. I tested this hypothesis using Escherichia coli/pB15 associations in laboratory serial culture. Contrary to most models of plasmid transfer kinetics, my data show that pB15 invades static (nonshaking) bacterial cultures only at intermediate densities. The results can be explained by phenotypic plasticity in traits governing plasmid transfer. As cells become more numerous, the plasmid's conjugative transfer unexpectedly declines, while the trade-off between transmission routes causes vertical transfer to increase. Thus, at intermediate densities the plasmid's horizontal transfer can offset selection against plasmid-bearing cells, but at high densities pB15 conjugates so poorly that it cannot invade. I discuss adaptive vs. nonadaptive causes for the phenotypic plasticity, as well as potential mechanisms that may lead to complex transfer dynamics of plasmids in liquid environments. PMID:15166133

  14. Plasmid acquisition in microgravity

    Science.gov (United States)

    Juergensmeyer, Margaret A.; Juergensmeyer, Elizabeth A.; Guikema, James A.

    1995-01-01

    In microgravity, bacteria often show an increased resistance to antibiotics. Bacteria can develop resistance to an antibiotic after transformation, the acquisition of DNA, usually in the form of a plasmid containing a gene for resistance to one or more antibiotics. In order to study the capacity of bacteria to become resistant to antibiotics in microgravity, we have modified the standard protocol for transformation of Escherichia coli for use in the NASA-flight-certified hardware package, The Fluid Processing Apparatus (FPA). Here we report on the ability of E. coli to remain competent for long periods of time at temperatures that are readily available on the Space Shuttle, and present some preliminary flight results.

  15. Plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.

    1982-01-01

    DNA recombination in exponential phase and competent Haemophilus influenzae was measured by an electron microscopic assay that relies on the conversion of plasmid RSF0885 monomers into multimeric forms. Dimer circles were present at a frequency of 2% in plasmid preparations from competent Rd (wild-type) cells; multimers were present at a frequency of 0.2% in preparations from exponential phase cells. Thus, plasmid recombination was stimulated in competent cells. Multimer formation occurred efficiently in cells of the transformation defective mutant rec2, implying that the rec2 gene product is not required for plasmid recombination. However, the absence of multimer plasmids in preparations from competent cells of the transformation defective mutant rec1 suggests that the rec1 gene product is required. Digestion of purified plasmids with restriction endonuclease PvuII, which makes a single cut in the monomer, revealed the presence of recombination intermediates composed of two linear plasmids joined to form two pairs of arms resembling the Greek letter chi. Length measurements of these arms taken from a population of recombination intermediates gave evidence that the plasmids were joined at sites of homology. The distributions of individual DNA strands, at the intersections of the four arms, could be resolved in some recombination intermediates and were of two types. The first type of junction appeared as a single-stranded arm appended to each corner. The second type of junction consisted of a single strand of DNA linking the two linear plasmids at a site of homology. The single-stranded linker was frequently situated at the edge of a short gap on one of the plasmids in the pair. The fine structures of the recombinational joints have been interpreted in terms of previously proposed models of recombination.

  16. Evolved plasmid-host interactions reduce plasmid interference cost.

    Science.gov (United States)

    Yano, Hirokazu; Wegrzyn, Katarznya; Loftie-Eaton, Wesley; Johnson, Jenny; Deckert, Gail E; Rogers, Linda M; Konieczny, Igor; Top, Eva M

    2016-09-01

    Antibiotic selection drives adaptation of antibiotic resistance plasmids to new bacterial hosts, but the molecular mechanisms are still poorly understood. We previously showed that a broad-host-range plasmid was poorly maintained in Shewanella oneidensis, but rapidly adapted through mutations in the replication initiation gene trfA1. Here we examined if these mutations reduced the fitness cost of TrfA1, and whether this was due to changes in interaction with the host's DNA helicase DnaB. The strains expressing evolved TrfA1 variants showed a higher growth rate than those expressing ancestral TrfA1. The evolved TrfA1 variants showed a lower affinity to the helicase than ancestral TrfA1 and were no longer able to activate the helicase at the oriV without host DnaA. Moreover, persistence of the ancestral plasmid was increased upon overexpression of DnaB. Finally, the evolved TrfA1 variants generated higher plasmid copy numbers than ancestral TrfA1. The findings suggest that ancestral plasmid instability can at least partly be explained by titration of DnaB by TrfA1. Thus under antibiotic selection resistance plasmids can adapt to a novel bacterial host through partial loss of function mutations that simultaneously increase plasmid copy number and decrease unfavorably high affinity to one of the hosts' essential proteins. PMID:27121483

  17. Effect of acute and chronic arsenic exposure on growth, structure and virulence of Aeromonas hydrophila isolated from fish.

    Science.gov (United States)

    Goswami, Ramansu; Ghosh, Debabrata; Saha, Dhira Rani; Padhy, Pratap Kumar; Mazumder, Shibnath

    2011-02-01

    Aeromonas hydrophila being a ubiquitous bacterium is prone to arsenic exposure. The present study was designed to determine the role of arsenic on growth and virulence of A. hydrophila. Exposure to arsenic (1 mg L(-1) and 2 mg L(-1)) had no effect on growth but significantly inhibited the hemolytic and cytotoxic potential of exposed bacteria. Transmission electron microscopy revealed loss of membrane integrity and presence of condensed cytoplasm suggestive of acute stress in bacteria exposed to arsenic. Arsenic-adapted bacteria were developed by repeated sub-culturing in presence of arsenic. Arsenic-adaptation led to significant recovery in hemolytic and cytotoxic potential. The arsenic-adapted bacteria exhibited normal membrane integrity, decreased cytoplasmic condensation and possessed scattered polysome like structures in the cytoplasm. A positive correlation was observed between arsenic tolerance and resistance to several antimicrobials. Arsenic-adaptation failed to confer cross-protection to mercury and cadmium stress. SDS-PAGE analysis revealed the expression of two new proteins of approximately 85 kDa and 79 kDa respectively in arsenic-adapted A. hydrophila. Plasmid-curing and transformation studies clearly indicate plasmid has no role on arsenic resistance trait of the bacteria. Our study, for the first time, reports a structure and function relationship of xenobiotics on bacteria.

  18. In vitro comparisons of the inhibitory activity of florfenicol copper sulfate and potassium permanganate towards Aeromonas hydrophila and Flavobacterium columnare

    Science.gov (United States)

    Aeromonas hydrophila and Flavobacterium columnare, the etiological agents of motile aeromonas septicemia (MAS) and columnaris disease, respectively, have been recently causing crippling moralities to the sunshine bass, Morone chrysops female X Morone saxatilis male (Percichthyidae), industry in the ...

  19. Screening and Characterization of a Bioflocculant Produced by Aeromonas sp.

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To isolate the bioflocculant-producing bacteria from activated sludge and investigate the flocculating characteristics of the newly isolated bioflocculant. Methods Bacteria were screened from activated sludge samples to isolate bioflocculant-producing bacteria. Flocculating activity was used as a measure of the flocculating capability of the bioflocculant.Results A novel bioflocculant-producing bacterium was isolated, which was identified to belong to genus Aeromonas and named as Aeromonas sp. N11. Flocculating activity increased in the presence of K+, Na+, or Ca2+. The highest flocculating activities for kaolin suspension were obtained in acidic pH ranges, and optimum pHs for it were 3.0, 4.0, and 5.0 with 1 mmol/L K+, Ca+, and Na+ present, respectively. The highest flocculating activities for soil suspension were observed at pH 8.0. The bioflocculant had a good flocculating activity and could achieve a flocculating activity of 92.4% for kaolin suspension at a Conclusion The bioflocculant produced by Aeromonas sp. N11 has strong flocculating activity and high stability, which affords high possibility of its practical use.

  20. Producción de ácido indol-3-acético por Pseudomonas veronii R4 y formación de raíces en hojas de vid “Thompson seedless” in vitro

    Directory of Open Access Journals (Sweden)

    María F. Peñafiel-Jaramillo

    2016-06-01

    Full Text Available Rizobacterias con actividad antagonista son empleadas a problemas patológicos y promueven el desarrollo de tejido radicular de especies vegetales El objetivo del estudio fue evaluar la capacidad en síntesis de ácido indol-3-acetico (AIA y la formación de raíces en hojas de vid “Thompson Seedless”, por el empleo de Pseudomonas veronii R4 in vitro. La bacteria, fue incubada en medio mínimo M9 y se verificó la producción de AIA a 24, 48 y 72 h de muestreo, los sobrenadantes fueron analizandos por HPLC. Peciolos de hojas de “Thompson Seedless” fueron sumergidos por 5 min, en el tampón PBS con P. veronii R4 a (1.5 x 109 células mL-1, y evaluados a los 32 días post-inoculación. Los tratamientos fueron sujetos al análisis de varianza por DCA. Los sobrenadantes muestreados a 72 h, del cultivo de líquido con P. veronii R4, se obtuvo los mayores niveles de AIA entre 0.0231 a 0.0245 mM, al emplear 5 a 10 mM del precursor L-triptofano, del sobrenadante obtenido de P. fluorescens CHA0 la producción de AIA con 0.0174 mM a 10 mM de L-triptofano. Se obtuvo la regeneración del sistema radicular en los peciolos de hojas de “Thompson Seedless”, con un promedio de 14 raíces/planta bajo la aplicación de P. veronii R4. Estos resultados preliminares realzan la importancia de la cepa nativa P. veronii R4, que se estaría proyectando para futuras investigaciones de su empleo en condición de invernadero al desarrollo del sistema radicular en “Thompson Seedless”.

  1. Diarrea del viajero asociada a Aeromonas hydrophila Traveller's diarrohea caused by Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    M. L. Pita Toledo

    2001-06-01

    Full Text Available Aeromonas hydrophila es una bacteria que se encuentra frecuentemente en aguas dulces y salobres. Desde hace varios años se conoce que determinadas cepas de la especie pueden causar enfermedad en pescados y en anfibios, existiendo cierta controversia sobre su patogeneicidad en humanos. No obstante, puede dar lugar a infección en personas, que la adquieren bien a través de heridas abiertas o bien por ingestión de alimentos o agua contaminados. Sin embargo, en prácticamente la totalidad de las publicaciones se hace referencia a casos aislados más que a brotes producidos por esta bacteria. Generalmente la infección por A. hydrophila da lugar a gastroenteritis en personas sanas, pudiendo generar una septicemia en individuos inmunodeprimidos.En 1998 se produjo una alarma en el sector hotelero por un incremento considerable de los casos de gastroenteritis entre los turistas que acudían durante sus vacaciones a determinadas zonas costeras de la isla de Tenerife.Tras tener conocimiento de la situación, la Dirección General de Salud Pública inició una investigación epidemiológica con el objetivo de conocer la magnitud real del problema detectado e identificar los factores asociados a la misma, para poner en marcha las medidas correctoras y de control necesarias. El análisis inicial permitió determinar que se trataba de un proceso de gran magnitud en su extensión aunque de carácter leve, que afectaba únicamente a los turistas en una zona costera en concreto, compatible por lo tanto con la diarrea del viajero, planteándose un origen probablemente hídrico.Aeromonas hydrophila is a bacteria frequently found in fresh water and saline water. Certain strains of the species have been known to cause disease in fish and amphibians for several years, although some controversy exists about its ability to cause illness in humans. Nevertheless, either through open wounds or contaminated food or water, it can give rise to infection. However, almost

  2. Classification of a hypervirulent Aeromonas hydrophila pathotype responsible for epidemic outbreaks in warm-water fishes

    Science.gov (United States)

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the P...

  3. Whole-Genome Sequence of Aeromonas hydrophila Strain AH-1 (Serotype O11).

    Science.gov (United States)

    Forn-Cuní, Gabriel; Tomás, Juan M; Merino, Susana

    2016-01-01

    Aeromonas hydrophila is an emerging pathogen of aquatic and terrestrial animals, including humans. Here, we report the whole-genome sequence of the septicemic A. hydrophila AH-1 strain, belonging to the serotype O11, and the first mesophilic Aeromonas with surface layer (S-layer) to be sequenced. PMID:27587829

  4. Aeromonas hydrophila urinary tract infection in pregnancy- Case report and literature review

    Directory of Open Access Journals (Sweden)

    Latha Ragunathan

    2012-03-01

    Full Text Available A case of a pregnant woman without previous or concomitant disease, who developed an Aeromonas hydrophila urinarytract infection (UTI at 12 weeks gestation, is reported. A brief review of the literature on the association and incidenceof Aeromonas spp in urinary infections and also in association with pregnancy is presented. J Microbiol Infect Dis 2012;2(1: 26-28

  5. Draft Genome Sequence of High-Melanin-Yielding Aeromonas media Strain WS

    OpenAIRE

    Chai, Baozhong; Wang, He; Chen, Xiangdong

    2012-01-01

    We sequenced the genome of the high-melanin-yielding Aeromonas media strain WS and then analyzed genes potentially involved in melanin formation. The 4.2-Mb draft genome carries multiple genes responsible for pyomelanin synthesis and other candidate genes identified in our separate study, which have no homolog in other strains of Aeromonas species.

  6. Effects of genes exerting growth inhibition and plasmid stability on plasmid maintenance.

    OpenAIRE

    Boe, L; Gerdes, K; Molin, S

    1987-01-01

    Plasmid stabilization mediated by the parA+ and parB+ genes of the R1 plasmid and the ccd+ and sop+ genes of the F plasmid was tested on a mini-R1 plasmid and a pBR322 plasmid derivative. The mini-R1 plasmid is thought to be unstably inherited owing to a low copy number and to random segregation of the plasmid at cell division, whereas cells harboring the pBR322 derivative used in this work are lost through competition with plasmid-free cells, mainly as a result of the shorter generation time...

  7. Effect of solar irradiation on extracellular enzymes of Aeromonas proteolytica

    Science.gov (United States)

    Foster, B. G.

    1973-01-01

    The bacterium Aeromonas proteolytica was selected for studying the effects of solar irradiation on extracellular enzymes because it produces an endopeptidase that is capable of degrading proteins and a hemolysin that is active in lysing human erythrocytes. Possible alterations in the rate of enzyme production in response to the test conditions are currently underway and are not available for this preliminary report. Completed viability studies are indicative that little difference exists among the survival curves derived for cells exposed to various components of ultraviolet irradiation in space.

  8. Multifocal Aeromonas Osteomyelitis in a Child with Leukemia

    Directory of Open Access Journals (Sweden)

    Dimitrios Doganis

    2016-01-01

    Full Text Available Aeromonas hydrophila is a Gram negative organism causing both intestinal and extraintestinal disease. The case of a 14-year-old girl with underlying immunodeficiency and leukemia who developed systemic A. hydrophila infection is described in this report. While in deep bone marrow aplasia she developed fever, severe pain in the lower extremities, and swelling of the left femur. Blood culture showed Escherichia coli and A. hydrophila whereas pus culture from the soft tissue swelling showed the presence of A. hydrophila. Imaging studies showed diffuse osteolytic lesions. Patient received 5 months of intravenous and oral antibiotics and she improved clinically whereas the radiology findings persisted.

  9. Genome sequencing and annotation of Aeromonas sp. HZM

    Directory of Open Access Journals (Sweden)

    Patric Chua

    2015-09-01

    Full Text Available We report the draft genome sequence of Aeromonas sp. strain HZM, isolated from tropical peat swamp forest soil. The draft genome size is 4,451,364 bp with a G + C content of 61.7% and contains 10 rRNA sequences (eight copies of 5S rRNA genes, single copy of 16S and 23S rRNA each. The genome sequence can be accessed at DDBJ/EMBL/GenBank under the accession no. JEMQ00000000.

  10. ISOLATION AND GENOTYPING OF AEROMONAS SPP. IN READY-TO-EAT FOODS. PRELIMINARY RESULTS

    Directory of Open Access Journals (Sweden)

    F. Montemurro

    2011-08-01

    Full Text Available The taxonomy of the genus Aeromonas is constantly changing and it is important that strains are identified and carefully differentiate. The aim of this preliminary investigation was to assess the presence of Aeromonas spp. in RTE foods from supermarkets and sushi Take Away and stored at refrigeration temperature. Particular attention was given to the choice of culture media in order to assess their ability to differentiate Aeromonas spp. and to perform subsequent phylogenetic analysis and the characterization of the species isolated.

  11. Bacterial Plasmids in Antarctic Natural Microbial Assemblages

    OpenAIRE

    Kobori, Hiromi; Sullivan, Cornelius W.; Shizuya, Hiroaki

    1984-01-01

    Samples of psychrophilic and psychrotrophic bacteria were collected from sea ice, seawater, sediments, and benthic or ice-associated animals in McMurdo Sound, Antarctica. A total of 155 strains were isolated and tested for the presence of plasmids by DNA agarose gel electrophoresis. Thirty-one percent of the isolates carried at least one kind of plasmid. Bacterial isolates taken from sediments showed the highest plasmid incidence (42%), and isolates from seawater showed the lowest plasmid inc...

  12. Plasmid required for virulence of Agrobacterium tumefaciens

    Energy Technology Data Exchange (ETDEWEB)

    Watson, B.; Currier, T.C.; Gordon, M.P.; Chilton, M.D.; Nester, E.W.

    1975-07-01

    The irreversible loss of crown gall-inducing ability of Agrobacterium tumefaciens strain C-58 during growth at 37/sup 0/C is shown to be due to loss of a large plasmid (1.2 x 10/sup 8/ daltons). The gene responsible for this high rate of plasmid loss at elevated temperatures seems to be located on the plasmid. In addition, another spontaneous avirulent variant, A. tumefaciens strain IIBNV6, is shown to lack the virulence plasmid which its virulent sibling strain, IIBV7, possesses. Deoxyribonucleic acid reassociation measurements prove that the plasmid is eliminated, not integrated into the chromosome, in both of the avirulent derivatives. Transfer of virulence from donor strain C-58 to avirulent recipient strain A136 results from the transfer of a plasmid, which appears identical to the donor plasmid by deoxyribonucleic acid reassociation measurements. The transfer of virulence in another cross, K27 x A136, was also shown to result from the transfer of a large plasmid. These findings establish unequivocally that the large plasmid determines virulence. Two additional genetic determinants have been located on the virulence plasmid of A. tumefaciens strain C-58: the ability to utilize nopaline and sensitivity to a bacteriocin produced by strain 84. The latter trait can be exploited for selection of avirulent plasmid-free derivatives of strain C-58. The trait of nopaline utilization appears to be on the virulence plasmid also in strains IIBV7 and K27.

  13. First case of Aeromonas schubertii infection in the freshwater cultured snakehead fish, Ophiocephalus argus (Cantor), in China.

    Science.gov (United States)

    Liu, J Y; Li, A H

    2012-05-01

    An epizootic in snakehead fish, Ophiocephalus argus, in earthen ponds in Xianning, Hubei Province, central China, from June to August 2009 was found to be caused by Aeromonas schubertii. The cumulative mortality within 40 days was 45%, and the diseased fish were 18 months old and 35-45 cm in length. Multiple, ivory-white, firm nodules, 0.5-1 mm in diameter, were scattered throughout the kidney. Blood clots, 3-5 mm in diameter, were found in the liver. This is a disease frequently found in cultured snakehead throughout China. Isolated bacteria were Gram negative, facultatively anaerobic, motile, short rod-shaped, with a length of 0.3-1.0 μm. Morphological and biochemical tests, as well as phylogenetic analysis derived from 16S rRNA, gyrB, rpoD and dnaJ gene sequencing all strongly indicated that these snakehead isolates are identical to A. schubertii. In addition, the isolates possessed two plasmids: 5.0 kb and 10.0 kb. Antibiotic sensitivity testing of the isolates was carried out by the standard Kirby-Bauer disc diffusion method. Experimental infection assays were conducted, and pathogenicity (by intraperitoneal injection) was demonstrated in snakehead fingerlings and zebrafish, Brachydanio rerio (Hamilton).

  14. In Silico Detection and Typing of Plasmids using PlasmidFinder and Plasmid Multilocus Sequence Typing

    DEFF Research Database (Denmark)

    Carattoli, Alessandra; Zankari, Ea; García-Fernández, Aurora;

    2014-01-01

    In the work presented here, we designed and developed two easy-to-use Web tools for in silico detection and characterization of whole-genome sequence (WGS) and whole-plasmid sequence data from members of the family Enterobacteriaceae. These tools will facilitate bacterial typing based on draft...

  15. Plasmid maintenance functions encoded on Dictyostelium discoideum nuclear plasmid Ddp1.

    OpenAIRE

    Hughes, J E; H. Kiyosawa; Welker, D L

    1994-01-01

    All of the plasmid-carried genes expressed during vegetative growth are essential for long-term maintenance of plasmid Ddp1 in the nucleus of Dictyostelium discoideum. Deletion of Ddp1 genes expressed only during development had no detectable effect on plasmid maintenance. Deletion of vegetatively expressed genes, either singly or in pairs, resulted in (i) a rapid loss of plasmid from cells grown in the absence of selection for plasmid retention, (ii) variation in the proportion of monomer to...

  16. Prevalence and diversity of Aeromonas and Vibrio spp. in coastal waters of Southern Italy

    DEFF Research Database (Denmark)

    Dumontet, S.; Krovacek, K.; Svenson, S.B.;

    2000-01-01

    % of samples were positive for Vibrio spp. It was interesting to note that 38% of the positive stations for both Aeromonas and Vibrio spp. showed a fecal coliform contamination of water at fecal......A survey was undertaken to examine sea water and sediment for the presence of Vibrio and Aeromonas spp. along approximately 900 km of coast in Southern Italy during early and late summer. A quantitative analysis was also done to evaluate the water fecal contamination at the stations examined....... The results indicate that all the investigated areas were submitted to a wide spatial fluctuation of fecal contamination and that Vibrio and Aeromonas spp. were present in both high and low fecal-contaminated stations. Sixty two percent of the investigated samples were positive for Aeromonas spp., while 42...

  17. Clonagem e expressão de uma potencial colagenase de Aeromonas hydrophila

    OpenAIRE

    Cavaleiro, Eliana Marisa dos Santos

    2008-01-01

    Aeromonas hydrophila é uma bactéria Gram-negativa que causa septicemia quer em peixes quer em humanos. Os principais patologias associados a infecções por Aeromonas são gastroenterites, infecção de feridas e doença sistémica. A patogenecidade deste organismo é multifactorial, possuindo factores na camada superficial proteica (camada S), diversos produtos extracelulares incluindo proteases, hemolisinas e enterotoxinas incluindo a acetilcolinesterase que contribui, significati...

  18. Impact of an Urban Effluent on Antibiotic Resistance of Riverine Enterobacteriaceae and Aeromonas spp.

    OpenAIRE

    Goñi-Urriza, Marisol; Capdepuy, Michèle; Arpin, Corinne; Raymond, Nathalie; Caumette, Pierre; Quentin, Claudine

    2000-01-01

    In order to evaluate the impact of an urban effluent on antibiotic resistance of freshwater bacterial populations, water samples were collected from the Arga river (Spain), upstream and downstream from the wastewater discharge of the city of Pamplona. Strains of Enterobacteriaceae (representative of the human and animal commensal flora) (110 isolates) and Aeromonas (typically waterborne bacteria) (118 isolates) were selected for antibiotic susceptibility testing. Most of the Aeromonas strains...

  19. Extended-spectrum -lactamase and carbapenemase-producing Aeromonas species in wild animals from Portugal

    OpenAIRE

    Dias, C; Serra, C. R.; Simões, Lúcia C.; Simões, M.; Martinez-Murcia, A.; Saavedra, M.J.

    2014-01-01

    Aeromonas are Gram-negative, facultative-anaerobic, non-spore-forming, glucose-fermenting, oxidase- and catalase-positive rods (Martin-Carnahan and Joseph 2005). Apart from fish, which are widely reported hosts for Aeromonads, insects, crustaceans, reptiles, birds and mammals were also found to harbour Aeromonas species, both in healthy and disease state (Pearson and others 2000, Turutoglu and others 2005, Evangelista-Barreto and others 2006, Ceylan and others 2009). An increase in resistance...

  20. Studies on the clinical characteristics of intestinal infection caused by Aeromonas and drug susceptibility of the strains%气单胞菌肠道感染的临床特点和耐药特征分析

    Institute of Scientific and Technical Information of China (English)

    刘志远; 马玉芝; 潘健; 张婷菊; 齐杰; 刘贵建

    2014-01-01

    Objective To analyze the clinical characteristics of intestinal infection caused by Aeromonas and their drug suscepti-bility .Methods The data of 52 patients infected with Aeromonas were analyzed retrospectively .Aeromonas strains were identified with Vitek Ⅱ Compact .Drug susceptibility was determined by disk diffusion methods .Cefoxitin combined with ceftazidime ,aztreo-nam ,cefotaxime and ceftriaxone were used to detect inducible expression of AmpC β-lactamase .Results Abdominal pain ,watery di-arrhea ,tenesmus occurred in 75 .0% ,48 .1% ,and 38 .0% of the patients ,respectively .White blood cell tests were positive in 50 .0%patients′stools .52 strains of Aeromonas were isolated ,including 38 strains of Aeromonas hydrophila ,13 strains of Aeromonas so-bria ,and 1 strain of Aeromonas veronii .All strains were sensitive to carbapenems ,the second ,third generation of cephalosporins , monobactam ,fluoroquinolone ,aminoglycosides .The susceptibility rates to chloramphenicol ,trimethoprim-sulfamethoxazole ,cefox-itin ,cefazolin ,and amoxicillin/clavulanic acid were 94 .2% ,92 .3% ,76 .9% ,51 .9% and 23 .1% ,respectively .75 .0% isolates exhibi-ted inducible expression of AmpC β-lactamas .Conclusion Diarrhea caused by A eromonas has different clinical manifestations .Ceph-alosporins ,monobactam ,fluoroquinolone ,aminoglycosides are available for empirical therapy of diarrhea caused by A eromonas .But the third generation of cephalosporins should be cautiously used because of high prevalence of inducible AmpC β-lactamase in A ero-monas .%目的:了解气单胞菌所致腹泻的临床特点及气单胞菌的耐药情况。方法回顾性分析52例气单胞菌感染性腹泻患者的临床特点,采用Vitek Ⅱ Compact微生物分析仪鉴定气单胞菌,药敏试验采用纸片琼脂扩散法,头孢西丁纸片联合头孢他啶、头孢噻肟、头孢曲松和氨曲南纸片检测诱导型AmpC酶。结果75.0%气单胞菌感

  1. High frequency of coinfecting enteropathogens in Aeromonas-associated diarrhea of hospitalized Peruvian infants.

    Science.gov (United States)

    Pazzaglia, G; Sack, R B; Salazar, E; Yi, A; Chea, E; Leon-Barua, R; Guerrero, C E; Palomino, J

    1991-06-01

    Rectal swabs from 391 infants less than 18 months of age who were hospitalized with acute diarrhea and from 138 similarly aged healthy infants were examined for the etiologic agents of diarrhea. Aeromonas spp. were recovered from 205 of 391 (52.4%) diarrheic patients, whereas they were recovered from 12 of 138 (8.7%) controls (P less than 10(-11). Among the 205 Aeromonas-positive diarrheic patients, 118 (57.6%) were found to be coinfected with other common enteropathogens. Of the 164 Aeromonas-positive initial diarrheic specimens, 82 (50.0%) had one or more other enteropathogens present; 30 patients were coinfected with rotavirus, 20 with enterotoxigenic Escherichia coli, 16 with Campylobacter spp., 14 with Shigella spp., 13 with enteropathogenic E. coli, 4 with Vibrio spp., 1 with Salmonella spp., and 1 with Plesiomonas spp. of Aeromonas strains from cases compared with that from controls supports an etiologic role for this organism. However, frequent concomitant infections with other well-recognized enteropathogens and a lack of disease correlation with common Aeromonas phenotypes suggest that only a subset of Aeromonas strains may be diarrhea causing and that such strains may be common to several of the existing species.

  2. Mortality of therapeutic fish Garra rufa caused by Aeromonas sobria

    Institute of Scientific and Technical Information of China (English)

    Juraj Majtn; Jaroslaverny; Alena Ofkan; Peter Tak; Milan Koznek

    2012-01-01

    Objective:To investigate a case of mass mortality of Garra rufa (G. rufa) from a fish hatchery farm in Slovakia. Methods: Causative bacterial agent was swabbing out of affected fish skin area and subsequently identified using commercial test system. Antibiotic susceptibility was determined by the disk diffusion method. Results: Infected G. rufa was characterized by abnormal swimming behaviour, bleeding of skin lesions and local haemorrhages. Despite of using recommended aquatic antibiotic treatment no improvement was achieved and Aeromonas sobria (A. sobria) was identified as a causative agent of fish mortality. Due to massive fish mortality, antibiotic susceptibility of pure isolated culture of A. sobria was evaluated employing eight antibiotics against human infections. A. sobria was resistant only against one antibiotic, namely ampicilin. Conclusions: These results indicate that A. sobria can act as a primary pathogen of G. rufa and may be a potential risk factor for immunodeficient or immunoincompetent patients during the ichthyotherapy.

  3. Comparação da sensibilidade de Danio rerio, Poecilia reticulata e Xiphophorus helleri a Aeromonas hydrophila e Aeromonas aquariorum

    OpenAIRE

    Chambel Martins, João

    2011-01-01

    Dissertação de Mestrado em Aquacultura apresentada à ESTM - Escola Superior de Turismo e Tecnologia do Mar do Instituto Politécnico de Leiria Este trabalho teve como objectivo comparar a sensibilidade de três das principais espécies de peixes ornamentais de água doce, Danio rerio, Poecilia reticulata e Xiphophorus helleri, a duas bactérias potencialmente zoonóticas para o Homem, Aeromonas hydprophila (SM 30016) e Aeromonas aquariorum (DSM 18362). De forma a efectuar a injecção intraperi...

  4. Homemade Site Directed Mutagenesis of Whole Plasmids

    Science.gov (United States)

    Laible, Mark; Boonrod, Kajohn

    2009-01-01

    Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned target gene can be altered by substitution, deletion or insertion of a few bases directly into a plasmid. It works by simply amplifying the whole plasmid, in a non PCR-based thermocycling reaction. During the reaction mutagenic primers, carrying the desired mutation, are integrated into the newly synthesized plasmid. In this video tutorial we demonstrate an easy and cost effective way to introduce base substitutions into a plasmid. The protocol works with standard reagents and is independent from commercial kits, which often are very expensive. Applying this protocol can reduce the total cost of a reaction to an eighth of what it costs using some of the commercial kits. In this video we also comment on critical steps during the process and give detailed instructions on how to design the mutagenic primers. PMID:19488024

  5. pLS010 plasmid vector

    Science.gov (United States)

    Lacks, Sanford A.; Balganesh, Tanjore S.

    1988-01-01

    Disclosed is recombinant plasmid pLS101, consisting essentially of a 2.0 Kb malM gene fragment ligated to a 4.4 Kb T.sub.c r DNA fragment, which is particularly useful for transforming Gram-positive bacteria. This plasmid contains at least four restriction sites suitable for inserting exogeneous gene sequences. Also disclosed is a method for plasmid isolation by penicillin selection, as well as processes for enrichment of recombinant plasmids in Gram-positive bacterial systems.

  6. ANALYSIS OF A MODEL OF PLASMID-BEARING, PLASMID-FREE COMPETITION IN A PULSED CHEMOSTAT

    OpenAIRE

    XIANGYUN SHI; XINYU SONG; XUEYONG ZHOU

    2006-01-01

    We introduce and study a chemostat model with plasmid-bearing, plasmid-free competition and impulsive effect. According to the stability analysis of the boundary periodic solution, we obtain the invasion threshold of the plasmid-free organism and plasmid-bearing organism. Furthermore, by using standard techniques of bifurcation theory, we prove the system has a positive τ-periodic solution, which shows that the impulsive effect destroys the equilibria of the unforced continuous system and ini...

  7. Effect of chromosome homology an plasmid transformation and plasmid conjugal transfer in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1984-05-14

    The pairing between plasmid and the homologous part of the chromosome associated with plasmid establishment may differ from the pairing which results from integration of a homologous region of the plasmid into the chromosome. Thus the rate of novobiocin transformation decreases with duplication of the chromosomal portion in pMB2, but the rate of establishment of the plasmid increases with this duplication. A model to explain these data is given. 17 references, 5 figures, 4 tables.

  8. The expression of a plasmid-specified exported protein causes structural plasmid instability in Bacillus subtilis

    NARCIS (Netherlands)

    Cordes, C.; Meima, R; Twiest, B; Kazemier, B; Venema, G; vanDijl, JM; Bron, S

    1996-01-01

    The rolling-circle plasmid pGP1 was used to study the effects of the expression of a plasmid-specified exported protein on structural plasmid stability in Bacillus subtilis. pGP1 contains a fusion between the Bacillus licheniformis penP gene, encoding a C-terminally truncated penicillinase, and the

  9. Plasmid ColVBtrp maintenance in Erwinia carotovora.

    OpenAIRE

    Schukin, N N

    1981-01-01

    Plasmid ColVBtrp maintenance in Erwinia carotovora cells was followed by measuring kinetics of elimination of plasmid genetic markers and loss of plasmid deoxyribonucleic acid. An E. carotovora mutant stably carrying plasmid ColVBtrp was isolated. Besides stable plasmid maintenance, the mutant showed altered sensitivity to male-specific phage MS2, sensitivity to drugs, and colony morphology.

  10. Identification and molecular characterization of the homogentisate pathway responsible for pyomelanin production, the major melanin constituents in Aeromonas media WS.

    Directory of Open Access Journals (Sweden)

    He Wang

    Full Text Available The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate. Gene products of phhA (encodes phenylalanine hydroxylase, tyrB and aspC (both encode aromatic amino acid aminotransferase, and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species.

  11. A prospective study on Aeromonas in outpatients with diarrhea in the central region of Rio Grande do Sul State

    Directory of Open Access Journals (Sweden)

    Karoline de Campos Prediger

    2012-09-01

    Full Text Available Aeromonas spp. were identified in five (2,7% of 182 diarrheal stool cultures, A. caviae was predominant, resistant mainly to ampicillin and cephalotin. This is the first study showing the presence of Aeromonas spp. in diarrheal stools of outpatients in the central region of Rio Grande do Sul State, Brazil.

  12. Molecular detection and antimicrobial resistance of Aeromonas from houseflies (Musca domestica in Iran

    Directory of Open Access Journals (Sweden)

    Davood Ommi

    2015-11-01

    Full Text Available Objective. This study aimed to report the molecular detection and antimicrobial resistance of Aeromonas among houseflies (Musca domestica in Shahrekord and Isfahan provinces of Iran. Materials and methods. Flies were caught from household kitchens, cattle farms, animal hospitals, human hospitals, slaughter house and poultry farms and put in collection separate sterile tubes. Isolation was accomplished by culture of flies in alkaline peptone water followed by identification with Aeromonas-specific Polymerase Chain Reaction (PCR. Results. Out of 600 houseflies 73 (12.2% were infected with Aeromonas spp. Significantly higher frequencies of Aeromonas were isolated in Shahrekord province (13.0%; 39/300 than in Isfahan province (11.3%; 34/300. The recovery frequencies of the organisms were significantly lower in kitchens as compared to those in cattle farms and hospital wards which were similar. Higher proportions of infected flies were obtained during summer whereas low proportions were obtained during winter. Conclusions. It is concluded that houseflies do harbor diarrheagenic pathogens, including Aeromonas especially during summer. The carried organisms are resistant to a number of antimicrobials at different levels. Thus, future plans aimed at stemming infections caused by these organisms should take flies into account. Control efforts of infections caused by this particular bacterium should therefore take into account Musca domestica.

  13. Ocorrência de bactérias do gênero Aeromonas em queijo-de-minas frescal artesanal Occurrence of the genus Aeromonas in minas frescal cheese

    Directory of Open Access Journals (Sweden)

    C.C.C. Bulhões

    2002-06-01

    Full Text Available Foram analisadas 160 amostras de queijo-de-minas frescal artesanal, adquiridas no comércio varejista dos municípios de Poços de Caldas - MG e Jaboticabal - SP, a fim de verificar a ocorrência de bactérias do gênero Aeromonas no produto. Oitenta e duas (51,2% encontravam-se contaminadas pelos microrganismos, com populações que variavam de 5,0×10³ a 4,0×10(5 UFC/grama. Foram identificadas as espécies Aeromonas hydrophila, Aeromonas caviae, Aeromonas schubertii, além de cepas consideradas atípicas. Os resultados evidenciam que bactérias do gênero Aeromonas podem ser veiculadas através do queijo tipo minas frescal artesanal e devem servir de alerta aos serviços de saúde pública.One hundred and sixty samples of homemade minas frescal cheese, obtained at the retail trade in the municipalities of Poços de Caldas-MG and Jaboticabal-SP, were analyzed for occurrence of the genus Aeromonas. Eighty two samples (51.2% were contaminated by these microorganisms with populations that ranged from 5.0 x 10³ to 4.0 x 10(5 CFU/g. The species Aeromonas hydrophila, Aeromonas caviae, Aeromonas schubertii, were identified, along with atypical strains. The results show that Aeromonas bacteria genus can be transmitted through the homemade minas frescal cheese and they should serve of alert to the public health services.

  14. Physiological control of amonabactin biosynthesis in Aeromonas hydrophila.

    Science.gov (United States)

    Barghouthi, S; Young, R; Arceneaux, J E; Byers, B R

    1989-01-01

    Amonabactin is a siderophore from Aeromonas hydrophila which is produced in two biologically active forms composed of the phenolate 2,3-dihydroxybenzoic acid (DHB), lysine, glycine, and either trytophan (amonabactin T) or phenylalanine (amonabactin P). Amonabactin biosynthetic mutants (generated by chemical mutagenesis) that either produced no amonabactin or overproduced the siderophore were isolated and identified on chrome azurol S siderophore detection agar. Amonabactin-negative mutants were of two categories. One type produced no phenolates and used exogenous DHB to synthesize amonabactin (both forms) while the other type excreted DHB but not amonabactin. This suggests an amonabactin biosynthetic pathway composed of two segments, one producing DHB and the other assembling amonabactin from DHB and the amino acids. Overproduction mutants used amonabactin poorly or not at all, indicating that they contained lesions in amonabactin utilization. Adding the analog D-tryptophan to wild-type A. hydrophila cultures reduced synthesis of both amonabactin T and amonabactin P and lengthened the lag phase in iron restricted medium. The tryptophan and phenylalanine forms of amonabactin may be synthesized by a single assembly pathway that contains a novel enzyme (sensitive to D-tryptophan) which inserts either tryptophan or phenylalanine into amonabactin.

  15. Virulence Factors of Aeromonas hydrophila: In the Wake of Reclassification.

    Science.gov (United States)

    Rasmussen-Ivey, Cody R; Figueras, Maria J; McGarey, Donald; Liles, Mark R

    2016-01-01

    The ubiquitous "jack-of-all-trades," Aeromonas hydrophila, is a freshwater, Gram-negative bacterial pathogen under revision in regard to its phylogenetic and functional affiliation with other aeromonads. While virulence factors are expectedly diverse across A. hydrophila strains and closely related species, our mechanistic knowledge of the vast majority of these factors is based on the molecular characterization of the strains A. hydrophila AH-3 and SSU, which were reclassified as A. piscicola AH-3 in 2009 and A. dhakensis SSU in 2013. Individually, these reclassifications raise important questions involving the applicability of previous research on A. hydrophila virulence mechanisms; however, this issue is exacerbated by a lack of genomic data on other research strains. Collectively, these changes represent a fundamental gap in the literature on A. hydrophila and confirm the necessity of biochemical, molecular, and morphological techniques in the classification of research strains that are used as a foundation for future research. This review revisits what is known about virulence in A. hydrophila and the feasibility of using comparative genomics in light of this phylogenetic revision. Conflicting data between virulence factors, secretion systems, quorum sensing, and their effect on A. hydrophila pathogenicity appears to be an artifact of inappropriate taxonomic comparisons and/or be due to the fact that these properties are strain-specific. This review audits emerging data on dominant virulence factors that are present in both A. dhakensis and A. hydrophila in order to synthesize existing data with the aim of locating where future research is needed. PMID:27610107

  16. Cloning, mutagenesis, and nucleotide sequence of a siderophore biosynthetic gene (amoA) from Aeromonas hydrophila.

    OpenAIRE

    Barghouthi, S; Payne, S M; Arceneaux, J E; Byers, B R

    1991-01-01

    Many isolates of the Aeromonas species produce amonabactin, a phenolate siderophore containing 2,3-dihydroxybenzoic acid (2,3-DHB). An amonabactin biosynthetic gene (amoA) was identified (in a Sau3A1 gene library of Aeromonas hydrophila 495A2 chromosomal DNA) by its complementation of the requirement of Escherichia coli SAB11 for exogenous 2,3-DHB to support siderophore (enterobactin) synthesis. The gene amoA was subcloned as a SalI-HindIII 3.4-kb DNA fragment into pSUP202, and the complete n...

  17. Bifurcation Analysis of a Chemostat Model of Plasmid-Bearing and Plasmid-Free Competition with Pulsed Input

    OpenAIRE

    Zhong Zhao; Baozhen Wang; Liuyong Pang; Ying Chen

    2014-01-01

    A chemostat model of plasmid-bearing and plasmid-free competition with pulsed input is proposed. The invasion threshold of the plasmid-bearing and plasmid-free organisms is obtained according to the stability of the boundary periodic solution. By use of standard techniques of bifurcation theory, the periodic oscillations in substrate, plasmid-bearing, and plasmid-free organisms are shown when some conditions are satisfied. Our results can be applied to control bioreactor aimed at producing co...

  18. Virulence Factors of Aeromonas hydrophila: In the Wake of Reclassification

    Science.gov (United States)

    Rasmussen-Ivey, Cody R.; Figueras, Maria J.; McGarey, Donald; Liles, Mark R.

    2016-01-01

    The ubiquitous “jack-of-all-trades,” Aeromonas hydrophila, is a freshwater, Gram-negative bacterial pathogen under revision in regard to its phylogenetic and functional affiliation with other aeromonads. While virulence factors are expectedly diverse across A. hydrophila strains and closely related species, our mechanistic knowledge of the vast majority of these factors is based on the molecular characterization of the strains A. hydrophila AH-3 and SSU, which were reclassified as A. piscicola AH-3 in 2009 and A. dhakensis SSU in 2013. Individually, these reclassifications raise important questions involving the applicability of previous research on A. hydrophila virulence mechanisms; however, this issue is exacerbated by a lack of genomic data on other research strains. Collectively, these changes represent a fundamental gap in the literature on A. hydrophila and confirm the necessity of biochemical, molecular, and morphological techniques in the classification of research strains that are used as a foundation for future research. This review revisits what is known about virulence in A. hydrophila and the feasibility of using comparative genomics in light of this phylogenetic revision. Conflicting data between virulence factors, secretion systems, quorum sensing, and their effect on A. hydrophila pathogenicity appears to be an artifact of inappropriate taxonomic comparisons and/or be due to the fact that these properties are strain-specific. This review audits emerging data on dominant virulence factors that are present in both A. dhakensis and A. hydrophila in order to synthesize existing data with the aim of locating where future research is needed. PMID:27610107

  19. Flagellar motility is necessary for Aeromonas hydrophila adhesion.

    Science.gov (United States)

    Qin, Yingxue; Lin, Guifang; Chen, Wenbo; Xu, Xiaojin; Yan, Qingpi

    2016-09-01

    Adhesion to host surface or cells is the initial step in bacterial pathogenesis, and the adhesion mechanisms of the fish pathogenic bacteria Aeromonas hydrophila were investigated in this study. First, a mutagenesis library of A. hydrophila that contained 332 random insertion mutants was constructed via mini-Tn10 Km mutagenesis. Four mutants displayed the most attenuated adhesion. Sequence analysis revealed that the mini-Tn10 insertion sites in the four mutant strains were flgC(GenBank accession numbers KX261880), cytb4(GenBank accession numbers JN133621), rbsR(GenBank accession numbers KX261881) and flgE(GenBank accession numbers JQ974982). To further study the roles of flgC and flgE in the adhesion of A. hydrophila, some biological characteristics of the wild-type strain B11, the mutants M121 and M240, and the complemented strains C121 and C240 were investigated. The results showed that the mutation in flgC or flgE led to the flagellar motility of A. hydrophila significant reduction or abolishment. flgC was not necessary for flagellar biosynthesis but was necessary for the full motility of A. hydrophila, flgE was involved in both flagellar biosynthesis and motility. The flagellar motility is necessary for A. hydrophila to adhere to the host mucus, which suggests flagellar motility plays crucial roles in the early infection process of this bacterium. PMID:27432325

  20. Immunomodulating effect of inositol hexaphosphate against Aeromonas hydrophila-endotoxin.

    Science.gov (United States)

    Abu-El-Saad, Abdel-Aziz S A

    2007-01-01

    The present study was carried out to evaluate the effect of inositol hexaphosphate (IP6) administration on endotoxemia as an example of the systemic inflammatory response. Mice were divided into three groups as follows: First group, remained as a naive group injected intraperitoneally (i.p.) with PBS (pH 7.4; 0.2 ml/mice) at intervals parallel to the treated groups. The second group was injected i.p. with the lipopolysaccharide (LPS) of Aeromonas hydrophila once a week for four weeks at a dose of LPS suspension: 20 mg/kg mice/week. The third group was injected with the same LPS dose and synergistically intubated with IP6 three times a week for four weeks at a total dose of 4 0mg/kg. At different experimental periods (1, 2, 3 and 4 weeks), six animals from each group were sacrificed under mild diethyl ether anesthesia. Blood and sera were taken for the estimation of phagocytic activity, electrophoretic pattern of proteins and immunoglobulin levels. Also, a slice of liver was homogenized to estimate the respiratory burst enzymes activities and nitric acid synthesis. Histopathological changes of hepatic tissues were investigated. In the LPS-treated group, marked increase in the phagocytic activities and nitric oxide synthesis, and a decrease in hepatocyte catalase, total peroxidase and superoxide dismutase activities were observed. The histopathological features revealed a degeneration and highly mitotic division within the hepatic nuclei in addition to some karyomegaly and nuclear pyknosis. During the treatment period, liver sections of the LPS+IP6 group showed somewhat regenerative features. Reduction in the toxicity of free radicals by IP6 was observed and the IP6 effect seemed to be responsible for the observed ameliorative influence.

  1. Plasmids as Tools for Containment.

    Science.gov (United States)

    García, José L; Díaz, Eduardo

    2014-10-01

    Active containment systems are a major tool for reducing the uncertainty associated with the introduction of monocultures, genetically engineered or not, into target habitats for a large number of biotechnological applications (e.g., bioremediation, bioleaching, biopesticides, biofuels, biotransformations, live vaccines, etc.). While biological containment reduces the survival of the introduced organism outside the target habitat and/or upon completion of the projected task, gene containment strategies reduce the lateral spread of the key genetic determinants to indigenous microorganisms. In fundamental research, suicide circuits become relevant tools to address the role of gene transfer, mainly plasmid transfer, in evolution and how this transfer contributes to genome plasticity and to the rapid adaptation of microbial communities to environmental changes. Many lethal functions and regulatory circuits have been used and combined to design efficient containment systems. As many new genomes are being sequenced, novel lethal genes and regulatory elements are available, e.g., new toxin-antitoxin modules, and they could be used to increase further the current containment efficiencies and to expand containment to other organisms. Although the current containment systems can increase the predictability of genetically modified organisms in the environment, containment will never be absolute, due to the existence of mutations that lead to the appearance of surviving subpopulations. In this sense, orthogonal systems (xenobiology) appear to be the solution for setting a functional genetic firewall that will allow absolute containment of recombinant organisms. PMID:26104372

  2. Enhancement of plasmid-mediated gene therapy for muscular dystrophy by directed plasmid integration

    OpenAIRE

    Bertoni, Carmen; Jarrahian, Sohail; Wheeler, Thurman M.; LI, YINING; Olivares, Eric C.; Michele P Calos; Rando, Thomas A.

    2005-01-01

    Plasmid-mediated gene therapy can restore dystrophin expression in skeletal muscle in the mdx mouse, a model of Duchenne muscular dystrophy. However, sufficient long-term expression and distribution of dystrophin remain a hurdle for translating this technology into a viable treatment for Duchenne muscular dystrophy. To improve plasmid-mediated gene therapy for muscle diseases, we studied the effects of targeted plasmid integration using a phage integrase (φC31) that can mediate the integratio...

  3. Genome Sequence of Aeromonas hydrophila Strain AH-3 (Serotype O34).

    Science.gov (United States)

    Forn-Cuní, Gabriel; Tomás, Juan M; Merino, Susana

    2016-01-01

    Aeromonas hydrophila is an emerging pathogen of poikilothermic animals, from fish to mammals, including humans. Here, we report the whole-genome sequence of the A. hydrophila AH-3 strain, isolated from a fish farm goldfish septicemia outbreak in Spain, with a characterized polar and lateral flagellum glycosylation pattern. PMID:27587828

  4. EFFECT OF ORAL IMMUNOSTIMULANT ANDROGRAPHIS PANICULATA AND RESISTANCE TO AEROMONAS HYDROPHILA IN CATLA CATLA

    OpenAIRE

    Innocent Xavier B.; Fathima Syed Ali M.; Sheeba; Sheeba S.

    2012-01-01

    The present study was designed to evaluate the immunostimulant potential of Andrographis paniculata in fish Catla catla, post challenged with Aeromonas hydrophila. Fishes were divided into 2 groups, one received control diet, and another received Andrographis paniculata incorporated diet for 14 days. The heamatological parameters and serum protein level were analyzed between control and experimental. There is a significant increase in TEC (P

  5. Draft genome sequences of four virulent aeromonas hydrophila strains from catfish aquaculture

    Science.gov (United States)

    Since 2009, a clonal group of virulent Aeromonas hydrophila (VAh) strains has been causing severe disease in the catfish aquaculture industry in the Southeastern United States. Here, we report draft genomes of four A. hydrophila isolates from catfish aquaculture that represent this clonal group....

  6. Antibiotic Susceptibility Profile of Aeromonas Species Isolated from Wastewater Treatment Plant

    Directory of Open Access Journals (Sweden)

    Isoken H. Igbinosa

    2012-01-01

    Full Text Available This study assessed the prevalence of antibiotic-resistant Aeromonas species isolated from Alice and Fort Beaufort wastewater treatment plant in the Eastern Cape Province of South Africa. Antibiotic susceptibility was determined using the disc diffusion method, and polymerase chain reaction (PCR assay was employed for the detection of antibiotics resistance genes. Variable susceptibilities were observed against ciprofloxacin, chloramphenicol, nalidixic acid, gentamicin, minocycline, among others. Aeromonas isolates from both locations were 100% resistant to penicillin, oxacillin, ampicillin, and vancomycin. Higher phenotypic resistance was observed in isolates from Fort Beaufort compared to isolates from Alice. Class A pse1 β-lactamase was detected in 20.8% of the isolates with a lower detection rate of 8.3% for blaTEM gene. Class 1 integron was present in 20.8% of Aeromonas isolates while class 2 integron and TetC gene were not detected in any isolate. The antibiotic resistance phenotypes observed in the isolates and the presence of β-lactamases genes detected in some isolates are of clinical and public health concern as this has consequences for antimicrobial chemotherapy of infections associated with Aeromonas species. This study further supports wastewater as potential reservoirs of antibiotic resistance determinants in the environment.

  7. Active Shiga-Like Toxin Produced by Some Aeromonas spp., Isolated in Mexico City

    Science.gov (United States)

    Palma-Martínez, Ingrid; Guerrero-Mandujano, Andrea; Ruiz-Ruiz, Manuel J.; Hernández-Cortez, Cecilia; Molina-López, José; Bocanegra-García, Virgilio; Castro-Escarpulli, Graciela

    2016-01-01

    Shiga-like toxins (Stx) represent a group of bacterial toxins involved in human and animal diseases. Stx is produced by enterohemorrhagic Escherichia coli, Shigella dysenteriae type 1, Citrobacter freundii, and Aeromonas spp.; Stx is an important cause of bloody diarrhea and hemolytic uremic syndrome (HUS). The aim of this study was to identify the stx1/stx2 genes in clinical strains and outer membrane vesicles (OMVs) of Aeromonas spp., 66 strains were isolated from children who live in Mexico City, and Stx effects were evaluated in Vero cell cultures. The capacity to express active Stx1 and Stx2 toxins was determined in Vero cell cultures and the concentration of Stx was evaluated by 50% lethal dose (LD50) assays, observing inhibition of damaged cells by specific monoclonal antibodies. The results obtained in this study support the hypothesis that the stx gene is another putative virulence factor of Aeromonas, and since this gene can be transferred horizontally through OMVs this genus should be included as a possible causal agents of gastroenteritis and it should be reported as part of standard health surveillance procedures. Furthermore, these results indicate that the Aeromonas genus might be a potential causative agent of HUS. PMID:27725813

  8. Draft Genome Sequences of Four Virulent Aeromonas hydrophila Strains from Catfish Aquaculture

    Science.gov (United States)

    Tekedar, Hasan C.; Kumru, Salih; Karsi, Attila; Waldbieser, Geoffrey C.; Sonstegard, Tad; Schroeder, Steven G.; Liles, Mark R.; Griffin, Matt J.

    2016-01-01

    Since 2009, a clonal group of virulent Aeromonas hydrophila strains has been causing severe disease in the catfish aquaculture industry in the southeastern United States. Here, we report draft genomes of four A. hydrophila isolates from catfish aquaculture that represent this clonal group. PMID:27540076

  9. Characterization of a Haemophilus ducreyi mobilizing plasmid.

    OpenAIRE

    McNicol, P J; Albritton, W L; Ronald, A R

    1986-01-01

    The OriV site of Haemophilus ducreyi mobilizing plasmid pHD147, determined by replication in Escherichia coli polA, is located close to the OriT site. The OriT site, located by recombination-proficient and -deficient cells, and the OriV site map in a region of pHD147 homologous to the beta-lactamase-specifying plasmids of H. ducreyi and Neisseria gonorrhoeae.

  10. Protein diversity confers specificity in plasmid segregation.

    Science.gov (United States)

    Fothergill, Timothy J G; Barillà, Daniela; Hayes, Finbarr

    2005-04-01

    The ParG segregation protein (8.6 kDa) of multidrug resistance plasmid TP228 is a homodimeric DNA-binding factor. The ParG dimer consists of intertwined C-terminal domains that adopt a ribbon-helix-helix architecture and a pair of flexible, unstructured N-terminal tails. A variety of plasmids possess partition loci with similar organizations to that of TP228, but instead of ParG homologs, these plasmids specify a diversity of unrelated, but similarly sized, partition proteins. These include the proteobacterial pTAR, pVT745, and pB171 plasmids. The ParG analogs of these plasmids were characterized in parallel with the ParG homolog encoded by the pseudomonal plasmid pVS1. Like ParG, the four proteins are dimeric. No heterodimerization was detectable in vivo among the proteins nor with the prototypical ParG protein, suggesting that monomer-monomer interactions are specific among the five proteins. Nevertheless, as with ParG, the ParG analogs all possess significant amounts of unordered amino acid residues, potentially highlighting a common structural link among the proteins. Furthermore, the ParG analogs bind specifically to the DNA regions located upstream of their homologous parF-like genes. These nucleoprotein interactions are largely restricted to cognate protein-DNA pairs. The results reveal that the partition complexes of these and related plasmids have recruited disparate DNA-binding factors that provide a layer of specificity to the macromolecular interactions that mediate plasmid segregation. PMID:15805511

  11. Plasmid and chromosome segregation in prokaryotes

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Bugge Jensen, Rasmus; Gerdes, Kenn

    2000-01-01

    Recent major advances in the understanding of prokaryotic DNA segregation have been achieved by using fluorescence microscopy to visualize the localization of cellular components. Plasmids and bacterial chromosomes are partitioned in a highly dynamic fashion, suggesting the presence of a mitotic......-like apparatus in prokaryotes. The identification of chromosomal homologues of the well-characterized plasmid partitioning genes indicates that there could be a general mechanism of bacterial DNA partitioning. Udgivelsesdato: July 1...

  12. Curing of plasmid pXO1 from Bacillus anthracis using plasmid incompatibility.

    Directory of Open Access Journals (Sweden)

    Xiankai Liu

    Full Text Available The large plasmid pXO1 encoding the anthrax toxin is important for the virulence of Bacillus anthracis. It is essential to cure pXO1 from B. anthracis to evaluate its role in the pathogenesis of anthrax infection. Because conventional methods for curing plasmids (e.g., curing agents or growth at elevated temperatures can induce mutations in the host chromosomal DNA, we developed a specific and reliable method to eliminate pXO1 from B. anthracis using plasmid incompatibility. Three putative replication origins of pXO1 were inserted into a temperature-sensitive plasmid to generate three incompatible plasmids. One of the three plasmids successfully eliminated the large plasmid pXO1 from B. anthracis vaccine strain A16R and wild type strain A16. These findings provided additional information about the replication/partitioning of pXO1 and demonstrated that introducing a small incompatible plasmid can generate plasmid-cured strains of B. anthracis without inducing spontaneous mutations in the host chromosome.

  13. Positive epistasis between co-infecting plasmids promotes plasmid survival in bacterial populations

    OpenAIRE

    San Millan, Alvaro; Heilbron, Karl; MacLean, R. Craig

    2014-01-01

    Plasmids have a key role in the horizontal transfer of genes among bacteria. Although plasmids are catalysts for bacterial evolution, it is challenging to understand how they can persist in bacterial populations over the long term because of the burden they impose on their hosts (the ‘plasmid paradox'). This paradox is especially perplexing in the case of ‘small' plasmids, which are unable to self-transfer by conjugation. Here, for the first time, we investigate how interactions between co-in...

  14. Mini-F plasmid genes that couple host cell division to plasmid proliferation.

    OpenAIRE

    Ogura, T; Hiraga, S

    1983-01-01

    A mechanism for stable maintenance of plasmids, besides the replication and partition mechanisms, has been found to be specified by genes of a mini-F plasmid. An oriC plasmid carrying both a mini-F segment necessary for partition [coordinates 46.4-49.4 kilobase pairs (kb) on the F map] and another segment (42.9-43.6 kb), designated ccd (coupled cell division), is more stably maintained than are oriC plasmids carrying only the partition segment; the stability is comparable to that of the paren...

  15. Adaptive Plasmid Evolution Results in Host-Range Expansion of a Broad-Host-Range Plasmid

    OpenAIRE

    De Gelder, Leen; Williams, Julia J.; Ponciano, José M; Sota, Masahiro; Eva M. Top

    2008-01-01

    Little is known about the range of hosts in which broad-host-range (BHR) plasmids can persist in the absence of selection for plasmid-encoded traits, and whether this “long-term host range” can evolve over time. Previously, the BHR multidrug resistance plasmid pB10 was shown to be highly unstable in Stenotrophomonas maltophilia P21 and Pseudomonas putida H2. To investigate whether this plasmid can adapt to such unfavorable hosts, we performed evolution experiments wherein pB10 was maintained ...

  16. Elimination of multicopy plasmid R6K by bleomycin.

    OpenAIRE

    Attfield, P V; Pinney, R. J.

    1985-01-01

    Bleomycin eliminated multicopy plasmid R6K from growing cells of Escherichia coli AB1157 but failed to cure either of the low-copy plasmids R1 or R46. Measurements of R6K-encoded beta-lactamase and of covalently closed plasmid DNA indicated that the drug causes a progressive reduction in plasmid copy number.

  17. Behavior of IncQ Plasmids in Agrobacterium tumefaciens

    NARCIS (Netherlands)

    Hille, Jacques; Schilperoort, Rob

    1981-01-01

    Inc-Q plasmids were introduced into Agrobacterium tumefuciens, by mobilization from Escherichia coli with an Inc-P plasmid, or by transformation with purified plasmid DNA. It was found that they were stably maintained. The presence of an Inc-Q plasmid did not influence tumorigenicity. These results

  18. Plasmid Evolution and Interaction between the Plasmid Addiction Stability Systems of Two Related Broad-Host-Range IncQ-Like Plasmids

    OpenAIRE

    Deane, Shelly M.; Rawlings, Douglas E

    2004-01-01

    Plasmid pTC-F14 contains a plasmid stability system called pas (plasmid addiction system), which consists of two proteins, a PasA antitoxin and a PasB toxin. This system is closely related to the pas of plasmid pTF-FC2 (81 and 72% amino acid identity for PasA and PasB, respectively) except that the pas of pTF-FC2 contains a third protein, PasC. As both pTC-F14 and pTF-FC2 are highly promiscuous broad-host-range plasmids isolated from bacteria that share a similar ecological niche, the plasmid...

  19. Aeromonas hydrophila subsp. ranae subsp. nov., isolated from septicaemic farmed frogs in Thailand.

    Science.gov (United States)

    Huys, Geert; Pearson, Marianne; Kämpfer, Peter; Denys, Rik; Cnockaert, Margo; Inglis, Valerie; Swings, Jean

    2003-05-01

    A group of seven sucrose-negative Aeromonas strains (referred to as group Au) isolated from the internal organs of septicaemic farmed frogs (Rana rugulosa) in Thailand was subjected to a polyphasic taxonomic study including fluorescent amplified fragment length polymorphism (FAFLP) and ERIC-PCR fingerprinting, 16S rDNA sequencing, microplate DNA-DNA hybridizations and extensive phenotypic characterization. Comparison of FAFLP and ERIC-PCR fingerprints indicated that the group Au isolates belonged to the species Aeromonas hydrophila DNA hybridization group (HG) 1 in which they represent a genotypic subgroup closely affiliated to A. hydrophila subsp. hydrophila and subsp. dhakensis. One representative of the Au group exhibited > or = 99.0% 16S rDNA sequence similarity with the type strains of the two A. hydrophila subspecies. DNA-DNA hybridization with type and reference strains of all known Aeromonas taxa revealed that the Au group represented a homogeneous taxon that exhibited the highest relatedness with members of the two A. hydrophila subspecies, ranging from 75 to 93%. Phenotypic characterization on the basis of 152 features further revealed that the Au group isolates differed from A. hydrophila subsp. hydrophila or subsp. dhakensis in a total of 13 biochemical properties. Of these, assimilation of L-glycine and isobutyrate as sole carbon source, acid production from salicin and D-sucrose, and aesculin hydrolysis were of diagnostic value. From the results of this study, it can be concluded that the Aeromonas frog isolates of the Au group represent a new subspecies of A. hydrophila, for which the name Aeromonas hydrophila subsp. ranae subsp. nov. is proposed. Its type strain is Au-1D12(T) (=LMG 19707(T) = CCUG 46211(T)). PMID:12807217

  20. Multi-Drug Resistance Mediated by Class 1 Integrons in Aeromonas Isolated from Farmed Freshwater Animals.

    Science.gov (United States)

    Deng, Yuting; Wu, Yali; Jiang, Lan; Tan, Aiping; Zhang, Ruiquan; Luo, Li

    2016-01-01

    Aeromonas is regarded as an important pathogen of freshwater animals but little is known about the genetics of its antimicrobial resistance in Chinese aquaculture. The aim of this study was to investigate the presence of integrons and characterize multidrug resistant Aeromonas spp. isolated from diseased farmed freshwater animals. These animal samples included fish, ornamental fish, shrimp, turtles, and amphibians which were collected from 64 farms in Guangdong province of South China. One hundred and twelve Aeromonas spp. isolates were examined for antimicrobial resistance phenotypes and the presence of class 1 integron sequences. Twenty-two (19.6%) of these isolates carried a class 1 integron comprising six different gene insertion cassettes including drfA12-orfF-aadA2, drfA12-orfF, aac(6')-II-bla OXA-21 -cat3, catB3, arr-3, and dfrA17. Among these, drfA12-orfF-aadA2 was the dominant gene cassette array (63.6%, 14/22) and this is the first report of aac(6')-II-bla OXA-21 -cat3 in an Aeromonas hydrophila isolate from a Chinese giant salamander (Andrias davidianus). All the integron-positive strains were resistant to more than five agents and 22 contained other resistance genes including bla CTX-M-3, bla TEM-1, aac(6')-Ib-cr, and tetA. All integron-positive isolates also contained mutations in the quinolone resistance determining regions (QRDR). Our investigation demonstrates that freshwater animals can serve as a reservoir for pathogenic Aeromonas strains containing multiple drug-resistance integrons. This data suggests that surveillance for antimicrobial resistance of animal origin and a prudent and responsible use of antimicrobials in aquaculture is necessary in these farms. PMID:27379065

  1. Historical Events That Spawned the Field of Plasmid Biology.

    Science.gov (United States)

    Kado, Clarence I

    2014-10-01

    This chapter revisits the historical development and outcome of studies focused on the transmissible, extrachromosomal genetic elements called plasmids. Early work on plasmids involved structural and genetic mapping of these molecules, followed by the development of an understanding of how plasmids replicate and segregate during cell division. The intriguing property of plasmid transmission between bacteria and between bacteria and higher cells has received considerable attention. The utilitarian aspects of plasmids are described, including examples of various plasmid vector systems. This chapter also discusses the functional attributes of plasmids needed for their persistence and survival in nature and in man-made environments. The term plasmid biology was first conceived at the Fallen Leaf Lake Conference on Promiscuous Plasmids, 1990, Lake Tahoe, California. The International Society for Plasmid Biology was established in 2004 (www.ISPB.org).

  2. Relationship between plasmid content and auxotype in Neisseria gonorrhoeae isolates.

    OpenAIRE

    Dillon, J R; Pauzé, M

    1981-01-01

    One hundred and forty strains of Neisseria gonorrhoeae, representing 12 different auxotype groups, were examined for differences in plasmid content. Most auxotype groups harbored a phenotypically cryptic 2,6-megadalton plasmid; a few groups also carried a 24.5-megadalton plasmid which has been previously characterized as a transfer plasmid. However, isolates of the proline-, citrulline-, and uracil-requiring (PCU-) auxotype were consistently free of plasmids. The correlation between auxotype ...

  3. Plasmid stability and maintenance of copy number using natural marker

    OpenAIRE

    Hamzah Basil Mohammed; Sudhakar Malla

    2015-01-01

    Present study was conducted to study the plasmid stability with the help of natural plasmid isolated from the bacteria which lodges the ink gland of the sea squid and emits bioluminescence. Isolated bacterial strain was identified by using 16srRNA sequencing and its plasmid DNA was used for the experimental studies. The plasmid is found to be responsible for the bioluminescence. The stability of this plasmid was studied in shake flask method using the different sugar sources (Gluc...

  4. An Asian Origin of Virulent Aeromonas hydrophila Responsible for Disease Epidemics in United States-Farmed Catfish

    OpenAIRE

    Hossain, Mohammad J.; Sun, Dawei; McGarey, Donald J.; Wrenn, Shannon; Alexander, Laura M.; Martino, Maria Elena; Xing, Ye; Terhune, Jeffery S.; Liles, Mark R.

    2014-01-01

    ABSTRACT Since 2009, catfish farming in the southeastern United States has been severely impacted by a highly virulent and clonal population of Aeromonas hydrophila causing motile Aeromonas septicemia (MAS) in catfish. The possible origin of this newly emerged highly virulent A. hydrophila strain is unknown. In this study, we show using whole-genome sequencing and comparative genomics that A. hydrophila isolates from diseased grass carp in China and catfish in the United States have highly si...

  5. Incidence of Aeromonas spp. infection in fish and chicken meat and its related public health hazards: A review

    Directory of Open Access Journals (Sweden)

    Praveen Kumar Praveen

    2016-01-01

    Full Text Available Aeromonas is recognized to cause a variety of diseases in man. In humans, they are associated with intestinal and extraintestinal infections. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors such as enterotoxins, hemolysins or cytotoxins, and antibiotic resistance against different antibiotics. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. Comprehensive enteric disease surveillance strategies, prevention and education are essential for meeting the challenges in the years ahead. It is important for us to promote the value of enteric cultures when patients have a gastrointestinal illness or bloody diarrhea or when multiple cases of enteric disease occur after a common exposure. With the growing importance of Aeromonas as an emerging pathogen, it is important to combat this organism. It is indisputable that Aeromonas strains may produce many different putative virulence factors, such as enterotoxins, hemolysins or cytotoxins. It has been established that aerolysin is a virulence factor contributing to the pathogenesis of Aeromonas hydrophila infection. Fish and chicken play an important role in the transmission of this pathogen to humans. In the present study, the high prevalence of toxin-producing strains was found among the Aeromonas isolates. The ability of these bacteria to grow competitively at 5°C may be indicative of their potential as a public health hazard. The present review was constructed with a view to highlight the zoonotic importance of Aeromonas pathogen in fish and chicken meat.

  6. Determination of the Viability of Aeromonas hydrophila in Different Types of Water by Flow Cytometry, and Comparison with Classical Methods

    OpenAIRE

    Pianetti, Anna; Falcioni, Tania; Bruscolini, Francesca; Sabatini, Luigia; Sisti, Elivio; Papa, Stefano

    2005-01-01

    The presence of Aeromonas spp. in water can represent a risk for human health. Therefore, it is important to know the physiological status of these bacteria and their survival in the environment. We studied the behavior of a strain of Aeromonas hydrophila in river water, spring water, brackish water, mineral water, and chlorinated drinking water, which had different physical and chemical characteristics. The bacterial content was evaluated by spectrophotometric and plate count techniques. Flo...

  7. White Mulberry (Morus alba) Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus)

    OpenAIRE

    Atefeh Sheikhlar; Abd Razk Alimon; Hassan Daud; Saad, Chee R.; Webster, Carl D.; Goh Yong Meng; Mahdi Ebrahimi

    2014-01-01

    Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diet...

  8. AEROMONAS SPP BACTEREMIA OF RAINBOW TROUT FRY (ONCORHYNCHUS MYKISS: BIOCHEMICAL CHARACTERIZATION OF THE CAUSATIVE ORGANISM AND ANTIMICROBIAL SUSCEPTIBILITY

    Directory of Open Access Journals (Sweden)

    Damir Kapetanović

    2004-09-01

    Full Text Available Aeromonas hydrophila and other members of Aeromonas genus are ubiquitus in aquatic environment and make part of normal bacterial flora of rainbow trout. Aeromonas spp. infections are worldwide registered. Here we present our experience and knowledge on Aeromonas bacteremia, which causes mortality of rainbow trout fry. Rainbow trout fry, 7 month old, started to die in November 2003. Fish samples (17 samples of dead and moribund fish were delivered to the Laboratory for aquaculture. With Api 20 NE tests Aeromonas hydrophila / caviae type I was identified with an average probability of 99.9 % (one test against, as well as Aeromonas hydrophila / caviae type II with an average probability of 99.5 % (one test against from liver, spleen, kidney, intestines and damaged eye. All of isolated and identified samples were tested for antibiotic susceptibility by disc diffusion method. The test showed that specimens were most sensitive on flumequin, and relatively less sensitive on chloramphenicol and enrofloxacin. Therapy was successfully applied with Flubactin®.

  9. PHENOTYPING, VIRULENCE CHARACTERISTICS OF AEROMONAS SPECIES AND THE EFFECTS OF ESSENTIAL PLANT OILS AS ANTIMICROBIAL AGENTS AGAINST PATHOGENIC ISOLATES FROM DIFFERENT SOURCES

    OpenAIRE

    Ahmed M.A. Mansour; Hoda M. Zaki; Nibal A. Hassan; Nihad A.M. El-Nashar

    2014-01-01

    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 frequentl...

  10. Production of a bioflocculant from Pseudomonas veronii L918 using the hydrolyzate of peanut hull and its application in the treatment of ash-flushing wastewater generated from coal fired power plant.

    Science.gov (United States)

    Liu, Weijie; Hao, Yan; Jiang, Jihong; Zhu, Aihua; Zhu, Jingrong; Dong, Zhen

    2016-10-01

    In this study, bioflocculant produced by Pseudomonas veronii L918 was applied to treat ash-flushing wastewater. The strain L918 could convert the hydrolyzate of peanut hull into bioflocculant, which can effectively reduce the production cost of bioflocculant. The yield of 3.39g/L bioflocculant MBF-L918 was achieved when 300mL/L peanut hull hydrolyzate was used as carbon source. The bioflocculant MBF-L918 contains 77.14% polysaccharides and 4.84% proteins, and the molecular weight (MW) of MBF-L918 is 24.77kDa. Furthermore, MBF-L918 showed good flocculating efficiency of 92.51% to ash-flushing wastewater when 2.83mg/L MBF-L918 was added, and thus achieved the recycling of ash-flushing wastewater. This study reported for the first time that the bioflocculant was produced using peanut hull hydrolyzate and effectively applied in the treatment of coal ash-flushing wastewater. PMID:27372012

  11. BioShuttle-mediated Plasmid Transfer

    Directory of Open Access Journals (Sweden)

    Klaus Braun, Leonie von Brasch, Ruediger Pipkorn, Volker Ehemann, Juergen Jenne, Herbert Spring, Juergen Debus, Bernd Didinger, Werner Rittgen, Waldemar Waldeck

    2007-01-01

    Full Text Available An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantification of the transfer efficiency was achieved by measurements of the sodium iodide symporter activity. EGFP gene expression was measured with Confocal Laser Scanning Microscopy and quantified with biostatistical methods by analysis of the frequency of the amplitude distribution in the CLSM images. The results demonstrate that the “BioShuttle”-Technology is an appropriate tool for an effective transfer of genetic material carried by a plasmid.

  12. BioShuttle-mediated Plasmid Transfer

    Science.gov (United States)

    Braun, Klaus; von Brasch, Leonie; Pipkorn, Ruediger; Ehemann, Volker; Jenne, Juergen; Spring, Herbert; Debus, Juergen; Didinger, Bernd; Rittgen, Werner; Waldeck, Waldemar

    2007-01-01

    An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantification of the transfer efficiency was achieved by measurements of the sodium iodide symporter activity. EGFP gene expression was measured with Confocal Laser Scanning Microscopy and quantified with biostatistical methods by analysis of the frequency of the amplitude distribution in the CLSM images. The results demonstrate that the “BioShuttle”-Technology is an appropriate tool for an effective transfer of genetic material carried by a plasmid. PMID:18026568

  13. Plasmid and chromosome partitioning: surprises from phylogeny

    DEFF Research Database (Denmark)

    Gerdes, Kenn; Møller-Jensen, Jakob; Bugge Jensen, Rasmus

    2000-01-01

    and chromosomes from prokaryotic organisms. All known plasmid-encoded par loci specify three components: a cis-acting centromere-like site and two trans-acting proteins that form a nucleoprotein complex at the centromere (i.e. the partition complex). The proteins are encoded by two genes in an operon...... that is autoregulated by the par-encoded proteins. In all cases, the upstream gene encodes an ATPase that is essential for partitioning. Recent cytological analyses indicate that the ATPases function as adaptors between a host-encoded component and the partition complex and thereby tether plasmids and chromosomal...... origin regions to specific subcellular sites (i.e. the poles or quarter-cell positions). Two types of partitioning ATPases are known: the Walker-type ATPases encoded by the par/sop gene family (type I partitioning loci) and the actin-like ATPase encoded by the par locus of plasmid R1 (type II...

  14. BioShuttle-mediated Plasmid Transfer

    OpenAIRE

    Braun, Klaus; von Brasch, Leonie; Pipkorn, Ruediger; Ehemann, Volker; Jenne, Juergen; Spring, Herbert; Debus, Juergen; Didinger, Bernd; Rittgen, Werner; Waldeck, Waldemar

    2007-01-01

    An efficient gene transfer into target tissues and cells is needed for safe and effective treatment of genetic diseases like cancer. In this paper, we describe the development of a transport system and show its ability for transporting plasmids. This non-viral peptide-based BioShuttle-mediated transfer system consists of a nuclear localization address sequence realizing the delivery of the plasmid phNIS-IRES-EGFP coding for two independent reporter genes into nuclei of HeLa cells. The quantif...

  15. EFFECT OF ORAL IMMUNOSTIMULANT ANDROGRAPHIS PANICULATA AND RESISTANCE TO AEROMONAS HYDROPHILA IN CATLA CATLA

    Directory of Open Access Journals (Sweden)

    Innocent Xavier B.

    2012-04-01

    Full Text Available The present study was designed to evaluate the immunostimulant potential of Andrographis paniculata in fish Catla catla, post challenged with Aeromonas hydrophila. Fishes were divided into 2 groups, one received control diet, and another received Andrographis paniculata incorporated diet for 14 days. The heamatological parameters and serum protein level were analyzed between control and experimental. There is a significant increase in TEC (P<0.05 and TLC (P<0.01 in immune-stimulant incorporated diet (IS. Remarkable increase in Hb content and serum protein was noticed as 2.42 to 3.2g% and 0.52 to 0.98g% respectively. The immunostimulant administered Cata catla when challenged with Aeromonas hydrophila showed decrease in TEC and drastic increase in TLC (P < 0.01 and lymphocyte as 58% when compared to control.

  16. Plasmid maintenance functions of the large virulence plasmid of Shigella flexneri.

    OpenAIRE

    Radnedge, L; Davis, M. A.; Youngren, B; Austin, S. J.

    1997-01-01

    The large virulence plasmid pMYSH6000 of Shigella flexneri contains a replicon and a plasmid maintenance stability determinant (Stb) on adjacent SalI fragments. The presence of a RepFIIA replicon on the SalI C fragment was confirmed, and the complete sequence of the adjacent SalI O fragment was determined. It shows homology to part of the transfer (tra) operon of the F plasmid. Stb stabilizes a partition-defective P1 miniplasmid in Escherichia coli. A 1.1-kb region containing a homolog of the...

  17. Plasmid transfer and plasmid-mediated genetic exchange in Brucella abortus.

    OpenAIRE

    Rigby, C E; Fraser, A.D.

    1989-01-01

    Naturally-occurring plasmids and gene transfer mechanisms have not yet been reported in brucellae. Here we show that Brucella abortus is capable of maintaining and transferring the broad-host-range plasmids pTH10 (IncP), pSa (IncW) and R751 (IncP), and describe pTH10-mediated transfer of B. abortus chromosomal genes to Escherichia coli. All three plasmids transferred by conjugation from E. coli to B. abortus S19, and from B. abortus S19 to B. abortus 292 (biovar 4). They were stably maintaine...

  18. Necrotizing Fasciitis Caused by Inconspicuous Infection of Aeromonas hydrophila in an Immunocompromised Host

    Directory of Open Access Journals (Sweden)

    Kuo-Chun Liao

    2010-09-01

    Full Text Available We present a case of serious necrotizing fasciitis due to Aeromonas hydrophila without a suggestive history of routes of pathogen invasion in a 60-year-old male. Despite prompt broad-spectrum antibiotic and extensive surgical therapies, the patient died within 72 hours following initial presentation. Our experience suggests clinicians be highly alert to the disease when an immunocompromised patient featured fulminant soft-tissue infection in the endemic area.

  19. Detection of multidrug resistance Aeromonas hydrophila in farm raised fresh water prawns

    OpenAIRE

    Md. Bakhtiar Lijon; Mst. Mousumi Khatun; Ariful Islam; Mst. Minara Khatun; Md. Ariful Islam

    2015-01-01

    This study was undertaken for isolation, identification and determination of antibiogram profile of Aeromonas hydrophila in farm raised fresh water prawn (Macrobrachium rosenbergii) on five commercial ghers. Fresh water prawns (n=25) were collected from five ghers located at Satkhira, Bagerhat and Khulna districts of Bangladesh. Brain (n=25), muscle (n=25) and intestine (n=25) samples were collected aseptically from fresh water prawn and inoculated into alkaline peptone (APW) water for enrich...

  20. Recombinant outer membrane protein C of Aeromonas hydrophila elicits mixed immune response and generates agglutinating antibodies.

    Science.gov (United States)

    Yadav, Sunita Kumari; Meena, Jitendra Kumar; Sharma, Mahima; Dixit, Aparna

    2016-08-01

    Aeromonas hydrophila is a gram-negative fish pathogenic bacterium, also responsible for causing opportunistic pathological conditions in humans. It causes a number of diseases in fish due to which the fish industry incurs huge economic losses annually. Due to problems of antibiotic resistance, and the rapidity with which the infection spreads among fishes, vaccination remains the most effective strategy to combat this infection in fish populations. Among various virulence factors associated with bacterial virulence, outer membrane proteins have been widely evaluated for their vaccine potential owing to their surface exposure and related role in pathogenicity. In the present study, we have investigated the immunogenic potential of a non-specific porin, outer membrane protein C (OmpC) whose expression is regulated by the two-component regulatory system and plays a major role in the survival of A. hydrophila under different osmolaric conditions. The full-length gene (~1 kb) encoding OmpC of A. hydrophila was cloned, characterized and expressed in E. coli. High yield (~112 mg/L at shake flask level) of the recombinant OmpC (rOmpC) (~40 kDa) of A. hydrophila was obtained upon purification from inclusion bodies using Ni(2+)-NTA affinity chromatography. Immunization with purified rOmpC in murine model generated high endpoint (>1:40,000) titers. IgG isotyping, ELISA and ELISPOT assay indicated mixed immune response with a TH2 bias. Also, the anti-rOmpC antibodies were able to agglutinate A. hydrophila in vitro and exhibited specific cross-reactivity with different Aeromonas strains, which will facilitate easy detection of different Aeromonas isolates in infected samples. Taken together, these data clearly indicate that rOmpC could serve as an effective vaccine against different strains of Aeromonas, a highly heterogenous group of bacteria. PMID:27328672

  1. Purification and Characterization of Aeromonas caviae ME-1 Xylanase V, Which Produces Exclusively Xylobiose from Xylan

    OpenAIRE

    Kubata, Bruno Kilunga; Suzuki, Tohru; Horitsu, Hiroyuki; Kawai, Keiichi; Takamizawa, Kazuhiro

    1994-01-01

    A xylanase, which produces exclusively xylobiose from oat spelt and birch xylans, was isolated from the culture medium of Aeromonas caviae ME-1. The enzyme (xylanase V) was purified by ammonium sulfate fractionation, hydrophobic interaction, and ion-exchange and gel filtration chromatographies. The homogeneity of the final preparation was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and agarose gel electrofocusing. The molecular mass and isoelectric point of the x...

  2. COMBINATION OF CONVENTIONAL AND IN-SILICO APPROACH FOR IDENTIFYING AN INDUSTRIALLY IMPORTANT ISOLATE OF AEROMONAS

    Directory of Open Access Journals (Sweden)

    Thungri Ghoshal

    2014-01-01

    Full Text Available Ammonia is required for various commercial applications while its production by standard process is immensely energy intense. Dairy industry on the other hand produces huge volume of effluent needing treatment before discharge. This study aims at developing an eco-friendly microbial approach for conversion of dairy industry effluent into ammonia under atmospheric pressure and 37°C temperature at laboratory scale. Thus the need for huge energy for ammonia production would be avoided. Moreover the dairy effluent would generate revenue while getting treated. This study is also an attempt to develop a bioinformatics approach for strain identification. Standard cultivation techniques and in-silico approach of analysis of draft genome was undertaken to decipher the identity of this industrially important strain. Aeromonas sp. MCC 2167 (isolated from active dairy sludge was capable of treating un-amended dairy effluent with 6% inoculum while producing ammonia as by product at a 11 folds higher rate than the modified Habers Process. Though the strain showed maximum identity with Aeromonas hydrophila, there were certain differences in terms of substrate utilization. Majority of the contigs showed identity with Aeromonas hydrophila, however there was significant difference among the two strains in terms of intergenomic distance (genome to genome distance calculator from DSMZ, inversion and frame shift (dot plot analysis as well as genome rearrangement (MAUVE analysis. As per the rapid annotation using subsystem technology analysis 53% of the contigs belong to the subsystem category while the remaining 47% do not match with existing sequences in the database. The above finding proposes bioinformatics validation of the wetlab based data pointing towards this industrially important strain to be a novel isolate of Aeromonas sp. This would lead to an energy efficient economically viable alternative for ammonia production while

  3. Diversity and antibiotic resistance of aeromonas spp. in drinking and waste water treatment plants

    OpenAIRE

    Figueira, Vânia; Vaz-Moreira, Ivone; Silva, Márcia; Manaia, Célia M

    2011-01-01

    The taxonomic diversity and antibiotic resistance phenotypes of aeromonads were examined in samples from drinking and waste water treatment plants (surface, ground and disinfected water in a drinking water treatment plant, and raw and treated waste water) and tap water. Bacteria identification and intra-species variation were determined based on the analysis of the 16S rRNA, gyrB and cpn60 gene sequences. Resistance phenotypes were determined using the disc diffusion method. Aeromonas ver...

  4. Bioremediation and Detoxification of Synthetic Wastewater Containing Triarylmethane Dyes by Aeromonas hydrophila Isolated from Industrial Effluent

    OpenAIRE

    Chimezie Jason Ogugbue; Thomas Sawidis

    2011-01-01

    Economical and bio-friendly approaches are needed to remediate dye-contaminated wastewater from various industries. In this study, a novel bacterial strain capable of decolorizing triarylmethane dyes was isolated from a textile wastewater treatment plant in Greece. The bacterial isolate was identified as Aeromonas hydrophila and was shown to decolorize three triarylmethane dyes tested within 24 h with color removal in the range of 72% to 96%. Decolorization efficiency of the bacterium was a f...

  5. Molecular detection and antimicrobial resistance of Aeromonas from houseflies (Musca domestica) in Iran

    OpenAIRE

    Davood Ommi; Seyed Mohammadreza Hashemian; Elahe Tajbakhsh; Faham Khamesipour

    2015-01-01

    ABSTRACT Objective. This study aimed to report the molecular detection and antimicrobial resistance of Aeromonas among houseflies (Musca domestica) in Shahrekord and Isfahan provinces of Iran. Materials and methods. Flies were caught from household kitchens, cattle farms, animal hospitals, human hospitals, slaughter house and poultry farms and put in collection separate sterile tubes. Isolation was accomplished by culture of flies in alkaline peptone water followed by identification with Aer...

  6. Antibiotic and heavy metal resistance of Aeromonas hydrophila isolated from charal (Chirostoma humboldtianum, Valenciannes, 1835)

    OpenAIRE

    Gloria Luz Paniagua; Eric Monroy; Magdalena Perches; Erasmo Negrete; Octavio García; Sergio Vaca

    2006-01-01

    Antibiotic and heavy metal susceptibilities of twenty Aeromonas hydrophila strains, isolated from the gastrointestinal tract of the charal (Chirostoma humboldtianum), an autochthonous Mexican fish, were analyzed. All strains produced #946;-lactamase and were resistant to penicillin and dicloxacillin, showing single peak for minimum inhibitory concentration (MIC) distributions at 2000-4000 µg/mL and 500-1000 µg/ml, respectively. Ampicillin MIC distribution was bimodal with 20% resistant strai...

  7. Phytoextracts-Synthesized Silver Nanoparticles Inhibit Bacterial Fish Pathogen Aeromonas hydrophila

    OpenAIRE

    Mahanty, Arabinda; Mishra, Snehasish; Bosu, Ranadhir; Maurya, UK; Netam, Surya Prakash; Sarkar, Biplab

    2013-01-01

    Fish disease is a major stumbling block towards sustainable growth of the fisheries sector. Aeromonas hydrophila, which is a major infectious aquatic pathogen is reportedly the causative agent of ulcers, fin-rot, tail-rot, hemorrhagic septicemia in fish, and has reportedly developed resistance against many of the available antibiotics. In this context, the inhibitory function of silver nanoparticles (AgNPs) against A. hydrophila was studied to evaluate its possible application in aquaculture ...

  8. Competition between Plasmid-Bearing and Plasmid-Free Organisms in a Chemostat with Pulsed Input and Washout

    Directory of Open Access Journals (Sweden)

    Sanling Yuan

    2009-01-01

    Full Text Available We consider a model of competition between plasmid-bearing and plasmid-free organisms in the chemostat with pulsed input and washout. We investigate the subsystem with nutrient and plasmid-free organism and study the stability of the boundary periodic solutions, which are the boundary periodic solutions of the system. The stability analysis of the boundary periodic solution yields the invasion threshold of the plasmid-bearing organism. By using the standard techniques of bifurcation theory, we prove that above this threshold there are periodic oscillations in substrate, plasmid-free, and plasmid-bearing organisms. Numerical simulations are carried out to illustrate our results.

  9. An investigation of the bactericidal activity of selected essential oils to Aeromonas spp.

    Directory of Open Access Journals (Sweden)

    Clifford E. Starliper

    2015-01-01

    Full Text Available Diseases of fishes caused by Aeromonas spp. are common, have broad host ranges and may cause high mortality. Treatments of captive-reared populations using antimicrobials are limited with concerns for bacterial resistance development and environmental dissemination. This study was done to determine whether selected plant-derived essential oils were bactericidal to Aeromonas spp. Initially, twelve essential oils were evaluated using a disk diffusion assay to an isolate of A. salmonicida subsp. salmonicida, cause of fish furunculosis. The greatest zones of inhibition were obtained with oils of cinnamon Cinnamomum cassia, oregano Origanum vulgare, lemongrass Cymbopogon citratus and thyme Thymus vulgaris. Minimum bactericidal concentrations (MBC’s were determined for these four oils, Allimed® (garlic extract, Allium sativum and colloidal silver to sixty-nine isolates representing nine Aeromonas spp. The lowest mean MBCs (0.02–0.04% were obtained with three different sources of cinnamon oil. MBCs for three sources of oregano and lemongrass oils ranged from 0.14% to 0.30% and 0.10% to 0.65%, respectively, and for two thyme oils were 2.11% and 2.22%. The highest concentration (5% of Allimed® tested resulted in MBCs to twelve isolates. A concentration of silver greater than 15 mg/L would be required to determine MBCs for all but one isolate.

  10. Influence of Limonia acidissima L. against the biofilm forming Aeromonas hydrophila isolated from fresh water fishes

    Directory of Open Access Journals (Sweden)

    Srinivasan Ponnuraj

    2015-04-01

    Full Text Available Limonia acidissima is a medicinal plant commonly used for multitude of ailments. In this context, the validation of traditionally used medicinal plant that the fruit extracts of L. acidissima exhibit antibiofilm activity against the predominant fish pathogen Aeromonas hydrophila was assessed. Among ten isolates of Aeromonas spp, KUAH1 showed strong biofilm formation and was studied further. The methanol (LA-M and ethyl acetate (LA-EA fraction of Limonia fruit extract clearly demonstrated significant (p ≤ 0.005 antibiofilm activity of 58 to 94% and 54 to 77% respectively. Furthermore, the potential of Limonia fruit extracts against some of the biofilm associated factors were tested by swimming and swarming motility assay, XTT and anti-haemolytic activity assay. Extracellular protein analysis revealed differential protein expression at molecular weight corresponding to 30-60 kDa. This is the first report on antibiofilm activity of L. acidissima fruit extracts, signifying the scope for development of complementary medicine to treat Aeromonas biofilm-associated infections.

  11. DAYA HAMBAT EKSTRAK METANOL RUMPUT LAUT (Euchema spinosum TERHADAP BAKTERI Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    Y Yunus

    2009-10-01

    Full Text Available Rumput laut secara tradisional telah lama dimanfaatkan sebagai bahan makanan, kosmetik dan obat-obatan, karena kaya akan mineral, elemen makro dan elemen  mikro lainnya, disamping itu rumput laut juga berfungsi sebagai bahan antimikroba seperti penyakit bakteri Aeromonas hydrophila yang sering menginfeksi ikan pada budidaya air tawar sehingga banyak merugikan petani. Oleh karena itu, maka perlu dilakukan penelitian tentang konsentrasi ekstrak metanol rumput laut yang efektif. Tujuan dari penelitian ini adalah untuk mengetahui konsentrasi ekstrak metanol rumput laut Eucheuma spinosum yang tepat untuk menghambat pertumbuhan bakteri Aeromonas hydrophyla. Metode penelitian yang digunakan adalah metode eksperimen, Pertama menggunakan Uji MIC  konsentrasi sebagai berikut : 25%; 12,5%; 6,25%; 3,125%; 1,56%; 0,78%; 0,39%; 0,195%; 0,098%; 0,049%; 0,025%; dan 0,0125%. Kedua menggunakan Uji Cakram konsentrasi 3%, 6%, 9%, 12%, dan 0%  sebagai kontrol. Hasil uji MIC setelah dibandingkan dengan kontrol positif sangat rendah yaitu sebesar 0,049%, hasil ini menunjukkan bahwa konsentrasi tersebut menghambat pertumbuhan bakteri Aeromnas hydrophila. Pada hasil uji cakram kemampuan ekstrak rumput laut pada konsentrasi 3%, 6% dan 9% dalam menghambat bakteri Aeromons hydrophila bersifat bakteriostatik, sedangkan pada konsentrasi 12% bersifat bakteriosidal.  Kata Kunci : Eucheuma spinosum, Polifenol, Aeromonas hydrophila

  12. Anti-biogram and resistogram profiling of Aeromonas species isolated from Malaysian aquatic sources

    Institute of Scientific and Technical Information of China (English)

    Olumide Adedokun Odeyemi; Asmat Ahmad

    2013-01-01

    Objective: To investigate antibiotics and heavy metals resistance profile of Aeromonas species isolated from Kuala Gula and Kuala Sepetang, Perak. Methods: Isolated Aeromonas species were subjected to 12 antibiotics: penicillin G 10 units, tetracycline 30 µg, novobiocin 5 µg, nitrofurantoin 100 µg, ampicillin 10 µg, chloramphenicol 10 µg, fluconazole 25 µg, gentamicin 10 µg, streptomycin 10 µg and 25 µg, kanamycin 30 µg, and vancomycin 30 µg and 5 heavy metals: MnSO4.H2O, Cr(NO3).9H2O, CuSO4, Pb(NO3)2, HgCl2. Results: Results from this study revealed that isolates from the two sources-displayed varied resistance. Isolates from Kuala Sepetang showed 91%, 66%, 61%, and 52% resistance to vancomycin, Novobiocin, streptomycin and gentamycin while Kuala Gula isolates showed 76%, 93%, 28% and 14% respectively. High rate of resistance was shown to be against lead (Pb), Chromium (Cr), copper (Cu) and Manganese (Mn) with minimum inhibitory concentration ranging from 2 560 µg/mL to>5120 µg/mL. Low resistance to mercury was observed among isolates from Kuala Gula. Conclusions: This study showed that there was widespread of both heavy metals and antibiotics resistance among Aeromonas isolates from the two marine sources with isolates from Kuala Sepetang showing more resistance thereby posing danger to both aquatic animals and human health.

  13. Occurrence of potential pathogenic Aeromonas species in tropical seafood, aquafarms and mangroves off Cochin coast in South India

    Directory of Open Access Journals (Sweden)

    Alphonsa Vijaya Joseph

    2013-12-01

    Full Text Available Background: The genus Aeromonas include gram-negative, motile, facultative anaerobic, rod shaped and oxidase positive bacteria comprising several species, associated with the aquatic environment. Aeromonas species have been implicated in human pathogenesis and are linked with gastroenteritis, muscle infections, septicemia, and skin diseases. In fish they are renowned as enteric pathogens causing haemorrhagic septicemia, fin rot, soft tissue rot and furunculosis resulting in major die-offs and fish kills. Aim: This study reports the occurrence of potential pathogenic Aeromonas sp. in tropical seafood (Squids, Prawns and Mussels, aquafarms and mangroves of Cochin, Kerala, South India. Materials and Methods :Tropical seafood (Squid, Prawn and Mussel, sediment and water samples from aquafarms and associated mangroves were screened for Aeromonas contamination. The isolates were identified by 16S rDNA sequence analysis and subjected to morphological and biochemical characterization. Haemolytic assay was used for determining pathogenicity of the organisms. Antibiotic susceptibility against 12 antibiotics were performed and the MAR index was calculated. Results: A total of 134 isolates were recovered from the samples of which 15 were identified as Aeromonas species by 16S rDNA sequence analysis and were assigned to 5 species namely, A. hydrophila, A. enteropelogenes, A. caviae, A. punctataand A. aquarorium. Morphological, biochemical and phylogenetic analyses revealed relatedness and variability among the strains. All the isolates were haemolytic on blood agar indicating their pathogenicity. The isolates exhibited varying degrees of resistance to vancomycin (86.66%, ampicillin (46.66%, nalidixic acid (20%, tetracycline (6.66%, co-trimaxozole (6.66% and rifampicin (6.66% and were susceptible to antibiotics like gentamycin, streptomycin, trimethoprim, azithromycin, cefixime and chloramphenicol. 20% of Aeromonas sp. showed MAR index > 0.2 indicative of the

  14. Plasmid mediated quinolone resistance in Enterobacteriaceae

    NARCIS (Netherlands)

    Veldman, K.T.

    2014-01-01

    This thesis describes the occurrence of Plasmid Mediated Quinolone Resistance (PMQR) in Salmonella and E. coli from The Netherlands and other European countries. Furthermore, the genetic background of these genes was characterized. Fluoroquinolones are widely used antibiotics in both human and veter

  15. Plasmid maintenance and protein overproduction in selective recycle bioreactors.

    Science.gov (United States)

    Ogden, K L; Davis, R H

    1991-02-20

    A new plasmid construct has been used in conjunction with selective recycle to successfully maintain otherwise unstable plasmid-bearing E. coli cells in a continuous bioreactor and to produce significant amounts of the plasmid-encoded protein beta-lactamase. The plasmid is constructed so that pilin expression, which leads to bacterial flocculation, is under control of the tac operon. The plasmid-bearing cells are induced to flocculate in the separator, whereas cell growth and product synthesis occur in the main fermentation vessel without the inhibiting effects of pilin production. Selective recycle allows for the maintenance of the plasmid-bearing cells by separating flocculent, plasmid-bearing cells from nonflocculent, segregant cells in an inclined settler, and recycling only the plasmid-bearing cells to the reactor. As a result, product expression levels are maintained that are more than ten times the level achieved without selective recycle. All experimental data agree well with theoretical predictions. PMID:18597374

  16. The influence of biofilms in the biology of plasmids

    OpenAIRE

    Cook, Laura C.C.; Dunny, Gary M.

    2014-01-01

    The field of plasmid biology has historically focused on bacteria growing in liquid culture. Surface attached communities of bacterial biofilms have recently been understood to be the normal environment of bacteria in the natural world. Thus, studies examining plasmid replication, maintenance, and transfer in biofilms are essential for a true understanding of bacterial plasmid biology. This chapter reviews the current knowledge of the interplay between bacterial biofilms and plasmids, focusin...

  17. The 2 micron plasmid purloins the yeast cohesin complex

    OpenAIRE

    Mehta, Shwetal; Yang, Xian Mei; Chan, Clarence S.; Dobson, Melanie J.; Jayaram, Makkuni; Velmurugan, Soundarapandian

    2002-01-01

    The yeast 2 micron plasmid achieves high fidelity segregation by coupling its partitioning pathway to that of the chromosomes. Mutations affecting distinct steps of chromosome segregation cause the plasmid to missegregate in tandem with the chromosomes. In the absence of the plasmid stability system, consisting of the Rep1 and Rep2 proteins and the STB DNA, plasmid and chromosome segregations are uncoupled. The Rep proteins, acting in concert, recruit the yeast cohesin complex to the STB locu...

  18. Postsegregational killing does not increase plasmid stability but acts to mediate the exclusion of competing plasmids

    OpenAIRE

    Cooper, Tim F; Heinemann, Jack A.

    2000-01-01

    Postsegregational killing (PSK) systems consist of a tightly linked toxin–antitoxin pair. Antitoxin must be continually produced to prevent the longer lived toxin from killing the cell. PSK systems on plasmids are widely believed to benefit the plasmid by ensuring its stable vertical inheritance. However, experimental tests of this “stability” hypothesis were not consistent with its predictions. We suggest an alternative hypothesis to explain the evolution of PSK: ...

  19. Compositional discordance between prokaryotic plasmids and host chromosomes

    NARCIS (Netherlands)

    M.W.J. van Passel; A. Bart; A.C.M. Luyf; A.H.C. van Kampen; A. van der Ende

    2006-01-01

    Background: Most plasmids depend on the host replication machinery and possess partitioning genes. These properties confine plasmids to a limited range of hosts, yielding a close and presumably stable relationship between plasmid and host. Hence, it is anticipated that due to amelioration the dinucl

  20. Plasmid Segregation: Spatial Awareness at the Molecular Level

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Gerdes, Kenn

    2007-01-01

    In bacteria, low-copy number plasmids ensure their stable inheritance by partition loci (par), which actively distribute plasmid replicates to each side of the cell division plane. Using time-lapse fluorescence microscopic tracking of segregating plasmid molecules, a new study provides novel...

  1. Cloning of Two Bacteriocin Genes from a Lactococcal Bacteriocin Plasmid

    NARCIS (Netherlands)

    Belkum, Marco J. van; Hayema, Bert Jan; Geis, Arnold; Kok, Jan; Venema, Gerard

    1989-01-01

    Lactococcus lactis subsp. cremoris 9B4 plasmid p9B4-6 (60 kilobases [kb]), which specifies bacteriocin production and immunity, was analyzed with restriction endonucleases, and fragments of this plasmid were cloned into shuttle vectors based on the broad-host-range plasmid pWVO1. Two regions on p9B4

  2. Competition between Plasmid-Bearing and Plasmid-Free Organisms in a Chemostat with Pulsed Input and Washout

    OpenAIRE

    Sanling Yuan; Yu Zhao; Anfeng Xiao

    2009-01-01

    We consider a model of competition between plasmid-bearing and plasmid-free organisms in the chemostat with pulsed input and washout. We investigate the subsystem with nutrient and plasmid-free organism and study the stability of the boundary periodic solutions, which are the boundary periodic solutions of the system. The stability analysis of the boundary periodic solution yields the invasion threshold of the plasmid-bearing organism. By using the standard techniques of bifurcation theory, w...

  3. Co-segregation of yeast plasmid sisters under monopolin-directed mitosis suggests association of plasmid sisters with sister chromatids

    OpenAIRE

    Liu, Yen-Ting; Ma, Chien-Hui; Jayaram, Makkuni

    2013-01-01

    The 2-micron plasmid, a high copy extrachromosomal element in Saccharomyces cerevisiae, propagates itself with nearly the same stability as the chromosomes of its host. Plasmid stability is conferred by a partitioning system consisting of the plasmid-coded proteins Rep1 and Rep2 and a cis-acting locus STB. Circumstantial evidence suggests that the partitioning system couples plasmid segregation to chromosome segregation during mitosis. However, the coupling mechanism has not been elucidated. ...

  4. Predicting Virulence of Aeromonas Isolates Based-on Changes in Transcription of c-jun and c-fos in Human Tissue Culture Cells

    Science.gov (United States)

    Aims: To assess virulence of Aeromonas isolates based on the change in regulation of c-jun and c-fos in the human intestinal tissue culture cell line Caco-2. Methods and Results: Aeromonas cells were added to Caco-2 cells at approximately a one to one ratio. After 1, 2 and 3 ...

  5. Mutation in the S-ribosylhomocysteinase (luxS) gene involved in quorum sensing affects biofilm formation and virulence in a clinical isolate of Aeromonas hydrophila

    Science.gov (United States)

    A diarrheal isolate SSU of Aeromonas hydrophila produces a cytotoxic enterotoxin (Act) with cytotoxic, enterotoxic, and hemolytic activities. Our laboratory has characterized from the above Aeromonas strain, in addition to Act, the type 3- and T6-secretion systems and their effec...

  6. THE INTEGRATED STATE OF THE ROLLING-CIRCLE PLASMID PTB913 IN THE COMPOSITE BACILLUS PLASMID PTB19

    NARCIS (Netherlands)

    OSKAM, L; HILLENGA, DJ; VENEMA, G; BRON, S

    1992-01-01

    pTB19, a 27 kb plasmid originating from a thermophilic Bacillus species, contains integrated copies of two rolling-circle type plasmids on a 10.6 kb DNA fragment. In the present study we analysed the part of pTB19 that contains the rolling-circle plasmid pTB913 and the region in between the two roll

  7. Stable inheritance of plasmid R1 requires two different loci.

    OpenAIRE

    Gerdes, K; Larsen, J E; Molin, S

    1985-01-01

    The largest EcoRI fragment from plasmid R1 mediates a stability phenotype which is required to ensure the stable inheritance of this low-copy-number plasmid. When covalently linked to small, unstable R1 derivatives, this fragment makes the plasmids as stable as the wild-type R1 plasmid. A genetic analysis showed that two independently acting stabilization functions are encoded by this EcoRI fragment, both of which have the potential of partial stabilization of mini-R1 plasmids. The two loci a...

  8. DNA restriction-modification systems mediate plasmid maintenance.

    OpenAIRE

    Kulakauskas, S; Lubys, A; Ehrlich, S. D.

    1995-01-01

    Two plasmid-carried restriction-modification (R-M) systems, EcoRI (from pMB1 of Escherichia coli) and Bsp6I (from pXH13 of Bacillus sp. strain RFL6), enhance plasmid segregational stability in E. coli and Bacillus subtilis, respectively. Inactivation of the endonuclease or the presence of the methylase in trans abolish the stabilizing activity of the R-M systems. We propose that R-M systems mediate plasmid segregational stability by postsegregational killing of plasmid-free cells. Plasmid-enc...

  9. Novo meio seletivo-indicador para detecção de Aeromonas e Plesiomonas: ágar UNISC New selective indicator medium for detection of Aeromonas and Plesiomonas: UNISC agar

    Directory of Open Access Journals (Sweden)

    Marion Pereira da Rocha

    2008-08-01

    Full Text Available Avaliou-se um novo meio seletivo-indicador (ágar UNISC para o isolamento de enteropatógenos clássicos e Aeromonas e Plesiomonas shigelloides. A capacidade de fermentação da xilose é indicada pela coloração amarela (fermentadores ou azul (não fermentadores que, aliada à prova da oxidase, constitui-se em indicador para a detecção de Aeromonas spp e Plesiomonas shigelloides. A produtividade e seletividade, avaliadas pelos índice de contagem absoluta e índice de contagem relativa indicam-no como uma alternativa aos coprocultivos clássicos porque permite, num só meio, o isolamento de Escherichia coli, Shigella spp, Salmonella spp, bem como, Aeromonas spp e Plesiomonas shigelloides, favorecendo o diagnóstico laboratorial das gastroenterites.We evaluated a new selective indicator medium (UNISC Agar for isolation of classical enteropathogens, Aeromonas spp and Plesiomonas shigelloides. The xylose fermentation capacity is indicated by a yellow color (fermenting agents or blue (no fermenting agent. This, together with the oxidase test, establishes it as an indicator for detecting Aeromonas and Plesiomonas shigelloides. Its productivity and selectivity, as assessed using the absolute count index and relative count index, indicate it as an alternative to the classical feces culturing media. This is because, in a single medium, it enables isolation of Escherichia coli, Shigella spp and Salmonella spp, in addition to Aeromonas and Plesiomonas shigelloides, thereby favoring the laboratory diagnosis of gastroenteritis.

  10. Role of Plasmid in Production of Acetobacter Xylinum Biofilms

    Directory of Open Access Journals (Sweden)

    Abbas Rezaee

    2005-01-01

    Full Text Available Acetobacter xylinum has the ability to produce cellulotic biofilms. Bacterial cellulose is expected to be used in many industrial or biomedical materials for its unique characteristics. A. xylinum contains a complex system of plasmid DNA molecules. A 44 kilobases (kb plasmid was isolated in wild type of A. xylinum. To improve the cellulose producing ability of A. xylinum, role of the plasmid in production of cellulose was studied. The comparisons between wild type and cured cells of A. xylinum showed that there is considerably difference in cellulose production. In order to study the relationship between plasmid and the rate of cellulose production, bacteria were screened for plasmid profile by a modified method for preparation of plasmid. This method yields high levels of pure plasmid DNA that can be used for common molecular techniques, such as digestion and transformation, with high efficiency.

  11. Deficient sumoylation of yeast 2-micron plasmid proteins Rep1 and Rep2 associated with their loss from the plasmid-partitioning locus and impaired plasmid inheritance.

    Directory of Open Access Journals (Sweden)

    Jordan B Pinder

    Full Text Available The 2-micron plasmid of the budding yeast Saccharomyces cerevisiae encodes copy-number amplification and partitioning systems that enable the plasmid to persist despite conferring no advantage to its host. Plasmid partitioning requires interaction of the plasmid Rep1 and Rep2 proteins with each other and with the plasmid-partitioning locus STB. Here we demonstrate that Rep1 stability is reduced in the absence of Rep2, and that both Rep proteins are sumoylated. Lysine-to-arginine substitutions in Rep1 and Rep2 that inhibited their sumoylation perturbed plasmid inheritance without affecting Rep protein stability or two-hybrid interaction between Rep1 and Rep2. One-hybrid and chromatin immunoprecipitation assays revealed that Rep1 was required for efficient retention of Rep2 at STB and that sumoylation-deficient mutants of Rep1 and Rep2 were impaired for association with STB. The normal co-localization of both Rep proteins with the punctate nuclear plasmid foci was also lost when Rep1 was sumoylation-deficient. The correlation of Rep protein sumoylation status with plasmid-partitioning locus association suggests a theme common to eukaryotic chromosome segregation proteins, sumoylated forms of which are found enriched at centromeres, and between the yeast 2-micron plasmid and viral episomes that depend on sumoylation of their maintenance proteins for persistence in their hosts.

  12. Caracterização de Aeromonas Spp. Isoladas de Águas Não Tratadas para Consumo Humano

    OpenAIRE

    Ribeiro, Maria Ermelinda de Aguiar

    2008-01-01

    O género Aeromonas forma uma unidade monofilética dentro do sub-grupo gama -3 das Proteobactérias. Bactérias do grupo das Aeromonas surgem em diversos ambientes aquáticos. Está também comprovada a presença de Aeromonas spp. em águas para consumo humano, tratadas e não tratadas. Os organismos deste grupo são frequentemente referidos como “patogénicos emergentes” sendo pesquisados obrigatóriamente no Canadá, Itália e Holanda. Assim, pretende-se com este estudo, fazer uma avaliação da presença d...

  13. Modeling sRNA-regulated Plasmid Maintenance

    CERN Document Server

    Gong, Chen Chris

    2016-01-01

    We study a theoretical model for the toxin-antitoxin (hok/sok) mechanism for plasmid maintenance in bacteria. Toxin-antitoxin systems enforce the maintenance of a plasmid through post-segregational killing of cells that have lost the plasmid. Key to their function is the tight regulation of expression of a protein toxin by an sRNA antitoxin. Here, we focus on the nonlinear nature of the regulatory circuit dynamics of the toxin-antitoxin mechanism. The mechanism relies on a transient increase in protein concentration rather than on the steady state of the genetic circuit. Through a systematic analysis of the parameter dependence of this transient increase, we confirm some known design features of this system and identify new ones: for an efficient toxin-antitoxin mechanism, the synthesis rate of the toxin's mRNA template should be lower that of the sRNA antitoxin, the mRNA template should be more stable than the sRNA antitoxin, and the mRNA-sRNA complex should be more stable than the sRNA antitoxin. Moreover, ...

  14. Identification and Molecular Characterization of the Homogentisate Pathway Responsible for Pyomelanin Production, the Major Melanin Constituents in Aeromonas media WS

    OpenAIRE

    He Wang; Yunqian Qiao; Baozhong Chai; Chenxi Qiu; Xiangdong Chen

    2015-01-01

    The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine) based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate). Gene products of phhA (encodes phenylalanine hydroxylase), ty...

  15. Impact of thermal loading and other water quality parameters on the epizootiology of Aeromonas hydrophila infections of centrarchids. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Esch, G.W.; Hazen, T.C.

    1983-01-01

    Red-sore disease is a rather recent problem for those involved in the sport and commercial fishing industries in the southeastern United States. The causative, or etiological, agent for red-sore disease is the widespread and commonly occurring bacterium, Aeromonas hydrophila. The disease is characterized by both external and internal lesions with serious damage to internal organs ultimately causing death of the host. Evidence points to the existence of several strains of Aeromonas hydrophila with some more virulent than others. The prevalence of the disease is related to elevated water temperature and body condition.

  16. A foodborne outbreak of Aeromonas hydrophila in a college, Xingyi City, Guizhou, China, 2012

    Directory of Open Access Journals (Sweden)

    Guang Zeng

    2012-12-01

    Full Text Available Background: On 12 May 2012, over 200 college students with acute diarrhoea were reported to the Guizhou Center for Disease Control and Prevention. We conducted an investigation to identify the agent and mode of transmission and to recommend control measures.Methods: A suspected case was a person at the college with onset of more than two of the following symptoms: diarrhoea (more than three loose stools in 24 hours, abdominal pain, vomiting or fever (> 37.5C between 6 and 15 May 2012. A confirmed case also had a positive Aeromonas hydrophila culture from a stool sample. A retrospective-cohort study of 902 students compared attack rates (AR by dining place, meals and food history. We reviewed the implicated premise, its processes and preparation of implicated food.Results: We identified 349 suspected cases (AR = 14% and isolated Aeromonas hydrophila from three stools of 15 cases. Students who ate in cafeteria A were more likely to be ill compared to those eating in other places (relative risk [RR]: 3.1, 95% confidence interval [CI]: 2.0–4.8. The cohort study implicated cold cucumber (RR: 2.6, 95% CI: 2.0–3.3 and houttuynia dishes (RR: 1.8, 95% CI: 1.4–2.3. Environmental investigation showed that vegetables were washed in polluted water from a tank close to the sewage ditch, then left at 30°C for two hours before serving. The Escherichia coli count of the tank was well above the standard for drinking water.Conclusion: This outbreak of Aeromonas hydrophila was most probably caused by salad ingredients washed in contaminated tank water. We recommended enhancing training of foodhandlers, ensuring tanks and sewerage systems comply with appropriate standards and adequate monitoring of drinking water sources.

  17. INHIBITION OF Aeromonas hydrophila BY PROBIOTIC STRAINS ISOLATED FROM THE DIGESTIVE TRACT OF Pterophyllum scalare

    OpenAIRE

    M. C. Monroy-Dosta; T. Castro-Barrera; F. J. Fernández-Perrino; L. Mayorga-Reyes

    2010-01-01

    The effect of three dierent probiotic strains of Bacillus (B1, B2 and B3) isolated from the digestive tract of Pterophyllum scalare against Aeromonas hydrophila was evaluated. All the sh that where fed with the dierent probiotic strains obtained higher resistance to the pathogen inoculation since they did not developed any signs of illness nor lesions and they stayed healthy until the end of the experiment, with an observed survival of 100 %. On the other hand, the sh where the probiotic bact...

  18. Purification and Properties of β-1, 4-Xylanase from Aeromonas caviae W-61

    OpenAIRE

    Viet, Dung Nguyen; Kamio, Yoshiyuki; Abe, Naoki; Kaneko, Jun; Izaki, Kazuo

    1991-01-01

    Aeromonas caviae W-61, which was isolated from water samples at the Faculty of Agriculture, Tohoku University, produced β-1, 4-xylanase (1,4-β-d-xylan xylanohydrolase; EC 3.2.1.8) extracellularly. The xylanase was purified to homogeneity by using DEAE-Sephadex A-50, CM-Sephadex C-50, and Sephadex G-100 column chromatographies. The molecular weight of the purified enzyme was estimated to be 22,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the enzyme...

  19. Molecular Cloning of the Tryptophan Operon from an Aeromonas hydrophila Freshwater Isolate

    OpenAIRE

    Anguita, Juan; Rivero, Octavio; Naharro, Germán

    1992-01-01

    A genomic library of Aeromonas hydrophila F9 was constructed by using pBR322 as a vector. From that, two DNA fragments (5.8 and 11.6 kb) were isolated containing genetic information to complement trpA and trpB defects (5.8-kb fragment) and to complement trpA, trpB, trpC, trpD, and trpE defects (11.6-kb fragment) in Escherichia coli mutants. Evidence of the existence of a secondary promoter is given.

  20. Diabetic Foot Due to Aeromonas Hydrophila and Pseudomonas Oryzihabitans: A Case Report

    Directory of Open Access Journals (Sweden)

    Aysel Talan

    2014-09-01

    Full Text Available Foot infections are the most common cause of amputations in diabetic patients. The main predisposing reasons for the development of foot ulcers and subsequent infection in diabetic patients are sensory neuropathy, vascular insufficiency and immune system abnormalities due to hyperglycemia. Limb-threatening infections are often polymicrobial. Staphylococcus aureus, group B streptococci, enterococci and facultative gram-negative bacilli are the most common pathogens. Here, we present two cases of diabetic foot infected by extremely rare microorganisms: Aeromonas hydrophila, and Pseudomonas (Flavimonas oryzihabitans. Turk Jem 2014; 18: 100-102

  1. Determination of Plasmid Segregational Stability in a Growing Bacterial Population.

    Science.gov (United States)

    Kramer, M Gabriela

    2016-01-01

    Bacterial plasmids are extensively used as cloning vectors for a number of genes for academic and commercial purposes. Moreover, attenuated bacteria carrying recombinant plasmids expressing genes with anti-tumor activity have shown promising therapeutic results in animal models of cancer. Equitable plasmid distribution between daughter cells during cell division, i.e., plasmid segregational stability, depends on many factors, including the plasmid copy number, its replication mechanism, the levels of recombinant gene expression, the type of bacterial host, and the metabolic burden associated with all these factors. Plasmid vectors usually code for antibiotic-resistant functions, and, in order to enrich the culture with bacteria containing plasmids, antibiotic selective pressure is commonly used to eliminate plasmid-free segregants from the growing population. However, administration of antibiotics can be inconvenient for many industrial and therapeutic applications. Extensive ongoing research is being carried out to develop stably-inherited plasmid vectors. Here, I present an easy and precise method for determining the kinetics of plasmid loss or maintenance for every ten generations of bacterial growth in culture. PMID:26846807

  2. Comparative analysis of plasmids in the genus Listeria.

    Directory of Open Access Journals (Sweden)

    Carsten Kuenne

    Full Text Available BACKGROUND: We sequenced four plasmids of the genus Listeria, including two novel plasmids from L. monocytogenes serotype 1/2c and 7 strains as well as one from the species L. grayi. A comparative analysis in conjunction with 10 published Listeria plasmids revealed a common evolutionary background. PRINCIPAL FINDINGS: All analysed plasmids share a common replicon-type related to theta-replicating plasmid pAMbeta1. Nonetheless plasmids could be broadly divided into two distinct groups based on replicon diversity and the genetic content of the respective plasmid groups. Listeria plasmids are characterized by the presence of a large number of diverse mobile genetic elements and a commonly occurring translesion DNA polymerase both of which have probably contributed to the evolution of these plasmids. We detected small non-coding RNAs on some plasmids that were homologous to those present on the chromosome of L. monocytogenes EGD-e. Multiple genes involved in heavy metal resistance (cadmium, copper, arsenite as well as multidrug efflux (MDR, SMR, MATE were detected on all listerial plasmids. These factors promote bacterial growth and survival in the environment and may have been acquired as a result of selective pressure due to the use of disinfectants in food processing environments. MDR efflux pumps have also recently been shown to promote transport of cyclic diadenosine monophosphate (c-di-AMP as a secreted molecule able to trigger a cytosolic host immune response following infection. CONCLUSIONS: The comparative analysis of 14 plasmids of genus Listeria implied the existence of a common ancestor. Ubiquitously-occurring MDR genes on plasmids and their role in listerial infection now deserve further attention.

  3. Unique type of plasmid maintenance function: postsegregational killing of plasmid-free cells.

    OpenAIRE

    Gerdes, K; Rasmussen, P. B.; Molin, S

    1986-01-01

    The stability locus parB+ of plasmid R1 has been found to specify a unique type of plasmid maintenance function. Two genes, hok (host killing) and sok (suppressor of killing), are required for the stabilizing activity. The hok gene encodes a highly toxic gene product, whose overexpression causes a rapid killing and a concomitant dramatic change in morphology of the host cell. The other gene, sok, was found to encode a product that counteracts the hok gene-mediated killing. The parB+ region wa...

  4. Molecular characterization of "plasmid-free" antibiotic-resistant Haemophilus influenzae.

    OpenAIRE

    Roberts, M C; Smith, A. L.

    1980-01-01

    We examined 14 multiresistant and 8 ampicillin- or tetracycline-resistant Haemophilus influenzae isolates and 4 ampicillin-resistant H. parainfluenzae isolates for plasmid deoxyribonucleic acid. Sixteen strains carried plasmids. Both "plasmid-free" and plasmid-carrying isolates transferred the antibiotic resistance by conjugation. All transconjugants carried plasmid deoxyribonucleic acid, suggesting that the apparent plasmid-free strains contained R plasmids encoding for antibiotic resistance.

  5. Permissiveness of soil microbial communities towards broad host range plasmids

    DEFF Research Database (Denmark)

    Klümper, Uli

    for plasmids carrying antibiotic resistance genes is increasingly suspected to majorly contribute to the emergence of multi-resistant pathogens. More specifically, I examined what fraction of a soil microbial community is permissive to plasmids, identified the phylogenetic identity of this fraction and studied......Horizontal transfer of mobile genetic elements facilitates adaptive and evolutionary processes in bacteria. Among the known mobile genetic elements, plasmids can confer their hosts with accessory adaptive traits, such as antibiotic or heavy metal resistances, or additional metabolic pathways....... Plasmids are implicated in the rapid spread of antibiotic resistance and the emergence of multi-resistant pathogenic bacteria, making it crucial to be able to quantify, understand, and, ideally, control plasmid transfer in mixed microbial communities. The fate of plasmids in microbial communities...

  6. Transformation of Haemophilus influenzae by plasmid RSF0885

    Energy Technology Data Exchange (ETDEWEB)

    Notani, N.K.; Setlow, J.K.; McCarthy, D.; Clayton, N.L.

    1981-12-01

    Plasmid RSF0885, which conferred ampicillin resistance, transformed competent Haemophilus influenzae cells with low efficiency (maximun, less than 0.01%). As judged by competition experiments and uptake of radioactivity, plasmid RSF0885 deoxyribonucleic acid was taken up into competent H. influenzae cells several orders of magnitude less efficiently than H. influenzae chromosomal deoxyribonucleic acid. Plasmid RSF0885 transformed cells with even lower efficiency than could be accounted for by the low uptake. Transformation was not affected by rec-1 and rec-2 mutations in the recipient, and strains cured of the plasmid did not show increased transformation. Plasmid molecules cut once with a restriction enzyme that made blunt ends did not transform. Transformation was favored by the closed circular form of the plasmid.

  7. Streamlined Purification of Plasmid DNA From Prokaryotic Cultures

    OpenAIRE

    Pueschel, Laura; Li, Hongshan; Hymes, Matthew

    2011-01-01

    We describe the complete process of AcroPrep Advance Filter Plates for 96 plasmid preparations, starting from prokaryotic culture and ending with high purity DNA. Based on multi-well filtration for bacterial lysate clearance and DNA purification, this method creates a streamlined process for plasmid preparation. Filter plates containing silica-based media can easily be processed by vacuum filtration or centrifuge to yield appreciable quantities of plasmid DNA. Quantitative analyses determine ...

  8. Processes for the production of pharmaceutical grade plasmid DNA

    OpenAIRE

    Voß, Carsten

    2008-01-01

    Plasmid DNA is currently used in gene therapy and genetic vaccination as a vector system for the delivery of therapeutic genes. Clinical trials as well as future therapeutics demand large amounts of high quality plasmid DNA that fulfils the specifications set by regulatory authorities. This thesis describes the development, analysis, and evaluation of pharmaceutical plasmid DNA production processes comprising cultivation, product isolation, and purification as well as stability assessment dur...

  9. Identification of plasmid partition function in coryneform bacteria.

    OpenAIRE

    Kurusu, Y; Satoh, Y.; Inui, M.; Kohama, K; Kobayashi, M.; Terasawa, M.; Yukawa, H

    1991-01-01

    We have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. ...

  10. Pathogenomics of the Virulence Plasmids of Escherichia coli

    OpenAIRE

    Johnson, Timothy J.; Lisa K. Nolan

    2009-01-01

    Summary: Bacterial plasmids are self-replicating, extrachromosomal elements that are key agents of change in microbial populations. They promote the dissemination of a variety of traits, including virulence, enhanced fitness, resistance to antimicrobial agents, and metabolism of rare substances. Escherichia coli, perhaps the most studied of microorganisms, has been found to possess a variety of plasmid types. Included among these are plasmids associated with virulence. Several types of E. col...

  11. Safety and efficacy of DNA vaccines: Plasmids vs. minicircles

    OpenAIRE

    Stenler, Sofia; Blomberg, Pontus; Smith, Ci Edvard

    2014-01-01

    While DNA vaccination using plasmid vectors is highly attractive, there is a need for further vector optimization regarding safety, stability, and efficiency. In this commentary, we review the minicircle vector (MC), which is an entity devoid of plasmid bacterial sequences, as an alternative to the traditional plasmid construct. The commentary highlights the recent discovery by Stenler et al. (2014) that the small size of an MC enables improved resistance to the shearing forces associated wit...

  12. Comparative genetic organization of incompatibility group P degradative plasmids.

    OpenAIRE

    Burlage, R S; Bemis, L A; Layton, A C; Sayler, G. S.; Larimer, F

    1990-01-01

    Plasmids that encode genes for the degradation of recalcitrant compounds are often examined only for characteristics of the degradative pathways and ignore regions that are necessary for plasmid replication, incompatibility, and conjugation. If these characteristics were known, then the mobility of the catabolic genes between species could be predicted and different catabolic pathways might be combined to alter substrate range. Two catabolic plasmids, pSS50 and pSS60, isolated from chlorobiph...

  13. Degradative Plasmid and Heavy Metal Resistance Plasmid Naturally Coexist in Phenol and Cyanide Assimilating Bacteria

    OpenAIRE

    Bahig E.  Deeb; Abdullah D. Altalhi

    2009-01-01

    Problem statement: Heavy metals are known to be powerful inhibitors of xenobiotics biodegradation activities. Alleviation the inhibitory effect of these metals on the phenol biodegradation activities in presence of heavy metals resistant plasmid was investigated. Approach: Combination of genetic systems of degradation of xenobiotic compound and heavy metal resistance was one of the approaches to the creation of polyfunctional strains for bioremediation of s...

  14. Photonic plasmid stability of transformed Salmonella typhimurium: A comparison of three unique plasmids

    Science.gov (United States)

    Acquiring a highly stable photonic plasmid in transformed Salmonella typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella typhimurium (S. typh-lux) u...

  15. Photonic plasmid stability of transformed Salmonella Typhimurium: A comparison of three unique plasmids

    Directory of Open Access Journals (Sweden)

    Lay Donald

    2009-07-01

    Full Text Available Abstract Background Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S. typh-lux using three different plasmids and characterize their respective photonic properties. Results In presence of ampicillin (AMP, S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 plasmids exhibited 100% photon-emitting colonies over a 10-d study period. Photon emitters of S. typh-lux with pCGLS-1, pAK1-lux and pXEN-1 without AMP selection decreased over time (P 7 to 1 × 109 CFU, P 0.05; although photonic emissions across a range of bacterial concentrations were not different (1 × 104 to 1 × 106 CFU, P > 0.05. For very low density bacterial concentrations imaged in 96 well plates photonic emissions were positively correlated with bacterial concentration (P 3 to 1 × 105 CFU low to high were different in the 96-well plate format (P Conclusion These data characterize photon stability properties for S. typh-lux transformed with three different photon generating plasmids that may facilitate real-time Salmonella tracking using in vivo or in situ biophotonic paradigms.

  16. Photonic Plasmid Stability of Transformed Salmonella Typhimurium: A Comparison of Three Unique Plasmids

    Science.gov (United States)

    Background: Acquiring a highly stable photonic plasmid in transformed Salmonella Typhimurium for use in biophotonic studies of bacterial tracking in vivo is critical to experimental paradigm development. The objective of this study was to determine stability of transformed Salmonella Typhimurium (S....

  17. Lysing activity of an indigenous algicidal bacterium Aeromonas sp. against Microcystis spp. isolated from Lake Taihu.

    Science.gov (United States)

    Yang, Fei; Li, Xiaoqin; Li, Yunhui; Wei, Haiyan; Yu, Guang; Yin, Lihong; Liang, Geyu; Pu, Yuepu

    2013-01-01

    This study aimed to isolate and characterize an indigenous algicidal bacterium named LTH-1 and its algae-lysing compounds active against three Microcystis aeruginosa strains (toxic TH1, nontoxic TH2 and standard FACHB 905). The LTH-1 isolated from Lake Taihu, near Wuxi City in China, was identified as Aeromonas sp. based on its morphological characteristic features and phylogenetic analysis by sequencing of 16S rDNA. Extracellular compounds produced by LTH-1 showed strong algaelysing activity, and they were water-soluble and heat-tolerant, with a molecular mass lower than 2 kDa. Two algae-lysing compounds were isolated and purified from extracellular filtrate using silica gel column chromatography. One of these was identified as phenylalanine (C9H11NO2, m/z 166.0862) and the other (C8H16N2O3, m/z 189.1232) was unidentified by hybrid ion trap/time-of-flight mass spectrometry coupled with a high-performance liquid chromatography (LC/MS-IT-TOF) system. The half maximal effective concentration (EC50) of phenylalanine produced by LTH-1 against FACHB 905 was 68.2 +/- 8.2 microg mL(-1) in 48h. These results suggest that the algicidal Aeromonas sp. LTH-1 could play a role in controlling Microcystis blooms, and its extracellular compounds are also potentially useful for regulating blooms of the harmful M. aeruginosa. PMID:24191475

  18. Detection and quantification of virulent Aeromonas hydrophila in channel catfish tissues following waterborne challenge.

    Science.gov (United States)

    Zhang, Dunhua; Moreira, Gabriel S A; Shoemaker, Craig; Newton, Joseph C; Xu, De-Hai

    2016-05-01

    The aim of this study was to understand the pathogenesis of motile aeromonas septicemia caused by an emergent, high virulent Aeromonas hydrophila (vAh) in channel catfish, Ictalurus punctatus Adipose fin clipped catfish were challenged with vAh using a waterborne challenge method, and the distribution of vAh over a time course was detected and quantified using real-time polymerase chain reaction. The results showed that 77.8% of fish died within 48 h post challenge with mean day to death of 1.5 days. At 2 h post challenge, vAh (inferred from genomic DNA copies or genome equivalents) was detected in all external and internal tissues sampled. Gill had the highest vAh cells at 1 h post challenge. Spleen harbored the most vAh cells among internal organs at 4 h post challenge. The tissues/organs with most vAh cells detected at 8 h post challenge were adipose fin, blood, intestine, kidney and skin, while liver showed the highest vAh cells at 24 h post challenge. These results suggest that vAh was able to rapidly proliferate and spread, following wound infection, through the fish blood circulation system and cause mortality within 8-24 h. PMID:27044300

  19. Bioremediation and Detoxification of Synthetic Wastewater Containing Triarylmethane Dyes by Aeromonas hydrophila Isolated from Industrial Effluent

    Directory of Open Access Journals (Sweden)

    Chimezie Jason Ogugbue

    2011-01-01

    Full Text Available Economical and bio-friendly approaches are needed to remediate dye-contaminated wastewater from various industries. In this study, a novel bacterial strain capable of decolorizing triarylmethane dyes was isolated from a textile wastewater treatment plant in Greece. The bacterial isolate was identified as Aeromonas hydrophila and was shown to decolorize three triarylmethane dyes tested within 24 h with color removal in the range of 72% to 96%. Decolorization efficiency of the bacterium was a function of operational parameters (aeration, dye concentration, temperature, and pH and the optimal operational conditions obtained for decolorization of the dyes were: pH 7-8, 35∘C and culture agitation. Effective color removal within 24 h was obtained at a maximum dye concentration of 50 mg/L. Dye decolorization was monitored using a scanning UV/visible spectrophotometer which indicated that decolorization was due to the degradation of dyes into non-colored intermediates. Phytotoxicity studies carried out using Triticum aestivum, Hordeum vulgare, and Lens esculenta revealed the triarylmethane dyes exerted toxic effects on plant growth parameters monitored. However, significant reduction in toxicity was obtained with the decolorized dye metabolites thus, indicating the detoxification of the dyes following degradation by Aeromonas hydrophila.

  20. The Aeromonas salmonicida Lipopolysaccharide Core from Different Subspecies: The Unusual subsp. pectinolytica.

    Science.gov (United States)

    Merino, Susana; Tomás, Juan M

    2016-01-01

    Initial hydridization tests using Aeromonas salmonicida typical and atypical strains showed the possibility of different lipopolysaccharide (LPS) outer cores among these strains. By chemical structural analysis, LPS-core SDS-PAGE gel migration, and functional and comparative genomics we demonstrated that typical A. salmonicida (subsp. salmonicida) strains and atypical subsp. masoucida and probably smithia strains showed the same LPS outer core. A. salmonicida subsp. achromogenes strains show a similar LPS outer core but lack one of the most external residues (a galactose linked α1-6 to heptose), not affecting the O-antigen LPS linkage. A. salmonicida subsp. pectinolytica strains show a rather changed LPS outer core, which is identical to the LPS outer core from the majority of the A. hydrophila strains studied by genomic analyses. The LPS inner core in all tested A. salmonicida strains, typical and atypical, is well-conserved. Furthermore, the LPS inner core seems to be conserved in all the Aeromonas (psychrophilic or mesophilic) strains studied by genomic analyses.

  1. Identification of atypical Aeromonas salmonicida : Inter-laboratory evaluation and harmonization of methods

    DEFF Research Database (Denmark)

    Dalsgaard, Inger; Gudmundsdottir, B.K.; Helgason, S.;

    1998-01-01

    The atypical isolates of Aeromonas salmonicida are becoming increasingly important as the frequency of isolation of bacteria belonging to this group continues to rise. The primary object of this study was to compare and evaluate the results obtained in various laboratories concerning the biochemi......The atypical isolates of Aeromonas salmonicida are becoming increasingly important as the frequency of isolation of bacteria belonging to this group continues to rise. The primary object of this study was to compare and evaluate the results obtained in various laboratories concerning the...... biochemical identification of atypical Aer. salmonicida before and after standardization of media and methods. Five laboratories examined 25 isolates of Aer. salmonicida from diverse fish species and geographical locations including the reference strains of Aer. salmonicida subsp, salmonicida (NCMB 1102) and...... reactions. After standardization, improvement was obtained with the identification; however, disagreement was still observed between the different laboratories. These findings demonstrate the difficulties involved in a proper identification of atypical Aer. salmonicida and also that data presented in the...

  2. Bioremediation and Detoxification of Synthetic Wastewater Containing Triarylmethane Dyes by Aeromonas hydrophila Isolated from Industrial Effluent.

    Science.gov (United States)

    Ogugbue, Chimezie Jason; Sawidis, Thomas

    2011-01-01

    Economical and bio-friendly approaches are needed to remediate dye-contaminated wastewater from various industries. In this study, a novel bacterial strain capable of decolorizing triarylmethane dyes was isolated from a textile wastewater treatment plant in Greece. The bacterial isolate was identified as Aeromonas hydrophila and was shown to decolorize three triarylmethane dyes tested within 24 h with color removal in the range of 72% to 96%. Decolorization efficiency of the bacterium was a function of operational parameters (aeration, dye concentration, temperature, and pH) and the optimal operational conditions obtained for decolorization of the dyes were: pH 7-8, 35°C and culture agitation. Effective color removal within 24 h was obtained at a maximum dye concentration of 50 mg/L. Dye decolorization was monitored using a scanning UV/visible spectrophotometer which indicated that decolorization was due to the degradation of dyes into non-colored intermediates. Phytotoxicity studies carried out using Triticum aestivum, Hordeum vulgare, and Lens esculenta revealed the triarylmethane dyes exerted toxic effects on plant growth parameters monitored. However, significant reduction in toxicity was obtained with the decolorized dye metabolites thus, indicating the detoxification of the dyes following degradation by Aeromonas hydrophila. PMID:21808740

  3. Nonribosomal peptide synthetase with a unique iterative-alternative-optional mechanism catalyzes amonabactin synthesis in Aeromonas.

    Science.gov (United States)

    Esmaeel, Qassim; Chevalier, Mickael; Chataigné, Gabrielle; Subashkumar, Rathinasamy; Jacques, Philippe; Leclère, Valérie

    2016-10-01

    Based on the exploration of data generated by genome sequencing, a bioinformatics approach has been chosen to identify the biosynthetic pathway of the siderophores produced by Aeromonas species. The amonabactins, considered as a virulence factor, represent a family of four variants of catechol peptidic siderophores containing Dhb, Lys, Gly, and an aromatic residue either Trp or Phe in a D-configuration. The synthesis operon is constituted of seven genes named amoCEBFAGH and is iron-regulated. The cluster includes genes encoding proteins involved in the synthesis and incorporation of the Dhb monomer, and genes encoding specific nonribosomal peptide synthetases, which are responsible for the building of the peptidic moiety. The amonabactin assembly line displays a still so far not described atypical mode of synthesis that is iterative, alternative, and optional. A disruption mutant in the adenylation domain of AmoG was unable to synthesize any amonabactin and to grow in iron stress conditions while a deletion of amoH resulted in the production of only two over the four forms. The amo cluster is widespread among most of the Aeromonas species, only few species produces the enterobactin siderophore. PMID:27531515

  4. Degradation of chitin and chitosan by a recombinant chitinase derived from a virulent Aeromonas hydrophila isolated from diseased channel catfish

    Science.gov (United States)

    A chitinase was identified in extracellular products of a virulent Aeromonas hydrophila isolated from diseased channel catfish (Ictalurus punctatus). Bioactive recombinant chitinase (rChi-Ah) was produced in Escherichia coli. Purified rChi-Ah had optimal activity at temperature of 42°C and pH 6.5. T...

  5. Polymerase chain reaction (PCR)-based typing analysis of atypical isolates of the fish pathogen Aeromonas salmonicida

    DEFF Research Database (Denmark)

    Høie, S.; Dalsgaard, Inger; Aase, I.L.;

    1999-01-01

    Two hundred and five isolates of atypical Aeromonas salmonicida, recovered from a wide range of hosts and countries were characterized by polymerase chain reaction (PCR) targeting four genes. The chosen genes were those encoding the extracellular A-layer protein (AP), the serine protease (Sprot...

  6. Draft genome sequence of Pseudomonas mosselii Gil3, isolated from catfish and antagonistic against hypervirulent Aeromonas hydrophila

    Science.gov (United States)

    Pseudomonas mosselii Gil3 was isolated from a catfish that survived from lethal challenge with hypervirulent Aeromonas hydrophila (vAh). When assayed in vitro, the bacterium showed antagonism against vAh. Sequence analysis revealed that the genome of P. mosselii Gil3 encodes numerous aromatic metabo...

  7. Bacteriophages Limit the Existence Conditions for Conjugative Plasmids

    Science.gov (United States)

    Wood, A. Jamie; Dytham, Calvin; Pitchford, Jonathan W.; Truman, Julie; Spiers, Andrew; Paterson, Steve; Brockhurst, Michael A.

    2015-01-01

    ABSTRACT Bacteriophages are a major cause of bacterial mortality and impose strong selection on natural bacterial populations, yet their effects on the dynamics of conjugative plasmids have rarely been tested. We combined experimental evolution, mathematical modeling, and individual-based simulations to explain how the ecological and population genetics effects of bacteriophages upon bacteria interact to determine the dynamics of conjugative plasmids and their persistence. The ecological effects of bacteriophages on bacteria are predicted to limit the existence conditions for conjugative plasmids, preventing persistence under weak selection for plasmid accessory traits. Experiments showed that phages drove faster extinction of plasmids in environments where the plasmid conferred no benefit, but they also revealed more complex effects of phages on plasmid dynamics under these conditions, specifically, the temporary maintenance of plasmids at fixation followed by rapid loss. We hypothesized that the population genetic effects of bacteriophages, specifically, selection for phage resistance mutations, may have caused this. Further mathematical modeling and individual-based simulations supported our hypothesis, showing that conjugative plasmids may hitchhike with phage resistance mutations in the bacterial chromosome. PMID:26037122

  8. Plasmid genes required for microcin B17 production.

    Science.gov (United States)

    San Millán, J L; Kolter, R; Moreno, F

    1985-09-01

    The production of the antibiotic substance microcin B17 (Mcc) is determined by a 3.5-kilobase DNA fragment from plasmid pMccB17. Several Mcc- mutations on plasmid pMccB17 were obtained by both transposon insertion and nitrosoguanidine mutagenesis. Plasmids carrying these mutations were tested for their ability to complement Mcc- insertion or deletion mutations on pMM102 (pMM102 is a pBR322 derivative carrying the region encoding microcin B17). Results from these experiments indicate that at least four plasmid genes are required for microcin production.

  9. Identification of plasmid partition function in coryneform bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Kurusu, Yasurou; Satoh, Yukie; Inui, Masayuki; Kohama, Keiko; Kobayashi, Miki; Terasawa, Masato; Yukawa, Hideaki (Mitsubishi Petrochemical Co., Ltd., Ibaraki (Japan))

    1991-03-01

    The authors have identified and characterized a partition function that is required for stable maintenance of plasmids in the coryneform bacteria Brevibacterium flavum MJ233 and Corynebacterium glutamicum ATCC 31831. This function is localized to a HindIII-NspV fragment (673 bp) adjacent to the replication region of the plasmid, named pBY503, from Brevibacterium stationis IFO 12144. The function was independent of copy number control and was not associated directly with plasmid replication functions. This fragment was able to stabilize the unstable plasmids in cis but not in trans.

  10. Plasmids foster diversification and adaptation of bacterial populations in soil.

    Science.gov (United States)

    Heuer, Holger; Smalla, Kornelia

    2012-11-01

    It is increasingly being recognized that the transfer of conjugative plasmids across species boundaries plays a vital role in the adaptability of bacterial populations in soil. There are specific driving forces and constraints of plasmid transfer within bacterial communities in soils. Plasmid-mediated genetic variation allows bacteria to respond rapidly with adaptive responses to challenges such as irregular antibiotic or metal concentrations, or opportunities such as the utilization of xenobiotic compounds. Cultivation-independent detection and capture of plasmids from soil bacteria, and complete sequencing have provided new insights into the role and ecology of plasmids. Broad host range plasmids such as those belonging to IncP-1 transfer a wealth of accessory functions which are carried by similar plasmid backbones. Plasmids with a narrower host range can be more specifically adapted to particular species and often transfer genes which complement chromosomally encoded functions. Plasmids seem to be an ancient and successful strategy to ensure survival of a soil population in spatial and temporal heterogeneous conditions with various environmental stresses or opportunities that occur irregularly or as a novel challenge in soil.

  11. Characterization of a plasmid from moderately halophilic eubacteria

    OpenAIRE

    Fernández Castillo, Rosario; Vargas, C.; Nieto Gutiérrez, Joaquín José; Ventosa Ucero, Antonio; Ruiz Berraquero, Francisco

    1992-01-01

    A plasmid has been isolated for the first time from moderately halophilic eubacteria. Halomonas elongata, Halomonrrs halmphila, Deleya halophila and Vibrio costkola were found to harbour an 11.5 kbp plasmid (pMH1). The plasmid was isolated and characterized after transformation into Escherichia coh'JM101 cells. A restriction map was constructed, and unique restriction sites for EcoRI, EcoRV and ClaI were detected. The occurrence of such a plasmid in the original halophilic strains was confirm...

  12. Degradative Plasmid and Heavy Metal Resistance Plasmid Naturally Coexist in Phenol and Cyanide Assimilating Bacteria

    Directory of Open Access Journals (Sweden)

    Bahig E.  Deeb

    2009-01-01

    Full Text Available Problem statement: Heavy metals are known to be powerful inhibitors of xenobiotics biodegradation activities. Alleviation the inhibitory effect of these metals on the phenol biodegradation activities in presence of heavy metals resistant plasmid was investigated. Approach: Combination of genetic systems of degradation of xenobiotic compound and heavy metal resistance was one of the approaches to the creation of polyfunctional strains for bioremediation of soil after co-contamination with organic pollutants and heavy metals. Results: A bacterial strain Pseudomonas putida PhCN (pPhCN1, pPhCN2 had been obtained. This bacterium contained two plasmids, a 120 Kb catabolic plasmid that encode for breakdown of phenol (pPhCN1 and pPhCN2 plasmid (100 Kb that code for cadmium and copper resistant. Cyanide assimilation by this bacterium was encoded by chromosomal genes. The inhibitory effect of cadmium (Cd2+ or copper (Cu2+ on the degradation of phenol and cyanide by P. putida strains PhCN and PhCN1 (contained pPhCN1 were investigated. The resistant strain PhCN showed high ability to degrade phenol and cyanide in presence of Cd2+ or Cu2+ comparing with the sensitive strain PhCN1. In addition, Cd2+ or Cu2+ was also found to exert a strong inhibitory effect on the C23O dioxygenase enzyme activity in the presence of cyanide as a nitrogen source. Conclusion: The presence of heavy metal resistance plasmid alleviated the inhibitory effect of metals on the phenol and cyanide assimilation by resistant strain.

  13. Hematologia e sinais clínicos de Piaractus mesopotamicus infectados experimentalmente com Aeromonas hydrophila = Hematology and clinical signs of Piaractus mesopotamicus experimentally infected with Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    Flávio Ruas Moraes

    2009-01-01

    Full Text Available Este ensaio teve como objetivo avaliar as respostas hematológicas e a ocorrência de sinais clínico-patológicos de Piaractus mesopotamicus infectados experimentalmente por Aeromonas hydrophila. Foram utilizados 324 pacus, P. mesopotamicus, com peso de 102,84 ±27,1 g, distribuídos em 27 caixas de 300 L de capacidade, sendo 14 peixes por caixa. Os peixes foram mantidos nas caixas durante um mês, recebendo dieta balanceada, período em que se evitou qualquer tipo de manejo estressante. Passado esse período, foram infectadoscom 6 x 106 UFC de A. hydrophila, injetada via intraperitoneal. As coletas e análises de sangue foram realizadas 24h antes da infecção experimental e 24h após a infecção. Os resultados permitem concluir que a infecção por A. hydrophila em P. mesopotamicus écaracterizada por escurecimento da derme e sinais clínicos-patológicos relacionados à hemorragia, anemia normocítica-hipocrômica, redução nos níveis de proteínas, globulinasplasmáticas e eosinófilos, leucopenia, linfopenia, trombocitopenia, neutrofilia e monocitose. The purpose of this study was to evaluate the hematological response and clinical sign of Piaractus mesopotamicus infected byAeromonas hydrophila. Were used 324 pacus, P. mesopotamicus weighing 102.84 ± 27.1 g distributed in 27 water tanks with 300 L of capacity, 14 fish per tank. The fish were kept in the water tanks during one month, receiving appropriate diet. In this period was avoidedstressing management. After this period, the fish were infected with 6 × 108 cfu of A. hydrophila fish-1, injected intraperitoneally. The blood collection was carried 24h before experimental infection and 24h after its. The A. hydrophila infection in P. mesopotamicus ischaracterized by dark skin and clinical signs related to hemorrhage, hypochromic normocytic anemia, decrease on plasmatic proteins and globulins levels, leucopenia, lymphopenia, thrombocytopenia, eosinophilia, neutrophilia and monocytosis.

  14. Homology of plasmids in strains of unicellular cyanobacteria

    NARCIS (Netherlands)

    Hondel, C.A.M.J.J. van den; Keegstra, W.; Borrias, W.E.; Arkel, G.A. van

    1979-01-01

    Six strains of unicellular cyanobacteria were examined for the presence of plasmids. Analysis of lysates of these strains by CsCl-ethidium bromide density centrifugation yielded a major chromosomal DNA band and a minor band containing covalently closed circular plasmid DNA, as shown by electron micr

  15. Purification of large plasmids with methacrylate monolithic columns.

    Science.gov (United States)

    Krajnc, Nika Lendero; Smrekar, Franci; Cerne, Jasmina; Raspor, Peter; Modic, Martina; Krgovic, Danijela; Strancar, Ales; Podgornik, Ales

    2009-08-01

    The rapid evolution of gene therapy and DNA vaccines results in an increasing interest in producing large quantities of pharmaceutical grade plasmid DNA. Most current clinical trials involve plasmids of 10 kb or smaller in size, however, future requirements for multigene vectors including extensive control regions may require the production of larger plasmids, e. g., 20 kb and bigger. The objective of this study was to examine certain process conditions for purification of large plasmids with the size of up to 93 kb. Since there is a lack of knowledge about production and purification of bigger plasmid DNA, cell lysis and storage conditions were investigated. The impact of chromatographic system and methacrylate monolithic column on the degradation of plasmid molecules under nonbinding conditions at different flow rates was studied. Furthermore, capacity measurements varying salt concentration in loading buffer were performed and the capacities up to 13 mg of plasmid per mL of the monolithic column were obtained. The capacity flow independence in the range from 130 to 370 cm/h was observed. Using high resolution monolithic column the separation of linear and supercoiled isoforms of large plasmids was obtained. Last but not least, since the baseline separation of RNA and pDNA was achieved, the one step purification on larger CIM DEAE 8 mL tube monolithic column was performed and the fractions were analyzed by CIM analytical monolithic columns. PMID:19598166

  16. Two novel conjugative plasmids from a single strain of Sulfolobus

    NARCIS (Netherlands)

    Erauso, G.; Stedman, K.M.; Werken, van de H.J.G.; Zillig, W.; Oost, van der J.

    2006-01-01

    Two conjugative plasmids (CPs) were isolated and characterized from the same 'Sulfolobus islandicus' strain, SOG2/4, The plasmids were separated from each other and transferred into Sulfolobus soltataricus. One has a high copy number and is not stable (pSOG1) whereas the other has a low copy number

  17. Functional analysis of three plasmids from Lactobacillus plantarum

    NARCIS (Netherlands)

    Kranenburg, R. van; Golic, N.; Bongers, R.; Leer, R.J.; Vos, W.M. de; Siezen, R.J.; Kleerebezem, M.

    2005-01-01

    Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism. Th

  18. Production and pharmaceutical formulation of plasmid DNA vaccines

    NARCIS (Netherlands)

    van der Heijden, I.

    2013-01-01

    Research leading to the thesis ‘Production and pharmaceutical formulation of plasmid DNA vaccines‘ can be divided into two parts. The first part describes the development of a Good Manufacturing Practice (GMP) compliant plasmid DNA production process of pDNA vaccines for the treatment of Human papil

  19. Examination of uropathogenic Escherichia coli strains conferring large plasmids

    Directory of Open Access Journals (Sweden)

    SUHARTONO

    2010-04-01

    Full Text Available Suhartono (2010 Examination of uropathogenic Escherichia coli strains conferring large plasmids. Biodiversitas 11: 59-64. Of major uropathogens, Escherichia coli has been widely known as a main pathogen of UTIs globally and has considerable medical and financial consequences. A strain of UPEC, namely E. coli ST131, confers a large plasmid encoding cephalosporinases (class C β-lactamase or AmpC that may be disseminated through horizontal transfer among bacterial populations. Therefore, it is worth examining such large plasmids by isolating, purifying, and digesting the plasmid with restriction enzymes. The examination of the large plasmids was conducted by isolating plasmid DNA visualized by agarose gel electrophoresis as well as by PFGE. The relationship of plasmids among isolates was carried out by HpaI restriction enzyme digestion. Of 36 isolates of E. coli ST 131, eight isolates possessed large plasmids, namely isolates 3, 9, 10, 12, 17, 18, 26 and 30 with the largest molecular size confirmed by agarose gel electrophoresis and PFGE was ~42kb and ~118kb respectively. Restriction enzyme analysis revealed that isolates 9, 10, 12, 17 and 18 have the common restriction patterns and those isolates might be closely related.

  20. Plasmid cloning vehicle for Haemophilus influenzae and Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Clayton, N.L.; Setlow, J.K.

    1982-09-01

    A new plasmid cloning vehicle (pDM2) was used to introduce a library of Haemophilus influenzae chromosomal fragments into H. influenzae. Transformants of the higly recombination-defective rec-1 mutant were more likely to contain exclusively recombinant plasmids after exposure to ligated DNA mixtures than was the wild type. pDM2 could replicate in Escherichia coli K-12.

  1. Construction and Application of R Prime Plasmids, Carrying Different Segments of an Octopine Ti Plasmid from Agrobacterium tumefaciens, for Complementation of vir Genes

    NARCIS (Netherlands)

    Hille, Jacques; Klasen, Ina; Schilperoort, Rob

    1982-01-01

    Several R prime plasmids have been obtained with high efficiency, by enclosing the R plasmid replicator, in an R::Ti cointegrate plasmid, between two copies of the transposon Tn1831, in the same orientation. These R primes carry different segments of an octopine Ti plasmid, and are compatible with T

  2. Classification of a Hypervirulent Aeromonas hydrophila Pathotype Responsible for Epidemic Outbreaks in Warm-Water Fishes

    Science.gov (United States)

    Rasmussen-Ivey, Cody R.; Hossain, Mohammad J.; Odom, Sara E.; Terhune, Jeffery S.; Hemstreet, William G.; Shoemaker, Craig A.; Zhang, Dunhua; Xu, De-Hai; Griffin, Matt J.; Liu, Yong-Jie; Figueras, Maria J.; Santos, Scott R.; Newton, Joseph C.; Liles, Mark R.

    2016-01-01

    Lineages of hypervirulent Aeromonas hydrophila (vAh) are the cause of persistent outbreaks of motile Aeromonas septicemia in warm-water fishes worldwide. Over the last decade, this virulent lineage of A. hydrophila has resulted in annual losses of millions of tons of farmed carp and catfish in the People's Republic of China and the United States (US). Multiple lines of evidence indicate US catfish and Asian carp isolates of A. hydrophila affiliated with sequence type 251 (ST251) share a recent common ancestor. To address the genomic context for the putative intercontinental transfer and subsequent geographic spread of this pathogen, we conducted a core genome phylogenetic analysis on 61 Aeromonas spp. genomes, of which 40 were affiliated with A. hydrophila, with 26 identified as epidemic strains. Phylogenetic analyses indicate all ST251 strains form a coherent lineage affiliated with A. hydrophila. Within this lineage, conserved genetic loci unique to A. hydrophila were identified, with some genes present in consistently higher copy numbers than in non-epidemic A. hydrophila isolates. In addition, results from analyses of representative ST251 isolates support the conclusion that multiple lineages are present within US vAh isolated from Mississippi, whereas vAh isolated from Alabama appear clonal. This is the first report of genomic heterogeneity within US vAh isolates, with some Mississippi isolates showing closer affiliation with the Asian grass carp isolate ZC1 than other vAh isolated in the US. To evaluate the biological significance of the identified heterogeneity, comparative disease challenges were conducted with representatives of different vAh genotypes. These studies revealed that isolate ZC1 yielded significantly lower mortality in channel catfish, relative to Alabama and Mississippi vAh isolates. Like other Asian vAh isolates, the ZC1 lineage contains all core genes for a complete type VI secretion system (T6SS). In contrast, more virulent US isolates

  3. Living in an Extremely Polluted Environment: Clues from the Genome of Melanin-Producing Aeromonas salmonicida subsp. pectinolytica 34melT

    OpenAIRE

    Pavan, María Elisa; Pavan, Esteban E.; López, Nancy I.; Levin, Laura; Pettinari, M. Julia

    2015-01-01

    Aeromonas salmonicida subsp. pectinolytica 34melT can be considered an extremophile due to the characteristics of the heavily polluted river from which it was isolated. While four subspecies of A. salmonicida are known fish pathogens, 34melT belongs to the only subspecies isolated solely from the environment. Genome analysis revealed a high metabolic versatility, the capability to cope with diverse stress agents, and the lack of several virulence factors found in pathogenic Aeromonas. The mos...

  4. Construction of a eukaryotic expression plasmid of Humanin

    Institute of Scientific and Technical Information of China (English)

    LUO Ben-yan; CHEN Xiang-ming; TANG Min; CHEN Feng; CHEN Zhi

    2005-01-01

    Objective: To construct a eukaryotic expression plasmid pcDNA3.1 (-)-Humanin. Methods: The recombinant plasm pGEMEX- 1-Humanin was digested with restriction endonucleases BamH I and Hind Ⅲ and the Humanin gene fragments, abo 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1 (-) and the recombinant plasmids pcDNA3. l(-)-Humanin were identified by sequencing. Results: Recombinant plasmid DNA succesfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence. Conclusions: A eukaryotic expression plasmid of Humanin was successfully constructed.

  5. Plasmid DNA induces dodecyl triethyl ammonium bromide to aggregate into vesicle

    Institute of Scientific and Technical Information of China (English)

    Xiang Mei Ran; Xia Guo; Jia Tong Ding

    2012-01-01

    Single-chained cationic surfactant dodecyl triethyl ammonium bromide and plasmid DNA together can form vesicles once the concentration of plasmid DNA reaches a critical value (Ccvc).Bigger the size of plasmid DNA,higher the value of Ccvc.

  6. Usefulness of Chromogenic CromoCen® AGN agar medium for the identification of the genus Aeromonas: Assessment of faecal samples.

    Science.gov (United States)

    Aguilera-Arreola, M G; Portillo-Muñoz, M I; Rodríguez-Martínez, C; Castro-Escarpulli, G

    2012-08-01

    Selective screening media for the detection and identification of Aeromonas strains are needed to guide primary isolation procedures in the clinical laboratory. This study compared the selective CromoCen® AGN chromogenic agar medium for the detection and identification of Aeromonas strains that were isolated from various samples against the conventional selective agar media that are commonly used for the isolation of this organism in food, environmental and clinical samples. The Miles and Misra and ecometric methods were used to evaluate the microbiological performance of CromoCen® AGN chromogenic agar medium, which was shown to be satisfactory. A total of 14 reference Aeromonas strains, 44 wild strains and 106 clinical stool specimens were examined using both non-chromogenic selective agars that are commonly used for Aeromonas isolation and CromoCen® AGN agar. The latter exhibited 94.73% sensitivity and 100% specificity for the various samples. On CromoCen® AGN agar medium, Aeromonas formed colonies with light green, greenish and salmon pigments with or without a surrounding wide transparent zone (halo) of 2-3mm in diameter around the entire border. This medium is recommended for the isolation and potential identification of the Aeromonas genus.

  7. Bacterial Mitosis: ParM of Plasmid R1 Moves Plasmid DNA by an Actin-like Insertional Polymerization Mechanism

    DEFF Research Database (Denmark)

    Møller-Jensen, Jakob; Borch, Jonas; Dam, Mette;

    2003-01-01

    Bacterial DNA segregation takes place in an active and ordered fashion. In the case of Escherichia coli plasmid R1, the partitioning system (par) separates paired plasmid copies and moves them to opposite cell poles. Here we address the mechanism by which the three components of the R1 par system...

  8. Bifurcation Analysis of a Chemostat Model of Plasmid-Bearing and Plasmid-Free Competition with Pulsed Input

    Directory of Open Access Journals (Sweden)

    Zhong Zhao

    2014-01-01

    to the stability of the boundary periodic solution. By use of standard techniques of bifurcation theory, the periodic oscillations in substrate, plasmid-bearing, and plasmid-free organisms are shown when some conditions are satisfied. Our results can be applied to control bioreactor aimed at producing commercial producers through genetically altered organisms.

  9. Plasmid stability in immobilized and free recombinant Escherichia coli JM105(pKK223-200): importance of oxygen diffusion, growth rate, and plasmid copy number.

    OpenAIRE

    de Taxis du Poët, P; Arcand, Y; Bernier, R.; Barbotin, J N; Thomas, D.

    1987-01-01

    Stability of the plasmid pKK223-200 in Escherichia coli JM105 was studied for both free and immobilized cells during continuous culture. The relationship between plasmid copy number, xylanase activity, which was coded for by the plasmid, and growth rate and culture conditions involved complex interactions which determined the plasmid stability. Generally, the plasmid stability was enhanced in cultured immobilized cells compared with free-cell cultures. This stability was associated with modif...

  10. Positive selection and compensatory adaptation interact to stabilize non-transmissible plasmids

    OpenAIRE

    Millan, A. San; Peña-Miller, R.; Toll-Riera, M.; Halbert, Z. V.; McLean, A R; Cooper, B. S.; Maclean, R. C.

    2014-01-01

    Plasmids are important drivers of bacterial evolution, but it is challenging to understand how plasmids persist over the long term because plasmid carriage is costly. Classical models predict that horizontal transfer is necessary for plasmid persistence, but recent work shows that almost half of plasmids are non-transmissible. Here we use a combination of mathematical modelling and experimental evolution to investigate how a costly, non-transmissible plasmid, pNUK73, can be maintained in popu...

  11. Transcriptome profiling of grass carp (Ctenopharyngodon idellus) infected with Aeromonas hydrophila.

    Science.gov (United States)

    Yang, Ying; Yu, Hui; Li, Hua; Wang, Anli

    2016-04-01

    Aeromonas hydrophila is the causative pathogen of intestinal hemorrhage which has caused great economic loss in grass carp aquaculture. In order to understand the immunological response of grass carp to infection by A. hydrophila, the transcriptomic profiles of the spleens from infected and non-infected grass carp groups were obtained using HiSeq™ 2500 (Illumina). An average of 63 million clean reads per library was obtained, and approximately 80% of these genes were successfully mapped to the reference genome. A total of 1591 up-regulated and 530 down-regulated genes were identified. Eight immune-related categories involving 105 differently expressed genes were scrutinized. 16 of the differently expressed genes involving immune response were further validated by qRT-PCR. Our results provide valuable information for further analysis of the mechanisms of grass carp defense against A. hydrophila invasion. PMID:26945937

  12. Aeromonas proteolyrica bacteria in aerospace environments. [possible genetic alterations and effects on man

    Science.gov (United States)

    Foster, B. G.

    1974-01-01

    Preflight studies on Aeromonas proteolytica are reported to investigate the possibility of genetic alterations resulting in increased proteolysis in spacecraft environments. This organism may be present on human tissue and could pose medical problems if its endopeptidase and a hemolysin were to be produced in ususually high quantities or altered in such a way as to be more effective in their activities. Considered are: (1) Development of a nutrative holding medium for suspension of organisms; (2) the establishment of baseline information for the standardization of the assay for endopeptidase levels and hemolytic titers; (3) formulation of a method by which intracutaneous hemorrhage could be quantitated in guinea pig tissue; and (4) the responses of these organisms to parameters of spaceflight and experimentation.

  13. Cylindrospermopsin Biodegradation Abilities of Aeromonas sp. Isolated from Rusałka Lake.

    Science.gov (United States)

    Dziga, Dariusz; Kokocinski, Mikolaj; Maksylewicz, Anna; Czaja-Prokop, Urszula; Barylski, Jakub

    2016-03-01

    The occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in freshwater reservoirs is a common phenomenon. However, the biodegradation of this toxin in environmental samples has been observed only occasionally. In this work the biodegradation ability of cylindrospermopsin was investigated based on isolates from lakes with previous cyanotoxin history. Bacterial strains were identified based on the 16S rDNA and rpoD gene comparison. CYN biodegradation was monitored using the HPLC method. The R6 strain identified as Aeromonas sp. was documented as being capable of CYN removal. This biodegradation was dependent on the pH and temperature. Additionally, the stimulation of the growth of the R6 strain in the presence of CYN was indicated. Our discovery supports the hypothesis that (in analogy to the well-known phenomenon of microcystin biodegradation) in lakes dominated by potential CYN-producing cyanobacteria, the processes of microbial utilization of this toxin may occur. PMID:26927173

  14. Production of Poly(3-Hydroxybutyrate-co-3-Hydroxyhexanoate) by Aeromonas hydrophila and Recombinant Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    丘远征; 刘力平; 陈国强

    2003-01-01

    Aeromonas hydrophila (A.hydrophila) 4AK4 produced poly(3-hydroxybutyrate-co-3- hydroxyhexanoate) (PHBHHx) with an almost constant 3-hydroxyhexanoate (3HHx) content of 10%-15% from lauric acid and/or soybean oil.Both A.hydrophila 4AK4 and recombinant Escherichia coli (E.coli) JMU193 (pBH32) produced PHBHHx with controllable 3HHx content when fed lauric acid and another co-substrate.With glucose or gluconate as the co-substrate, the 3HHx content in the copolyester produced by A.hydrophila 4AK4 was reduced slightly from 12% to 9%.However, the 3HHx content in the copolyester produced by E.coli JMU193 (pBH32) was significantly reduced from 9% to 2% with fructose as the co-substrate.These results show that regulation of 3HHx content in PHBHHx can be achieved using genetically engineered E.coli.

  15. A Report of Peritonitis from Aeromonas sobria in a Peritoneal Dialysis (PD) Patient with Necrotizing Fasciitis.

    Science.gov (United States)

    Janma, Jirayut; Linasmita, Patcharasarn; Changsirikulchai, Siribha

    2015-11-01

    A 70-years of age, male patient with underlying type 2 diabetes mellitus, hypertension, dyslipidemia and ischemic heart disease had undergone continuous ambulatory peritoneal dialysis (CAPD)for 3 years without any episodes of peritonitis. He was diagnosed with necrotizing fasciitis and later developed peritonitis after receiving a laceration from an aquatic injury suffered during the flood disaster of 2011. The blood culture, necrotic tissue and the clear dialysate collected upon admission had shown Aeromonas sobria. The route of peritonitis may be from the hematogenous spread of A. sobria resulting in necrotizing fasciitis. A. sobria should be considered as the pathogen of peritonitis in PD patients who have history of wounds from contaminated water. We suggest that the PD patients who present with septicemia and did not meet the criteria for peritonitis, the initial dialysate effluent should be sent for culture. The benefit of this is to allow early recognition and treatment of peritonitis.

  16. Cylindrospermopsin Biodegradation Abilities of Aeromonas sp. Isolated from Rusałka Lake

    Science.gov (United States)

    Dziga, Dariusz; Kokocinski, Mikolaj; Maksylewicz, Anna; Czaja-Prokop, Urszula; Barylski, Jakub

    2016-01-01

    The occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in freshwater reservoirs is a common phenomenon. However, the biodegradation of this toxin in environmental samples has been observed only occasionally. In this work the biodegradation ability of cylindrospermopsin was investigated based on isolates from lakes with previous cyanotoxin history. Bacterial strains were identified based on the 16S rDNA and rpoD gene comparison. CYN biodegradation was monitored using the HPLC method. The R6 strain identified as Aeromonas sp. was documented as being capable of CYN removal. This biodegradation was dependent on the pH and temperature. Additionally, the stimulation of the growth of the R6 strain in the presence of CYN was indicated. Our discovery supports the hypothesis that (in analogy to the well-known phenomenon of microcystin biodegradation) in lakes dominated by potential CYN-producing cyanobacteria, the processes of microbial utilization of this toxin may occur. PMID:26927173

  17. Transcriptome profiling of grass carp (Ctenopharyngodon idellus) infected with Aeromonas hydrophila.

    Science.gov (United States)

    Yang, Ying; Yu, Hui; Li, Hua; Wang, Anli

    2016-04-01

    Aeromonas hydrophila is the causative pathogen of intestinal hemorrhage which has caused great economic loss in grass carp aquaculture. In order to understand the immunological response of grass carp to infection by A. hydrophila, the transcriptomic profiles of the spleens from infected and non-infected grass carp groups were obtained using HiSeq™ 2500 (Illumina). An average of 63 million clean reads per library was obtained, and approximately 80% of these genes were successfully mapped to the reference genome. A total of 1591 up-regulated and 530 down-regulated genes were identified. Eight immune-related categories involving 105 differently expressed genes were scrutinized. 16 of the differently expressed genes involving immune response were further validated by qRT-PCR. Our results provide valuable information for further analysis of the mechanisms of grass carp defense against A. hydrophila invasion.

  18. In Vitro Antibacterial Effect of Effective Microorganisms( EM) on Aeromonas hydrophila

    Institute of Scientific and Technical Information of China (English)

    Zhu; Zhixiu; He; Houjun; Zhang; Wenbo; Wu; Xiangdong; Wan; Gen; Deng; Shunzhou; Liu; Songlin

    2014-01-01

    In order to understand in vitro antibacterial effect of antibiotics and EM on Aeromonas hydrophila,14 isolates of A. hydrophila were isolated from diseased fish and aquaculture in the study. PCR amplification results of aerolysin gene( Aero gene) of isolates indicated that 7 out of 14 isolates were pathogenic A.hydrophila. In vitro antibacterial test indicated that 14 isolates were resistance to penicillin G,ampicillin,cephradine,cefazolin,tetracycline,terimethoprim,lincomycin and cephalexin,but showed high sensitivity to cefotaxime and furazolidone. In vitro antibacterial effect of EM on 14 isolates of A. hydrophila was positively correlated with concentration,and EM with the concentration greater than 60% had antibacterial effect on 14 isolates.

  19. Protection of ornamental gold fish Carassius auratus against Aeromonas hydrophila by treating Ixora coccinea active principles.

    Science.gov (United States)

    Anusha, Paulraj; Thangaviji, Vijayaragavan; Velmurugan, Subramanian; Michaelbabu, Mariavincent; Citarasu, Thavasimuthu

    2014-02-01

    Herbals such as Ixora coccinea, Daemia extensa and Tridax procumbens were selected to screen in vitro antibacterial and immunostimulant activity against the freshwater fish pathogen Aeromonas hydrophila using different organic polar and non-polar solvents. Initial screening results revealed that, ethyl acetate extracts and its purified fraction of I. coccinea was able to suppress the A. hydrophila strains at more than 15 mm of zone of inhibition and positive immunostimulant activity. The purified active fraction, which eluted from H40: EA60 mobile phase was structurally characterized by GC-MS analysis. Two compounds such as Diethyl Phthalate (1,2-Benzene dicarboxylic acid, monobutyl ester) and Dibutyl Phthalate were characterized using NIST database search. In order to study the in vivo immunostimulant influence of the compounds, the crude extracts (ICE) and purified fractions (ICF) were incorporated to the artificial diets at the concentration of 400 mg kg⁻¹ and fed to the ornamental gold fish Carassius auratus for 30 days. After termination of feeding experiment, they were challenged with highly virulent A. hydrophila AHV-1 which was isolated from infected gold fish and studied the survival, specific bacterial load reduction, serum biochemistry, haematology, immunology and histological parameters. The control diet fed fishes succumbed to death within five days at 100% mortality whereas ICE and ICF fed groups survived 60 and 80% respectively after 10 days. The diets also helped to decrease the Aeromonas load after challenge and significantly (P ≤ 0.01) improved the serum albumin, globulin and protein. The diets also helped to increase the RBC and haemoglobin level significantly (P ≤ 0.05) from the control group. Surprisingly the immunological parameters like phagocytic activity, serum bactericidal activity and lysozyme activity were significantly increased (P ≤ 0.001) in the experimental diets. Macrophages and erythrocytes were abundantly expressed in the

  20. Chemical modification of polyvinyl chloride and silicone elastomer in inhibiting adhesion of Aeromonas hydrophila.

    Science.gov (United States)

    Kregiel, Dorota; Berlowska, Joanna; Mizerska, Urszula; Fortuniak, Witold; Chojnowski, Julian; Ambroziak, Wojciech

    2013-07-01

    Disease-causing bacteria of the genus Aeromonas are able to adhere to pipe materials, colonizing the surfaces and forming biofilms in water distribution systems. The aim of our research was to study how the modification of materials used commonly in the water industry can reduce bacterial cell attachment. Polyvinyl chloride and silicone elastomer surfaces were activated and modified with reactive organo-silanes by coupling or co-crosslinking silanes with the native material. Both the native and modified surfaces were tested using the bacterial strain Aeromonas hydrophila, which was isolated from the Polish water distribution system. The surface tension of both the native and modified surfaces was measured. To determine cell viability and bacterial adhesion two methods were used, namely plate count and luminometry. Results were expressed in colony-forming units (c.f.u.) and in relative light units (RLU) per cm(2). Almost all the chemically modified surfaces exhibited higher anti-adhesive and anti-microbial properties in comparison to the native surfaces. Among the modifying agents examined, poly[dimethylsiloxane-co-(N,N-dimethyl-N-n-octylammoniopropyl chloride) methylsiloxane)] terminated with hydroxydimethylsilyl groups (20 %) in silicone elastomer gave the most desirable results. The surface tension of this modifier, was comparable to the non-polar native surface. However, almost half of this value was due to the result of polar forces. In this case, in an adhesion analysis, only 1 RLU cm(-2) and less than 1 c.f.u. cm(-2) were noted. For the native gumosil, the results were 9,375 RLU cm(-2) and 2.5 × 10(8) c.f.u. cm(-2), respectively. The antibacterial activity of active organo-silanes was associated only with the carrier surface because no antibacterial compounds were detected in liquid culture media, in concentrations that were able to inhibit cell growth.

  1. The occurrence of Aeromonas spp. in the bottled mineral water, well water and tap water from the municipal supplies

    Directory of Open Access Journals (Sweden)

    Denise de Oliveira Scoaris

    2008-10-01

    Full Text Available The aim of this work was to study the occurrence of Aeromonas sp in the bottled mineral water, well water and tap water from the municipal supplies. Positive samples were found for Aeromonas spp. 12.7% from the mineral water, 8.3% from the artesian water and 6.5% from the tap water. The recovery of Aeromonas spp. was significantly higher in the bottled mineral and artesian water than in the tap water from municipal supplies. The occurrence of the Aeromonas spp. did not correlate significantly with the contamination indicator bacteria (i.e. total coliforms in the artesian water samples. However, a significant correlation was found between Aeromonas spp. and total coliforms in the both mineral water and tap water samples. The presence or absence of a correlation between the indicator bacteria and Aeromonas could reflect the occasional appearance of the pathogen in the drinking water and the different rates of survival and recovery of these agents compared with those fecal indicators. The finding that 41.6, 14.8 and 9.0 % of the artesian water, bottled mineral water and tap water, respectively, sampled in the current study failed to meet the Brazilian standard for total coliforms in the drinking water should therefore be of concern.A porcentagem de amostras positivas para Aeromonas foi de 12.7% para água mineral, 8.3% para água de poço artesiano e 6.5% para água do sistema público de abastecimento. O isolamento de Aeromonas spp. foi significativamente maior em água mineral e água de poço artesiano do que em água do sistema público. A ocorrência de Aeromonas spp. não teve correlação significativa com os indicadores de contaminação tradicionalmente utilizados (coliformes totais em amostras de água de poço artesiano. No entanto, esta correlação foi positiva e significativa em água mineral e água do sistema público. A presença ou ausência de correlação entre bactérias indicadoras e a presença de Aeromonas pode refletir o

  2. Plasmid Copy Number Determination by Quantitative Polymerase Chain Reaction.

    Science.gov (United States)

    Anindyajati; Artarini, A Anita; Riani, Catur; Retnoningrum, Debbie S

    2016-01-01

    Recombinant therapeutic proteins are biopharmaceutical products that develop rapidly for years. Recombinant protein production in certain hosts requires vector expression harboring the gene encoding the corresponding protein. Escherichia coli is the prokaryote organism mostly used in recombinant protein production, commonly using a plasmid as the expression vector. Recombinant protein production is affected by plasmid copy number harboring the encoded gene, hence the determination of plasmid copy number also plays an important role in establishing a recombinant protein production system. On the industrial scale, a low copy number of plasmids are more suitable due to their better stability. In the previous study we constructed pCAD, a plasmid derived from the low copy number pBR322 plasmid. This study was aimed to confirm pCAD's copy number by quantitative polymerase chain reaction (qPCR). Plasmid copy number was determined by comparing the quantification signal from the plasmid to those from the chromosome. Copy number was then calculated by using a known copy number plasmid as a standard. Two pairs of primers, called tdk and ori, were designed for targeting a single gene tdk in the chromosome and a conserved domain in the plasmid's ori, respectively. Primer quality was analyzed in silico using PrimerSelect DNASTAR and PraTo software prior to in vitro evaluation on primer specificity and efficiency as well as optimization of qPCR conditions. Plasmid copy number determination was conducted on E. coli lysates harboring each plasmid, with the number of cells ranging from 10(2)-10(5) cells/μL. Cells were lysed by incubation at 95ºC for 10 minutes, followed by immediate freezing at -4°C. pBR322 plasmid with the copy number of ~19 copies/cell was used as the standard, while pJExpress414-sod plasmid possessing the high copy number pUC ori was also determined to test the method being used. In silico analysis based on primer-primer and primer-template interactions showed

  3. An updated view of plasmid conjugation and mobilization in Staphylococcus.

    Science.gov (United States)

    Ramsay, Joshua P; Kwong, Stephen M; Murphy, Riley J T; Yui Eto, Karina; Price, Karina J; Nguyen, Quang T; O'Brien, Frances G; Grubb, Warren B; Coombs, Geoffrey W; Firth, Neville

    2016-01-01

    The horizontal gene transfer facilitated by mobile genetic elements impacts almost all areas of bacterial evolution, including the accretion and dissemination of antimicrobial-resistance genes in the human and animal pathogen Staphylococcus aureus. Genome surveys of staphylococcal plasmids have revealed an unexpected paucity of conjugation and mobilization loci, perhaps suggesting that conjugation plays only a minor role in the evolution of this genus. In this letter we present the DNA sequences of historically documented staphylococcal conjugative plasmids and highlight that at least 3 distinct and widely distributed families of conjugative plasmids currently contribute to the dissemination of antimicrobial resistance in Staphylococcus. We also review the recently documented "relaxase-in trans" mechanism of conjugative mobilization facilitated by conjugative plasmids pWBG749 and pSK41, and discuss how this may facilitate the horizontal transmission of around 90% of plasmids that were previously considered non-mobilizable. Finally, we enumerate unique sequenced S. aureus plasmids with a potential mechanism of mobilization and predict that at least 80% of all non-conjugative S. aureus plasmids are mobilizable by at least one mechanism. We suggest that a greater research focus on the molecular biology of conjugation is essential if we are to recognize gene-transfer mechanisms from our increasingly in silico analyses. PMID:27583185

  4. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    International Nuclear Information System (INIS)

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores

  5. Investigation of plasmid-induced growth defect in Pseudomonas putida.

    Science.gov (United States)

    Mi, Jia; Sydow, Anne; Schempp, Florence; Becher, Daniela; Schewe, Hendrik; Schrader, Jens; Buchhaupt, Markus

    2016-08-10

    Genetic engineering in bacteria mainly relies on the use of plasmids. But despite their pervasive use for physiological studies as well as for the design and optimization of industrially used production strains, only limited information about plasmid induced growth defects is available for different replicons and organisms. Here, we present the identification and characterization of such a phenomenon for Pseudomonas putida transformants carrying the pBBR1-derived plasmid pMiS1. We identified the kanamycin resistance gene and the transcription factor encoding rhaR gene to be causal for the growth defect in P. putida. In contrast, this effect was not observed in Escherichia coli. The plasmid-induced growth defect was eliminated after introduction of a mutation in the plasmid-encoded rep gene, thus enabling construction of the non-toxic variant pMiS4. GFP reporters construct analyses and qPCR experiments revealed a distinctly lowered plasmid copy number for pMiS4, which is probably the reason for alleviation of the growth defect by this mutation. Our work expands the knowledge about plasmid-induced growth defects and provides a useful low-copy pBBR1 replicon variant. PMID:27287537

  6. Plasmid-associated sensitivity of Bacillus thuringiensis to UV light

    Energy Technology Data Exchange (ETDEWEB)

    Benoit, T.G.; Wilson, G.R.; Bull, D.L.; Aronson, A.I. (Department of Agriculture, College Station, TX (USA))

    1990-08-01

    Spores and vegetative cells of Bacillus thuringiensis were more sensitive to UV light than were spores or cells of plasmid-cured B. thuringiensis strains or of the closely related Bacillus cereus. Introduction of B. thuringiensis plasmids into B. cereus by cell mating increased the UV sensitivity of the cells and spores. Protoxins encoded by one or more B. thuringiensis plasmids were not involved in spore sensitivity, since a B. thuringiensis strain conditional for protoxin accumulation was equally sensitive at the permissive and nonpermissive temperatures. In addition, introduction of either a cloned protoxin gene, the cloning vector, or another plasmid not containing a protoxin gene into a plasmid-cured strain of B. thuringiensis all increased the UV sensitivity of the spores. Although the variety of small, acid-soluble proteins was the same in the spores of all strains examined, the quantity of dipicolinic acid was about twice as high in the plasmid-containing strains, and this may account for the differences in UV sensitivity of the spores. The cells of some strains harboring only B. thuringiensis plasmids were much more sensitive than cells of any of the other strains, and the differences were much greater than observed with spores.

  7. Multilocus sequence typing of IncN plasmids

    DEFF Research Database (Denmark)

    García-Fernández, Aurora; Villa, Laura; Moodley, Arshnee;

    2011-01-01

    OBJECTIVES: Incompatibility group N (IncN) plasmids have been associated with the dissemination of antimicrobial resistance and are a major vehicle for the spread of blaVIM-1 in humans and blaCTX-M-1 in animals. A plasmid multilocus sequence typing (pMLST) scheme was developed for rapid categoriz......OBJECTIVES: Incompatibility group N (IncN) plasmids have been associated with the dissemination of antimicrobial resistance and are a major vehicle for the spread of blaVIM-1 in humans and blaCTX-M-1 in animals. A plasmid multilocus sequence typing (pMLST) scheme was developed for rapid...... in different countries from both animals and humans belonged to ST1, suggesting dissemination of an epidemic plasmid through the food chain. Fifteen of 17 plasmids carrying blaVIM-1 from Klebsiella pneumoniae and Escherichia coli, isolated during a 5year period in Greece were assigned to ST10, suggesting...... that spread and persistence of this particular IncN-carrying blaVIM-1 lineage in Greece. CONCLUSIONS: This study proposes the use of pMLST as a suitable and rapid method for identification of IncN epidemic plasmid lineages. The recent spread of blaCTX-M-1 among humans and animals seems to be associated...

  8. Construction and Characterization of DNA Vaccine for Major Outer Membrane Protein Gene of Aeromonas hydrophila%嗜水气单胞菌外膜蛋白基因 DNA 疫苗载体的构建及分析

    Institute of Scientific and Technical Information of China (English)

    李盼; 李素一; 吴唯维; 林天龙; 林晨韬; 陈叙

    2013-01-01

    为构建嗜水气单胞菌的 DNA 疫苗载体,根据已发表的该菌外膜蛋白基因 momp 的核苷酸序列设计一对特异性引物,应用 PCR 技术,扩增到嗜水气单胞菌 L316的主要外膜蛋白基因,并插入到真核表达载体 pcDNA3上,构建成 DNA 疫苗,命名为 pcDNA3-POMP 。以纯化的原核表达蛋白 GST-POMP 免疫 SD 大鼠制备抗血清,用 ELISA 测定抗体效价达到1∶100000以上;用 pcDNA3-POM P 质粒转染293细胞,转染48 h 后收集细胞,提取细胞的 RNA 进行 RT-PCR ,检测到外源基因的表达。至此,初步完成嗜水气单胞菌 DNA 疫苗表达载体的构建,为进一步疫苗免疫和效价的检测奠定基础。%For constructing the DNA vaccine , a pair of specific PCR primers were designed according to the nucleotide sequence of Aeromonas hydrophila major outer membrane protein gene published by our lab previously . The PCR products were inserted into the eukaryotic expression vector pcDNA 3 to construct a DNA vaccine named pcDNA3-POMP . SD rats were immunized with purified prokaryotic-expressed protein GST-POMP to make antiserum ,whose antibody titer ,measured by ELISA ,was over 1 ∶ 100 000 .293 cells were transfected with pcDNA3-POMP ,48 hours after transfection ,the cells were collected for total RNA extraction and RT-PCR ,and the expression of exogenous momp were detected .Therefore ,we have constructed a plasmid of DNA vaccine for Aeromonas hydrophila ,vaccination and evaluation of this DNA vaccine will be studied in the future .

  9. Separation of plasmid DNA topoisomers by multimodal chromatography.

    Science.gov (United States)

    Silva-Santos, A Rita; Alves, Cláudia P A; Prazeres, Duarte Miguel F; Azevedo, Ana M

    2016-06-15

    The ability to analyze the distribution of topoisomers in a plasmid DNA sample is important when evaluating the quality of preparations intended for gene therapy and DNA vaccination or when performing biochemical studies on the action of topoisomerases and gyrases. Here, we describe the separation of supercoiled (sc) and open circular (oc) topoisomers by multimodal chromatography. A medium modified with the ligand N-benzyl-N-methyl ethanolamine and an elution scheme with increasing NaCl concentration are used to accomplish the baseline separation of sc and oc plasmid. The utility of the method is demonstrated by quantitating topoisomers in a purified plasmid sample. PMID:27033004

  10. Quorum sensing signals are produced by Aeromonas salmonicida and quorum sensing inhibitors can reduce production of a potential virulence factor

    DEFF Research Database (Denmark)

    Rasch, Maria; Kastbjerg, Vicky Gaedt; Bruhn, Jesper Bartholin;

    2007-01-01

    . Pigment production was only observed in broth under highly aerated conditions. Quorum sensing inhibitors (QSIs) are compounds that specifically block QS systems without affecting bacterial growth and 2 such compounds, sulphur-containing AHL-analogues, reduced production of protease in a typical strain...... of Aeromonas salmonicida strains. All 31 typical strains were AHL producers as were 21 of 26 atypical strains, but on a strain population basis, production of virulence factors such as protease, lipase, A-layer or pigment did not correlate with the production and accumulation of AHLs in the growth medium...... of Aeromonas salmonicida. The most efficient compound N-(heptylsulfanylacetyl)-L-homoserine lactone (HepS-AHL), reduced protease production by a factor of 10. Five extracellular proteases were detected on gelatin-containing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gels and 3...

  11. Protection against atypical Aeromonas salmonicida infection in carp (Cyprinus carpio L.) by oral administration of humus extract.

    Science.gov (United States)

    Kodama, Hiroshi; Denso; Nakagawa, Tsuyoshi

    2007-04-01

    Humic substances are formed during the decomposition of organic matter in humus, and are found in many natural environments in which organic materials and microorganisms have been present. In the present study, oral administration of humus extract to common carp (Cyprinus carpio L.) induced effective protection against experimental atypical Aeromonas salmonicida infection. Mortality of fish and development of skin lesions such as hemorrhages and ulcers were significantly suppressed in carp treated with 10%, 5% or 1% humus extract adsorbed on dry feeding pellets. The median surviving days was also greater in fish treated with 10% or 5% humus extract than in untreated fish. Atypical A. salmonicida was isolated from ulcerative lesions of part of dead fish, but Aeromonas hydrophila and Flavobacterium sp. were also isolated from these fish, verifying bacterial population changes during the progression of skin lesions. These results clearly show that treatment of fish with humus extract is effective in preventing A. salmonicida disease.

  12. Ekstrak Daun Sambiloto (Andrographis paniculata) Dalam Mengendalikan Pertumbuhan Bakteri Aeromonas hydrophila, Edwarsiella tarda dan Jamur Saprolegnia sp. Secara In Vitro

    OpenAIRE

    Sinaga, Lihardo

    2016-01-01

    This study was aimed to know antimicrobial potential of extract of sambiloto’s Leaf (Andrographis paniculata) against bacteria Aeromonas hydrophila, Edwardsiella tarda and Saprolegnia sp. and to know the extract toxicity to brine shrimp (Artemia salina). Sambiloto’s leaf was extracted using methanol, ethyl acetate and n-Hexane. Antimicrobial activity test was done using diffusion discs method to know the compunds contained in the extract phytochemical test was conducted. The chemical compound...

  13. Influenza Plasmid DNA Vaccines: Progress and Prospects.

    Science.gov (United States)

    Bicho, Diana; Queiroz, João António; Tomaz, Cândida Teixeira

    2015-01-01

    Current influenza vaccines have long been used to fight flu infectious; however, recent advances highlight the importance of produce new alternatives. Even though traditional influenza vaccines are safe and usually effective, they need to be uploaded every year to anticipate circulating flu viruses. This limitation together with the use of embryonated chicken eggs as the substrate for vaccine production, is time-consuming and could involve potential biohazards in growth of new virus strains. Plasmid DNA produced by prokaryote microorganisms and encoding foreign proteins had emerged as a promising therapeutic tool. This technology allows the expression of a gene of interest by eukaryotic cells in order to induce protective immune responses against the pathogen of interest. In this review, we discuss the strategies to choose the best DNA vaccine to be applied in the treatment and prevention of influenza. Specifically, we give an update of influenza DNA vaccines developments, all involved techniques, their main characteristics, applicability and technical features to obtain the best option against influenza infections.

  14. Use of Aeromonas spp. as General Indicators of Antimicrobial Susceptibility among Bacteria in Aquatic Environments in Thailand.

    Science.gov (United States)

    Usui, Masaru; Tagaki, Chie; Fukuda, Akira; Okubo, Torahiko; Boonla, Chanchai; Suzuki, Satoru; Seki, Kanako; Takada, Hideshige; Tamura, Yutaka

    2016-01-01

    Antimicrobials are widely used, not only for treating human infections, but also for treatment of livestock and in fish farms. Human habitats in Southeastern Asian countries are located in close proximity to aquatic environments. As such, the human populations within these regions are at risk of exposure to antimicrobial resistant bacteria, and thereby disseminating antimicrobial resistance genes (ARGs). In this study, we collected water samples from 15 sites (5 sites in Chao Phraya River, 2 sites at the mouth of Chao Phraya River, 3 sites in Ta Chin River, and 5 sites at city canals) and 12 sites (6 sites at city canals; 2 sites at chicken farms; 2 sites at pig farms; and 2 samples from sites at pig farms, which were subsequently treated at a biogas plant) in Thailand in 2013 and 2014, respectively. In total, 117 Aeromonas spp. were isolated from the water samples, and these organisms exhibited various antimicrobial susceptibility profiles. Notably, there was a significant correlation between the environmental concentration of tetracyclines and the rates of tetracycline resistance in the isolated Aeromonas spp.; however, both the concentration and rates of tetracycline resistance in samples derived from pig farms were higher than those of samples harvested from other aquatic environments. These findings suggest that the high concentrations of antimicrobials observed in these aquatic environments likely select for ARGs. Furthermore, they indicate that Aeromonas spp. comprise an effective marker for monitoring antimicrobial resistance in aquatic environments. PMID:27433156

  15. Use of Aeromonas spp. as General Indicators of Antimicrobial Susceptibility among Bacteria in Aquatic Environments in Thailand

    Science.gov (United States)

    Usui, Masaru; Tagaki, Chie; Fukuda, Akira; Okubo, Torahiko; Boonla, Chanchai; Suzuki, Satoru; Seki, Kanako; Takada, Hideshige; Tamura, Yutaka

    2016-01-01

    Antimicrobials are widely used, not only for treating human infections, but also for treatment of livestock and in fish farms. Human habitats in Southeastern Asian countries are located in close proximity to aquatic environments. As such, the human populations within these regions are at risk of exposure to antimicrobial resistant bacteria, and thereby disseminating antimicrobial resistance genes (ARGs). In this study, we collected water samples from 15 sites (5 sites in Chao Phraya River, 2 sites at the mouth of Chao Phraya River, 3 sites in Ta Chin River, and 5 sites at city canals) and 12 sites (6 sites at city canals; 2 sites at chicken farms; 2 sites at pig farms; and 2 samples from sites at pig farms, which were subsequently treated at a biogas plant) in Thailand in 2013 and 2014, respectively. In total, 117 Aeromonas spp. were isolated from the water samples, and these organisms exhibited various antimicrobial susceptibility profiles. Notably, there was a significant correlation between the environmental concentration of tetracyclines and the rates of tetracycline resistance in the isolated Aeromonas spp.; however, both the concentration and rates of tetracycline resistance in samples derived from pig farms were higher than those of samples harvested from other aquatic environments. These findings suggest that the high concentrations of antimicrobials observed in these aquatic environments likely select for ARGs. Furthermore, they indicate that Aeromonas spp. comprise an effective marker for monitoring antimicrobial resistance in aquatic environments. PMID:27433156

  16. Presence of Vibrios and Aeromonas strains and total psychrotrophs in hexagonal spotted grouper and Spanish Mackerel Fish

    International Nuclear Information System (INIS)

    This work was conducted on different parts of hexagonal-spotted grouper and Spanish mackerel fish from 10 major fishery shops in Riyadh city to assess the presence of vibrios and aeromonas using the API20E and Biolog system. Also, pH and total psychotrophs were assessed as two indexes of quality. Similar data were obtained from both identification systems except for Aeromonas media-like and Vibrio anguillarum which are not included in the API20E database. Aeromonas hydrophila, A. media-like, V. alginolyticus, V.anguillarum, V. damsel and V. fluvialis were recovered from fish samples A. medial-like and V. damsel most predominant. Unexpectedly, V. cholera and V. parahemolyticus were not recovered. The numbers of skin samples of grouper having more than 7 log CFU/gm were much higher than that of mackerel skin samples. Similar psychrotrophic counts were noticed in gut samples of the two fish types, indicating similarly in growth habitat. Most of the grouper fish samples had a pH in the range of >6.40-7.02, whereas 95% of mackerel samples had a pH at 6.40 or below. (author)

  17. Research Progress on Aeromonas hydrophila%嗜水气单胞菌的研究进展

    Institute of Scientific and Technical Information of China (English)

    姜艳丽; 李丽丽

    2015-01-01

    为有效预防嗜水气单胞菌引起传染病的发生,以减少对人畜的损害,通过回顾嗜水气单胞菌的分类地位(嗜温性和运动性的气单胞菌属的模式菌)、生物学性质,对其主要毒力因子(外毒素及胞外蛋白酶)和鉴定技术(生化鉴定、免疫血清技术和分子生物学技术)的研究成果进行了综述。%In order to prevent the infectious dieases caused by Aeromonas hydrophila ,and reduce the damage to human and animals ,the taxonomic status and biological character of Aeromonas hydrophila were reviewed ,the research of virulence factor (exotoxin and extracellular protease)and identification of Aeromonas hydrophila were summarized ,in cluding biochenrical identification ,immune serum and molecular biology techniques .

  18. Cloning and Analysis of a Large Plasmid pBMB165 from Bacillus thuringiensis Revealed a Novel Plasmid Organization

    OpenAIRE

    Yueying Wang; Donghai Peng; Zhaoxia Dong; Lei Zhu; Suxia Guo; Ming Sun

    2013-01-01

    In this study, we report a rapid cloning strategy for large native plasmids via a contig linkage map by BAC libraries. Using this method, we cloned a large plasmid pBMB165 from Bacillus thuringiensis serovar tenebrionis strain YBT-1765. Complete sequencing showed that pBMB165 is 77,627 bp long with a GC-content of 35.36%, and contains 103 open reading frames (ORFs). Sequence analysis and comparison reveals that pBMB165 represents a novel plasmid organization: it mainly consists of a pXO2-like...

  19. Plasmid pGA1 from Corynebacterium glutamicum codes for a gene product that positively influences plasmid copy number.

    OpenAIRE

    Nesvera, J; Pátek, M; Hochmannová, J; Abrhámová, Z; Becvárová, V; Jelínkova, M; Vohradský, J

    1997-01-01

    The complete nucleotide sequence (4,826 bp) of the cryptic plasmid pGA1 from Corynebacterium glutamicum was determined. DNA sequence analysis revealed four putative coding regions (open reading frame A [ORFA], ORFA2, ORFB, and ORFC). ORFC was identified as a rep gene coding for an initiator of plasmid replication (Rep) according to the high level of homology of its deduced amino acid sequence with the Rep proteins of plasmids pSR1 (from C. glutamicum) and pNG2 (from Corynebacterium diphtheria...

  20. Characterization of the Double-Partitioning Modules of R27: Correlating Plasmid Stability with Plasmid Localization

    OpenAIRE

    Trevor D Lawley; Taylor, Diane E.

    2003-01-01

    Plasmid R27 contains two independent partitioning modules, designated Par1 and Par2, within transfer region 2. Par1 is member of the type I partitioning family (Walker-type ATPase), and Par2 is a member of the type II partitioning family (actin-type ATPase). Stability tests of cloned Par1 and Par2 and insertional disruptions of Par1 and Par2 within R27 demonstrated that Par1 is the major stability determinant whereas Par2 is the minor stability determinant. Creation of double-partitioning mut...

  1. A series of template plasmids for Escherichia coli genome engineering.

    Science.gov (United States)

    Deb, Shalini S; Reshamwala, Shamlan M S; Lali, Arvind M

    2016-06-01

    Metabolic engineering strategies often employ multi-copy episomal vectors to overexpress genes. However, chromosome-based overexpression is preferred as it avoids the use of selective pressure and reduces metabolic burden on the cell. We have constructed a series of template plasmids for λ Red-mediated Escherichia coli genome engineering. The template plasmids allow construction of genome integrating cassettes that can be used to integrate single copies of DNA sequences at predetermined sites or replace promoter regions. The constructed cassettes provide flexibility in terms of expression levels achieved and antibiotics used for selection, as well as allowing construction of marker-free strains. The modular design of the template plasmids allows replacement of genetic parts to construct new templates. Gene integration and promoter replacement using the template plasmids are illustrated. PMID:27071533

  2. Construction and Identification of Plasmid pTA-TUB2

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    An about 1.40 Kb target gene fragment was yielded by PCR amplification with the plasmid pRB 129,which was identified by restriction enzyme digestion that the PCR product was TU B2 gene.The gene was digested by the restriction enzyme and was linked with pTA plasmid to construct pTA-TU B2 plasmid.The plasmid was transformed into Chaetomium spp.by PEG method and the transformation rate was 27/(2×105) and it is nine times higher than that of pRB 129.The transformants can grow on the PDA containing 1 000 μg*mL-1 carbendazim,which is 1 000 times higher than the original Chaetomium spp.The resistance was stable after 10 times transfer on non-selective medium.

  3. Characterization of Nocardia Plasmid pXT107

    Institute of Scientific and Technical Information of China (English)

    Hai-Yang XIA; Yong-Qiang TIAN; Ran ZHANG; Kai-Chun LIN; Zhong-Jun QIN

    2006-01-01

    Nocardia, Rhodococcus and Streptornyces, all members of the actinomycetes family, are Gram-positive eubacteria with high G+C content and able to form mycelium. We report here a newly identified plasmid pXT107 of Nocardia sp. 107, one of the smallest circular plasmids found in Nocardia.The complete nucleotide sequence of pXT107 consisted of 4335 bp with 65% G+C content, and encoded one replication extragenic palindromic (Rep) and six hypothetical proteins. The Rep, double-strand origin and single-strand origin of pXT107 resembled those of typical rolling-circle-replication plasmids, such as pNI100 of Nocardia, pRE8424 of Rhodococcus and plJ101 of Streptomyces. The Escherichia coli-Nocardia shuttle plasmid pHAQ22, containing thc rep gene of pXT107, is able to propagate in Nocardia but not in Streptomyces.

  4. Pharmaceutical development of the plasmid DNA vaccine pDERMATT

    OpenAIRE

    Quaak, S.G.L.

    2009-01-01

    The discovery of tumor specific antigens and self tolerance mechanisms against these antigens led to the assumption that antigens circulating at sufficient concentration levels could break this self tolerance mechanism and evoke immunological antitumor effects. pDERMATT (plasmid DNA encoding recombinant MART-1 and tetanus toxin fragment-c) is a plasmid that encodes for MART-1, a melanoma associated antigen that is expressed in a large fraction of melanomas. In animal models administration of ...

  5. Isolation and properties of plasmids from Deinococcus radiodurans Sark

    International Nuclear Information System (INIS)

    Radioresistant bacterium, Deinococcus radiodurans, can repair completely almost all of DNA damages including double strand breaks induced by gamma-rays up to about 5 kGy. In order to reveal the repair mechanism, it is necessary to develop a cloning vector available for the genetic analysis. We tried to isolate plasmids from D.radiodurans Sark strain. In the present paper the isolation and properties of plasmids were described. (author)

  6. Plasmid transfer and genetic recombination by protoplast fusion in staphylococci.

    OpenAIRE

    Götz, F; Ahrné, S.; Lindberg, M

    1981-01-01

    The experimental conditions for plasmid transfer and genetic recombination in Staphylococcus aureus and some coagulase-negative staphylococci by protoplast fusion are described. Protoplasts were prepared by treatment with lysostaphin and lysozyme in a buffered medium with 0.7 to 0.8 M sucrose. Regeneration of cell walls was accomplished on a hypertonic agar medium containing succinate and bovine serum albumin. Transfer of plasmids occurred after treatment of the protoplast mixtures with polye...

  7. Transfer of conjugative plasmids among bacteria under environmentally relevant conditions

    DEFF Research Database (Denmark)

    Musovic, Sanin

    at spredningskapacitet af en konjugerbare plasmid, der koder for kviksølv resistens via merA genet, finder sted under substrat begrænsede forhold til syntetisk bakterielt samfund. Plasmid overførsel var meget forhøjet ved kontinuert udsættelse af mikrokosms for en høj koncentration af kviksølv. De forskellige vækstrater...

  8. Construction of a bioluminescence reporter plasmid for Francisella tularensis

    OpenAIRE

    Bina, Xiaowen R.; Miller, Mark A.; James E Bina

    2010-01-01

    A Francisella tularensis shuttle vector that constitutively expresses the Photorhabdus luminescens lux operon in type A and type B strains of F. tularensis was constructed. The bioluminescence reporter plasmid was introduced into the live vaccine strain of F. tularensis and used to follow F. tularensis growth in a murine intranasal challenge model in real time by bioluminescence imaging. The results show that the new bioluminescence reporter plasmid represents a useful tool for tularemia rese...

  9. Plasmid vector with temperature-controlled gene expression

    International Nuclear Information System (INIS)

    In plasmid pBR327, a fragment 169 b.p. long including promotor p3 of the bla gene has been deleted. The deletional derivative so obtained (pSP2) has been used to construct a recombinant plasmid bearing a fragment of phage λ DNA with the p/sub R/ promotor and the gene of the temperature-sensitive repressor cI. It has been shown that the plasmid vector so constructed (pCE119) with promotor cR performs repressor-cI-controlled transcription of the bla gene, as a result of which induction for an hour at 420C leads to an almost 100-fold increase in the amount of product of the bla gene as compared with that at 320C. The possibility of the use of plasmid cPE119 for the expression of other genes has been demonstrated for the case of the semisynthetic β-galactosidase gene of E. coli. In this case, on induction of the cells with recombinant plasmid pCEZ12 for 3 hours at 420C, a 300-fold increase in the amount of active β-galactosidase, as compared with that at 320C, was observed. It is important to point out that under these conditions (at 420C), at least 99% of the cells containing the plasmid retain the phenotype lacZ+, which indicates the stability of the proposed vector system

  10. Plasmid copy number noise in monoclonal populations of bacteria

    Science.gov (United States)

    Wong Ng, Jérôme; Chatenay, Didier; Robert, Jérôme; Poirier, Michael Guy

    2010-01-01

    Plasmids are extra chromosomal DNA that can confer to their hosts’ supplementary characteristics such as antibiotic resistance. Plasmids code for their copy number through their own replication frequency. Even though the biochemical networks underlying the plasmid copy number (PCN) regulation processes have been studied and modeled, no measurement of the heterogeneity in PCN within a whole population has been done. We have developed a fluorescent-based measurement system, which enables determination of the mean and noise in PCN within a monoclonal population of bacteria. Two different fluorescent protein reporters were inserted: one on the chromosome and the other on the plasmid. The fluorescence of these bacteria was measured with a microfluidic flow cytometry device. We show that our measurements are consistent with known plasmid characteristics. We find that the partitioning system lowers the PCN mean and standard deviation. Finally, bacterial populations were allowed to grow without selective pressure. In this case, we were able to determine the plasmid loss rate and growth inhibition effect.

  11. Plasmid-free T7-based Escherichia coli expression systems.

    Science.gov (United States)

    Striedner, Gerald; Pfaffenzeller, Irene; Markus, Luchner; Nemecek, Sabine; Grabherr, Reingard; Bayer, Karl

    2010-03-01

    In order to release host cells from plasmid-mediated increases in metabolic load and high gene dosages, we developed a plasmid-free, T7-based E. coli expression system in which the target gene is site-specifically integrated into the genome of the host. With this system, plasmid-loss, a source of instability for conventional expression systems, was eliminated. At the same time, system leakiness, a challenging problem with recombinant systems, was minimized. The efficiency of the T7 RNA polymerase compensates for low gene dosage and provides high rates of recombinant gene expression without fatal consequences to host metabolism. Relative to conventional pET systems, this system permits improved process stability and increases the host cell's capacity for recombinant gene expression, resulting in higher product yields. The stability of the plasmid-free system was proven in chemostat cultivation for 40 generations in a non-induced and for 10 generations in a fully induced state. For this reason plasmid-free systems benefit the development of continuous production processes with E. coli. However, time and effort of the more complex cloning procedure have to be considered in relation to the advantages of plasmid-free systems in upstream-processing. PMID:19891007

  12. Construction and Use of Flow Cytometry Optimized Plasmid-Sensor Strains

    DEFF Research Database (Denmark)

    Bahl, Martin Iain; Oregaard, Gunnar; Sørensen, Søren Johannes;

    2009-01-01

    stability of the plasmid is high. The method presented here relies on a phenotypic (green fluorescence protein) marker, which is switched on if the host bacteria loses the residing plasmid. The incorporation of flow cytometry for single-cell detection and discrimination between plasmid-free and plasmid...

  13. Comparative analysis of conjugative plasmids mediating gentamicin resistance in Staphylococcus aureus.

    OpenAIRE

    Goering, R. V.; Ruff, E A

    1983-01-01

    Five gentamicin-resistant clinical isolates of Staphylococcus aureus were found to contain self-transmissible plasmids of 32 to 37 megadaltons in size. Restriction endonuclease digests of the plasmids were markedly similar to those of reference plasmids of unrelated geographical origin, thus suggesting the significant contribution of common conjugal plasmids to the emergence of gentamicin resistance in S. aureus populations.

  14. Genetic markers for detection of Escherichia coli K-12 harboring ampicillin-resistance plasmid from an industrial wastewater treatment effluent pond.

    Science.gov (United States)

    Simões, G A R; Xavier, M A S; Oliveira, D A; Menezes, E V; Magalhães, S S G; Gandra, J A C D; Xavier, A R E O

    2016-01-01

    Biotechnology industries that use recombinant DNA technology are potential sources for release of genetically modified organisms to the environment. Antibiotic-resistance marker genes are commonly used for recombinant bacteria selection. One example is the marker gene coding for β-lactamase (bla) in plasmids found in Escherichia coli K-12. The aim of this study was to provide an approach to develop a molecular method for genetic marker detection in E. coli K-12 harboring bla genes from an industrial wastewater treatment effluent pond (IWTEP). For the detection of bla and Achromobacter lyticus protease I (api) genes in samples from IWTEP, we employed multiplex polymerase chain reaction (PCR) using E. coli K-12 genetic marker detection primers, previously described in the literature, and primers designed in our laboratory. The microbiological screening method resulted in 22 bacterial colony-forming units isolated from three different IWTEP harvesting points. The multiplex PCR amplicons showed that five isolates were positive for the bla gene marker and negative for the E. coli K-12 and api genes. The 16S rRNA regions of positive microorganisms carrying the bla gene were genotyped by the MicroSeq®500 system. The bacteria found were Escherichia spp (3/5), Chromobacterium spp (1/5), and Aeromonas spp (1/5). None of the 22 isolated microorganisms presented the molecular pattern of E. coli K-12 harboring the bla gene. The presence of microorganisms positive for the bla gene and negative for E. coli K-12 harboring bla genes at IWTEP suggests that the ampicillin resistance found in the isolated bacteria could be from microorganisms other than the E. coli K-12 strain harboring plasmid. PMID:27323199

  15. Transfer of chimeric plasmids among Salmonella typhimurium strains by P22 transduction.

    OpenAIRE

    Orbach, M J; Jackson, E N

    1982-01-01

    Salmonella typhimurium bacteriophage P22 transduced plasmids having P22 sequences inserted in the vector pBR322 with high frequency. Analysis of the structure of the transducing particle DNA and the transduced plasmids indicates that this plasmid transduction involves two homologous recombination events. In the donor cell, a single recombination between the phage and the homologous sequences on the plasmid inserted the plasmid into the phage chromosome, which was then packaged by headfuls int...

  16. Tyrosine Partners Coordinate DNA Nicking by the Salmonella typhimurium Plasmid pCU1 Relaxase Enzyme

    OpenAIRE

    Nash, Rebekah P.; Niblock, Franklin C.; Redinbo, Matthew R.

    2011-01-01

    Conjugative plasmid transfer results in the spread of antibiotic resistance genes and virulence factors between bacterial cells. Plasmid transfer is dependent upon the DNA nicking activity of a plasmid-encoded relaxase enzyme. Tyrosine residues within the relaxase cleave the DNA plasmid nic site in a highly sequence-specific manner. The conjugative resistance plasmid pCU1 encodes a relaxase with four tyrosine residues surrounding its active site (Y18,19,26,27). We use activity assays to demon...

  17. Linear Plasmid SLP2 Is Maintained by Partitioning, Intrahyphal Spread, and Conjugal Transfer in Streptomyces▿

    OpenAIRE

    Hsu, Chin-Chen; Chen, Carton W.

    2009-01-01

    Low-copy-number plasmids generally encode a partitioning system to ensure proper segregation after replication. Little is known about partitioning of linear plasmids in Streptomyces. SLP2 is a 50-kb low-copy-number linear plasmid in Streptomyces lividans, which contains a typical parAB partitioning operon. In S. lividans and Streptomyces coelicolor, a parAB deletion resulted in moderate plasmid loss and growth retardation of colonies. The latter was caused by conjugal transfer from plasmid-co...

  18. Plasmids and rickettsial evolution: insight from Rickettsia felis.

    Directory of Open Access Journals (Sweden)

    Joseph J Gillespie

    Full Text Available BACKGROUND: The genome sequence of Rickettsia felis revealed a number of rickettsial genetic anomalies that likely contribute not only to a large genome size relative to other rickettsiae, but also to phenotypic oddities that have confounded the categorization of R. felis as either typhus group (TG or spotted fever group (SFG rickettsiae. Most intriguing was the first report from rickettsiae of a conjugative plasmid (pRF that contains 68 putative open reading frames, several of which are predicted to encode proteins with high similarity to conjugative machinery in other plasmid-containing bacteria. METHODOLOGY/PRINCIPAL FINDINGS: Using phylogeny estimation, we determined the mode of inheritance of pRF genes relative to conserved rickettsial chromosomal genes. Phylogenies of chromosomal genes were in agreement with other published rickettsial trees. However, phylogenies including pRF genes yielded different topologies and suggest a close relationship between pRF and ancestral group (AG rickettsiae, including the recently completed genome of R. bellii str. RML369-C. This relatedness is further supported by the distribution of pRF genes across other rickettsiae, as 10 pRF genes (or inactive derivatives also occur in AG (but not SFG rickettsiae, with five of these genes characteristic of typical plasmids. Detailed characterization of pRF genes resulted in two novel findings: the identification of oriV and replication termination regions, and the likelihood that a second proposed plasmid, pRFdelta, is an artifact of the original genome assembly. CONCLUSION/SIGNIFICANCE: Altogether, we propose a new rickettsial classification scheme with the addition of a fourth lineage, transitional group (TRG rickettsiae, that is unique from TG and SFG rickettsiae and harbors genes from possible exchanges with AG rickettsiae via conjugation. We offer insight into the evolution of a plastic plasmid system in rickettsiae, including the role plasmids may have played in

  19. Detection of multidrug resistance Aeromonas hydrophila in farm raised fresh water prawns

    Directory of Open Access Journals (Sweden)

    Md. Bakhtiar Lijon

    2015-12-01

    Full Text Available This study was undertaken for isolation, identification and determination of antibiogram profile of Aeromonas hydrophila in farm raised fresh water prawn (Macrobrachium rosenbergii on five commercial ghers. Fresh water prawns (n=25 were collected from five ghers located at Satkhira, Bagerhat and Khulna districts of Bangladesh. Brain (n=25, muscle (n=25 and intestine (n=25 samples were collected aseptically from fresh water prawn and inoculated into alkaline peptone (APW water for enrichment at 37℃ for 8 h. Enriched cultured was streaked into Thiosulfate Citrate Bile Salt Sucrose (TCBS agar to isolate bacteria. Bacteria were identified by cultural, Gram staining, biochemical properties and polymerase chain reaction (PCR assay. The antibiogram profiles of bacteria were investigated against 5 commonly used antibiotics (Gentamicin, Cefalexin, Ampicillin, Azithromycin and Ciprofloxacin by disc diffusion method. Thirteen A. hydrophila isolates were identified and the prevalence of the A. hydrophila in fresh water prawn was 17.33%. All 13 (100% isolates were sensitive to Ciprofloxacin, Gentamicin, Azithromycin and resistant to Ampicillin and Cefalexin. The results of this study indicate that farm raised fresh water prawn harbor multidrug resistant A. hydrophila which might causes public health problem if enter into human food chain.

  20. Reduction of Aeromonas hidrophyla biofilm on stainless stell surface by essential oils

    Directory of Open Access Journals (Sweden)

    Alessandra Farias Millezi

    2013-01-01

    Full Text Available This study demonstrates the possibility of using sanitizing detergents based on natural products for the elimination and/or reduction of Aeromonas hydrophila biofilm formed on stainless steel surfaces. The goal of this work was to determine the reduction effect of sanitizing detergents containing essential oils of Thymus vulgaris (thyme and Cymbopogon citratus (lemongrass on biofilm formed by A. hydrophila on AISI 304 stainless steel coupons, using UHT skimmed milk as substratum. There was adhesion and biofilm formation by A. hydrophila at 28 ºC, presenting 7.60 log cfu.cm-2 after the fourth day of cultivation. There was no significant difference between the lemongrass treatment and that of the thyme oil (p 0.05. The treatment with lemongrass solution reduced the biofilm by 4.51 log cfu cm-2 at 25 ºC. The thyme detergent also reduced the number of cfu cm-2 by 3.84 log cycles at 25 ºC. The use of the lemongrass and thyme solutions efficiently reduced the A. hydrophila biofilm.

  1. Antibacterial activities of bioactive compounds extracted from Marine algae Gracilaria salicornia against Aeromonas hydrophila

    Directory of Open Access Journals (Sweden)

    Somayeh Rasooli

    2015-04-01

    Full Text Available Herbal medicinal products have attracted significant research interest in recent years. Considering the efficiency of algae products in controlling pathogenic bacteria and also easy access to large resources of algae, this study was conducted to evaluate the effects of methanolic, chloroformic and aqueous extracts of Gracilaria salicornia against Aeromonas hydrophila, a heterotrophic, Gram-negative, rod-shaped bacterium found mainly in warm climate. Algae samples were collected from Qeshm Island coastlines and transferred to the laboratory. Standard methods were used to obtain the algae extract. Antibacterial activities of various extracts were tested against the bacterium using well diffusion assay method. Significant differences were observed in antibacterial activities of different extracts (P<0.05. The diameter of zone of growth inhibition varied in correlation with concentration of the extracts (50, 100, 200 and 300 mg.ml-1. The best inhibition zone was observed at 100, 200 and 300 mg.ml-1 methanolic and 300 mg.ml-1 aqueous extracts.

  2. Non-specific immune response of bullfrog Rana catesbeiana to intraperitoneal injection of bacterium Aeromonas hydrophila

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally (i.p.) with 0.2 ml bacterial suspension at a density of 5.2 × 106 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution (0.85%, w/v). Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection (dpi) for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase (SOD) activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.

  3. Inactivation of Pseudomonas aeruginosa and Aeromonas hydrophila by silver in tap water.

    Science.gov (United States)

    Silvestry-Rodriguez, Nadia; Bright, Kelly R; Uhlmann, Donald R; Slack, Donald C; Gerba, Charles P

    2007-09-01

    This study was conducted to assess the efficacy of silver as a secondary disinfectant to replace or reduce the level of chlorine utilized in water distribution systems. Pseudomonas aeruginosa and Aeromonas hydrophila are opportunistic pathogens present in drinking water and have been associated with waterborne disease. After 8 hours of exposure to 100 microg/L of silver, there was a >6-log10 reduction in P. aeruginosa in tap water at room temperature at pH7 and a 5.55-log10 reduction in the presence of 3 mg/L humic acid. Similar reductions were observed at pH9. At 4 degrees C, reductions greater than 4-log10 were observed after 24 hours. For A. hydrophila, a >6-log10 reduction occurred at both pH7 and pH9 within nine hours. The World Health Organization has determined that this amount of silver could be used for water disinfection without health risks. Furthermore, silver shows promise as a secondary disinfectant, even in the presence of organic matter in concentrations that would reduce the effectiveness of free chlorine.

  4. Anti-biofouling property of vanillin on Aeromonas hydrophila initial biofilm on various membrane surfaces.

    Science.gov (United States)

    Ponnusamy, K; Kappachery, S; Thekeettle, M; Song, J H; Kweon, J H

    2013-09-01

    Biofouling is a serious problem on filter membranes of water purification systems due to formation of bacterial biofilms, which can be detrimental to the membrane performance. Biofouling occurs on membrane surface and therefore greatly influences the physical and chemical aspects of the surface. Several membranes including microfiltration, ultrafiltration, and reverse osmosis (RO) membranes were used to learn about the anti-biofouling properties of vanillin affecting the membrane performances. Vanillin has been recognized as a potential quorum quenching compound for Aeromonas hydrophila biofilms. The initial attachment and dynamics of biofilm growth were monitored using scanning electron microscopy and confocal laser scanning microscopy. Biofilm quantities were measured using a plate count method and total protein determinations. Vanillin addition was effective in the prevention of biofilm formation on the tested membrane surfaces. Among the membranes, RO membranes made with cellulose acetate showed the most substantial reduction of biofilm formation by addition of vanillin. The biofilm reduction was confirmed by the results of surface coverage, biomass and protein accumulation. The HPLC spectrum of the spent culture with vanillin addition showed that vanillin may interfere with quorum sensing molecules and thus prevent the formation of the biofilms.

  5. Effect of a phytogenic feed additive on the susceptibility of Onchorhynchus mykiss to Aeromonas salmonicida.

    Science.gov (United States)

    Menanteau-Ledouble, S; Krauss, I; Santos, G; Fibi, S; Weber, B; El-Matbouli, M

    2015-06-29

    In recent years, feed additives have increasingly been adopted by the aquaculture industry. These supplements not only offer an alternative to antibiotics but have also been linked to enhanced growth performance. However, the literature is still limited and provides contradictory information on their effectiveness. This is mainly due to the wide variety of available products and their complex mechanisms of action. Phytogenic feed additives have been shown to have antimicrobial effects and can improve growth performance. In the present study, we investigated the susceptibility of several fish pathogenic bacteria to a phytogenic essential oil product in vitro. In addition, we determined the protective effect of a commercial phytogenic feed additive containing oregano, anis and citrus oils on the resistance of rainbow trout Oncorhynchus mykiss to infection by Aeromonas salmonicida. The bacterium was administered through 3 different routes: intra-peritoneal injection, immersion in a bacterial solution and cohabitation with infected fish. Mortality rates were significantly lower in infected rainbow trout that had received the feed additive: the overall mortality rate across all routes of infection was 18% in fish fed a diet containing the additive compared to 37% in fish that received unsupplemented feed. The route of infection also significantly impacted mortality, with average mortality rates of 60, 17.5 and 5% for intra-peritoneal injection, immersion and cohabitation, respectively. In general, fish were better protected against infection by immersion than infection by injection. PMID:26119300

  6. Isolation and characterization of Aeromonas schubertii from diseased snakehead, Channa maculata (Lacepède).

    Science.gov (United States)

    Chen, Y F; Liang, R S; Zhuo, X L; Wu, X T; Zou, J X

    2012-06-01

    Pure bacterial cultures were isolated from diseased snakeheads, Channa maculata (Lacepède), suffering high mortality in a farm in Zhongshan, southern China. Three isolates, namely ZS20100725, ZS20100725-1 and ZS20100725-2, were identified as Aeromonas schubertii. All the isolates showed high 16S rRNA sequence similarities with A. schubertii. The isolates exhibited strong virulence to snakeheads in experimental challenges with LD(50) ranging between 1.4 × 10(4) and 6.4 × 10(6) CFU g(-1). Two of the isolates were positive for haemolysin, elastase, lipase and lecithinase by phenotypic determination, which was further confirmed by PCR amplification of the haemolysin and elastase genes. In sterile liquid medium, the best growth conditions of strain ZS20100725 were 30 °C, pH 7 and 0.5% salinity (w/v). Antibiotic susceptibility tests showed that strain ZS20100725 was susceptible to cefoxitin, cefoperazone and chloramphenicol. Furthermore, histopathology of diseased snakeheads infected with A. schubertii showed necrosis and congestion in liver, kidney and spleen and also damage to the cardiac muscle, intestine and gills.

  7. The LBP Gene and Its Association with Resistance to Aeromonas hydrophila in Tilapia

    Directory of Open Access Journals (Sweden)

    Gui Hong Fu

    2014-12-01

    Full Text Available Resistance to pathogens is important for the sustainability and profitability of food fish production. In immune-related genes, the lipopolysaccharide-binding protein (LBP gene is an important mediator of the inflammatory reaction. We analyzed the cDNA and genomic structure of the LBP gene in tilapia. The full-length cDNA (1901 bp of the gene contained a 1416 bp open reading frame, encoding 471 amino acid residues. Its genomic sequence was 5577 bp, comprising 15 exons and 14 introns. Under normal conditions, the gene was constitutively expressed in all examined tissues. The highest expression was detected in intestine and kidney. We examined the responses of the gene to challenges with two bacterial pathogens Streptcoccus agalactiae and Aeromonas hydrophila. The gene was significantly upregulated in kidney and spleen post-infection with S. agalactiae and A. hydrophila, respectively. However, the expression profiles of the gene after the challenge with the two pathogens were different. Furthermore, we identified three SNPs in the gene. There were significant associations (p < 0.05 of two of the three SNPs with the resistance to A. hydrophila, but not with the resistance to S. agalactiae or growth performance. These results suggest that the LBP gene is involved in the acute-phase immunologic response to the bacterial infections, and the responses to the two bacterial pathogens are different. The two SNPs associated with the resistance to A. hydrophila may be useful in the selection of tilapia resistant to A. hydrophila.

  8. An experimental vaccine against Aeromonas hydrophila can induce protection in rainbow trout, Oncorhynchus mykiss (Walbaum)

    Science.gov (United States)

    LaPatra, S.E.; Plant, K.P.; Alcorn, S.; Ostland, V.; Winton, J.

    2010-01-01

    A candidate vaccine against Aeromonas hydrophila in rainbow trout, Oncorhynchus mykiss, was developed using a bacterial lysate. To test the strength of protection, A. hydrophila challenge models were compared using injection into both the intraperitoneal (IP) cavity and the dorsal sinus (DS) with selected doses of live bacteria washed in saline or left untreated. Unlike the IP route, injection into the DS with either saline washed or unwashed cells resulted in consistent cumulative mortality and a dose response that could be used to establish a standard challenge having an LD50 of approximately 3 × 107 colony forming units per fish. Survivors of the challenge suffered significantly lower mortality upon re-challenge than naïve fish, suggesting a high level of acquired resistance was elicited by infection. Passive immunization using serum from hyper-immunized fish also resulted in significantly reduced mortality indicating protection can be transferred and that some portion of resistance may be antibody mediated. Vaccination of groups of rainbow trout with A. hydrophila lysate resulted in significant protection against a high challenge dose but only when injected along with Freund’s complete adjuvant. At a low challenge dose, mortality in all groups was low, but the bacterial lysate alone appeared to offer some protection.

  9. Aeromonas hydrophila-associated skin lesions and septicaemia in a Nile crocodile (Crocodylus niloticus : clinical communication

    Directory of Open Access Journals (Sweden)

    H. Turutoglu

    2005-06-01

    Full Text Available Aeromonas hydrophila is one of the most common bacteria associated with the aquatic environment. There are , however, limited data on A. hydrophila infection in crocodilians. The aim of this report is to describe a case of skin lesions and septicaemia associated with A. hydrophila in a Nile crocodile (Crocodylus niloticus. A captive male crocodile in the Zoological Park of Antalya (Turkey was found dead without showing signs of any disease. Gross examination showed brown or red-spotted skin lesions of varying size. These lesions were mostly scattered over the abdomen and occasionally on the tail and feet. At necropsy, numerous white, multifocal and randomly distributed areas were seen on the liver. Gram-stained smears from skin and liver lesions showed Gram-negative bacilli arranged in clusters. Pure cultures of A. hydrophila were recovered from skin, internal organs and blood. Isolates were found to be susceptible to ceftiofur, amoxicillin + clavulanic acid, oxytetracycline, enrofloxacin, danofloxacin, neomycin, gentamicin, and lincomycin + neomycin. A pathogenicity test was performed using this isolate on 4 male 2-year-old New Zealand white rabbits. Local abscesses formed in 2 rabbits injected subcutaneously and the 2 that were injected intraperitoneally died as a result of septicaemia. In conclusion, this report has shown that A. hydrophila may cause skin lesions and even death due to septicaemia in crocodiles.

  10. Roles of phagocytosis activating protein (PAP) in Aeromonas hydrophila infected Cyprinus carpio.

    Science.gov (United States)

    Wonglapsuwan, Monwadee; Kongmee, Pataraporn; Suanyuk, Naraid; Chotigeat, Wilaiwan

    2016-06-01

    Cyprinus carpio (koi) is one of the most popular ornamental fish. A major problem for C. carpio farming is bacterial infections especially by Aeromonas hydrophila. Previously studies had shown that the Phagocytosis Activating Protein (PAP) gene was involved in the innate immune response of animals. Therefore, we attempted to identify a role for the PAP gene in the immunology of C. carpio. The expression of the PAP was found in C. carpio whole blood and increased when the fish were stimulated by inactivated A. hydrophila. In addition, PAP-phMGFP DNA was injected as an immunostimulant. The survival rate and the phagocytic index were significantly increased in the A. hydrophila infected fish that received the PAP-phMGFP DNA immunostimulant. A chitosan-PAP-phMGFP nanoparticle was then developed and feeded into fish which infected with A. hydrophila. These fish had a significantly lower mortality rate than the control. Therefore, this research confirmed a key role for PAP in protection fish from bacterial infection and the chitosan-PAP-phMGFP nanoparticle could be a good prototype for fish immunostimulant in the future. PMID:26748248

  11. Novel application of Mahua (Madhuca sp.) flowers for augmented protease production from Aeromonas sp. S1.

    Science.gov (United States)

    Bhattacharya, Amrik; Saini, Vandana; Gupta, Anshu

    2012-10-01

    The present study explored the utilization of Mahua (Madhuca sp.) flowers, a major non-timber forest product (NTFP) of India, as a low-cost, natural substrate for protease production under submerged fermentation. Bacterial strain Aeromonas sp. Si1, previously reported by us, was used as the protease producer. Using Mahua flower extract (MFE) as the medium additive, the protease production could successfully be enhanced by 5.6-fold (564.5 UmL-1) after 24 h of fermentation under optimized conditions compared with initial production of 99.9 UmL' in the absence of MFE. The cultural parameters for optimum production of protease were determined to be: incubation time-24 h; pH-7.0; MFE concentration-5% (v/v); inoculum size-0.3% (v/v) and agitation rate-200 rpm. The results obtained demonstrate the potential of cheaper and abundantly available Mahua flowers for induction of proteases, and thus offer a new approach for value addition to this biomass through industrial enzyme production. PMID:23157010

  12. Surface protein composition of Aeromonas hydrophila strains virulent for fish: identification of a surface array protein

    Energy Technology Data Exchange (ETDEWEB)

    Dooley, J.S.G.; Trust, T.J.

    1988-02-01

    The surface protein composition of members of a serogroup of Aeromonas hydrophila was examined. Immunoblotting with antiserum raised against formalinized whole cells of A. hydrophila TF7 showed a 52K S-layer protein to be the major surface protein antigen, and impermeant Sulfo-NHS-Biotin cell surface labeling showed that the 52K S-layer protein was the only protein accessible to the Sulfo-NHS-Biotin label and effectively masked underlying outer membrane (OM) proteins. In its native surface conformation the 52K S-layer protein was only weakly reactive with a lactoperoxidase /sup 125/I surface iodination procedure. A UV-induced rough lipopolysaccharide (LPS) mutant of TF7 was found to produce an intact S layer, but a deep rough LPS mutant was unable to maintain an array on the cell surface and excreted the S-layer protein into the growth medium, indicating that a minimum LPS oligosaccharide size required for A. hydrophila S-layer anchoring. The native S layer was permeable to /sup 125/I in the lactoperoxidase radiolabeling procedure, and two major OM proteins of molecular weights 30,000 and 48,000 were iodinated. The 48K species was a peptidoglycan-associated, transmembrane protein which exhibited heat-modifiable SDS solubilization behavior characteristic of a porin protein. A 50K major peptidoglycan-associated OM protein which was not radiolabeled exhibited similar SDS heat modification characteristics and possibly represents a second porin protein.

  13. Plasma-activated air mediates plasmid DNA delivery in vivo.

    Science.gov (United States)

    Edelblute, Chelsea M; Heller, Loree C; Malik, Muhammad A; Bulysheva, Anna; Heller, Richard

    2016-01-01

    Plasma-activated air (PAA) provides a noncontact DNA transfer platform. In the current study, PAA was used for the delivery of plasmid DNA in a 3D human skin model, as well as in vivo. Delivery of plasmid DNA encoding luciferase to recellularized dermal constructs was enhanced, resulting in a fourfold increase in luciferase expression over 120 hours compared to injection only (P plasmid DNA encoding green fluorescent protein (GFP) was confirmed in the epidermal layers of the construct. In vivo experiments were performed in BALB/c mice, with skin as the delivery target. PAA exposure significantly enhanced luciferase expression levels 460-fold in exposed sites compared to levels obtained from the injection of plasmid DNA alone (P plasmid DNA encoding GFP to mouse skin was confirmed by immunostaining, where a 3-minute exposure at a 10 mm distance displayed delivery distribution deep within the dermal layers compared to an exposure at 3 mm where GFP expression was localized within the epidermis. Our findings suggest PAA-mediated delivery warrants further exploration as an alternative approach for DNA transfer for skin targets. PMID:27110584

  14. Expansion of the IncX plasmid family for improved identification and typing of novel plasmids in drug-resistant Enterobacteriaceae

    DEFF Research Database (Denmark)

    Johnson, Timothy J.; Bielak, Eliza Maria; Fortini, Daniela;

    2012-01-01

    and biofilm formation. Previous plasmid-based replicon typing procedures have indicated that the prevalence of IncX plasmids is low among members of the Enterobacteriaceae. However, examination of a number of IncX-like plasmid sequences and their occurrence in various organisms suggests that IncX plasmid......, based upon these sequences and subtypes, was then developed. Use of this revised typing procedure revealed that IncX plasmid occurrence among bacterial populations is much more common than had previously been acknowledged. Thus, this revised procedure can be used to better discern the occurrence of Inc...

  15. Recombinogenic engineering of conjugative plasmids with fluorescent marker cassettes

    DEFF Research Database (Denmark)

    Reisner, A.; Molin, Søren; Zechner, E.L.

    2002-01-01

    An efficient approach for the insertion of fluorescent marker genes with sequence specificity into conjugative plasmids in Escherichia coli is described. For this purpose, homologous recombination of linear double-stranded targeting DNA was mediated by the bacteriophage lambda recombination...... functions using very short regions of homology. Initial manipulation of the IncFII target plasmids R1 and R1drd19 indicated that the linear targeting DNA should be devoid of all extraneous homologies to. the target molecule for optimal insertion specificity. Indeed, a simple recombination assay proved...... resistance genes and fluorescent markers. The choice of 5' non-homologous extensions in primer pairs used for amplifying the marker cassettes determines the site specificity of the targeting DNA. This methodology is applicable to the modification of all plasmids that replicate in E coli and is not restricted...

  16. A novel plasmid pEA68 of Erwinia amylovora and the description of a new family of plasmids.

    Science.gov (United States)

    Ismail, Emadeldeen; Blom, Jochen; Bultreys, Alain; Ivanović, Milan; Obradović, Aleksa; van Doorn, Joop; Bergsma-Vlami, Maria; Maes, Martine; Willems, Anne; Duffy, Brion; Stockwell, Virginia O; Smits, Theo H M; Puławska, Joanna

    2014-12-01

    Recent genome analysis of Erwinia amylovora, the causal agent of fire blight disease on Rosaceae, has shown that the chromosome is highly conserved among strains and that plasmids are the principal source of genomic diversity. A new circular plasmid, pEA68, was found in E. amylovora strain 692 (LMG 28361), isolated in Poland from Sorbus (mountain ash) with fire blight symptoms. Annotation of the 68,763-bp IncFIIa-type plasmid revealed that it contains 79 predicted CDS, among which two operons (tra, pil) are associated with mobility. The plasmid is maintained stably in E. amylovora and does not possess genes associated with antibiotic resistance or known virulence genes. Curing E. amylovora strain 692 of pEA68 did not influence its virulence in apple shoots nor amylovoran synthesis. Of 488 strains of E. amylovora from seventeen countries, pEA68 was only found in two additional strains from Belgium. Although the spread of pEA68 is currently limited to Europe, pEA68 comprises, together with pEA72 and pEA78 both found in North America, a new plasmid family that spans two continents.

  17. Melanization and pathogenicity in the insect, Tenebrio molitor, and the crustacean, Pacifastacus leniusculus, by Aeromonas hydrophila AH-3.

    Directory of Open Access Journals (Sweden)

    Chadanat Noonin

    Full Text Available Aeromonas hydrophila is the most common Aeromonas species causing infections in human and other animals such as amphibians, reptiles, fish and crustaceans. Pathogenesis of Aeromonas species have been reported to be associated with virulence factors such as lipopolysaccharides (LPS, bacterial toxins, bacterial secretion systems, flagella, and other surface molecules. Several mutant strains of A. hydrophila AH-3 were initially used to study their virulence in two animal species, Pacifastacus leniusculus (crayfish and Tenebrio molitor larvae (mealworm. The AH-3 strains used in this study have mutations in genes involving the synthesis of flagella, LPS structures, secretion systems, and some other factors, which have been reported to be involved in A. hydrophila pathogenicity. Our study shows that the LPS (O-antigen and external core is the most determinant A. hydrophila AH-3 virulence factor in both animals. Furthermore, we studied the immune responses of these hosts to infection of virulent or non-virulent strains of A. hydrophila AH-3. The AH-3 wild type (WT containing the complete LPS core is highly virulent and this bacterium strongly stimulated the prophenoloxidase activating system resulting in melanization in both crayfish and mealworm. In contrast, the ΔwaaE mutant which has LPS without O-antigen and external core was non-virulent and lost ability to stimulate this system and melanization in these two animals. The high phenoloxidase activity found in WT infected crayfish appears to result from a low expression of pacifastin, a prophenoloxidase activating enzyme inhibitor, and this gene expression was not changed in the ΔwaaE mutant infected animal and consequently phenoloxidase activity was not altered as compared to non-infected animals. Therefore we show that the virulence factors of A. hydrophila are the same regardless whether an insect or a crustacean is infected and the O-antigen and external core is essential for activation of the

  18. Mitochondrial pAL2-1 plasmid homologs are senescence factors in Podospora anserina independent of intrinsic senescence

    NARCIS (Netherlands)

    van Diepeningen, Anne D; Debets, Alfons J M; Slakhorst, S Marijke; Hoekstra, Rolf F

    2008-01-01

    Since the first description of a linear mitochondrial plasmid in Podospora anserina, pAL2-1, and homologous plasmids have gone from being considered beneficial longevity plasmids, via neutral genetic elements, toward mutator plasmids causing senescence. The plasmid has an invertron structure, with t

  19. Anion exchange purification of plasmid DNA using expanded bed adsorption.

    Science.gov (United States)

    Ferreira, G N; Cabral, J M; Prazeres, D M

    2000-01-01

    Recent developments in gene therapy with non-viral vectors and DNA vaccination have increased the demand for large amounts of pharmaceutical-grade plasmid DNA. The high viscosity of process streams is of major concern in the purification of plasmids, since it can cause high back pressures in column operations, thus limiting the throughput. In order to avoid these high back pressures, expanded bed anion exchange chromatography was evaluated as an alternative to fixed bed chromatography. A Streamline 25 column filled with 100 ml of Streamline QXL media, was equilibrated with 0.5 M NaCl in TE (10 mM Tris, 1 mM EDTA, pH = 8.0) buffer at an upward flow of 300 cmh-1, E. coli lysates (obtained from up to 3 liters of fermentation broth) were injected in the column. After washing out the unbound material, the media was allowed to sediment and the plasmid was eluted with 1 M NaCl in TE buffer at a downward flow of 120 cmh-1. Purification factors of 36 +/- 1 fold, 26 +/- 0.4 plasmid purity, and close to 100% yields were obtained when less than one settled column volume of plasmid feed was injected. However, both recovery yield and purity abruptly decreased when larger amounts were processed-values of 35 +/- 2 and 5 +/- 0.7 were obtained for the recovery yield and purity, respectively, when 250 ml of feedstock were processed. In these cases, gel clogging and expansion collapse were observed. The processing of larger volumes, thus larger plasmid quantities, was only possible by performing an isopropanol precipitation step prior to the chromatographic step. This step led to an enhancement of the purification step.

  20. The development of plasmid-free strains of Agrobacterium tumefaciens by using incompatibility with a Rhizobium meliloti plasmid to eliminate pAtC58.

    Science.gov (United States)

    Hynes, M F; Simon, R; Pühler, A

    1985-03-01

    Agrobacterium tumefaciens strains LBA275 and LBA290 were cured of their cryptic plasmid pAtC58 by the introduction of the Rhizobium meliloti plasmid pRme41a, which is incompatible with pAtC58. pRme41a and pTiC58, the resident Ti plasmid of LBA275, were subsequently eliminated by growth at supraoptimal temperature (40 degrees C). The resulting plasmid-free Agrobacterium strains, UBAPF1 and UBAPF2, have proved extremely useful for the study of Rhizobium plasmids. The loss of the cryptic plasmid pAtC58 has no effect on the tumor-forming ability of the Agrobacterium strains; when the Ti plasmid is present, normal tumors are formed on Kalanchoe daigremontiana. PMID:4001194

  1. Ascophyllum nodosum in the diet of tilapia (Oreochromis niloticus) and its effect after inoculation of Aeromonas hydrophila

    OpenAIRE

    Samira T.L. Oliveira; Gisele Veneroni-Gouveia; Augusto C. Santos; Silia M.N. Sousa; Marcelo L. Veiga; Cristina C. Krewer; Mateus M. Costa

    2014-01-01

    The objective of this study was to evaluate the effect of Ascophyllum nodosum brown seaweed meal (FAM) on the health of Nile tilapia submitted to inoculation with Aeromonas hydrophila. The experiment was conducted for a period of 40 days using 120 Nile tilapia fingerlings, with age of 40 days, distributed in 20 tanks. A diet including Ascophyllum nodosum seaweed meal at 20g.kg-1 and a control diet (without FAM) were provided which constituted the treatments. Thirty days after beginning the ex...

  2. Characterisation of potential virulence markers in Aeromonas caviae isolated from polluted and unpolluted aquatic environments in Morocco

    DEFF Research Database (Denmark)

    Imziln, Boujama; Krovacek, Karel; Baloda, Suraj B.;

    1998-01-01

    A total of 100 Aeromonas caviae strains isolated from river waters (38 isolates), raw sewage (30 isolates) and effluents of stabilisation ponds (i.e. treated sewage; 32 isolates) in Marrakech, Morocco, were tested for the presence of putative virulence factors to delineate differences, if any.......406) than those isolated from raw sewage (PVI = 0.175) or those from river waters (PVI = 0.09). These results suggest that stabilisation pond systems used for sewage purification under arid climate conditions in Marrakech, may have selected potentially enteropathogenic A. caviae strains....

  3. Skin and subcutaneous mycoses in tilapia (Oreochromis niloticus) caused by Fusarium oxysporum in coinfection with Aeromonas hydrophila.

    Science.gov (United States)

    Cutuli, M Teresa; Gibello, Alicia; Rodriguez-Bertos, Antonio; Blanco, M Mar; Villarroel, Morris; Giraldo, Alejandra; Guarro, Josep

    2015-09-01

    Subcutaneous mycoses in freshwater fish are rare infections usually caused by oomycetes of the genus Saprolegnia and some filamentous fungi. To date, Fusarium infections in farmed fish have only been described in marine fish. Here, we report the presence of Fusarium oxysporum in subcutaneous lesions of Nile tilapia (Oreochromis niloticus). Histopathologic evaluation revealed granuloma formation with fungal structures, and the identity of the etiological agent was demonstrated by morphological and molecular analyses. Some of the animals died as a result of systemic coinfection with Aeromonas hydrophila. PMID:26155462

  4. A novel challenge method with Aeromonas salmonicida in rainbow trout for evaluation of furunculosis vaccines

    DEFF Research Database (Denmark)

    Marana, Moonika Haahr; Skov, Jakob; Chettri, Jiwan Kumar;

    Challenge methods used to induce furunculosis in rainbow trout when testing the potency of vaccines may include exposure to Aeromonas salmonicida by intraperitoneal injection, cohabitation or bath immersion. Intraperitoneal injection is effective but will not reflect systemic immunity because...... inflammatory cells at the vaccine injection site may combat injected bacteria fast. The cohabitation and bath immersion methods both mimic the natural infection route but are less effective in inducing the disease. We have tested a new challenge method mimicking that rainbow trout in fish farms might...

  5. Resistencia a antibioticos y presencia de plasmidos en: Aeromonas hydrophila, Vibrio fluvialis y Vibrio furnissii, aislados de Carassius auratus auratus

    OpenAIRE

    Pilar Negrete Redondo; Jorge Romero Jarero; José Luis Arredondo Figueroa

    2004-01-01

    Se aislaron e identificaron 70 cepas bacterianas del riñón de peces de ornato Carassius auratus con signos y lesiones de infección. Las especies bacterianas aisladas e identificadas, Aeromonas hydrophila, Vibrio fluvialis y Vibrio furnissii, han sido descritas como causantes de diarrea en humano y como patógeno muy agresivo en cultivos de Ciprinus carpio. Por esta razón se ha hecho mal uso y abuso de antibióticos para evitar y controlar la presencia de esta bacteria en los cultivos. El uso fu...

  6. Effect of electron irradiation and packaging atmosphere on the survival of Aeromonas hydrophila in minced poultry meat

    Energy Technology Data Exchange (ETDEWEB)

    Stecchini, M.L.; Sarais, I.; Del Torre, M. [Dipartimento di Scienze degli Alimenti, Udine (Italy); Fuochi, P.G. [Istituto FRAE-CNR, Bologna (Italy)

    1995-10-01

    Resistance to electron irradiation of Aeromonas hydrophila inoculated in minced poultry meat packed in presence of air or under vacuum was examined. Surviving bacteria were counted on starch ampicillin agar containing 100 {mu}g/ml of ampicillin. Radiation resistance, expressed as D{sub 10} values, was calculated from the survival curves and found to be 0.12 and 0.14 kGy in poultry meat packed in air or under vacuum respectively. Storage at 2{sup o}C of meat samples irradiated at 0.5 kGy further reduced the number of A. hydrophila. (Author).

  7. Effect of electron irradiation and packaging atmosphere on the survival of Aeromonas hydrophila in minced poultry meat

    International Nuclear Information System (INIS)

    Resistance to electron irradiation of Aeromonas hydrophila inoculated in minced poultry meat packed in presence of air or under vacuum was examined. Surviving bacteria were counted on starch ampicillin agar containing 100 μg/ml of ampicillin. Radiation resistance, expressed as D10 values, was calculated from the survival curves and found to be 0.12 and 0.14 kGy in poultry meat packed in air or under vacuum respectively. Storage at 2oC of meat samples irradiated at 0.5 kGy further reduced the number of A. hydrophila. (Author)

  8. DIARRHEA OUTBREAK IN PERNAMBUCO, BRAZIL, ASSOCIATED WITH A HEAT-STABLE CYTOTOXIC ENTEROTOXIN PRODUCED BY Aeromonas caviae.

    Science.gov (United States)

    Lopes, Ana Carolina Amaral; Martins, Luciano Moura; Gatti, Maria Silvia Viccari; Falavina Dos Reis, Cristhiane Moura; Hofer, Ernesto; Yano, Tomomasa

    2015-01-01

    In the present study enterotoxic and cytotoxic activities of twenty Aeromonas caviae strains were examined. They originated from fecal specimens of patients with acute diarrhea during an outbreak in Brazil in 2004. Culture supernatants of fourteen strains (70%) caused fluid accumulation in rabbit ileal intestinal loops and in suckling mice assays, and also showed a cytotoxic activity in Vero and Caco-2 cells. The enterotoxic and cytotoxic factors were heat-stable after culture supernatants treatment at 100 ºC. The results revealed that A. caviae strains produce a putative diarrheagenic virulence factor, a heat-stable cytotoxic enterotoxin that could be linked to the diarrhea outbreak that took place in Brazil.

  9. Restriction Fragment Length Polymorphisms of Virulence Plasmids in Rhodococcus equi

    OpenAIRE

    Takai, Shinji; Shoda, Masato; Sasaki, Yukako; Tsubaki, Shiro; Fortier, Guillaume; Pronost, Stephane; Rahal, Karim; Becu, Teotimo; Begg, Angela; Browning, Glenn; Nicholson, Vivian M.; Prescott, John F.

    1999-01-01

    Virulent Rhodococcus equi, which is a well-known cause of pyogranulomatous pneumonia in foals, possesses a large plasmid encoding virulence-associated 15- to 17-kDa antigens. Foal and soil isolates from five countries—Argentina, Australia, Canada, France, and Japan—were investigated for the presence of 15- to 17-kDa antigens by colony blotting, using the monoclonal antibody 10G5, and the gene coding for 15- to 17-kDa antigens by PCR. Plasmid DNAs extracted from positive isolates were digested...

  10. Sustained plasmid DNA release from dissolving mineral coatings

    OpenAIRE

    Choi, Siyoung; Murphy, William L.

    2010-01-01

    Calcium phosphate (Ca-P) minerals such as hydroxyapatite are able to bind a diverse range of biological molecules due to the presence of anions and cations in their crystal structure. The well-characterized ability of Ca-P minerals to bind and release plasmid DNA, coupled with the ability of biodegradable Ca-P coatings to form on the surface of common biomaterials, provides a potential mechanism for controlled release of plasmid DNA from various biomaterials. In this study we hypothesized tha...

  11. Ecofriendly biosorption of dyes and metals by bacterial biomass of Aeromonas hydrophila RC1.

    Science.gov (United States)

    Busi, Siddhardha; Chatterjee, Rahul; Rajkumari, Jobina; Hnamte, Sairengpuii

    2016-03-01

    The ability of dried bacterial biomass in azo dye and heavy metal removal from aqueous solution was explored. Biosorption of three textile dyes, Eriochrome black T (EBT), Acid Red 26 (AR) and Trypan blue (TB) and heavy metals (Pb and Cr) by dried biomass of Aeromonas hydrophila RC1, was investigated in a batch system under various parameters such as dye concentration, contact time, concentration of biomass, pH, and temperature. The experimental results showed that the extent of biosorption for dyes increased with increase in initial concentration of dyes, biomass concentration, contact time, temperature and decreased with increase in pH. The experimental isotherms data were analyzed using Langmuir and Freundlich isotherm equations. The Langmuir model yielded good fit to the experimental data (R² approximately 0.794, 0.844 and 0.969 for the dyes, EBT, AR and TB, respectively) with maximum monolayer adsorption capacity of 58.8 mg g⁻¹ for AR. Similarly results were obtained for heavy metals and the data fit in Langmuir model (R² value of 0.849 and 0.787) with q(m) value of 40 mg g⁻¹ for Pb. The results fit in pseudo first order kinetics with removal upto 96.67 % for Pb. Involvement of the surface characteristics of the biomass in biosorption was studied using scanning electron micrographs, FTIR, EDX and XRD analysis. Thus, use ofA. hydrophila RC1 biomass can be extensively employed in water treatment plants in order to get desired water quality in the most economical way. PMID:27097447

  12. Biological approach to synthesize TiO2 nanoparticles using Aeromonas hydrophila and its antibacterial activity.

    Science.gov (United States)

    Jayaseelan, Chidambaram; Rahuman, Abdul Abdul; Roopan, Selvaraj Mohana; Kirthi, Arivarasan Vishnu; Venkatesan, Jayachandran; Kim, Se-Kwon; Iyappan, Moorthy; Siva, Chinnadurai

    2013-04-15

    Nanosized materials have been an important subject in basic and applied sciences. A novel, low-cost, green and reproducible bacteria, Aeromonas hydrophila mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs) was reported. The resulting nanoparticles were characterized by FTIR, XRD, AFM and FESEM with EDX. FTIR showed characteristic bands (1643 and 3430 cm(-1)) finds the role of carboxyl group OH stretching amine NH stretch in the formation of TiO2 NPs. The XRD spectrum confirmed that the synthesized TiO2 NPs were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.47°, 31.77°, 36.11°, 41.25°, 54.39°, 56.64° and 69.54° were identified as 110, 100, 101, 111, 211, 220 and 301 reflections, respectively. The crystallite sizes were calculated using Scherrer's formula applied to the major intense peaks and found to be the size of 40.50 nm. The morphological characterization was analyzed by FESEM and the analysis showed the NPs smooth shaped, spherical and uneven. GC-MS analysis showed the main compounds found in A. hydrophila were uric acid (2.95%), glycyl-L-glutamic acid (6.90%), glycyl-L-proline (74.41%) and L-Leucyl-D-leucine (15.74%). The potential glycyl-L-proline could have played an important role as a capping agent. A possible mechanism for the biosynthesis of TiO2 NPs has been proposed. The antibacterial activity of the synthesized TiO2 NPs was assessed by well diffusion method toward A. hydrophila, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis and showed effective inhibitory activity against S. aureus (33 mm) and S. pyogenes (31 mm). PMID:23416912

  13. Biological approach to synthesize TiO2 nanoparticles using Aeromonas hydrophila and its antibacterial activity

    Science.gov (United States)

    Jayaseelan, Chidambaram; Rahuman, Abdul Abdul; Roopan, Selvaraj Mohana; Kirthi, Arivarasan Vishnu; Venkatesan, Jayachandran; Kim, Se-Kwon; Iyappan, Moorthy; Siva, Chinnadurai

    2013-04-01

    Nanosized materials have been an important subject in basic and applied sciences. A novel, low-cost, green and reproducible bacteria, Aeromonas hydrophila mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs) was reported. The resulting nanoparticles were characterized by FTIR, XRD, AFM and FESEM with EDX. FTIR showed characteristic bands (1643 and 3430 cm-1) finds the role of carboxyl group Osbnd H stretching amine Nsbnd H stretch in the formation of TiO2 NPs. The XRD spectrum confirmed that the synthesized TiO2 NPs were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.47°, 31.77°, 36.11°, 41.25°, 54.39°, 56.64° and 69.54° were identified as 110, 100, 101, 111, 211, 220 and 301 reflections, respectively. The crystallite sizes were calculated using Scherrer's formula applied to the major intense peaks and found to be the size of 40.50 nm. The morphological characterization was analyzed by FESEM and the analysis showed the NPs smooth shaped, spherical and uneven. GC-MS analysis showed the main compounds found in A. hydrophila were uric acid (2.95%), glycyl-L-glutamic acid (6.90%), glycyl-L-proline (74.41%) and l-Leucyl-d-leucine (15.74%). The potential glycyl-L-proline could have played an important role as a capping agent. A possible mechanism for the biosynthesis of TiO2 NPs has been proposed. The antibacterial activity of the synthesized TiO2 NPs was assessed by well diffusion method toward A. hydrophila, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis and showed effective inhibitory activity against S. aureus (33 mm) and S. pyogenes (31 mm).

  14. EVALUATION OF IMMUNOSTIMULATORY POTENTIAL OF PHYLLANTHUS AMARUS IN LABEO ROHITA INFECTED WITH AEROMONAS HYDROPHILA: HAEMATOLOGICAL ASSESSMENT

    Directory of Open Access Journals (Sweden)

    T.Annalakshmi

    2013-02-01

    Full Text Available Outbreak of disease is an important limiting factor in aquaculture. Incorporation of herbal immunostimulants in diet is one of the widely followed methods to improve the general resistance in fish. The present experiment was carried out to evaluate the Immunostimulatory potential of the medicinal herb Phyllanthus amarus in fish. The experiment was carried out in two sets. In experiment-I, one group of Labeo rohita was fed with control diet (‘C’ diet and the other group was fed with Phyllanthus amarus incorporated diet (‘P’ diet for 14 days and the haematological parameters were analysed on 1st, 3rd, 7th and 14th day of the experiment. Significant increase in Total Erythrocyte Count (TEC, Haemoglobin (Hb g%, Total Leucocyte Count (TLC, lymphocytes, monocytes and neutrophils were observed in ‘P’ diet fed fishes with increase in the duration of the experiment. As there is positive response in the first experiment hence experiment-II was carried out. In experiment-II, One group of Labeo rohita were fed with ‘C’ diet and other group received ‘P’ diet for 14 days, post-infected with Aeromonas hydrophila and haematological parameters were analyzed on 1st, 3rd, 7th & 14th days after infection. Significant increase in TEC, Hb, TLC, lymphocytes and a steady population of monocytes and neutrophils were observed in ‘P’ diet fed fishes. On the other hand basophils and eosinophil count exhibited a fluctuating trend. Thus dietary incorporation of Phyllanthus amarus helps to improve the general health and resistance of the fish.

  15. Phytoextracts-Synthesized Silver Nanoparticles Inhibit Bacterial Fish Pathogen Aeromonas hydrophila.

    Science.gov (United States)

    Mahanty, Arabinda; Mishra, Snehasish; Bosu, Ranadhir; Maurya, Uk; Netam, Surya Prakash; Sarkar, Biplab

    2013-12-01

    Fish disease is a major stumbling block towards sustainable growth of the fisheries sector. Aeromonas hydrophila, which is a major infectious aquatic pathogen is reportedly the causative agent of ulcers, fin-rot, tail-rot, hemorrhagic septicemia in fish, and has reportedly developed resistance against many of the available antibiotics. In this context, the inhibitory function of silver nanoparticles (AgNPs) against A. hydrophila was studied to evaluate its possible application in aquaculture as alternative to antibiotics. AgNPs were synthesized using the leaf extracts of subtropical plants Mangifera indica (Mango), Eucalyptus terticornis (Eucalyptus), Carica papaya (Papaya) and Musa paradisiaca (Banana). The absorbance maxima, size range and shape of the AgNPs as characterized by the UV-Vis spectroscopy, high resolution transmission electron microscopy (HR-TEM), and energy dispersive X-ray spectroscopy (EDX) were, Mangifera-442, 50-65 nm, ovular; Eucalyptus-465, 60-150 nm, oval; Carica-442, 25-40 nm, round, irregular; and Musa-454, 10-50 nm, round, irregular, respectively. Well-diffusion of these AgNPs for their antimicrobial characteristics exhibited that, the papaya leaf extract synthesized AgNPs had maximum antimicrobial activity at 153.6 μg/ml concentrations, and that from the eucalyptus leaves was least effective. As observed, the potency of the nanoparticles enhanced with the decrease in particle size, from 60-150 nm in eucalyptus to 25-40 nm in papaya. Due to its purely natural sourcing, phytosynthesized AgNPs can be applied as alternative to antibiotics and other biocides as a cost-effective and eco-friendly therapeutic agent against A. hydrophila stimulated diseases in aquatic animals. PMID:24426148

  16. ANTIBIOTICS RESISTANCE AND PUTATIVE VIRULENCE FACTORS OF AEROMONAS HYDROPHILA ISOLATED FROM ESTUARY

    Directory of Open Access Journals (Sweden)

    Olumide Adedokun Odeyemi

    2012-06-01

    Full Text Available This study aim to investigate antibiotics resistance profile and putative virulence factors of Aeromonas hydrophila isolated from estuary. Bacteria used for this study were isolated from water and sediment samples obtained from Sungai Melayu, Johor, Malaysia. Serially diluted 100 µL water and 1g sediment were inoculated on modified Rimler - Shott (mRS agar. Colonies with distinct cultural characteristics were picked for further studies. Isolates were tested for biofilm productions, protease enzyme and antibiotics resistance profile using agar well diffusion method against 10 commercial antibiotics. Congo Red Agar (CRA, Microplate and Standard Tube (ST methods were used for assessment of biofilm formation among the isolates while Skim Milk Agar was used for protease production. Sw.KMJ 3 and Sw.KMJ 9 produced black crystalline colonies on CRA. Six of the isolates were biofilm producers in ST method. Result of Microplate method, helped in grouping the isolates into weak (n = 8, moderate (n = 3 and strong producers (n = 4 at 540 nm wavelength. All the isolates were classified as weak ODc  ODi 0.1, moderate ODi = 0.1  0.12 and strong producers ODi  0.12 respectively at 540 nm wavelength. Antibiotics susceptibility test also revealed that all the isolates were resistant to between 6 and 10 antibiotics. Two isolates each were resistant to 6 (60 %, 7 (70 % and 9 (90 % antibiotics respectively. Eight of the isolates showed resistance to 8 (80 % antibiotics while only isolate Sw.KMJ-7 showed resistance to all the tested antibiotics. Sw.KMJ-3, Sw.KMJ-8 and Sw.KMJ-9 produced protease enzyme on SMA. The isolates were also found to be resistant to both antibiotics and heavy metals.

  17. A highly selectable and highly transferable Ti plasmid to study conjugal host range and Ti plasmid dissemination in complex ecosystems.

    Science.gov (United States)

    Teyssier-Cuvelle, S; Oger, P; Mougel, C; Groud, K; Farrand, S K; Nesme, X

    2004-07-01

    A conjugal donor system, ST2, was constructed to study the conjugal dissemination of a Ti plasmid to wild-type recipient bacteria in vitro and in situ. The system consisted of a polyauxotrophic derivative of C58 harboring a hyperconjugative and highly selectable Ti plasmid, pSTiEGK, which was constructed by inserting a multiple antibiotic resistance cassette in the traM- mcpA region of pTiC58Delta accR. ST2 transfers pSTiEGK constitutively at frequencies up to 10(-1) to plasmidless Agrobacterium recipients. The host range of pSTiEGK includes all the known genomic species of Agrobacterium, indigenous soil agrobacteria and some Rhizobium and Phyllobacterium spp. All transconjugants became pathogenic upon acquisition of the Ti plasmid and were also able to transfer pSTiEGK by conjugation. This host range was indistinguishable from that of its wild-type parent pTiC58, and therefore pSTiEGK constitute a valid proxy to study the dissemination of Ti plasmids directly in the environment. Transconjugants can be selected on a combination of four antibiotics, which efficiently prevents the growth of the indigenous microbiota present in complex environments. The transfer of pSTiEGK to members of the genus Agrobacterium was affected primarily by the plasmid content of the recipient strain (10(3)- to 10(5)-fold reduction), e.g., the presence of incompatible plasmids. As a consequence, a species should be considered permissive to Ti transfer whenever one permissive isolate is found. PMID:15164241

  18. DNA sequence analysis of plasmids from multidrug resistant Salmonella enterica serotype Heidelberg isolates.

    Directory of Open Access Journals (Sweden)

    Jing Han

    Full Text Available Salmonella enterica serovar Heidelberg is among the most detected serovars in swine and poultry, ranks among the top five serotypes associated with human salmonellosis and is disproportionately associated with invasive infections and mortality in humans. Salmonella are known to carry plasmids associated with antimicrobial resistance and virulence. To identify plasmid-associated genes in multidrug resistant S. enterica serovar Heidelberg, antimicrobial resistance plasmids from five isolates were sequenced using the 454 LifeSciences pyrosequencing technology. Four of the isolates contained incompatibility group (Inc A/C multidrug resistance plasmids harboring at least eight antimicrobial resistance genes. Each of these strains also carried a second resistance plasmid including two IncFIB, an IncHI2 and a plasmid lacking an identified Inc group. The fifth isolate contained an IncI1 plasmid, encoding resistance to gentamicin, streptomycin and sulfonamides. Some of the IncA/C plasmids lacked the full concert of transfer genes and yet were able to be conjugally transferred, likely due to the transfer genes carried on the companion plasmids in the strains. Several non-IncA/C resistance plasmids also carried putative virulence genes. When the sequences were compared to previously sequenced plasmids, it was found that while all plasmids demonstrated some similarity to other plasmids, they were unique, often due to differences in mobile genetic elements in the plasmids. Our study suggests that Salmonella Heidelberg isolates harbor plasmids that co-select for antimicrobial resistance and virulence, along with genes that can mediate the transfer of plasmids within and among other bacterial isolates. Prevalence of such plasmids can complicate efforts to control the spread of S. enterica serovar Heidelberg in food animal and human populations.

  19. Genetic Characterization of ExPEC-Like Virulence Plasmids among a Subset of NMEC.

    Science.gov (United States)

    Nicholson, Bryon A; West, Aaron C; Mangiamele, Paul; Barbieri, Nicolle; Wannemuehler, Yvonne; Nolan, Lisa K; Logue, Catherine M; Li, Ganwu

    2016-01-01

    Neonatal Meningitis Escherichia coli (NMEC) is one of the most common causes of neonatal bacterial meningitis in the US and elsewhere resulting in mortality or neurologic deficits in survivors. Large plasmids have been shown experimentally to increase the virulence of NMEC in the rat model of neonatal meningitis. Here, 9 ExPEC-like plasmids were isolated from NMEC and sequenced to identify the core and accessory plasmid genes of ExPEC-like virulence plasmids in NMEC and create an expanded plasmid phylogeny. Results showed sequenced virulence plasmids carry a strongly conserved core of genes with predicted functions in five distinct categories including: virulence, metabolism, plasmid stability, mobile elements, and unknown genes. The major functions of virulence-associated and plasmid core genes serve to increase in vivo fitness by adding multiple iron uptake systems to the genetic repertoire to facilitate NMEC's survival in the host's low iron environment, and systems to enhance bacterial resistance to host innate immunity. Phylogenetic analysis based on these core plasmid genes showed that at least two lineages of ExPEC-like plasmids could be discerned. Further, virulence plasmids from Avian Pathogenic E. coli and NMEC plasmids could not be differentiated based solely on the genes of the core plasmid genome. PMID:26800268

  20. Genetic Characterization of ExPEC-Like Virulence Plasmids among a Subset of NMEC.

    Directory of Open Access Journals (Sweden)

    Bryon A Nicholson

    Full Text Available Neonatal Meningitis Escherichia coli (NMEC is one of the most common causes of neonatal bacterial meningitis in the US and elsewhere resulting in mortality or neurologic deficits in survivors. Large plasmids have been shown experimentally to increase the virulence of NMEC in the rat model of neonatal meningitis. Here, 9 ExPEC-like plasmids were isolated from NMEC and sequenced to identify the core and accessory plasmid genes of ExPEC-like virulence plasmids in NMEC and create an expanded plasmid phylogeny. Results showed sequenced virulence plasmids carry a strongly conserved core of genes with predicted functions in five distinct categories including: virulence, metabolism, plasmid stability, mobile elements, and unknown genes. The major functions of virulence-associated and plasmid core genes serve to increase in vivo fitness by adding multiple iron uptake systems to the genetic repertoire to facilitate NMEC's survival in the host's low iron environment, and systems to enhance bacterial resistance to host innate immunity. Phylogenetic analysis based on these core plasmid genes showed that at least two lineages of ExPEC-like plasmids could be discerned. Further, virulence plasmids from Avian Pathogenic E. coli and NMEC plasmids could not be differentiated based solely on the genes of the core plasmid genome.

  1. Pelacakan Gen Aerolysin dari Aeromonas hydrophila pada Ikan Mas yang Diberi Pakan Ekstrak Bawang Putih (DETECTION OF AEROLYSIN GEN FROM AEROMONAS HYDROPHILA IN COMMON CARP FED WITH GARLIC EXTRACT

    Directory of Open Access Journals (Sweden)

    Iesje Lukistyowati

    2014-04-01

    Full Text Available Aeromonas hydrophila is a gram negative and opportunistic bacteria, which could cause fish mortalityin a short time from 80%-100%. One virulent factor of A. hydrophila on common carp (Cyprinus carpio Lthat could cause fish mortality is aerolysin. This research used a synthetic primers of oligonukleotide todetect aerolysin, a specific genomes of A. hydrophila on common carp (Cyprinus carpio L. The commoncarps have been feed a woof that contain garlic extract during 30 days before they challenged with A.hydrophila. Polymerase Chain Reaction (PCR was used to detect an aerolysin gen from A. hydrophila. Theelectrophoresis result showed aerolysin gene of Aeromonas hydrophila from Veterinary Faculty of GadjahMada University (FKH-UGM isolate was amplified with 462 bp of molecule weight. While the aerolysingen was detected in the fish kidney with 900 bp of molecule weight. Further, DNA sequence analysis of thePCR product of A. hydrophila from FKH – UGM isolate showed homolog with isolate A. hydrophila subsphydrophila ATCC 7966 complete genome with score 55.4 (71%.

  2. Plasmid-determined copper resistance in Pseudomonas syringae from impatiens

    Energy Technology Data Exchange (ETDEWEB)

    Cooksey, D.A. (Univ. of California, Riverside (USA))

    1990-01-01

    A strain of Pseudomonas syringae was recently identified as the cause of a new foliar blight of impatiens. The bacterium was resistant to copper compounds, which are used on a variety of crops for bacterial and fungal disease control. The bacterium contained a single 47-kilobase plasmid (pPSI1) that showed homology to a copper resistance operon previously cloned and characterized from P. syringae pv. tomato plasmid pPT23D (D. Cooksey, Appl. Environ. Microbiol. 53:454-456, 1987). pPSI1 was transformed by electroporation into a copper-sensitive P. syringae strain, and the resulting transformants were copper resistant. A physical map of pPSI1 was constructed, and the extent of homology to pPT23D outside the copper resistance operon was determined in Southern hybridizations. The two plasmids shared approximately 20 kilobases of homologous DNA, with the remainder of each plasmid showing no detectable homology. The homologous regions hybridized strongly, but there was little or no conservation of restriction enzyme recognition sites.

  3. Studying plasmid horizontal transfer in situ: a critical review

    DEFF Research Database (Denmark)

    Sørensen, Søren Johannes; Bailey, Mark; Hansen, Lars Hestbjerg;

    2005-01-01

    This review deals with the prospective, experimental documentation of horizontal gene transfer (HGT) and its role in real-time, local adaptation. We have focused on plasmids and their function as an accessory and/or adaptive gene pool. Studies of the extent of HGT in natural environments have...

  4. New tetracycline resistance determinant on R plasmids from Vibrio anguillarum.

    OpenAIRE

    Aoki, T.; Satoh, T.; Kitao, T.

    1987-01-01

    Two classes of tetracycline resistance determinants on R plasmids were detected in Vibrio anguillarum strains isolated from ayu (sweat fish; Plecoglossus altivelis) farms in Japan. Tetracycline resistance genes categorized as class B were prevalent from 1973 to 1977; however, a new tetracycline resistance gene, which was not classified into tetracycline resistance determinant class A, B, C, or D, has been prevalent since 1981.

  5. Use of plasmid DNA for induction of protective immunity

    DEFF Research Database (Denmark)

    Lorenzen, Niels

    2004-01-01

    Vaccines based on plasmid DNA have been tested for a number of fish pathogens but so far it is only in case of the rhabdoviruses, where the technology has been a real break through in vaccine research. Aspects of dose, time-course and mechanisms of protection, as well as practical use are discussed....

  6. Pharmaceutical development of the plasmid DNA vaccine pDERMATT

    NARCIS (Netherlands)

    Quaak, S.G.L.

    2009-01-01

    The discovery of tumor specific antigens and self tolerance mechanisms against these antigens led to the assumption that antigens circulating at sufficient concentration levels could break this self tolerance mechanism and evoke immunological antitumor effects. pDERMATT (plasmid DNA encoding recombi

  7. Stability of Integrated Plasmids in the Chromosome of Lactococcus lactis

    NARCIS (Netherlands)

    Leenhouts, Kees J.; Kok, Jan; Venema, Gerhardus

    1990-01-01

    Derivatives of plasmids pBR322, pUB110, pSC101, and pTB19, all containing an identical fragment of lactococcal chromosomal DNA, were integrated via a Campbell-like mechanism into the same chromosomal site of Lactococcus lactis MG1363, and the transformants were analyzed for the stability of the inte

  8. Plasmid containing a DNA ligase gene from Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Griffin, K.; Setlow, J.K.

    1984-05-01

    A ligase gene from Haemophilus influenzae was cloned into the shuttle vector pDM2. Although the plasmid did not affect X-ray sensitivity, it caused an increase in UV sensitivity of the wild-type but not excision-defective H. influenzae and a decrease in UV sensitivity of the rec-1 mutant. 14 references, 2 figures.

  9. Characterization of the Lactobacillus plantarum plasmid pCD033 and generation of the plasmid free strain L. plantarum 3NSH.

    Science.gov (United States)

    Heiss, Silvia; Grabherr, Reingard; Heinl, Stefan

    2015-09-01

    Lactobacillus plantarum CD033, a strain isolated from grass silage in Austria, harbors a 7.9 kb plasmid designated pCD033. Sequence analysis identified 14 open reading frames and 8 of these were supposed to be putative coding sequences. Gene annotation revealed no putative essential genes being plasmid encoded, but a plasmid addiction system based on a PemI/PemK-like toxin-antitoxin system, able to stabilize plasmid maintenance. Absence of a replication initiation protein, a double strand origin as well as a single strand origin on plasmid pCD033 suggests replication via a new type of theta mechanism, whereby plasmid replication is potentially initiated and regulated by non-coding RNA. Detailed examination of segregational stability of plasmid vectors consisting of pCD033-fragments, combined with a selection marker, resulted in definition of a stably maintained minimal replicon. A gene encoding a RepB/OrfX-like protein was found to be not essential for plasmid replication. Alignment of the amino acid sequence of this protein with related proteins unveiled a highly conserved amino acid motif (LLDQQQ). L. plantarum CD033 was cured of pCD033 resulting in the novel plasmid free strain L. plantarum 3NSH. Plasmid curing demonstrated that no essential features are provided by pCD033 under laboratory conditions.

  10. Survival and evolution of a large multidrug resistance plasmid in new clinical bacterial hosts

    DEFF Research Database (Denmark)

    Porse, Andreas; Schønning, Kristian; Munck, Christian;

    2016-01-01

    and population sequencing to show that the long-term persistence and molecular integrity of the plasmid is highly influenced by multiple factors within a 25 kb plasmid region constituting a host-dependent burden. In the E. coli hosts investigated here, improved plasmid stability readily evolves via IS......Large conjugative plasmids are important drivers of bacterial evolution and contribute significantly to the dissemination of antibiotic resistance. Although plasmid borne multidrug resistance is recognized as one of the main challenges in modern medicine, the adaptive forces shaping the evolution...... of these plasmids within pathogenic hosts are poorly understood. Here we study plasmid-host adaptations following transfer of a 73 kb conjugative multidrug resistance plasmid to naïve clinical isolates of Klebsiella pneumoniae and Escherichia coli We use experimental evolution, mathematical modelling...

  11. Design of expanded bed supports for the recovery of plasmid DNA by anion exchange adsorption

    DEFF Research Database (Denmark)

    Theodossiou, Irini; Søndergaard, M.; Thomas, Owen R. T.

    2001-01-01

    In this study we detail the rational design of new chromatographic adsorbents tailored for the capture of plasmid DNA. Features present on current chromatographic supports that can significantly enhance plasmid binding capacity have been identified in packed bed chromatography experiments...

  12. Conservation of plasmids among plant-pathogenic Pseudomonas syringae isolates of diverse origins.

    Science.gov (United States)

    von Bodman, S B; Shaw, P D

    1987-05-01

    Thirty isolates of Pseudomonas syringae pv. tabaci, pv. angulata (pathogens on tobacco), pv. coronafaciens, and pv. striafaciens (pathogens on oats) were examined for plasmid DNAs. The strains were obtained from plants throughout the world, some over 50 years ago. Of the 22 tobacco pathogens, 16 contain predominantly one type of plasmid, the pJP27.00 type. The remaining six tobacco-specific strains do not harbor detectable plasmids. The oat pathogens contain one, two, or three plasmids. DNA homology studies indicate that the plasmid DNAs are highly conserved. More importantly, the plasmids harbored by strains isolated from one host plant are conserved most stringently; e.g., the plasmids from the tobacco pathogens are, with one exception, indistinguishable by restriction endonuclease digestion and Southern hybridization. There is also extensive homology among plasmids indigenous to the oat-specific P. syringae pv. coronafaciens and pv. striafaciens strains. PMID:3628554

  13. White Mulberry (Morus alba Foliage Methanolic Extract Can Alleviate Aeromonas hydrophila Infection in African Catfish (Clarias gariepinus

    Directory of Open Access Journals (Sweden)

    Atefeh Sheikhlar

    2014-01-01

    Full Text Available Two experiments were simultaneously conducted with Morus alba (white mulberry foliage extract (MFE as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp. in African catfish (Clarias gariepinus. In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC, albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM. Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.

  14. White mulberry (Morus alba) foliage methanolic extract can alleviate Aeromonas hydrophila infection in African catfish (Clarias gariepinus).

    Science.gov (United States)

    Sheikhlar, Atefeh; Alimon, Abd Razk; Daud, Hassan; Saad, Chee R; Webster, Carl D; Meng, Goh Yong; Ebrahimi, Mahdi

    2014-01-01

    Two experiments were simultaneously conducted with Morus alba (white mulberry) foliage extract (MFE) as a growth promoter and treatment of Aeromonas hydrophila infection in separate 60 and 30 days trail (Experiments 1 and 2, resp.) in African catfish (Clarias gariepinus). In Experiment 1, four diets, control and control supplemented with 2, 5, or 7 g MFE/kg dry matter (DM) of diet, were used. In Experiment 2, fish were intraperitoneally infected with Aeromonas hydrophila and fed the same diets as experiment 1 plus additional two diets with or without antibiotic. Results of experiment 1 showed that growth was unaffected by dietary levels of MFE. Treatments with the inclusion of MFE at the levels of 5 and 7 g/Kg DM had no mortality. Red blood cells (RBC), albumin, and total protein were all higher for the treatments fed MFE (5 and 7 g/Kg DM). Results of experiment 2 showed RBC, hemoglobin, hematocrit, globulin, albumin, and total protein improved with the increase in MFE in the infected fish. The dietary MFE at the level of 7 g/kg DM reduced mortality rate. In conclusion, MFE at the level of 7 g/kg DM could be a valuable dietary supplement to cure the infected fish.

  15. Determination of the viability of Aeromonas hydrophila in different types of water by flow cytometry, and comparison with classical methods.

    Science.gov (United States)

    Pianetti, Anna; Falcioni, Tania; Bruscolini, Francesca; Sabatini, Luigia; Sisti, Elivio; Papa, Stefano

    2005-12-01

    The presence of Aeromonas spp. in water can represent a risk for human health. Therefore, it is important to know the physiological status of these bacteria and their survival in the environment. We studied the behavior of a strain of Aeromonas hydrophila in river water, spring water, brackish water, mineral water, and chlorinated drinking water, which had different physical and chemical characteristics. The bacterial content was evaluated by spectrophotometric and plate count techniques. Flow cytometric determination of viability was carried out using a dual-staining technique that enabled us to distinguish viable bacteria from damaged and membrane-compromised bacteria. The traditional methods showed that the bacterial content was variable and dependent on the type of water. The results obtained from the plate count analysis correlated with the absorbance data. In contrast, the flow cytometric analysis results did not correlate with the results obtained by traditional methods; in fact, this technique showed that there were viable cells even when the optical density was low or no longer detectable and there was no plate count value. According to our results, flow cytometry is a suitable method for assessing the viability of bacteria in water samples. Furthermore, it permits fast detection of bacteria that are in a viable but nonculturable state, which are not detectable by conventional methods.

  16. Resistance of genetically different common carp, Cyprinus carpio L., families against experimental bacterial challenge with Aeromonas hydrophila.

    Science.gov (United States)

    Jeney, G; Ardó, L; Rónyai, A; Bercsényi, M; Jeney, Z

    2011-01-01

    The objective of this study was to determine the differences in disease resistance against artificial infection with Aeromonas hydrophila between genetically different common carp families. Four strains differing in their origin and breeding history were selected from the live gene bank of common carp maintained at the Research Institute for Fisheries, Aquaculture and Irrigation (HAKI, Szarvas, Hungary) to establish families with wide genetic background: Szarvas 15 (15), an inbred mirror line; Tata (T) scaly noble carp; Duna (D), a Hungarian wild carp and Amur (A), an East Asian wild carp. A diallele mating structure was used to allow the assessment of genetic variation within and between the tested 96 families for a variety of traits. The existing technologies of fertilization and incubation of carp eggs, as well as larval and fingerling rearing had been modified because of the large number of baseline populations. Two challenge trials of the 96 families of carp with Aeromonas hydrophila were done. The 10 most resistant and 10 most susceptible families to A. hydrophila were identified from these two challenges. The crosses that produced the most resistant families were mainly those having parents from Tata and Szarvas 15 domesticated strains, while the most susceptible families were from the wild strains Duna and Amur.

  17. Hemolysin, Protease, and EPS Producing Pathogenic Aeromonas hydrophila Strain An4 Shows Antibacterial Activity against Marine Bacterial Fish Pathogens

    Directory of Open Access Journals (Sweden)

    Anju Pandey

    2010-01-01

    Full Text Available A pathogenic Aeromonas hydrophila strain An4 was isolated from marine catfish and characterized with reference to its proteolytic and hemolytic activity along with SDS-PAGE profile (sodium dodecyl sulphate-Polyacrylamide gel electrophoresis of ECPs (extracellular proteins showing hemolysin (approximately 50 kDa. Agar well diffusion assay using crude cell extract of the bacterial isolate clearly demonstrated antibacterial activity against indicator pathogenic bacteria, Staphylococcus arlettae strain An1, Acinetobacter sp. strain An2, Vibrio parahaemolyticus strain An3, and Alteromonas aurentia SE3 showing inhibitory zone >10 mm well comparable to common antibiotics. Further GC-MS analysis of crude cell extract revealed several metabolites, namely, phenolics, pyrrolo-pyrazines, pyrrolo-pyridine, and butylated hydroxytoluene (well-known antimicrobials. Characterization of EPS using FTIR indicated presence of several protein-related amine and amide groups along with peaks corresponding to carboxylic and phenyl rings which may be attributed to its virulent and antibacterial properties, respectively. Besides hemolysin, EPS, and protease, Aeromonas hydrophila strain An4 also produced several antibacterial metabolites.

  18. Molecular identfication and virulence of three Aeromonas hydrophila isolates cultured from infected channel catfish during a disease outbreak in West Alabama (USA) in 2009

    Science.gov (United States)

    Three isolates (AL09-71, AL09-72, and AL09-73) of Aeromonas hydrophila were cultured from infected channel catfish during a disease outbreak in West Alabama in August 2009. Sequence analysis of 16S-23S rDNA intergenic spacer region (ISR), cpn60, gyrB, and rpoD genes of the three strains revealed tha...

  19. Complete Genome Sequence of the Highly Virulent Aeromonas schubertii Strain WL1483, Isolated from Diseased Snakehead Fish (Channa argus) in China.

    Science.gov (United States)

    Liu, Lihui; Li, Ningqiu; Zhang, Defeng; Fu, Xiaozhe; Shi, Cunbin; Lin, Qiang; Hao, Guijie

    2016-01-21

    We sequenced the complete genome of the highly virulent Aeromonas schubertii strain WL1483, which was isolated from diseased snakehead fish (Channa argus) in China. The full genome sequence of A. schubertii WL1483 is 4,400,034 bp, which encodes 4,376 proteins and contains 195 predicted RNA genes.

  20. Aeromonas hydrophila as an agent of infection in alligators. Phase I, final report. Progress report, September 1, 1976--September 30, 1977, Part I

    Energy Technology Data Exchange (ETDEWEB)

    Gorden, R.W.; Esch, G.W.

    1977-01-01

    Experimental alligators were exposed to various concentrations of young, washed cells of Aeromonas hydrophila under controlled conditions. Responses of all alligators were monitored on the basis of: observations of external lesions; immunoglobulin production; blood chemistry and hematology; bacteriology, parasitology, and pathology of internal organs, skeletal muscle and external lesions at necropsy. The findings are summarized.

  1. A study on genetic variability of pathogenic Aeromonas hydrophila strains and the varied responses of the strains towards phyto-extracts.

    Science.gov (United States)

    Balasundaram, A; Kumari, P Rathna; Kolanchinathan, P; Masilamani, V; John, George

    2013-11-01

    The present study evaluated genetic variation in Aeromonas hydrophila strains using PCR-RAPD and their varied susceptibility to phyto-extract. Four strains of Aeromonas hydrophila isolated from skin infections of common freshwater fish, Cyprinus carpio were characterized by various biochemical methods, physiological tests and PCR- RAPD. Antimicrobial activity of the leaf extracts of three medicinal plants, Ocimum sanctum, Adathoda vasica and Calendula officinalis were tested against the four strains of A. hydrophila by disc diffusion (Kirby-Bauer) method. Antagonistic effects of leaf extracts against A. hydrophila strains were assessed by co-culture method. RAPD analysis showed that all the microbes isolated from skin infection belong to the same species but there was no 100% genetic similarity among them Dendrogram constructed by UPGMA clearly supported the PCR pattern of genetic variability among the strains. This study revealed that Aeromonas hydophila exhibits genetic variability and varied susceptibility towards phyto-extracts. Results indicated that phyto-extracts offers a promising alternative to the use of antibiotics in controlling Aeromonas hydrophila. PMID:24511738

  2. THE IDENTIFICATION OF THE WATER-BORNE PATHOGEN AEROMONAS USING WHOLE CELL ANALYSIS BY MATRIX ASSISTED LASER DESORPTION/IONIZATION-MASS

    Science.gov (United States)

    MALDI-MS has long been established as a tool by which microorganisms can be characterized and identified. The U.S. Environmental Protection Agency (EPA) is investigating the potential of using this technique as a way to rapidly identify Aeromonas species in drinking water. A nu...

  3. DIFFERENTIATION OF AEROMONAS ISOLATES OBTAINED FROM DRINKING WATER DISTRIBUTION SYSTEM USING MATRIX-ASSISTED LASER DESCRIPTION/IONIZATION-MASS SPECTROMETRY (MALDI-MS)

    Science.gov (United States)

    The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the...

  4. THE USE OF MATRIX-ASSISTED LASER DESORPTION/IONIZATION-MASS SPECTROMETRY FOR THE IDENTIFICATION OF AEROMONAS ISOLATES OBTAINED FROM WATER DISTRIBUTION SYSTEMS

    Science.gov (United States)

    Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) has long been established as a tool by which microorganisms can be characterized and identified. EPA is investigating the potential of using this technology as a way to rapidly identify Aeromonas species fo...

  5. Assessment of Aquaflor (c), copper sulfate and potassium permanganate for control of Aeromonas hydrophila and Flavobacterium columnare infection in sunshine bass, Morone chrysops female x Morone saxatilis male

    Science.gov (United States)

    Two experiments were conducted to assess different therapeutants against a mixed infection of Aeromonas hydrophila and Flavobacterium columnare in sunshine bass (SB) (Morone chrysops female x Morone saxatilis male). Experiment 1 assessed the efficacy of copper sulfate (CuSO4), florfenicol-medicated...

  6. Identification, virulence, and mass spectrometry of toxic ECP fractons of West Alabama isolates of Aeromonas hydrophila obtained from a 2010 disease outbreak

    Science.gov (United States)

    In West Alabama, disease outbreaks in 2009 caused by Aeromonas hydrophila have led to an estimated loss of more than $3 million. In 2010, disease outbreak occurred again in West Alabama, causing losses of hundreds of thousands of pounds of market size channel catfish. During the 2010 disease outbrea...

  7. Transcription of ColE1Ap mbeC induced by conjugative plasmids from twelve different incompatibility groups.

    OpenAIRE

    Selvaratnam, S; Gealt, M A

    1993-01-01

    Although nonconjugative mobilizable plasmids require helping functions of conjugative plasmids in order to be mobilized into recipients, at least some genes from the nonconjugative plasmids may be induced to assist in the DNA transfer process. Conjugative plasmids from 12 different incompatibility groups mobilized the nonconjugative plasmid ColE1Ap between Escherichia coli strains. Introduction of any of the conjugative plasmids into the ColE1Ap-containing strain resulted in an induction of m...

  8. Plasmid profiling of bacterial isolates from confined environments

    Science.gov (United States)

    van Houdt, Rob; Provoost, Ann; Coninx, Ilse; Leys, Natalie; Mergeay, Max

    Plasmid profiling of bacterial isolates from confined environments R. Van Houdt, I. Coninx, A. Provoost, N. Leys, and M. Mergeay Expertise group for Molecular and Cellular Biology, Institute for Environment, Health and Safety, Belgian Nuclear Research Centre (SCK•CEN), Boeretang 200, B-2400 Mol, Belgium. Human exploration of extreme and isolated hostile environments such as space requires special confined small volume habitats to protect and house the crew. However, human confinement in such small volume habitats has restrictions on waste disposal and personal hygiene and inevitably generates a particular community of microorganisms within the habitat. These microorganisms are mainly originating from the crew (skin, mucous membranes, upper respiratory tract, mouth, and gastrointestinal tract) but also include the residing environmental microorganisms. Earth-based confined habitats such as the Antarctic Research Station Concordia are used as test beds for long-duration spaceflights to study the physiologic and psychological adaptation to isolated environments. The dynamics of the environmental microbial population in such a test bed could render additional insights in assessing the potential health risks in long-duration space missions. Not only total bacterial contamination levels are important, but it is essential to identify also the predominant microbial taxa and their mobile genetic elements (MGE). These MGEs could be exchanged between bacteria by horizontal gene transfer and may alter the pathogenic potential since they often carry antibiotic resistance or more in general adaptation-enhancing traits. In this study several bacterial strains isolated in the Concordia research station were examined for their plasmid content. An optimized protocol for extraction of large plasmids showed the present of at least one plasmid in 50% of the strains. For all strains the minimal inhibitory concentration of a range of antibiotics was determined indicating resistance to

  9. Comparison of electrically mediated and liposome-complexed plasmid DNA delivery to the skin

    OpenAIRE

    Heller, Loree C.; Jaroszeski, Mark J; Coppola, Domenico; Heller, Richard

    2008-01-01

    Background Electroporation is an established technique for enhancing plasmid delivery to many tissues in vivo, including the skin. We have previously demonstrated efficient delivery of plasmid DNA to the skin utilizing a custom-built four-plate electrode. The experiments described here further evaluate cutaneous plasmid delivery using in vivo electroporation. Plasmid expression levels are compared to those after liposome mediated delivery. Methods Enhanced electrically-mediated delivery, and ...

  10. Identification of tetracycline-resistant R-plasmids in Streptococcus agalactiae (group B).

    OpenAIRE

    Burdett, V

    1980-01-01

    In this report, 30 tetracycline-resistant clinical isolates of group B Streptococcus were examined to assess the extent to which tetracycline resistance is plasmid mediated. Of these, 27 showed no physical or genetic evidence of plasmid-mediated resistance; however, one conjugative and two small (3.5 X 10(6)-dalton) multicopy non-self-transmissible tetracycline resistance plasmids were identified. The conjugative plasmid was transmissible to Streptococcus faecalis as well as to Streptococcus ...

  11. Broad host range plasmids can invade an unexpectedly diverse fraction of a soil bacterial community

    DEFF Research Database (Denmark)

    Klümper, Uli; Riber, Leise; Dechesne, Arnaud;

    2014-01-01

    Conjugal plasmids can provide microbes with full complements of new genes and constitute potent vehicles for horizontal gene transfer. Conjugal plasmid transfer is deemed responsible for the rapid spread of antibiotic resistance among microbes. While broad host range plasmids are known to transfer...... bacteria and can, therefore, directly connect large proportions of the soil bacterial gene pool. This finding reinforces the evolutionary and medical significances of these plasmids....

  12. Incidence of Plasmids in Thermus spp. Isolated in Yellowstone National Park

    OpenAIRE

    Munster, Michael J.; Munster, Ann P.; Sharp, Richard J.

    1985-01-01

    Forty-eight strains of Thermus spp. were isolated from thermal sites in Yellowstone National Park, Wyo., and 62.5% showed evidence of plasmid DNA. Attempts to assign function to the plasmid DNA were unsuccessful, and the presence of plasmid DNA could not be correlated with antibiotic or heavy metal resistance. A number of these cryptic plasmids are now being investigated for their potential as vectors for molecular cloning in Thermus spp.

  13. Conservation of Plasmid-Encoded Traits among Bean-Nodulating Rhizobium Species

    OpenAIRE

    Brom, Susana; Girard, Lourdes; García-de los Santos, Alejandro; Sanjuan-Pinilla, Julio M.; Olivares, José; Sanjuan, Juan

    2002-01-01

    Rhizobium etli type strain CFN42 contains six plasmids. We analyzed the distribution of genetic markers from some of these plasmids in bean-nodulating strains belonging to different species (Rhizobium etli, Rhizobium gallicum, Rhizobium giardinii, Rhizobium leguminosarum, and Sinorhizobium fredii). Our results indicate that independent of geographic origin, R. etli strains usually share not only the pSym plasmid but also other plasmids containing symbiosis-related genes, with a similar organi...

  14. A DNA polymerase mutation that suppresses the segregation bias of an ARS plasmid in Saccharomyces cerevisiae.

    OpenAIRE

    Houtteman, S W; Elder, R T

    1993-01-01

    Yeast autonomously replicating sequence (ARS) plasmids exhibit an unusual segregation pattern during mitosis. While the nucleus divides equally into mother and daughter cells, all copies of the ARS plasmid will often remain in the mother cell. A screen was designed to isolate mutations that suppress this segregation bias. A plasmid with a weak ARS (wARS) that displayed an extremely high segregation bias was constructed. When cells were grown under selection for the wARS plasmid, the resulting...

  15. Modulation of ColE1-like Plasmid Replication for Recombinant Gene Expression

    OpenAIRE

    Camps, Manel

    2010-01-01

    ColE1-like plasmids constitute the most popular vectors for recombinant protein expression. ColE1 plasmid replication is tightly controlled by an antisense RNA mechanism that is highly dynamic, tuning plasmid metabolic burden to the physiological state of the host. Plasmid homeostasis is upset upon induction of recombinant protein expression because of non-physiological levels of expression and because of the frequently biased amino acid composition of recombinant proteins. Disregulation of p...

  16. Participation of the lytic replicon in bacteriophage P1 plasmid maintenance.

    OpenAIRE

    Yarmolinsky, M B; Hansen, E B; Jafri, S; Chattoraj, D K

    1989-01-01

    P1 bacteriophage carries at least two replicons: a plasmid replicon and a viral lytic replicon. Since the isolated plasmid replicon can maintain itself stably at the low copy number characteristic of intact P1 prophage, it has been assumed that this replicon is responsible for driving prophage replication. We provide evidence that when replication from the plasmid replicon is prevented, prophage replication continues, albeit at a reduced rate. The residual plasmid replication is due to incomp...

  17. Stability in Escherichia coli of an antibiotic resistance plasmid from Bacteroides fragilis.

    OpenAIRE

    Rashtchian, A; Booth, S J

    1981-01-01

    A Bacteroides fragilis strain resistant to penicillin G, tetracycline, and clindamycin was screened for the presence of plasmid deoxyribonucleic acid (DNA). Agarose gel electrophoresis of ethanol-precipitated DNA from cleared lysates of this strain revealed two plasmid DNA bands. The molecular weights of the plasmids were estimated by their relative mobility in agarose gel and compared with standard plasmids with known molecular weights. The molecular weights were 3.40 +/- 0.20 x 10(6) and 1....

  18. Properties of R1162, a broad-host-range, high-copy-number plasmid.

    OpenAIRE

    R. MEYER; Hinds, M; Brasch, M.

    1982-01-01

    Regions of plasmid DNA encoding characteristic properties of the IncQ (P-4) group plasmid R1162 were identified by mutagenesis and in vitro cloning. Coding sequences sufficient for expression of incompatibility and efficient conjugal mobilization by plasmid R751 were found to be linked to the origin of DNA replication. In contrast, there was a region remote from the origin, and active in trans, that was required for plasmid maintenance. A derivative that was temperature sensitive for stabilit...

  19. Characterization of different plasmid-borne dihydropteroate synthases mediating bacterial resistance to sulfonamides.

    OpenAIRE

    Swedberg, G; Sköld, O

    1980-01-01

    Plasmid-borne resistance to sulfonamides was studied in both newly isolated and earlier characterized R plasmids. Two different classes of drug-resistant dihydropteroate synthases were found to be responsible for most cases of plasmid-mediated sulfonamide resistance. The plasmid-coded enzymes could be completely separated from their chromosomal counterpart and also showed differences in heat stability and molecular size. The resistant and chromosomal enzymes could bind the normal substrate, p...

  20. The critical role of the linear plasmid lp36 in the infectious cycle of Borrelia burgdorferi

    OpenAIRE

    Mollie W Jewett; Lawrence, Kevin; Bestor, Aaron C; Tilly, Kit; Grimm, Dorothee; Shaw, Pamela; VanRaden, Mark; Gherardini, Frank; Rosa, Patricia A.

    2007-01-01

    Borrelia burgdorferi, the aetiological agent of Lyme disease, follows a life cycle that involves passage between the tick vector and the mammalian host. To investigate the role of the 36 kb linear plasmid, lp36 (also designated the B. burgdorferi K plasmid), in the infectious cycle of B. burgdorferi, we examined a clone lacking this plasmid, but containing all other plasmids known to be required for infectivity. Our results indicated that lp36 was not required for spirochete survival in the t...

  1. PHENOTYPING, VIRULENCE CHARACTERISTICS OF AEROMONAS SPECIES AND THE EFFECTS OF ESSENTIAL PLANT OILS AS ANTIMICROBIAL AGENTS AGAINST PATHOGENIC ISOLATES FROM DIFFERENT SOURCES

    Directory of Open Access Journals (Sweden)

    Ahmed M.A. Mansour

    2014-01-01

    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

  2. Plasmid vectors for Xylella fastidiosa utilizing a toxin-antitoxin system for plasmid stability in the absence of antibiotic selection

    Science.gov (United States)

    The phytopathogen Xylella fastidiosa causes disease in a variety of important crop and landscape plants. Functional genetic studies have led to a broader understanding of virulence mechanisms used by this pathogen in the grapevine host. Plasmid shuttle vectors are important tools in studies of bacte...

  3. Studies on the expression of plasmid-borne genes in the endosymbiotic state of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Krol, A.J.M.

    1982-01-01

    The subject matter of the research reported in this thesis is the role of plasmid-borne genes of Rhizobium in symbiosis and nitrogen fixation. Plasmid DNA was isolated from Rhizobium leguminosarum strain PRE and the expression of plasmid DNA in nitrogen fixing nodules was investigated by hybridizati

  4. Presence and analysis of plasmids in human and animal associated Arcobacter species

    DEFF Research Database (Denmark)

    Douidah, Laid; De Zutter, Lieven; Van Nieuwerburgh, Filip;

    2014-01-01

    In this study, we report the screening of four Arcobacter species for the presence of small and large plasmids. Plasmids were present in 9.9% of the 273 examined strains. One Arcobacter cryaerophilus and four Arcobacter butzleri plasmids were selected for further sequencing. The size of three sma...

  5. Novel plasmid conferring kanamycin and tetracycline resistance in turkey-derived Campylobacter jejuni strain 11601MD

    Science.gov (United States)

    In Campylobacter spp., resistance to the antibiotics kanamycin and tetracycline is frequently associated with plasmid-borne genes. However, relatively few plasmids of Campylobacter jejuni have been fully characterized to date. A novel plasmid (p11601MD; 44,095 bp.) harboring tet(O) was identified in...

  6. Expansion of a Plasmid Classification System for Gram-Positive Bacteria and Determination of the Diversity of Plasmids in Staphylococcus aureus Strains of Human, Animal, and Food Origins

    OpenAIRE

    Lozano, Carmen; García-Migura, Lourdes; Aspiroz, Carmen; Zarazaga, Myriam; Torres, Carmen; Aarestrup, Frank Møller

    2012-01-01

    An expansion of a previously described plasmid classification was performed and used to reveal the plasmid content of a collection of 92 Staphylococcus aureus strains of different origins. rep genes of other genera were detected in Staphylococcus. S1 pulsed-field gel electrophoresis (PFGE) hybridizations were performed with 18 representative S. aureus strains, and a high number of plasmids of different sizes and organizations were detected.

  7. Determination of plasmid copy number reveals the total plasmid DNA amount is greater than the chromosomal DNA amount in Bacillus thuringiensis YBT-1520.

    Directory of Open Access Journals (Sweden)

    Chunying Zhong

    Full Text Available Bacillus thuringiensis is the most widely used bacterial bio-insecticide, and most insecticidal crystal protein-coding genes are located on plasmids. Most strains of B. thuringiensis harbor numerous diverse plasmids, although the plasmid copy numbers (PCNs of all native plasmids in this host and the corresponding total plasmid DNA amount remains unknown. In this study, we determined the PCNs of 11 plasmids (ranging from 2 kb to 416 kb in a sequenced B. thuringiensis subsp. kurstaki strain YBT-1520 using real-time qPCR. PCNs were found to range from 1.38 to 172, and were negatively correlated to plasmid size. The amount of total plasmid DNA (∼8.7 Mbp was 1.62-fold greater than the amount of chromosomal DNA (∼5.4 Mbp at the mid-exponential growth stage (OD(600 = 2.0 of the organism. Furthermore, we selected three plasmids with different sizes and replication mechanisms to determine the PCNs over the entire life cycle. We found that the PCNs dynamically shifted at different stages, reaching their maximum during the mid-exponential growth or stationary phases and remaining stable and close to their minimum after the prespore formation stage. The PCN of pBMB2062, which is the smallest plasmid (2062 bp and has the highest PCN of those tested, varied in strain YBT-1520, HD-1, and HD-136 (172, 115, and 94, respectively. These findings provide insight into both the total plasmid DNA amount of B. thuringiensis and the strong ability of the species to harbor plasmids.

  8. Role of the 85-Kilobase Plasmid and Plasmid-Encoded Virulence-Associated Protein A in Intracellular Survival and Virulence of Rhodococcus equi

    OpenAIRE

    Giguère, Steeve; Hondalus, Mary K.; Yager, Julie A.; Darrah, Patricia; Mosser, David M.; Prescott, John F.

    1999-01-01

    Rhodococcus equi is a facultative intracellular pathogen of macrophages and a cause of pneumonia in young horses (foals) and immunocompromised people. Isolates of R. equi from pneumonic foals typically contain large, 85- or 90-kb plasmids encoding a highly immunogenic virulence-associated protein (VapA). The objective of this study was to determine the role of the 85-kb plasmid and VapA in the intracellular survival and virulence of R. equi. Clinical isolates containing the plasmid and expres...

  9. QTL detection for Aeromonas salmonicida resistance related traits in turbot (Scophthalmus maximus

    Directory of Open Access Journals (Sweden)

    Martínez Paulino

    2011-11-01

    Full Text Available Abstract Background Interactions between fish and pathogens, that may be harmless under natural conditions, often result in serious diseases in aquaculture systems. This is especially important due to the fact that the strains used in aquaculture are derived from wild strains that may not have had enough time to adapt to new disease pressures. The turbot is one of the most promising European aquaculture species. Furunculosis, caused by the bacterium Aeromonas salmonicida, produces important losses to turbot industry. An appealing solution is to achieve more robust broodstock, which can prevent or diminish the devastating effects of epizooties. Genomics strategies have been developed in turbot to look for candidate genes for resistance to furunculosis and a genetic map with appropriate density to screen for genomic associations has been also constructed. In the present study, a genome scan for QTL affecting resistance and survival to A. salmonicida in four turbot families was carried out. The objectives were to identify consistent QTL using different statistical approaches (linear regression and maximum likelihood and to locate the tightest associated markers for their application in genetic breeding strategies. Results Significant QTL for resistance were identified by the linear regression method in three linkage groups (LGs 4, 6 and 9 and for survival in two LGs (6 and 9. The maximum likelihood methodology identified QTL in three LGs (5, 6 and 9 for both traits. Significant association between disease traits and genotypes was detected for several markers, some of them explaining up to 17% of the phenotypic variance. We also identified candidate genes located in the detected QTL using data from previously mapped markers. Conclusions Several regions controlling resistance to A. salmonicida in turbot have been detected. The observed concordance between different statistical methods at particular linkage groups gives consistency to our results. The

  10. Exposure to pairs of Aeromonas strains enhances virulence in the Caenorhabditis elegans infection model

    Directory of Open Access Journals (Sweden)

    Thomas eMosser

    2015-11-01

    Full Text Available Aeromonad virulence remains poorly understood, and is difficult to predict from strain characteristics. In addition, infections are often polymicrobial (i.e., are mixed infections, and 5-10% of such infections include two distinct aeromonads, which has an unknown impact on virulence. In this work, we studied the virulence of aeromonads recovered from human mixed infections. We tested them individually and in association with other strains with the aim of improving our understanding of aeromonosis. Twelve strains that were recovered in pairs from six mixed infections were tested in a virulence model of the worm Caenorhabditis elegans. Nine isolates were weak worm killers (median time to death, TD50, ≥7 days when administered alone. Two pairs showed enhanced virulence, as indicated by a significantly shortened TD50 after co-infection versus infection with a single strain. Enhanced virulence was also observed for five of the 14 additional experimental pairs, and each of these pairs included one strain from a natural synergistic pair. These experiments indicated that synergistic effects were frequent and were limited to pairs that were composed of strains belonging to different species. The genome content of virulence-associated genes failed to explain virulence synergy, although some virulence-associated genes that were present in some strains were absent from their companion strain (e.g., T3SS. The synergy observed in virulence when 2 Aeromonas isolates were co-infected stresses the idea that consideration should be given to the fact that infection does not depend only on single strain virulence but is instead the result of a more complex interaction between the microbes involved, the host and the environment. These results are of interest for other diseases in which mixed infections are likely and in particular for water-borne diseases (e.g., legionellosis, vibriosis, in which pathogens may display enhanced virulence in the presence of the

  11. Plasmid-Chromosome Recombination of Irradiated Shuttle Vector DNA in African Green Monkey Kidney Cells.

    Science.gov (United States)

    Mudgett, John Stuart

    1987-09-01

    An autonomously replicating shuttle vector was used to investigate the enhancement of plasmid-chromosome recombination in mammalian host cells by ultraviolet light and gamma radiation. Sequences homologous to the shuttle vector were stably inserted into the genome of African Green Monkey kidney cells to act as the target substrate for these recombination events. The SV40- and pBR322-derived plasmid DNA was irradiated with various doses of radiation before transfection into the transformed mammalian host cells. The successful homologous transfer of the bacterial ampicillin resistance (amp^{rm r}) gene from the inserted sequences to replace a mutant amp^->=ne on the shuttle vector was identified by plasmid extraction and transformation into E. coli host cells. Ultraviolet light (UV) was found not to induce homologous plasmid-chromosome recombination, while gamma radiation increased the frequency of recombinant plasmids detected. The introduction of specific double -strand breaks in the plasmid or prolonging the time of plasmid residence in the mammalian host cells also enhanced plasmid-chromosome recombination. In contrast, plasmid mutagenesis was found to be increased by plasmid UV irradiation, but not to change with time. Plasmid survival, recombination, and mutagenesis were not affected by treating the mammalian host cells with UV light prior to plasmid transfection. The amp^{rm r} recombinant plasmid molecules analyzed were found to be mostly the result of nonconservative exchanges which appeared to involve both homologous and possibly nonhomologous interactions with the host chromosome. The observation that these recombinant structures were obtained from all of the plasmid alterations investigated suggests a common mechanistic origin for plasmid -chromosome recombination in these mammalian cells.

  12. The stb operon balances the requirements for vegetative stability and conjugative transfer of plasmid R388.

    Directory of Open Access Journals (Sweden)

    Catherine Guynet

    2011-05-01

    Full Text Available The conjugative plasmid R388 and a number of other plasmids carry an operon, stbABC, adjacent to the origin of conjugative transfer. We investigated the role of the stbA, stbB, and stbC genes. Deletion of stbA affected both conjugation and stability. It led to a 50-fold increase in R388 transfer frequency, as well as to high plasmid loss. In contrast, deletion of stbB abolished conjugation but provoked no change in plasmid stability. Deletion of stbC showed no effect, neither in conjugation nor in stability. Deletion of the entire stb operon had no effect on conjugation, which remained as in the wild-type plasmid, but led to a plasmid loss phenotype similar to that of the R388ΔstbA mutant. We concluded that StbA is required for plasmid stability and that StbA and StbB control conjugation. We next observed the intracellular positioning of R388 DNA molecules and showed that they localize as discrete foci evenly distributed in live Escherichia coli cells. Plasmid instability of the R388ΔΔstbA mutant correlated with aberrant localization of the plasmid DNA molecules as clusters, either at one cell pole, at both poles, or at the cell center. In contrast, plasmid molecules in the R388ΔΔstbB mutant were mostly excluded from the cell poles. Thus, results indicate that defects in both plasmid maintenance and transfer are a consequence of variations in the intracellular positioning of plasmid DNA. We propose that StbA and StbB constitute an atypical plasmid stabilization system that reconciles two modes of plasmid R388 physiology: a maintenance mode (replication and segregation and a propagation mode (conjugation. The consequences of this novel concept in plasmid physiology will be discussed.

  13. Coupling between the Basic Replicon and the Kis-Kid Maintenance System of Plasmid R1: Modulation by Kis Antitoxin Levels and Involvement in Control of Plasmid Replication

    OpenAIRE

    Juan López-Villarejo; Damián Lobato-Márquez; Ramón Díaz-Orejas

    2015-01-01

    kis-kid, the auxiliary maintenance system of plasmid R1 and copB, the auxiliary copy number control gene of this plasmid, contribute to increase plasmid replication efficiency in cells with lower than average copy number. It is thought that Kis antitoxin levels decrease in these cells and that this acts as the switch that activates the Kid toxin; activated Kid toxin reduces copB-mRNA levels and this increases RepA levels that increases plasmid copy number. In support of this model we now repo...

  14. Coupling between the Basic Replicon and the Kis-Kid Maintenance System of Plasmid R1: Modulation by Kis Antitoxin Levels and Involvement in Control of Plasmid Replication

    Directory of Open Access Journals (Sweden)

    Juan López-Villarejo

    2015-02-01

    Full Text Available kis-kid, the auxiliary maintenance system of plasmid R1 and copB, the auxiliary copy number control gene of this plasmid, contribute to increase plasmid replication efficiency in cells with lower than average copy number. It is thought that Kis antitoxin levels decrease in these cells and that this acts as the switch that activates the Kid toxin; activated Kid toxin reduces copB-mRNA levels and this increases RepA levels that increases plasmid copy number. In support of this model we now report that: (i the Kis antitoxin levels do decrease in cells containing a mini-R1 plasmid carrying a repA mutation that reduces plasmid copy number; (ii kid-dependent replication rescue is abolished in cells in which the Kis antitoxin levels or the CopB levels are increased. Unexpectedly we found that this coordination significantly increases both the copy number of the repA mutant and of the wt mini-R1 plasmid. This indicates that the coordination between plasmid replication functions and kis-kid system contributes significantly to control plasmid R1 replication.

  15. Studies on the plasmid stability, plasmid copy number and endo(1, 3)(1, 4) b-glucanase production by free and alginate immobilised recombinant saccharomyces cerevisiae cells

    OpenAIRE

    Canavan, Peter D.

    1994-01-01

    A recombinant yeast strain, Saccharomyces cerevisiae DBY746, containing the plasmid pJG317, was grown in a variety of fermentation modes including batch, serial batch and chemostat culture incorporating a wide range of media types Plasmid pJG317 consists of a 2^-denved yeast episomal plasmid containing the gene which encodes for the bacterial enzyme endo (1,3)(1,4) P-glucanase. The concentration of enzyme produced appears to be proportional to the number of plasmid copies per cell. Specific e...

  16. Presence of Glycopeptide-Encoding Plasmids in Enterococcal Isolates from Food and Humans in Denmark

    DEFF Research Database (Denmark)

    Migura, Lourdes Garcia; Valenzuela, Antonio Jesus Sanchez; Jensen, Lars Bogø

    2011-01-01

    developed techniques for classification of plasmids. Replicons associated with sex pheromone-inducible plasmids were detected in all GR E. faecalis, whereas GR Enterococcus faecium contained plasmids known to be widely distributed among enterococci. vanA resistance is common in E. faecium isolates from meat...... elements (MGE) such as plasmids and transposons. Presence of MGE was tested in all GRE isolated from food in Denmark in 2005–2007 including the first vanA mediated Enterococcus faecalis isolated from food. The ability of these plasmids to transfer and persist among enterococci was investigated using newly...

  17. Enterococcus faecalis hemolysin-bacteriocin plasmids belong to the same incompatibility group.

    OpenAIRE

    Colmar, I; Horaud, T

    1987-01-01

    Plasmid pair coexistence was studied both among nine Enterococcus faecalis hemolysin-bacteriocin (Hly-Bcn) plasmids, including pJH2, pAD1, pAM gamma 1, and pIP964, and between pIP964 and five R plasmids. Some of the Hly-Bcn plasmids used were derivatives encoding resistance to erythromycin or tetracycline. The Hly-Bcn plasmids were incompatible with each other; 40 to 100% displacement was observed bilaterally for eight pairs and unilaterally for one pair. In contrast, pIP964 stably coexisted ...

  18. [Plasmids of streptomycetes strains isolated from soils of Ukraine with different anthropogenic loading].

    Science.gov (United States)

    Luk'ianchuk, V V; Polishchuk, L V; Matseliukh, B P

    2010-01-01

    Screening of plasmid DNA was carried out among 94 streptomycetes cultures which were isolated from the samples of Ukrainian soils with different anthropogenic contamination. Seventeen streptomycetes strains containing plasmid DNA were found. It is established that some cultures contain more than one plasmid (Streptomyces sp.M15, S.sp.T8, S.sp.T19). Depending on a molecular sizes the found plasmids were divided in 2 groups: 3 kb-15 kb, and 30 kb-70 kb. Research of a few morphological and physiological properties of plasmid strains of streptomycetes was carried out. The paper is presented in Ukrainian. PMID:21117293

  19. Investigation of diversity of plasmids carrying the blaTEM-52 gene

    DEFF Research Database (Denmark)

    Bielak, Eliza Maria; Bergenholtz, Rikke D.; Jørgensen, Mikael Skaanning;

    2011-01-01

    OBJECTIVES: To investigate the diversity of plasmids that carry blaTEM-52 genes among Escherichia coli and Salmonella enterica originating from animals, meat products and humans. METHODS: A collection of 22 blaTEM-52-encoding plasmids was characterized by restriction fragment length polymorphism...... of self-transfer to a plasmid-free E. coli recipient. CONCLUSIONS: The blaTEM-52 gene found in humans could have been transmitted on transferable plasmids originating from animal sources. Some of the blaTEM-52 plasmids carry replicons that differ from the classical ones. Two novel replicons were detected...

  20. Structural and genetic analyses of a par locus that regulates plasmid partition in Bacillus subtilis.

    OpenAIRE

    Chang, S.; Chang, S Y; Gray, O

    1987-01-01

    The Bacillus plasmid pLS11 partitions faithfully during cell division. Using a partition-deficient plasmid vector, we randomly cloned DNA fragments of plasmid pLS11 and identified the locus that regulates plasmid partition (par) by cis complementation in Bacillus subtilis. The cloned par gene conferred upon the vector plasmid a high degree of segregational stability. The par locus was mapped to a 167-base-pair segment on pLS11, and its nucleotide sequence was determined. The cloned par fragme...

  1. cmp, a cis-acting plasmid locus that increases interaction between replication origin and initiator protein.

    OpenAIRE

    Gennaro, M L; Novick, R P

    1986-01-01

    pT181, a 4.4-kilobase multicopy plasmid of Staphylococcus aureus, encodes a trans-acting initiator protein, RepC, which was rate limiting for replication. Deletions in a 500-base-pair region of the plasmid external to the minimal replicon decreased the ability of the plasmid to compete with a coexisting incompatible plasmid. These deletions, which define a region called cmp (for competition), appeared to affect the interaction of RepC and the plasmid origin of replication. However, in the hom...

  2. Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3

    OpenAIRE

    Al-Allaf, Faisal A.; Tolmachov, Oleg E.; Zambetti, Lia Paola; Tchetchelnitski, Viktoria; Mehmet, Huseyin

    2012-01-01

    Large-scale production of plasmid DNA to prepare therapeutic gene vectors or DNA-based vaccines requires a suitable bacterial host, which can stably maintain the plasmid DNA during industrial cultivation. Plasmid loss during bacterial cell divisions and structural changes in the plasmid DNA can dramatically reduce the yield of the desired recombinant plasmid DNA. While generating an HIV-based gene vector containing a bicistronic expression cassette 5′-Olig2cDNA-IRES-dsRed2-3′, we encountered ...

  3. The incC Sequence Is Required for R27 Plasmid Stability

    OpenAIRE

    Tassinari, Eleonora; Aznar, Sonia; Urcola, Imanol; Prieto, Alejandro; Hüttener, Mário; Juárez, Antonio

    2016-01-01

    IncHI plasmids account for multiple antimicrobial resistance in Salmonella and other enterobacterial genera. These plasmids are generally very stable in their bacterial hosts. R27 is the archetype of IncHI1 plasmids. A high percentage of the R27-encoded open reading frames (ORFs) (66.7%) do not show similarity to any known ORFs. We performed a deletion analysis of all non-essential R27 DNA sequences to search for hitherto non-identified plasmid functions that might be required for plasmid sta...

  4. Prevalence of plasmid-mediated multidrug resistance determinants in fluoroquinolone-resistant bacteria isolated from sewage and surface water.

    Science.gov (United States)

    Osińska, Adriana; Harnisz, Monika; Korzeniewska, Ewa

    2016-06-01

    Fluoroquinolones (FQs) are fully synthetic broad-spectrum antibacterial agents that are becoming increasingly popular in the treatment of clinical and veterinary infections. Being excreted during treatment, mostly as active compounds, their biological action is not limited to the therapeutic site, but it is moved further as resistance selection pressure into the environment. Water environment is an ideal medium for the aggregation and dissemination of antibiotics, antibiotic-resistant bacteria (ARB), and antibiotic resistance genes (ARGs), which can pose a serious threat to human health. Because of this, the aim of this study was to determine the number of fluoroquinolone-resistant bacteria (FQRB) and their share in total heterotrophic plate counts (HPC) in treated wastewater (TWW), and upstream and downstream river water (URW, DRW) samples where TWW is discharged. The spread of plasmid-mediated quinolone resistance (PMQR) determinants and the presence/absence of resistance genes to other most popular antibiotic groups (against tetracyclines and beta-lactams) in selected 116 multiresistant isolates were investigated. The share of FQRB in total HPC in all samples was rather small and ranged from 0.7 % in URW samples to 7.5 % in TWW. Bacteria from Escherichia (25.0 %), Acinetobacter (25.0 %), and Aeromonas (6.9 %) genera were predominant in the FQRB group. Fluoroquinolone resistance was mostly caused by the presence of the gene aac(6')-1b-cr (91.4 %). More rarely reported was the occurrence of qnrS, qnrD, as well as oqxA, but qnrA, qnrB, qepA, and oqxB were extremely rarely or never noted in FQRB. The most prevalent bacterial genes connected with beta-lactams' resistance in FQRB were bla TEM, bla OXA, and bla CTX-M. The bla SHV was less common in the community of FQRB. The occurrence of bla genes was reported in almost 29.3 % of FQRB. The most abundant tet genes in FQRB were tet(A), tet(L), tet(K), and tet(S). The prevalence of tet genes was observed in 41.4

  5. Conjugative plasmids: Vessels of the communal gene pool

    DEFF Research Database (Denmark)

    Norman, Anders; Hansen, Lars H.; Sørensen, Søren Johannes

    2009-01-01

    Comparative whole-genome analyses have demonstrated that horizontal gene transfer (HGT) provides a significant contribution to prokaryotic genome innovation. The evolution of specific prokaryotes is therefore tightly linked to the environment in which they live and the communal pool of genes...... available within that environment. Here we use the term supergenome to describe the set of all genes that a prokaryotic ‘individual' can draw on within a particular environmental setting. Conjugative plasmids can be considered particularly successful entities within the communal pool, which have enabled HGT...... to the hosts and, potentially, other resident prokaryotes within specific environmental niches. Insight into the evolution of plasmid modules therefore contributes to our knowledge of gene dissemination and evolution within prokaryotic communities. This communal pool provides the prokaryotes with an important...

  6. Plasmids and packaging cell lines for use in phage display

    Science.gov (United States)

    Bradbury, Andrew M.

    2012-07-24

    The invention relates to a novel phagemid display system for packaging phagemid DNA into phagemid particles which completely avoids the use of helper phage. The system of the invention incorporates the use of bacterial packaging cell lines which have been transformed with helper plasmids containing all required phage proteins but not the packaging signals. The absence of packaging signals in these helper plasmids prevents their DNA from being packaged in the bacterial cell, which provides a number of significant advantages over the use of both standard and modified helper phage. Packaged phagemids expressing a protein or peptide of interest, in fusion with a phage coat protein such as g3p, are generated simply by transfecting phagemid into the packaging cell line.

  7. Cationic lipids delay the transfer of plasmid DNA to lysosomes.

    Science.gov (United States)

    Wattiaux, R; Jadot, M; Laurent, N; Dubois, F; Wattiaux-De Coninck, S

    1996-10-14

    Plasmid 35S DNA, naked or associated with different cationic lipid preparations was injected to rats. Subcellular distribution of radioactivity in the liver one hour after injection, was established by centrifugation methods. Results show that at that time, 35S DNA has reached lysosomes. On the contrary, when 35S DNA was complexed with lipids, radioactivity remains located in organelles whose distribution after differential and isopycnic centrifugation, is clearly distinct from that of arylsulfatase, lysosome marker enzyme. Injection of Triton WR 1339, a specific density perturbant of lysosomes, four days before 35S DNA injection causes a density decrease of radioactivity bearing structures, apparent one hour after naked 35S DNA injection but visible only after more than five hours, when 35S DNA associated with a cationic lipid is injected. These observations show that cationic lipids delay the transfer to lysosomes, of plasmid DNA taken up by the liver.

  8. Polymerase chain reaction-based gene removal from plasmids

    Directory of Open Access Journals (Sweden)

    Vishnu Vardhan Krishnamurthy

    2015-09-01

    Full Text Available This data article contains supplementary figures and methods to the research article entitled, “Multiplex gene removal by two-step polymerase chain reactions” (Krishnamurthy et al., Anal. Biochem., 2015, doi:http://dx.doi.org/10.1016/j.ab.2015.03.033, which presents a restriction-enzyme free method to remove multiple DNA segments from plasmids. Restriction-free cloning methods have dramatically improved the flexibility and speed of genetic manipulation compared to conventional assays based on restriction enzyme digestion (Lale and Valla, 2014. DNA Cloning and Assembly Methods, vol. 1116. Here, we show the basic scheme and characterize the success rate for single and multiplex gene removal from plasmids. In addition, we optimize experimental conditions, including the amount of template, multiple primers mixing, and buffers for DpnI treatment, used in the one-pot reaction for multiplex gene removal.

  9. Current trends in separation of plasmid DNA vaccines: a review.

    Science.gov (United States)

    Ghanem, Ashraf; Healey, Robert; Adly, Frady G

    2013-01-14

    Plasmid DNA (pDNA)-based vaccines offer more rapid avenues for development and production if compared to those of conventional virus-based vaccines. They do not rely on time- or labour-intensive cell culture processes and allow greater flexibility in shipping and storage. Stimulating antibodies and cell-mediated components of the immune system are considered as some of the major advantages associated with the use of pDNA vaccines. This review summarizes the current trends in the purification of pDNA vaccines for practical and analytical applications. Special attention is paid to chromatographic techniques aimed at reducing the steps of final purification, post primary isolation and intermediate recovery, in order to reduce the number of steps necessary to reach a purified end product from the crude plasmid.

  10. Replisome Assembly at Bacterial Chromosomes and Iteron Plasmids.

    Science.gov (United States)

    Wegrzyn, Katarzyna E; Gross, Marta; Uciechowska, Urszula; Konieczny, Igor

    2016-01-01

    The proper initiation and occurrence of DNA synthesis depends on the formation and rearrangements of nucleoprotein complexes within the origin of DNA replication. In this review article, we present the current knowledge on the molecular mechanism of replication complex assembly at the origin of bacterial chromosome and plasmid replicon containing direct repeats (iterons) within the origin sequence. We describe recent findings on chromosomal and plasmid replication initiators, DnaA and Rep proteins, respectively, and their sequence-specific interactions with double- and single-stranded DNA. Also, we discuss the current understanding of the activities of DnaA and Rep proteins required for replisome assembly that is fundamental to the duplication and stability of genetic information in bacterial cells. PMID:27563644

  11. Rapid Construction of Recombinant Plasmids by QuickStep-Cloning.

    Science.gov (United States)

    Jajesniak, Pawel; Wong, Tuck Seng

    2017-01-01

    QuickStep-Cloning is a novel molecular cloning technique that builds upon the concepts of asymmetric PCR and megaprimer-based amplification of whole plasmid. It was designed specifically to address the major drawbacks of previously reported cloning methods. The fully optimized protocol allows for a seamless integration of a long DNA fragment into any position within a plasmid of choice, in a time-efficient and cost-effective manner, without the need of a tedious DNA gel purification, a restriction digestion, and an enzymatic ligation. QuickStep-Cloning can be completed in less than 6 h, significantly faster than most of the existing cloning methods, while retaining high efficiency. PMID:27671943

  12. Characterization of two novel plasmids from Geobacillus sp. 610 and 1121 strains.

    Science.gov (United States)

    Kananavičiūtė, Rūta; Butaitė, Elena; Citavičius, Donaldas

    2014-01-01

    We describe two cryptic low molecular weight plasmids, pGTD7 (3279bp) and pGTG5 (1540bp), isolated from Geobacillus sp. 610 and 1121 strains, respectively. Homology analysis of the replication protein (Rep) sequences and detection of ssDNA indicate that both of them replicate via rolling circle mechanism. As revealed by sequence similarities of dso region and Rep protein, plasmid pGTD7 belongs to pC194/pUB110 plasmid family. The replicon of pGTD7 was proved to be functional in another Geobacillus host. For this purpose, a construct pUCK7, containing a replicon of the analyzed plasmid, was created and transferred to G. stearothermophilus NUB3621R strain by electroporation. Plasmid pGTG5, based on Rep protein sequence similarity, was found to be related mostly to some poorly characterized bacterial plasmids. Rep proteins encoded by these plasmids contain conservative motifs that are most similar to those of Microviridae phages. This feature suggests that pGTG5, together with other plasmids containing the same motifs, could constitute a new family of bacterial plasmids. To date, pGTG5 is the smallest plasmid identified in bacteria belonging to the genus Geobacillus. The two plasmids described in this study can be used for the construction of new vectors suitable for biotechnologically important bacteria of the genus Geobacillus.

  13. A Rare Case of Aeromonas Hydrophila Catheter Related Sepsis in a Patient with Chronic Kidney Disease Receiving Steroids and Dialysis: A Case Report and Review of Aeromonas Infections in Chronic Kidney Disease Patients

    Directory of Open Access Journals (Sweden)

    Muhammad Abdul Mabood Khalil

    2013-01-01

    Full Text Available Aeromonas hydrophila (AH is an aquatic bacterium. We present a case of fifty-five-year-old gentleman with chronic kidney disease (CKD due to crescentic IgA nephropathy who presented to us with fever. He was recently pulsed with methyl prednisolone followed by oral prednisolone and discharged on maintenance dialysis through a double lumen dialysis catheter. Blood culture from peripheral vein and double lumen dialysis catheter grew AH. We speculate low immunity due to steroids and uremia along with touch contamination of dialysis catheter by the patient or dialysis nurse could have led to this rare infection. Dialysis catheter related infection by AH is rare. We present our case here and take the opportunity to give a brief review of AH infections in CKD patients.

  14. Differences in the stability of the plasmids of Yersinia pestis cultures in vitro: impact on virulence

    Directory of Open Access Journals (Sweden)

    TC Leal-Balbino

    2004-11-01

    Full Text Available Plasmid and chromosomal genes encode determinants of virulence for Yersinia pestis, the causative agent of plague. However, in vitro, Y. pestis genome is very plastic and several changes have been described. To evaluate the alterations in the plasmid content of the cultures in vitro and the impact of the alterations to their pathogenicity, three Y. pestis isolates were submitted to serial subculture, analysis of the plasmid content, and testing for the presence of characteristic genes in each plasmid of colonies selected after subculture. Different results were obtained with each strain. The plasmid content of one of them was shown to be stable; no apparent alteration was produced through 32 subcultures. In the other two strains, several alterations were observed. LD50 in mice of the parental strains and the derived cultures with different plasmid content were compared. No changes in the virulence plasmid content could be specifically correlated with changes in the LD50.

  15. STABILITY OF PLASMIDS IN 5 STRAINS OF SALMONELLA MAINTAINED IN STAB CULTURE AT DIFFERENT TEMPERATURES

    DEFF Research Database (Denmark)

    Olsen, J. E.; Brown, D. J.; Baggesen, Dorte Lau;

    1994-01-01

    Four strains of Salmonella berta and one of Salm. enteritidis were stored as stab cultures in sugar-free agar at 5 degrees, 22 degrees and 30 degrees C and in 15% glycerol at -80 degrees C. The stability of the plasmid profiles in each of the strains was monitored over a period of 2.5 years....... Plasmid profiles were stable in all strains stored at -80 degrees C, and only six of 450 colonies examined from strains kept in sugar-free agar at 5 degrees C had lost plasmid molecules. Seventy of 440 colonies from stab cultures that were kept at 22 degrees C, and 71 of 440 colonies at 30 degrees C...... showed changed plasmid profiles. The total number of plasmids lost increased with time, and occasionally, more than one plasmid molecule was lost in the same strain. The virulence associated plasmid of Salm. enteritidis was remarkably stable as it was maintained in all colonies examined at all...

  16. Efficient transformation of Bacillus thuringiensis requires nonmethylated plasmid DNA.

    OpenAIRE

    Macaluso, A; Mettus, A M

    1991-01-01

    The transformation efficiency of Bacillus thuringiensis depends upon the source of plasmid DNA. DNA isolated from B. thuringiensis, Bacillus megaterium, or a Dam- Dcm- Escherichia coli strain efficiently transformed several B. thuringiensis strains, B. thuringiensis strains were grouped according to which B. thuringiensis backgrounds were suitable sources of DNA for transformation of other B. thuringiensis strains, suggesting that B. thuringiensis strains differ in DNA modification and restri...

  17. Reconstruction of the yeast 2 micron plasmid partitioning mechanism.

    OpenAIRE

    Dobson, M J; Yull, F E; Molina, M.; Kingsman, S M; Kingsman, A J

    1988-01-01

    The effect of the yeast 2 micron circle encoded REP1 and REP2 gene products on plasmid partitioning and copy number control was analyzed by removing the open reading frames from their normal sequence context and transcriptional control regions and directing their expression using heterologous promoters in [cir0] host strains. Both the REP1 and REP2 gene products are directly required at appropriate levels of expression to reconstitute the 2 microns circle partitioning system in conjunction wi...

  18. Plasmid Mediated Antibiotic Resistance in Isolated Bacteria From Burned Patients

    OpenAIRE

    Beige, Fahimeh; Baseri Salehi, Majid; Bahador, Nima; Mobasherzadeh, Sina

    2014-01-01

    Background: Nowadays, the treatment of burned patients is difficult because of the high frequency of infection with antibiotic resistance bacteria. Objectives: This study was conducted to evaluate the level of antibiotic resistance in Gram-negative bacteria and its relation with the existence of plasmid. Materials and Methods: The samples were collected from two hundred twenty hospitalized burned patients in Isfahan burn hospital during a three-month period (March 2012 to June 2012). The samp...

  19. Enhancer-activated plasmid transcription complexes contain constrained supercoiling.

    OpenAIRE

    Bonilla, P J; Freytag, S O; Lutter, L C

    1991-01-01

    It has been proposed that transcriptionally active chromatin contains totally unconstrained supercoiling. The results of recent studies have raised the possibility that this topological state is the property of highly transcribed genes. Since the transcription rate of RNA polymerase II genes can be dramatically increased by the presence of an enhancer, we have determined if the transcription complex of an enhancer-activated plasmid contains totally unconstrained supercoils. Following transfec...

  20. Plasmid addiction systems: perspectives and applications in biotechnology

    OpenAIRE

    Kroll, Jens; Klinter, Stefan; Schneider,Cornelia; Voß, Isabella; Steinbüchel, Alexander

    2010-01-01

    Summary Biotechnical production processes often operate with plasmid‐based expression systems in well‐established prokaryotic and eukaryotic hosts such as Escherichia coli or Saccharomyces cerevisiae, respectively. Genetically engineered organisms produce important chemicals, biopolymers, biofuels and high‐value proteins like insulin. In those bioprocesses plasmids in recombinant hosts have an essential impact on productivity. Plasmid‐free cells lead to losses in the entire product recovery a...

  1. Spaceflight Effects on Genetics and Plasmids of Streptomycetes

    Science.gov (United States)

    Voeikova, T. A.; Emelyanova, L. K.; Tyaglov, B. V.; Novikova, L. M.; Goins, T. L.; Pyle, B. H.

    2008-06-01

    In 2007, experiments with streptomycetes were conducted during a 12-day flight of the Russian Foton-M3 spacecraft. The flight (F), synchronous control (SC) and laboratory control (LC) specimens were kept at 30°C. The objective of the experiments was to study spaceflight effects on the streptomycetes growth, differentiation, pigmentation, enzyme formation, genetic stability of plasmid and crossing between strains. It was found that the frequency of strain Streptomyces lividans segregation, the enzyme synthesis, pigmentation, and the level of sporulation were higher in F than in SC organisms. The study of pIJ702 plasmid inheritance in S. lividans showed that the frequency of plasmid loss in F and LC was similar and lower than that in SC specimens. The study of melanin synthesis in S. lividans (pIJ702) strain demonstrated decreased melanin specific yield and increased biomass accumulation in F microorganisms. HPTLC analysis of melanin showed that the number, molecular mass and the percentage of fractions were similar in SC and LC but different in F organisms. The study of spaceflight effects on genetic recombination in crosses between Streptomyces coelicolor A3(2) auxotrophic mutants showed that the frequency of various recombinant classes in F specimens differed from that in SC and LC. The frequency of a distal donor marker entry to the recipient in F was higher than in SC and LC.

  2. DNA immunization with plasmids expressing hCGbeta-chimeras.

    Science.gov (United States)

    Terrazzini, Nadia; Hannesdóttir, Sólveig; Delves, Peter J; Lund, Torben

    2004-06-01

    Human chorionic gonadotropin has been used as an anti-fertility vaccine and as a target for cancer immunotherapy. We have explored the use of DNA immunization with the aim of improving the immunogenicity of this hormone. Stimulating the muscle with electric pulses following intramuscular injection of plasmids expressing hCGbeta resulted in higher levels of human chorionic gonadotropin (hCG)-specific antibodies, which could be further enhanced following a protein boost with hCG mixed with adjuvant. DNA vaccines encoding a membrane attached or a secreted form of hCGbeta produced similar-albeit relatively modest-antibody responses. Providing hCGbeta with additional T cell help by vaccinating with a plasmid encoding a hCGbeta-hFc fusion protein did not further increase the antibody levels in the immunized animals. However, immunization of mice with a construct encoding hCGbeta fused to C3d(3) produced significantly lower antibody levels relative to mice immunized with the hCGbeta-alone expression plasmid, even though the hCGbeta-C3d(3) chimera was expected to facilitate cross-linking of the antigen-specific B-cell receptor and CR2 thereby lowering the threshold of activation. Thus the limiting factor determining the antibody levels following hCGbeta immunization, at least for DNA immunization, is related to the amount of protein available rather than the form of protein produced or lack of T cell epitopes. PMID:15149771

  3. The use of eugenol against Aeromonas hydrophila and its effect on hematological and immunological parameters in silver catfish (Rhamdia quelen).

    Science.gov (United States)

    Sutili, Fernando Jonas; Kreutz, Luiz Carlos; Noro, Mirela; Gressler, Leticia Trevisan; Heinzmann, Berta Maria; de Vargas, Agueda Castagna; Baldisserotto, Bernardo

    2014-02-15

    The aim of this study was to evaluate the activity of eugenol against the fish pathogen Aeromonas hydrophila and eugenol's effect on hematological and natural immune parameters in silver catfish (Rhamdia quelen). In vitro, eugenol showed weak activity against A. hydrophila, but in vivo, at a subinhibitory concentration (10 mg L(-1)), it promoted survival in infected silver catfish. Eugenol (50 μg mL(-1)) reduced the hemolytic activity of A. hydrophila supernatant in vitro in fish erythrocytes. Subjecting catfish to eugenol baths (5 and 10 mg L(-1)) for five days did not alter the hematological and immunological parameters studied in this work. Based on these results, eugenol can be used to treat or prevent bacterial diseases in fish.

  4. Multiple drug resistance of Aeromonas hydrophila isolates from Chicken samples collected from Mhow and Indore city of Madhyapradesh

    Directory of Open Access Journals (Sweden)

    Kaskhedikar

    2009-02-01

    Full Text Available Fourteen antibacterial agents belonging to 9 different groups of antibiotics viz. aminoglycosides, cephalosporins, nitrofurantoin, fluroquinolones, chloramphenicol, sulphonamides, tetracyclines, penicillin and polymixin were used for in vitro sensitivity testing of Aeromonas hydrophila isolated from fifteen samples of chicken collected from retail shops in Mhow city. The sensitivity (100% was attributed to ciprofloxacin, cefuroxime, ceftriaxone, cephotaxime, chloramphenicol, gentamycin, kanamycin, nitrofurantoin, nalidixic acid and ofloxacin followed by oxytetracycline (50%. All the isolates were resistant to ampicillin and colistin antibiotics. That means, none of the isolates were found to be sensitive for penicillin and polymixin group of antibiotics. Multiple drug resistance was also observed in all A. hydrophila isolates. Out of total isolates, 100% were resistant to two antimicrobial drugs and 50% to three drugs. [Vet. World 2009; 2(1.000: 31-32

  5. Comparative evaluation of infection methods and environmental factors on challenge success: Aeromonas salmonicida infection in vaccinated rainbow trout

    DEFF Research Database (Denmark)

    Chettri, Jiwan Kumar; Skov, Jakob; Jaafar, Rzgar M.;

    2015-01-01

    When testing vaccine-induced protection an effective and reliable challenge method is a basic requirement and we here present a comparative study on different challenge methods used for infection of rainbow trout Oncorhynchus mykiss with Aeromonas salmonicida, a bacterial pathogen eliciting...... furunculosis. Fish were vaccinated with three different adjuvanted trivalent vaccines containing formalin killed A. salmonicida, Vibrio anguillarum O1 and O2a. These were 1) the commercial vaccine Alpha Ject 3000, 2) an experimental vaccine with water in paraffin oil adjuvant, 3) an experimental vaccine...... differentiation of vaccine types compared to i.p. injection of live bacteria. The latter infection mode is less successful probably due to the intra-abdominal inflammatory reactions (characterized in this study according to the Speilberg scale) induced by i.p. vaccination whereby injected live bacteria more...

  6. Pathogenicity of Aeromonas hydrophila isolated from the Malaysian Sea against coral (Turbinaria sp.) and sea bass (Lates calcarifer).

    Science.gov (United States)

    Hamid, Rahimi; Ahmad, Asmat; Usup, Gires

    2016-09-01

    A study was carried out to determine the pathogenicity (hemolytic activity) on corals (Turbinaria sp.) and sea bass (Lates calcarifer) of Aeromonas hydrophila from water, sediment, and coral. Samples were collected from coastal water and coral reef areas. One hundred and sixty-two isolates were successfully isolated. Out of 162, 95 were from seawater, 49 from sediment, and 18 from coral. Sixteen isolates were picked and identified. Isolates were identified using a conventional biochemical test, the API 20NE kit, and 16S rRNA nucleotide sequences. Hemolytic activity was determined. Out of 16 isolates, 14 isolates were β-hemolytic and two isolates were non-hemolytic. Corals infected with A. hydrophila suffered bleaching. Similar effect was observed for both hemolytic and non-hemolytic isolates. Intramuscular injection of A. hydrophila into sea bass resulted in muscular bleeding and death. Higher infection rates were obtained from hemolytic compared to non-hemolytic strains of A. hydrophila isolates. PMID:27221587

  7. Occurrence and significance of atypical Aeromonas salmonicida in non-salmonid and salmonid fish species : A review

    DEFF Research Database (Denmark)

    Wiklund, T.; Dalsgaard, Inger

    1998-01-01

    Bacterial strains of Aeromonas salmonicida included in the recognized subsp. achromogenes, subsp, masoucida, and subsp. smithia in addition to the large number of strains not included in any of the described subspecies are referred to as atypical A. salmonicida. The atypical strains form a very...... heterogeneous group with respect to biochemical characteristics, growth conditions, and production of extracellular proteases. Consequently, the present taxonomy of the species A. salmonicida is rather ambiguous. Atypical A. salmonicida has been isolated from a wide range of cultivated and wild fish species...... variation. An extracellular A-layer has been detected in different atypical strains, but virulence mechanisms different from those described for (typical) A. salmonicida subsp. salmonicida, for example an extracellular metallo-protease and a different iron utilization mechanism, have been described. Limited...

  8. Role of Aeromonas hydrophila Flagella Glycosylation in Adhesion to Hep-2 Cells, Biofilm Formation and Immune Stimulation

    Directory of Open Access Journals (Sweden)

    Susana Merino

    2014-11-01

    Full Text Available Polar flagellin proteins from Aeromonas hydrophila strain AH-3 (serotype O34 were found to be O-glycosylated with a heterogeneous heptasaccharide glycan. Two mutants with altered (light and strong polar flagella glycosylation still able to produce flagella were previously obtained, as well as mutants lacking the O34-antigen lipopolysaccharide (LPS but with unaltered polar flagella glycosylation. We compared these mutants, altogether with the wild type strain, in different studies to conclude that polar flagella glycosylation is extremely important for A. hydrophila adhesion to Hep-2 cells and biofilm formation. Furthermore, the polar flagella glycosylation is an important factor for the immune stimulation of IL-8 production via toll receptor 5 (TLR5.

  9. Two domains at the origin are required for replication and maintenance of broad-host-range plasmid R1162.

    OpenAIRE

    Kim, Y.J.; Lin, L. S.; Meyer, R. J.

    1987-01-01

    Two domains at the replicative origin of broad-host-range plasmid R1162 are required in cis for plasmid maintenance in Escherichia coli and for plasmid DNA replication in cell extracts. Increasing the distance between the domains reduces replication in vitro, without substantially changing plasmid DNA content or stability in vivo.

  10. Bacillus anthracis Virulent Plasmid pX02 Genes Found in Large Plasmids of Two Other Bacillus Species

    OpenAIRE

    Luna, Vicki A.; King, Debra S.; Peak, K. Kealy; Reeves, Frank; Heberlein-Larson, Lea; Veguilla, William; Heller, L.; Duncan, Kathleen E; Cannons, Andrew C.; Amuso, Philip; Cattani, Jacqueline

    2006-01-01

    In order to cause the disease anthrax, Bacillus anthracis requires two plasmids, pX01 and pX02, which carry toxin and capsule genes, respectively, that are used as genetic targets in the laboratory detection of the bacterium. Clinical, forensic, and environmental samples that test positive by PCR protocols established by the Centers for Disease Control and Prevention for B. anthracis are considered to be potentially B. anthracis until confirmed by culture and a secondary battery of tests. We ...

  11. Concerted transfer of the virulence Ti plasmid and companion At plasmid in the Agrobacterium tumefaciens-induced plant tumour.

    Science.gov (United States)

    Lang, Julien; Planamente, Sara; Mondy, Samuel; Dessaux, Yves; Moréra, Solange; Faure, Denis

    2013-12-01

    The plant pathogen Agrobacterium tumefaciens C58 harbours three independent type IV secretion (T4SS) machineries. T4SST-DNA promotes the transfer of the T-DNA to host plant cells, provoking tumour development and accumulation of opines such as nopaline and agrocinopines. T4SSpTi and T4SSpAt control the bacterial conjugation of the Ti and At plasmids respectively. Expression of T4SSpTi is controlled by the agrocinopine-responsive transcriptional repressor AccR. In this work, we compared the genome-wide transcriptional profile of the wild-type A. tumefaciens strain C58 with that of its accR KO-mutant to delineate the AccR regulon. In addition to the genes that encode agrocinopine catabolism and T4SSpTi , we found that AccR also regulated genes coding for nopaline catabolism and T4SSpAt . Further opine detection and conjugation assays confirmed the enhancement of nopaline consumption and At plasmid conjugation frequency in accR. Moreover, co-regulation of the T4SSpTi and T4SSpAt correlated with the co-transfer of the At and Ti plasmids both in vitro and in plant tumours. Finally, unlike T4SSpTi , T4SSpAt activation does not require quorum-sensing. Overall this study highlights the regulatory interplays between opines, At and Ti plasmids that contribute to a concerted dissemination of the two replicons in bacterial populations colonizing the plant tumour. PMID:24118167

  12. Type 3 Fimbriae Encoded on Plasmids Are Expressed from a Unique Promoter without Affecting Host Motility, Facilitating an Exceptional Phenotype That Enhances Conjugal Plasmid Transfer

    Science.gov (United States)

    Madsen, Jonas Stenløkke; Riber, Leise; Kot, Witold; Basfeld, Alrun; Burmølle, Mette; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2016-01-01

    Horizontal gene transfer (HGT), the transmission of genetic material to a recipient that is not the progeny of the donor, is fundamental in bacterial evolution. HGT is often mediated by mobile genetic elements such as conjugative plasmids, which may be in conflict with the chromosomal elements of the genome because they are independent replicons that may petition their own evolutionary strategy. Here we study differences between type 3 fimbriae encoded on wild type plasmids and in chromosomes. Using known and newly characterized plasmids we show that the expression of type 3 fimbriae encoded on plasmids is systematically different, as MrkH, a c-di-GMP dependent transcriptional activator is not needed for strong expression of the fimbriae. MrkH is required for expression of type 3 fimbriae of the Klebsiella pneumoniae chromosome, wherefrom the fimbriae operon (mrkABCDF) of plasmids is believed to have originated. We find that mrkABCDFs of plasmids are highly expressed via a unique promoter that differs from the original Klebsiella promoter resulting in fundamental behavioral consequences. Plasmid associated mrkABCDFs did not influence the swimming behavior of the host, that hereby acquired an exceptional phenotype being able to both actively swim (planktonic behavior) and express biofilm associated fimbriae (sessile behavior). We show that this exceptional phenotype enhances the conjugal transfer of the plasmid. PMID:27627107

  13. Type 3 fimbriae encoded on plasmids are expressed from a unique promoter without affecting host motility, facilitating an exceptional phenotype that enhances conjugal plasmid transfer

    DEFF Research Database (Denmark)

    Madsen, Jonas Stenløkke; Riber, Leise; Kot, Witold Piotr;

    2016-01-01

    of the genome because they are independent replicons that may petition their own evolutionary strategy. Here we study differences between type 3 fimbriae encoded on wild type plasmids and in chromosomes. Using known and newly characterized plasmids we show that the expression of type 3 fimbriae encoded...

  14. A Site-Specific Integrative Plasmid Found in Pseudomonas aeruginosa Clinical Isolate HS87 along with A Plasmid Carrying an Aminoglycoside-Resistant Gene.

    Directory of Open Access Journals (Sweden)

    Dexi Bi

    Full Text Available Plasmids play critical roles in bacterial fitness and evolution of Pseudomonas aeruginosa. Here two plasmids found in a drug-resistant P. aeruginosa clinical isolate HS87 were completely sequenced. The pHS87b plasmid (11.2 kb carries phage-related genes and function-unknown genes. Notably, pHS87b encodes an integrase and has an adjacent tRNAThr-associated attachment site. A corresponding integrated form of pHS87b at the tRNAThr locus was identified on the chromosome of P. aeruginosa, showing that pHS87b is able to site-specifically integrate into the 3'-end of the tRNAThr gene. The pHS87a plasmid (26.8 kb displays a plastic structure containing a putative replication module, stability factors and a variable region. The RepA of pHS87a shows significant similarity to the replication proteins of pPT23A-family plasmids. pHS87a carries a transposon Tn6049, a truncated insertion sequence ΔIS1071 and a Tn402-like class 1 integron which contains an aacA4 cassette that may confer aminoglycoside resistance. Thus, pHS87b is a site-specific integrative plasmid whereas pHS87a is a plastic antibiotic resistance plasmid. The two native plasmids may promote the fitness and evolution of P. aeruginosa.

  15. A Site-Specific Integrative Plasmid Found in Pseudomonas aeruginosa Clinical Isolate HS87 along with A Plasmid Carrying an Aminoglycoside-Resistant Gene.

    Science.gov (United States)

    Bi, Dexi; Xie, Yingzhou; Tai, Cui; Jiang, Xiaofei; Zhang, Jie; Harrison, Ewan M; Jia, Shiru; Deng, Zixin; Rajakumar, Kumar; Ou, Hong-Yu

    2016-01-01

    Plasmids play critical roles in bacterial fitness and evolution of Pseudomonas aeruginosa. Here two plasmids found in a drug-resistant P. aeruginosa clinical isolate HS87 were completely sequenced. The pHS87b plasmid (11.2 kb) carries phage-related genes and function-unknown genes. Notably, pHS87b encodes an integrase and has an adjacent tRNAThr-associated attachment site. A corresponding integrated form of pHS87b at the tRNAThr locus was identified on the chromosome of P. aeruginosa, showing that pHS87b is able to site-specifically integrate into the 3'-end of the tRNAThr gene. The pHS87a plasmid (26.8 kb) displays a plastic structure containing a putative replication module, stability factors and a variable region. The RepA of pHS87a shows significant similarity to the replication proteins of pPT23A-family plasmids. pHS87a carries a transposon Tn6049, a truncated insertion sequence ΔIS1071 and a Tn402-like class 1 integron which contains an aacA4 cassette that may confer aminoglycoside resistance. Thus, pHS87b is a site-specific integrative plasmid whereas pHS87a is a plastic antibiotic resistance plasmid. The two native plasmids may promote the fitness and evolution of P. aeruginosa. PMID:26841043

  16. Type 3 Fimbriae Encoded on Plasmids Are Expressed from a Unique Promoter without Affecting Host Motility, Facilitating an Exceptional Phenotype That Enhances Conjugal Plasmid Transfer.

    Science.gov (United States)

    Madsen, Jonas Stenløkke; Riber, Leise; Kot, Witold; Basfeld, Alrun; Burmølle, Mette; Hansen, Lars Hestbjerg; Sørensen, Søren Johannes

    2016-01-01

    Horizontal gene transfer (HGT), the transmission of genetic material to a recipient that is not the progeny of the donor, is fundamental in bacterial evolution. HGT is often mediated by mobile genetic elements such as conjugative plasmids, which may be in conflict with the chromosomal elements of the genome because they are independent replicons that may petition their own evolutionary strategy. Here we study differences between type 3 fimbriae encoded on wild type plasmids and in chromosomes. Using known and newly characterized plasmids we show that the expression of type 3 fimbriae encoded on plasmids is systematically different, as MrkH, a c-di-GMP dependent transcriptional activator is not needed for strong expression of the fimbriae. MrkH is required for expression of type 3 fimbriae of the Klebsiella pneumoniae chromosome, wherefrom the fimbriae operon (mrkABCDF) of plasmids is believed to have originated. We find that mrkABCDFs of plasmids are highly expressed via a unique promoter that differs from the original Klebsiella promoter resulting in fundamental behavioral consequences. Plasmid associated mrkABCDFs did not influence the swimming behavior of the host, that hereby acquired an exceptional phenotype being able to both actively swim (planktonic behavior) and express biofilm associated fimbriae (sessile behavior). We show that this exceptional phenotype enhances the conjugal transfer of the plasmid. PMID:27627107

  17. Expansion of a plasmid classification system for Gram-positive bacteria and determination of the diversity of plasmids in Staphylococcus aureus strains of human, animal, and food origins

    DEFF Research Database (Denmark)

    Lozano, C.; Garcia-Migura, L.; Aspiroz, C.;

    2012-01-01

    An expansion of a previously described plasmid classification was performed and used to reveal the plasmid content of a collection of 92 Staphylococcus aureus strains of different origins. rep genes of other genera were detected in Staphylococcus. S1 pulsed-field gel electrophoresis (PFGE...

  18. Genetic transformation of a clinical (genital tract, plasmid-free isolate of Chlamydia trachomatis: engineering the plasmid as a cloning vector.

    Directory of Open Access Journals (Sweden)

    Yibing Wang

    Full Text Available Our study had three objectives: to extend the plasmid-based transformation protocol to a clinical isolate of C. trachomatis belonging to the trachoma biovar, to provide "proof of principle" that it is possible to "knock out" selected plasmid genes (retaining a replication competent plasmid and to investigate the plasticity of the plasmid. A recently developed, plasmid-based transformation protocol for LGV isolates of C. trachomatis was modified and a plasmid-free, genital tract C. trachomatis isolate from Sweden (SWFP- was genetically transformed. Transformation of this non-LGV C. trachomatis host required a centrifugation step, but the absence of the natural plasmid removed the need for plaque purification of transformants. Transformants expressed GFP, were penicillin resistant and iodine stain positive for accumulated glycogen. The transforming plasmid did not recombine with the host chromosome. A derivative of pGFP::SW2 carrying a deletion of the plasmid CDS5 gene was engineered. CDS5 encodes pgp3, a protein secreted from the inclusion into the cell cytoplasm. This plasmid (pCDS5KO was used to transform C. trachomatis SWFP-, and established that pgp3 is dispensable for plasmid function. The work shows it is possible to selectively delete segments of the chlamydial plasmid, and this is the first step towards a detailed molecular dissection of the role of the plasmid. The 3.6 kb β-galactosidase cassette was inserted into the deletion site of CDS5 to produce plasmid placZ-CDS5KO. Transformants were penicillin resistant, expressed GFP and stained for glycogen. In addition, they expressed β-galactosidase showing that the lacZ cassette was functional in C. trachomatis. An assay was developed that allowed the visualisation of individual inclusions by X-gal staining. The ability to express active β-galactosidase within chlamydial inclusions is an important advance as it allows simple, rapid assays to measure directly chlamydial infectivity without

  19. RK2 plasmid dynamics in Caulobacter crescentus cells--two modes of DNA replication initiation.

    Science.gov (United States)

    Wegrzyn, Katarzyna; Witosinska, Monika; Schweiger, Pawel; Bury, Katarzyna; Jenal, Urs; Konieczny, Igor

    2013-06-01

    Undisturbed plasmid dynamics is required for the stable maintenance of plasmid DNA in bacterial cells. In this work, we analysed subcellular localization, DNA synthesis and nucleoprotein complex formation of plasmid RK2 during the cell cycle of Caulobacter crescentus. Our microscopic observations showed asymmetrical distribution of plasmid RK2 foci between the two compartments of Caulobacter predivisional cells, resulting in asymmetrical allocation of plasmids to progeny cells. Moreover, using a quantitative PCR (qPCR) method, we estimated that multiple plasmid particles form a single fluorescent focus and that the number of plasmids per focus is approximately equal in both swarmer and predivisional Caulobacter cells. Analysis of the dynamics of TrfA-oriV complex formation during the Caulobacter cell cycle revealed that TrfA binds oriV primarily during the G1 phase, however, plasmid DNA synthesis occurs during the S and G2 phases of the Caulobacter cell cycle. Both in vitro and in vivo analysis of RK2 replication initiation in C. crescentus cells demonstrated that it is independent of the Caulobacter DnaA protein in the presence of the longer version of TrfA protein, TrfA-44. However, in vivo stability tests of plasmid RK2 derivatives suggested that a DnaA-dependent mode of plasmid replication initiation is also possible.

  20. A polymerization-based method to construct a plasmid containing clustered DNA damage and a mismatch.

    Science.gov (United States)

    Takahashi, Momoko; Akamatsu, Ken; Shikazono, Naoya

    2016-10-01

    Exposure of biological materials to ionizing radiation often induces clustered DNA damage. The mutagenicity of clustered DNA damage can be analyzed with plasmids carrying a clustered DNA damage site, in which the strand bias of a replicating plasmid (i.e., the degree to which each of the two strands of the plasmid are used as the template for replication of the plasmid) can help to clarify how clustered DNA damage enhances the mutagenic potential of comprising lesions. Placement of a mismatch near a clustered DNA damage site can help to determine the strand bias, but present plasmid-based methods do not allow insertion of a mismatch at a given site in the plasmid. Here, we describe a polymerization-based method for constructing a plasmid containing clustered DNA lesions and a mismatch. The presence of a DNA lesion and a mismatch in the plasmid was verified by enzymatic treatment and by determining the relative abundance of the progeny plasmids derived from each of the two strands of the plasmid. PMID:27449134

  1. Plasmids of the pRM/pRF family occur in diverse Rickettsia species.

    Science.gov (United States)

    Baldridge, Gerald D; Burkhardt, Nicole Y; Felsheim, Roderick F; Kurtti, Timothy J; Munderloh, Ulrike G

    2008-02-01

    The recent discoveries of the pRF and pRM plasmids of Rickettsia felis and R. monacensis have contravened the long-held dogma that plasmids are not present in the bacterial genus Rickettsia (Rickettsiales; Rickettsiaceae). We report the existence of plasmids in R. helvetica, R. peacockii, R. amblyommii, and R. massiliae isolates from ixodid ticks and in an R. hoogstraalii isolate from an argasid tick. R. peacockii and four isolates of R. amblyommii from widely separated geographic locations contained plasmids that comigrated with pRM during pulsed-field gel electrophoresis and larger plasmids with mobilities similar to that of pRF. The R. peacockii plasmids were lost during long-term serial passage in cultured cells. R. montanensis did not contain a plasmid. Southern blots showed that sequences similar to those of a DnaA-like replication initiator protein, a small heat shock protein 2, and the Sca12 cell surface antigen genes on pRM and pRF were present on all of the plasmids except for that of R. massiliae, which lacked the heat shock gene and was the smallest of the plasmids. The R. hoogstraalii plasmid was most similar to pRM and contained apparent homologs of proline/betaine transporter and SpoT stringent response genes on pRM and pRF that were absent from the other plasmids. The R. hoogstraalii, R. helvetica, and R. amblyommii plasmids contained homologs of a pRM-carried gene similar to a Nitrobacter sp. helicase RecD/TraA gene, but none of the plasmids hybridized with a probe derived from a pRM-encoded gene similar to a Burkholderia sp. transposon resolvase gene.

  2. Effect of excessive cadmium chloride on the plasmids of E. coli HB 101 in vivo

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    After Escherichia coli HB101 with plasmid pWH58, pWH98, or pTBa5 were cultered respectively in amp LB broth which contained 50 mg/L CdCl2 constantly for 24 h, these plasmids were isolated from E. coli, and the effect of excessive CdCl2 on the E. coli HB101 and plasmid DNA was studied by surveying the growth of E. coli HB101 and plasmid, argarose gel electrophoresis and analysis of restriction fragment length polymorphism (RFLP) of plasmids, and plasmid transformation. The results showed that 50 mg/L CdCl2 treatment lagged the growth of E. coli HB101 for at least 4h, but after grown for 24h there were not significant differences in the growths of E. coli HB101s and the productions of plasmids between the treatment and control. These results implified that E. coli HB101 have induced adaptability to cadmium stress and excessive CdCl2 did not inhibit the replication and amp+ gene's expression of plasmid DNA in vivo of E. coli significantly. 50 mg/L CdCl2 treatment for 24 hours might cause the sequence's change of plasmid DNA, but could not lead to the random breakage of plasmid DNA strands. Moreover, after 50 mg/L of CdCl2 treatment in vivo the transformation activities of plasmid did not altered, implied excessive CdCl2 could not affect the superhelical structure of plasmid and also not break the loop of plasmid DNA evidently.

  3. Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

    Directory of Open Access Journals (Sweden)

    Mark Eppinger

    Full Text Available Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

  4. Plasmid DNA Supercoiling and Gyrase Activity in Escherichia coli Wild-Type and rpoS Stationary-Phase Cells

    Science.gov (United States)

    Reyes-Domínguez, Yazmid; Contreras-Ferrat, Gabriel; Ramírez-Santos, Jesús; Membrillo-Hernández, Jorge; Gómez-Eichelmann, M. Carmen

    2003-01-01

    Stationary-phase cells displayed a distribution of relaxed plasmids and had the ability to recover plasmid supercoiling as soon as nutrients became available. Preexisting gyrase molecules in these cells were responsible for this recovery. Stationary-phase rpoS cells showed a bimodal distribution of plasmids and failed to supercoil plasmids after the addition of nutrients, suggesting that rpoS plays a role in the regulation of plasmid topology during the stationary phase. PMID:12533486

  5. Multiple drug resistant carbapenemases producing Acinetobacter baumannii isolates harbours multiple R-plasmids

    Directory of Open Access Journals (Sweden)

    Rajagopalan Saranathan

    2014-01-01

    Full Text Available Background & objectives: The nosocomial human pathogen Acinetobacter baumannii has high propensity to develop resistance to antimicrobials and to become multidrug resistant (MDR, consequently complicating the treatment. This study was carried out to investigate the presence of resistant plasmids (R-plasmids among the clinical isolates of A. baumannii. In addition, the study was performed to check the presence of common β-lactamases encoding genes on these plasmids. Methods: A total of 55 clinical isolates of A. baumannii were included in the study and all were subjected to plasmid DNA isolation, followed by PCR to check the presence of resistance gene determinants such as blaOXA-23 , blaOXA-51, blaOXA-58 and blaIMP-1 on these plasmids that encode for oxacillinase (OXA and metallo-β-lactamase (MBL type of carbapenemases. Plasmid curing experiments were carried out on selected isolates using ethidium bromide and acridine orange as curing agents and the antibiotic resistance profiles were evaluated before and after curing. Results: All the isolates were identified as A. baumannii by 16SrDNA amplification and sequencing. Plasmid DNA isolated from these isolates showed the occurrence of multiple plasmids with size ranging from 500bp to ≥ 25 kb. The percentage of blaOXA-51 and blaOXA-23 on plasmids were found to be 78 and 42 per cent, respectively and 20 isolates (36% carried blaIMP-1 gene on plasmids. Significant difference was observed in the antibiograms of plasmid cured isolates when compared to their parental ones. The clinical isolates became susceptible to more than two antibiotic classes after curing of plasmids indicating plasmid borne resistance. Interpretation & conclusions: Our study determined the plasmid mediated resistance mechanisms and occurrence of different resistance genes on various plasmids isolated from MDR A. baumannii. The present findings showed the evidence for antibiotic resistance mediated through multiple plasmids in

  6. TLR21's agonists in combination with Aeromonas antigens synergistically up-regulate functional TLR21 and cytokine gene expression in yellowtail leucocytes.

    Science.gov (United States)

    Reyes-Becerril, Martha; Ascencio-Valle, Felipe; Hirono, Ikuo; Kondo, Hidehiro; Jirapongpairoj, Walissara; Esteban, Maria Angeles; Alamillo, Erika; Angulo, Carlos

    2016-08-01

    The purpose of this study was to characterize the TLR21 gene from yellowtail (Seriola lalandi) and its functional activity using TLR agonist stimulation and Aeromonas antigens. The TLR21 nucleotide sequence from yellowtail was obtained using the whole-genome shotgun sequencing method and bioinformatics tools. Basal TLR21 gene expression was analyzed in several tissues. Subsequently, the gene expression of TLR21 and cytokines IL-1β and TNF-α was evaluated in TLR agonist (CpG-ODN2006, LPS, and Poly I:C) exposing head kidney leucocytes, which were then subjected to Aeromonas antigen stimulation. The yellowtail full-length cDNA sequence of SlTLR21 was 3615 bp (980 aa) showing a high degree of similarity with the counterparts of other fish species and sharing the common structural architecture of the TLR family, including LRR domains, one C-terminal LRR region, and a TIR domain. Gene expression studies revealed the constitutive expression of TLR21 mRNA in all the analyzed tissues; the highest levels were observed in spleen and head kidney where they play an important role in the fish immune system. Transcripts of TLR21 and the downstream IL-1β and TNF-α cytokine genes were most strongly up-regulated after exposure to the TLR agonists following Aeromonas antigen stimulation, suggesting they are involved in immune response. The results indicated that TLR agonists, in combination with Aeromonas antigens in head kidney leucocytes, synergistically enhance TLR21 and cytokines in yellowtail. PMID:26987525

  7. Characterization of a Cryptic and Intriguing Low Molecular Weight Plasmid.

    Science.gov (United States)

    Carneiro, Lilian C; Mendes, Paulo Vinicius C; Silva, Silvana P; Souza, Guilherme R L; Bataus, Luiz Artur M

    2016-03-01

    The complete nucleotide sequence of cryptic plasmid pVCM04 isolated from Salmonella enterica serovar Enteritidis was determined and analyzed. pVCM04 contains 3853 bp with 53.6 % GC content and has twelve ORFs with more than 50 amino acids. Five of these sequences showed homology with replication and mobilization proteins. ORF1 and ORF2 showed homology with replication proteins, while ORFs 3-5 showed homology with mobilization proteins. The pVCM04 possesses a region associated with the theta-type replication mechanism. BLASTn search analysis revealed unexpectedly no similarity with sequences deposited in GenBank. The nucleotide sequence of pVCM04 can be divided into two arms: the region between nucleotides 552-1774 (encoding RepA and RepB) and the region between nucleotides 1775-3853 (encoding MobA, MobB and MobC). Codon bias pattern is distinct between mobA and repA, so the program Modeltest was used to select the best evolutionary model to study these genes. The result of ModelTest (model GTR+G for mobA and model HKY+G for repA) suggests that these genes would be subject to different selective pressures. Considering the differences in the codon usage, the selection of two different evolutionary models, and the absence of plasmids with homology to pVCM04 in GenBank, we believe that pVCM04 is a chimeric molecule and represents a new plasmid lineage. PMID:26670037

  8. Strategies and approaches in plasmidome studies – uncovering plasmid diversity disregarding of linear elements?

    Directory of Open Access Journals (Sweden)

    Julian Rafael Dib

    2015-05-01

    Full Text Available The term plasmid was originally coined for circular, extrachromosomal genetic elements. Today, plasmids are widely recognised not only as important factors facilitating genome restructuring but also as vehicles for the dissemination of beneficial characters within bacterial communities. Plasmid diversity has been uncovered by means of culture–dependent or –independent approaches, such as endogenous or exogenous plasmid isolation as well as PCR-based detection or transposon-aided capture, respectively. High-throughput-sequencing made possible to cover total plasmid populations in a given environment, i.e. the plasmidome, and allowed to address the quality and significance of self-replicating genetic elements. Since such efforts were and still are rather restricted to circular molecules, here we put equal emphasis on the linear plasmids which – despite their frequent occurrence in a large number of bacteria – are largely neglected in prevalent plasmidome conceptions.

  9. Strategies and approaches in plasmidome studies—uncovering plasmid diversity disregarding of linear elements?

    Science.gov (United States)

    Dib, Julián R.; Wagenknecht, Martin; Farías, María E.; Meinhardt, Friedhelm

    2015-01-01

    The term plasmid was originally coined for circular, extrachromosomal genetic elements. Today, plasmids are widely recognized not only as important factors facilitating genome restructuring but also as vehicles for the dissemination of beneficial characters within bacterial communities. Plasmid diversity has been uncovered by means of culture-dependent or -independent approaches, such as endogenous or exogenous plasmid isolation as well as PCR-based detection or transposon-aided capture, respectively. High-throughput-sequencing made possible to cover total plasmid populations in a given environment, i.e., the plasmidome, and allowed to address the quality and significance of self-replicating genetic elements. Since such efforts were and still are rather restricted to circular molecules, here we put equal emphasis on the linear plasmids which—despite their frequent occurrence in a large number of bacteria—are largely neglected in prevalent plasmidome conceptions. PMID:26074886

  10. Long- term manure exposure increases soil bacterial community potential for plasmid uptake

    DEFF Research Database (Denmark)

    Musovic, Sanin; Klümper, Uli; Dechesne, Arnaud;

    2014-01-01

    Microbial communities derived from soils subject to different agronomic treatments were challenged with three broad host range plasmids, RP4, pIPO2tet and pRO101, via solid surface filter matings to assess their permissiveness. Approximately 1 in 10 000 soil bacterial cells could receive and main......Microbial communities derived from soils subject to different agronomic treatments were challenged with three broad host range plasmids, RP4, pIPO2tet and pRO101, via solid surface filter matings to assess their permissiveness. Approximately 1 in 10 000 soil bacterial cells could receive...... and maintain the plasmids. The community permissiveness increased up to 100% in communities derived from manured soil. While the plasmid transfer frequency was significantly influenced by both the type of plasmid and the agronomic treatment, the diversity of the transconjugal pools was purely plasmid dependent...

  11. Survival and evolution of a large multidrug resistance plasmid in new clinical bacterial hosts

    DEFF Research Database (Denmark)

    Porse, Andreas; Schønning, Kristian; Munck, Christian;

    2016-01-01

    Large conjugative plasmids are important drivers of bacterial evolution and contribute significantly to the dissemination of antibiotic resistance. Although plasmid borne multidrug resistance is recognized as one of the main challenges in modern medicine, the adaptive forces shaping the evolution...... sequencing to show that the long-term persistence and molecular integrity of the plasmid is highly influenced by multiple factors within a 25 kb plasmid region constituting a host-dependent burden. In the E. coli hosts investigated here, improved plasmid stability readily evolves via IS26 mediated deletions...... consistently followed by all evolved E. coli lineages exposes a trade-off between horizontal and vertical transmission that may ultimately limit the dissemination potential of clinical multidrug resistance plasmids in these hosts....

  12. Plasmid-associated transfer of tetracycline resistance in Bacteroides ruminicola.

    OpenAIRE

    Flint, H J; Thomson, A. M.; Bisset, J

    1988-01-01

    Tetracycline resistance was transferred at frequencies between 10(-7) and 10(-6) per recipient cell in anaerobic matings between two strains of the strictly anaerobic rumen bacterium Bacteroides ruminicola. The donor strain, 223/M2/7, was a multiple-plasmid-bearing tetracycline-resistant strain from the ovine rumen, and the recipient, F101, was a rifampin-resistant mutant of B14, a bovine strain belonging to B. ruminicola subsp. brevis. Resistance transfer could occur in the presence of DNase...

  13. Diversity of Clavulanic Acid-Inhibited Extended-Spectrum β-Lactamases in Aeromonas spp. from the Seine River, Paris, France▿

    Science.gov (United States)

    Girlich, Delphine; Poirel, Laurent; Nordmann, Patrice

    2011-01-01

    Environmental Aeromonas sp. isolates resistant to ceftazidime were recovered during an environmental survey performed with water samples from the Seine River, in Paris, France, in November 2009. Selected isolates were identified by sequencing of the 16S rRNA and rpoB genes. PCR and cloning experiments were used to identify broad-spectrum-β-lactamase-encoding genes and their genetic context. Clavulanic acid-inhibited extended-spectrum-β-lactamase (ESBL) genes were identified in 71% of the Aeromonas sp. isolates. A variety of ESBL genes were detected, including blaVEB-1a, blaSHV-12, blaPER-1, blaPER-6, blaTLA-2, and blaGES-7, suggesting an aquatic reservoir of those ESBL genes. Moreover, the repeated elements and different insertion sequences were identified in association with the blaPER-6 and the blaVEB-1a genes, respectively, indicating a wide diversity of mobilization events, making Aeromonas spp. a vehicle for ESBL dissemination. PMID:21149627

  14. Role of plasmids in Lactobacillus brevis BSO 464 hop tolerance and beer spoilage.

    Science.gov (United States)

    Bergsveinson, Jordyn; Baecker, Nina; Pittet, Vanessa; Ziola, Barry

    2015-02-01

    Specific isolates of lactic acid bacteria (LAB) can grow in the harsh beer environment, thus posing a threat to brew quality and the economic success of breweries worldwide. Plasmid-localized genes, such as horA, horC, and hitA, have been suggested to confer hop tolerance, a trait required for LAB survival in beer. The presence and expression of these genes among LAB, however, do not universally correlate with the ability to grow in beer. Genome sequencing of the virulent beer spoilage organism Lactobacillus brevis BSO 464 revealed the presence of eight plasmids, with plasmids 1, 2, and 3 containing horA, horC, and hitA, respectively. To investigate the roles that these and the other five plasmids play in L. brevis BSO 464 growth in beer, plasmid curing with novobiocin was used to derive 10 plasmid variants. Multiplex PCRs were utilized to determine the presence or absence of each plasmid, and how plasmid loss affected hop tolerance and growth in degassed (noncarbonated) beer was assessed. Loss of three of the eight plasmids was found to affect hop tolerance and growth in beer. Loss of plasmid 2 (horC and 28 other genes) had the most dramatic effect, with loss of plasmid 4 (120 genes) and plasmid 8 (47 genes) having significant, but smaller, impacts. These results support the contention that genes on mobile genetic elements are essential for bacterial growth in beer and that beer spoilage ability is not dependent solely on the three previously described hop tolerance genes or on the chromosome of a beer spoilage LAB isolate.

  15. Characterization and molecular cloning in Escherichia coli of a plasmid from the mollicute Spiroplasma citri.

    OpenAIRE

    Mouches, C; Barroso, G.; Bové, J M

    1983-01-01

    Two plasmids, pMH1 with 7 kilobase pairs and pM41 with 8 kilobase pairs, were purified from the plant pathogen Spiroplasma citri and characterized by restriction mapping. Upon in vitro DNA recombination with plasmid pBR328 as a vector, we have cloned pMH1 in Escherichia coli. A radioactive probe obtained upon nick translation of the recombinant plasmid was used to further characterize and compare pMH1 and pM41.

  16. Cloning in Streptococcus lactis of plasmid-mediated UV resistance and effect on prophage stability

    International Nuclear Information System (INIS)

    Plasmid pIL7 (33 kilobases) from Streptococcus lactis enhances UV resistance and prophage stability. A 5.4-kilobase pIL7 fragment carrying genes coding for both characters was cloned into S. lactis, using plasmid pHV1301 as the cloning vector. The recombinant plasmid was subsequently transferred to three other S. lactis strains by transformation or protoplast fusion. Cloned genes were expressed in all tested strains

  17. Reporter gene expression in dendritic cells after gene gun administration of plasmid DNA

    OpenAIRE

    Watkins, Craig; Hopkins, John; Harkiss, Gordon

    2005-01-01

    Dendritic cells (DC) play an integral role in plasmid DNA vaccination. However, the interaction between plasmid DNA and DC in vivo is incompletely understood. In this report, we utilise the sheep pseudoafferent cannulation model to examine the interaction between plasmid DNA encoding enhanced green fluorescent protein (pEGFP) and afferent lymph DC (ALDC) following gene gun administration. The results show that peaks of fluorescent ALDC tended to appear around days 1-4 and 9-13, then erratical...

  18. A stable luciferase reporter plasmid for in vivo imaging in murine models of Staphylococcus aureus infections.

    Science.gov (United States)

    Bacconi, Marta; Haag, Andreas F; Torre, Antonina; Castagnetti, Andrea; Chiarot, Emiliano; Delany, Isabel; Bensi, Giuliano

    2016-04-01

    In vivo imaging of bioluminescent bacteria permits their visualization in infected mice, allowing spatial and temporal evaluation of infection progression. Most available bioluminescent strains were obtained by integration of the luciferase genes into the bacterial chromosome, a challenging and time-consuming approach. Recently, episomal plasmids were used, which were introduced in bacteria and expressed all genes required for bioluminescence emission. However, the plasmid was progressively lost in vitro and in vivo, if bacteria were not maintained under antibiotic selective pressure. Increased stability could be obtained inserting into the plasmid backbone sequences that assured plasmid partition between daughter bacterial cells, or caused death of bacteria that had lost the plasmid. So far, no detailed analysis was performed of either plasmid stability in vivo or contribution of different stabilizing sequence types. Here we report the construction of a plasmid, which includes the Photorhabdus luminescens lux cassette expressed under the control of a Staphylococcus aureus specific gene promoter, and toxin/antitoxin (T/A) and partition sequences (Par) conferring stability and transmissibility of the plasmid. Following infection of mice with S. aureus carrying this plasmid, we demonstrated that the promoter-lux fusion was functional in vivo, that the plasmid was retained by 70-100% of bacterial cells 7 days post-infection, and that both stabilizing sequence types were required to maximize plasmid retention. These data suggest that the plasmid can be a valuable tool to study gene expression and bacterial spread in small laboratory animals infected with S. aureus or possibly other Gram-positive human pathogens. PMID:26685857

  19. A New Extant Respirometric Assay to Estimate Intrinsic Growth Parameters Applied to Study Plasmid Metabolic Burden

    DEFF Research Database (Denmark)

    Seoane, Jose Miguel; Sin, Gürkan; Lardon, Laurent;

    2010-01-01

    burden caused by the carriage of a pWW0 TOL plasmid in the model organism Pseudomonas putida KT2440; The metabolic,burden associated was manifested as a reduction in the yield and the specific growth rate of the host, with both plasmid maintenance and the over-expression of recombinant proteins from...... the plasmid contributing equally to the overall effect. Biotechnol. Bioeng. 2010;105: 141-149. (C) 2009 Wiley Periodicals, Inc....

  20. Physical characterization of plasmids determining synthesis of a microcin which inhibits methionine synthesis in Escherichia coli.

    OpenAIRE

    Perez-Diaz, J C; Clowes, R C

    1980-01-01

    Plasmid deoxyribonucleic acid (DNA) isolated from each of three antibiotic-resistant clinical strains of Escherichia coli producing the same microcin showed multiple bands upon agarose gel electrophoresis. Transformants selected either for microcin resistance or ampicillin resistance yielded plasmid DNA corresponding in size to only one of the multiple bands. Plasmids, isolated from all three hosts, which determined microcin resistance and microcin production measured about 4 megadaltons by s...

  1. Parallel Compensatory Evolution Stabilizes Plasmids across the Parasitism-Mutualism Continuum

    OpenAIRE

    Harrison, Ellie; Guymer, David; Spiers, Andrew J.; Paterson, Steve; Brockhurst, Michael A.

    2015-01-01

    Plasmids drive genomic diversity in bacteria via horizontal gene transfer [1, 2]; nevertheless, explaining their survival in bacterial populations is challenging [3]. Theory predicts that irrespective of their net fitness effects, plasmids should be lost: when parasitic (costs outweigh benefits), plasmids should decline due to purifying selection [4-6], yet under mutualism (benefits outweigh costs), selection favors the capture of beneficial accessory genes by the chromosome and loss of the c...

  2. Stability of plasmid content in Salmonella wien in late phases of the epidemic history.

    OpenAIRE

    Casalino, M.; Comanducci, A; Nicoletti, M; Maimone, F

    1984-01-01

    Prevalence, genetic characteristics, and EcoRI cleavage analysis of plasmids identified in clinical strains of Salmonella wien isolated in recent years showed that the plasmid content in this serotype has remained uniform and stable over more than a decade and also late in the epidemic history. No correlation between decrease in S. wien isolations and naturally occurring systematic changes in the DNA of its most common FIme plasmid was structurally detectable.

  3. Differences in the stability of the plasmids of Yersinia pestis cultures in vitro: impact on virulence

    OpenAIRE

    TC Leal-Balbino; NC Leal; CV Lopes; AMP de Almeida

    2004-01-01

    Plasmid and chromosomal genes encode determinants of virulence for Yersinia pestis, the causative agent of plague. However, in vitro, Y. pestis genome is very plastic and several changes have been described. To evaluate the alterations in the plasmid content of the cultures in vitro and the impact of the alterations to their pathogenicity, three Y. pestis isolates were submitted to serial subculture, analysis of the plasmid content, and testing for the presence of characteristic genes in each...

  4. Adaptation of model genetically engineered microorganisms to lake water: growth rate enhancements and plasmid loss.

    OpenAIRE

    Sobecky, P A; Schell, M A; Moran, M. A.; Hodson, R. E.

    1992-01-01

    When a genetically engineered microorganism (GEM) is released into a natural ecosystem, its survival, and hence its potential environmental impact, depends on its genetic stability and potential for growth under highly oligotrophic conditions. In this study, we compared plasmid stability and potential for growth on low concentrations of organic nutrients of strains of Pseudomonas putida serving as model GEMs. Plasmid-free and plasmid-bearing (NAH7) prototrophic isogenic strains and two amino-...

  5. Cloning in Streptococcus lactis of plasmid-mediated UV resistance and effect on prophage stability.

    OpenAIRE

    Chopin, M C; Chopin, A; Rouault, A.; Simon, D.

    1986-01-01

    Plasmid pIL7 (33 kilobases) from Streptococcus lactis enhances UV resistance and prophage stability. A 5.4-kilobase pIL7 fragment carrying genes coding for both characters was cloned into S. lactis, using plasmid pHV1301 as the cloning vector. The recombinant plasmid was subsequently transferred to three other S. lactis strains by transformation or protoplast fusion. Cloned genes were expressed in all tested strains.

  6. Replication and segregational stability of Bacillus plasmid pBAA1.

    OpenAIRE

    DEVINE, KEVIN; MC CONNELL, DAVID JOHN

    1989-01-01

    PUBLISHED A cryptic plasmid, pBAA1, was identified in an industrial Bacillus strain. The plasmid is 6.8 kilobases in size and is present in cells at a copy number of approximately 5 per chromosome equivalent. The plasmid has been maintained under industrial fermentation conditions without apparent selective pressure and so is assumed to be partition proficient. The minimal replicon was localized to a 1.4-kilobase fragment which also contains the functions required for copy number control. ...

  7. A cell engineering strategy to enhance supercoiled plasmid DNA production for gene therapy

    OpenAIRE

    Hassan, S.; Keshavarz-Moore, E.; J. Ward

    2016-01-01

    With the recent revival of the promise of plasmid DNA vectors in gene therapy, a novel synthetic biology approach was used to enhance the quantity, (yield), and quality of the plasmid DNA. Quality was measured by percentage supercoiling and supercoiling density, as well as improving segregational stability in fermentation. We examined the hypothesis that adding a Strong Gyrase binding Site (SGS) would increase DNA gyrase-mediated plasmid supercoiling. SGS from 3 different replicons, (the Mu b...

  8. Cefotaxime resistant Escherichia coli collected from a healthy volunteer; characterisation and the effect of plasmid loss.

    Directory of Open Access Journals (Sweden)

    Miranda Kirchner

    Full Text Available In this study 6 CTX-M positive E. coli isolates collected during a clinical study examining the effect of antibiotic use in a human trial were analysed. The aim of the study was to analyse these isolates and assess the effect of full or partial loss of plasmid genes on bacterial fitness and pathogenicity. A DNA array was utilised to assess resistance and virulence gene carriage. Plasmids were characterised by PCR-based replicon typing and addiction system multiplex PCR. A phenotypic array and insect virulence model were utilised to assess the effect of plasmid-loss in E. coli of a large multi-resistance plasmid. All six E. coli carrying bla CTX-M-14 were detected from a single participant and were identical by pulse field gel electrophoresis and MLST. Plasmid profiling and arrays indicated absence of a large multi-drug resistance (MDR F-replicon plasmid carrying blaTEM, aadA4, strA, strB, dfrA17/19, sul1, and tetB from one isolate. Although this isolate partially retained the plasmid it showed altered fitness characteristics e.g. inability to respire in presence of antiseptics, similar to a plasmid-cured strain. However, unlike the plasmid-cured or plasmid harbouring strains, the survival rate for Galleria mellonella infected by the former strain was approximately 5-times lower, indicating other possible changes accompanying partial plasmid loss. In conclusion, our results demonstrated that an apparently healthy individual can harbour bla CTX-M-14 E. coli strains. In one such strain, isolated from the same individual, partial absence of a large MDR plasmid resulted in altered fitness and virulence characteristics, which may have implications in the ability of this strain to infect and any subsequent treatment.

  9. Selective Conditions for a Multidrug Resistance Plasmid Depend on the Sociality of Antibiotic Resistance

    OpenAIRE

    Bottery, Michael; Wood, A. Jamie; Brockhurst, Michael

    2016-01-01

    Multidrug resistance (MDR) plasmids frequently carry antibiotic resistance genes conferring qualitatively different mechanisms of resistance. We show here that the antibiotic concentrations selecting for the RK2 plasmid in Escherichia coli depend upon the sociality of the drug resistance: the selection for selfish drug resistance (efflux pump) occurred at very low drug concentrations, just 1.3% of the MIC of the plasmid-free antibiotic-sensitive strain, whereas selection for cooperative drug ...

  10. Plasmids which make their host bacteria mutable as well as resistant to ultraviolet irradiation

    International Nuclear Information System (INIS)

    Some of the naturally occurring Iα, I zeta, M, N, O and T group plasmids increase both the mutability and UV resistance of their host bacteria, while group H and S plasmids only increase mutability. This suggests that these two plasmid-mediated repair functions are separable. The two functions have no direct relation to their restriction-modification systems and nitrofuran resistant functions. In addition, the close linking between the restriction-modification genes and these repair function genes was suggested in group N plasmids. (author)

  11. Modular construction of plasmids by parallel assembly of linear vector components.

    Science.gov (United States)

    Gao, XinZheng; Yan, Pu; Shen, Wentao; Li, Xiaoying; Zhou, Peng; Li, Yuenan

    2013-06-15

    Construction of plasmids is the basic and pivotal technology in molecular biology. The common method for constructing plasmids is to cut DNA fragments by restriction enzymes and then join the resulting fragments using ligase. We present here a modified Golden Gate cloning method for modular construction of plasmids. Unlike the original Golden Gate cloning system for cloning from entry vector to expression vector, this method can be used to construct plasmids immediately from linear DNA fragments. After polymerase chain reaction (PCR) amplification for flanking with BsaI sites, multiple linear DNA components (modules) can be parallel assembled into a circle plasmid by a single restriction-ligation reaction using the method. This method is flexible to construct different types of plasmids because the modules can be freely selected and assembled in any combination. This method was applied successfully to construct a prokaryotic expression plasmid from four modules and a plant expression plasmid from five modules (fragments). The results suggest that this method provides a simple and flexible platform for modular construction of plasmids.

  12. Effects of medium composition on the production of plasmid DNA vector potentially for human gene therapy

    Institute of Scientific and Technical Information of China (English)

    XU Zhi-nan; SHEN Wen-he; CHEN Hao; CEN Pei-lin

    2005-01-01

    Plasmid vector is increasingly applied to gene therapy or gene vaccine. The production of plasmid pCMV-AP3 for cancer gene therapy was conducted in a modified MBL medium using a recombinant E. coli BL21 system. The effects of different MMBL components on plasmid yield, cell mass and specific plasmid DNA productivity were evaluated on shake-flask scale. The results showed that glucose was the optimal carbon source. High plasmid yield (58.3 mg/L) was obtained when 5.0 g/L glucose was added to MMBL. Glycerol could be chosen as a complementary carbon source because of the highest specific plasmid productivity (37.9 mg DNA/g DCW). After tests of different levels of nitrogen source and inorganic phosphate, a modified MMBL medium was formulated for optimal plasmid production. Further study showed that the initial acetate addition (less than 4.0 g/L) in MMBL improved plasmid production significantly, although it inhibited cell growth. The results will be useful for large-scale plasmid production using recombinant E. coli system.

  13. Peaceful coexistence amongst Borrelia plasmids: getting by with a little help from their friends?

    OpenAIRE

    Chaconas, George; Norris, Steven J

    2013-01-01

    Borrelia species comprise a unique genus of bacterial pathogens. These organisms contain a segmented genome with up to two dozen plasmids ranging in size from 5kb up to about 200 kb. The plasmids have also been referred to as mini-chromosomes or essential genetic elements, as some of them carry information important for infection of vertebrates or for survival in the tick vector. Most of the plasmids are linear with covalently closed hairpin telomeres and these linear plasmids are in a consta...

  14. Postsymbiotic plasmid acquisition and evolution of the repA1-replicon in Buchnera aphidicola

    Science.gov (United States)

    Van Ham, Roeland C. H. J.; González-Candelas, Fernando; Silva, Francisco J.; Sabater, Beatriz; Moya, Andrés; Latorre, Amparo

    2000-01-01

    Buchnera aphidicola is an obligate, strictly vertically transmitted, bacterial symbiont of aphids. It supplies its host with essential amino acids, nutrients required by aphids but deficient in their diet of plant phloem sap. Several lineages of Buchnera show adaptation to their nutritional role in the form of plasmid-mediated amplification of key-genes involved in the biosynthesis of tryptophan (trpEG) and leucine (leuABCD). Phylogenetic analyses of these plasmid-encoded functions have thus far suggested the absence of horizontal plasmid exchange among lineages of Buchnera. Here, we describe three new Buchnera plasmids, obtained from species of the aphid host families Lachnidae and Pemphigidae. All three plasmids belong to the repA1 family of Buchnera plasmids, which is characterized by the presence of a repA1-replicon responsible for replication initiation. A comprehensive analysis of this family of plasmids unexpectedly revealed significantly incongruent phylogenies for different plasmid and chromosomally encoded loci. We infer from these incongruencies a case of horizontal plasmid transfer in Buchnera. This process may have been mediated by secondary endosymbionts, which occasionally undergo horizontal transmission in aphids. PMID:10984505

  15. High-frequency transformation of Brevibacterium lactofermentum protoplasts by plasmid DNA.

    OpenAIRE

    Santamaria, R I; Gil, J.A.; Martin, J. F.

    1985-01-01

    An efficient polyethylene glycol-assisted method for transformation of Brevibacterium lactofermentum protoplasts that uses plasmid vectors has been developed. Two small plasmids, pUL330 (5.2 kilobases) and pUL340 (5.8 kilobases), both containing the kanamycin resistance gene from transposon Tn5 and the replication origin of the natural plasmid pBL1 of B. lactofermentum, were selected as vectors. Supercoiled forms of the plasmids yielded a 100-fold higher transformation frequency than did line...

  16. Quantifying and visualizing the transfer of exogenous plasmids to environmental microbial communities

    DEFF Research Database (Denmark)

    Dechesne, Arnaud

    2015-01-01

    Plasmid transfer is deemed responsible for the rapid spread of antibiotic resistance among microbes. While broad host range plasmids are known to transfer to diverse hosts in pure culture, their transfer potential to complex communities has not been comprehensively studied. The ability...... take up a model plasmid?) and in term of diversity (what type of bacteria take up the plasmid?). The method takes advantage of fluorescent marker genes, image analysis, flow cytometry and next generation sequencing. We revealed that an unexpectedly high diversity of soil microbes can take up broad host...

  17. Replication of the R6K plasmid during the Escherichia coli cell cycle.

    OpenAIRE

    Keasling, J.D.; Palsson, B O; Cooper, S.

    1992-01-01

    The cell-cycle replication pattern of the R6K plasmid has been investigated by using the membrane-elution technique to produce cells labelled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The high-copy plasmid R6K replicates exponentially in a cell-cycle-independent manner. A mini-R6K plasmid deleted for the ori alpha origin of replication also replicates, exponentially in a cell-cycle-independent manner.

  18. The 2-micron plasmid as a nonselectable, stable, high copy number yeast vector

    Science.gov (United States)

    Ludwig, D. L.; Bruschi, C. V.

    1991-01-01

    The endogenous 2-microns plasmid of Saccharomyces cerevisiae has been used extensively for the construction of yeast cloning and expression plasmids because it is a native yeast plasmid that is able to be maintained stably in cells at high copy number. Almost invariably, these plasmid constructs, containing some or all 2-microns sequences, exhibit copy number levels lower than 2-microns and are maintained stably only under selective conditions. We were interested in determining if there was a means by which 2-microns could be utilized for vector construction, without forfeiting either copy number or nonselective stability. We identified sites in the 2-microns plasmid that could be used for the insertion of genetic sequences without disrupting 2-microns coding elements and then assessed subsequent plasmid constructs for stability and copy number in vivo. We demonstrate the utility of a previously described 2-microns recombination chimera, pBH-2L, for the manipulation and transformation of 2-microns as a pure yeast plasmid vector. We show that the HpaI site near the STB element in the 2-microns plasmid can be utilized to clone yeast DNA of at least 3.9 kb with no loss of plasmid stability. Additionally, the copy number of these constructs is as high as levels reported for the endogenous 2-microns.

  19. Selective Conditions for a Multidrug Resistance Plasmid Depend on the Sociality of Antibiotic Resistance.

    Science.gov (United States)

    Bottery, Michael J; Wood, A Jamie; Brockhurst, Michael A

    2016-04-01

    Multidrug resistance (MDR) plasmids frequently carry antibiotic resistance genes conferring qualitatively different mechanisms of resistance. We show here that the antibiotic concentrations selecting for the RK2 plasmid inEscherichia colidepend upon the sociality of the drug resistance: the selection for selfish drug resistance (efflux pump) occurred at very low drug concentrations, just 1.3% of the MIC of the plasmid-free antibiotic-sensitive strain, whereas selection for cooperative drug resistance (modifying enzyme) occurred at drug concentrations exceeding the MIC of the plasmid-free strain. PMID:26787694

  20. Simian virus 40 promoters direct expression of the tetracycline gene in plasmid pACYC184.

    OpenAIRE

    Jenkins, F J; Howett, M K; Rapp, F

    1983-01-01

    Insertion of HindIII DNA fragments into the HindIII site of plasmid pACYC184 destroys the promoter of the plasmid tetracycline resistance gene and causes Escherichia coli cells harboring recombinant plasmids to be tetracycline sensitive and chloramphenicol resistant. The HindIII-C DNA fragment of simian virus 40 contains the two virus promoters and the virus origin of replication. We report the isolation of recombinant plasmids that contained the simian virus 40 HindIII-C DNA fragment at the ...