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Sample records for stenotrophomonas maltophilia cgmcc

  1. [Three cases of Stenotrophomonas maltophilia pneumonia].

    PubMed

    Fujino, Satoru; Hisatomi, Keiko; Iida, Tetsuya; Ohe, Nobuharu; Hirakata, Yoichi; Hara, Kohei

    2003-07-01

    We encountered 3 cases of pneumonia caused by Stenotrophomonas maltophilia between January and June 2001. S. maltophilia is resistant to broad-spectrum antibiotics including carbapenem. Reported studies indicate that excessive use of broad-spectrum antibiotics may induce resistance in this organism. However, our data showed that there was no clear correlation between the amounts of carbapenems used in our hospital and the isolation of the organism. If broad-spectrum antibiotics are ineffective or even actually worsen a case of pneumonia, S. maltophilia may be the sole causative organism, and a potent double- (or triple-) combination therapy consisting of minocyclin and one or two other potent antimicrobial agents should be considered.

  2. Stenotrophomonas maltophilia: an Emerging Global Opportunistic Pathogen

    PubMed Central

    2012-01-01

    Summary: Stenotrophomonas maltophilia is an emerging multidrug-resistant global opportunistic pathogen. The increasing incidence of nosocomial and community-acquired S. maltophilia infections is of particular concern for immunocompromised individuals, as this bacterial pathogen is associated with a significant fatality/case ratio. S. maltophilia is an environmental bacterium found in aqueous habitats, including plant rhizospheres, animals, foods, and water sources. Infections of S. maltophilia can occur in a range of organs and tissues; the organism is commonly found in respiratory tract infections. This review summarizes the current literature and presents S. maltophilia as an organism with various molecular mechanisms used for colonization and infection. S. maltophilia can be recovered from polymicrobial infections, most notably from the respiratory tract of cystic fibrosis patients, as a cocolonizer with Pseudomonas aeruginosa. Recent evidence of cell-cell communication between these pathogens has implications for the development of novel pharmacological therapies. Animal models of S. maltophilia infection have provided useful information about the type of host immune response induced by this opportunistic pathogen. Current and emerging treatments for patients infected with S. maltophilia are discussed. PMID:22232370

  3. Stenotrophomonas maltophilia: an emerging global opportunistic pathogen.

    PubMed

    Brooke, Joanna S

    2012-01-01

    Stenotrophomonas maltophilia is an emerging multidrug-resistant global opportunistic pathogen. The increasing incidence of nosocomial and community-acquired S. maltophilia infections is of particular concern for immunocompromised individuals, as this bacterial pathogen is associated with a significant fatality/case ratio. S. maltophilia is an environmental bacterium found in aqueous habitats, including plant rhizospheres, animals, foods, and water sources. Infections of S. maltophilia can occur in a range of organs and tissues; the organism is commonly found in respiratory tract infections. This review summarizes the current literature and presents S. maltophilia as an organism with various molecular mechanisms used for colonization and infection. S. maltophilia can be recovered from polymicrobial infections, most notably from the respiratory tract of cystic fibrosis patients, as a cocolonizer with Pseudomonas aeruginosa. Recent evidence of cell-cell communication between these pathogens has implications for the development of novel pharmacological therapies. Animal models of S. maltophilia infection have provided useful information about the type of host immune response induced by this opportunistic pathogen. Current and emerging treatments for patients infected with S. maltophilia are discussed.

  4. Polysorbate 80 and polymyxin B inhibit Stenotrophomonas maltophilia biofilm.

    PubMed

    Malinowski, Adam M; McClarty, Bryan M; Robinson, Carolyn; Spear, William; Sanchez, Maria; Sparkes, Timothy C; Brooke, Joanna S

    2017-02-01

    Stenotrophomonas maltophilia is an opportunistic multiple-drug-resistant human pathogen that forms biofilms on implanted medical devices. We examined the potential inhibitory activity of polysorbate 80 and polymyxin B against S. maltophilia. A combination of subMIC polymyxin B and polysorbate 80 was the most effective inhibitor of growth and biofilm formation.

  5. Microbiological and Clinical Aspects of Infection Associated with Stenotrophomonas maltophilia

    PubMed Central

    Denton, Miles; Kerr, Kevin G.

    1998-01-01

    The gram-negative bacterium Stenotrophomonas maltophilia is increasingly recognized as an important cause of nosocomial infection. Infection occurs principally, but not exclusively, in debilitated and immunosuppressed individuals. Management of S. maltophilia-associated infection is problematic because many strains of the bacterium manifest resistance to multiple antibiotics. These difficulties are compounded by methodological problems in in vitro susceptibility testing for which there are, as yet, no formal guidelines. Despite its acknowledged importance as a nosocomial pathogen, little is known of the epidemiology of S. maltophilia, and although it is considered an environmental bacterium, its sources and reservoirs are often not readily apparent. Molecular typing systems may contribute to our knowledge of the epidemiology of S. maltophilia infection, thus allowing the development of strategies to interrupt the transmission of the bacterium in the hospital setting. Even less is known of pathogenic mechanisms and putative virulence factors involved in the natural history of S. maltophilia infection and this, coupled with difficulties in distinguishing colonization from true infection, has fostered the view that the bacterium is essentially nonpathogenic. This article aims to review the current taxonomic status of S. maltophilia, and it discusses the laboratory identification of the bacterium. The epidemiology of the organism is considered with particular reference to nosocomial outbreaks, several of which have been investigated by molecular typing techniques. Risk factors for acquisition of the bacterium are also reviewed, and the ever-expanding spectrum of clinical syndromes associated with S. maltophilia is surveyed. Antimicrobial resistance mechanisms, pitfalls in in vitro susceptibility testing, and therapy of S. maltophilia infections are also discussed. PMID:9457429

  6. Stenotrophomonas maltophilia: An Emerging Pathogen in Paediatric Population

    PubMed Central

    Nayyar, Charu; Thakur, Preeti; Saigal, Karnika

    2017-01-01

    Introduction Stenotrophomonas maltophilia (formerly Pseudomonas maltophilia/Xanthomonas maltophilia), a Gram- negative, non-fermenting bacillus, is being increasingly recognized as a threatening nosocomial pathogen, associated with significant mortality. Aim To determine the prevalence of infection, antimicrobial susceptibility pattern and clinical outcome of S. maltophilia in a paediatric population. Materials and Methods This was a retrospective study conducted over a period of eight months, i.e., October 2015 to May 2016. All clinical samples received in the microbiology laboratory during the study period were processed using standard microbiological procedures. S. maltophilia isolates were selected. Antibiotic susceptibility was performed for levofloxacin and trimethoprim-sulphamethoxazole by Vitek 2C system (Biomerieux, France). Average length of stay and mortality caused by S. maltophilia infection was compared with age and sex matched controls without S. maltophilia infection. Results A total of 16,234 clinical specimens were received in the microbiology laboratory in the study period, with 2,734 pathogenic bacteria isolated. A total of 1,339 (1.7% of total isolates) Gram-negative bacteria were isolated, out of which 414 were non-fermenters. Among the non-fermenters, 23 (5.5%) were S. maltophilia. Out of the 23 isolates, 15 (65.2%) were isolated from blood, 4 (17.3%) were isolated from urine and tracheal aspirate each. A total of 91.3% of strains were susceptible and 8.6% were resistant to trimethoprim-sulphamethoxazole. Total 80% of strains were sensitive and 20% had intermediate susceptibility for levofloxacin. None of the strains were resistant to levofloxacin. Average length of stay of patients with S. maltophilia infection was found out to be 23.3 days as compared to 44.8 days in controls. The average mortality of patients with S. maltophilia infection was found to be same as that of controls (35.2%). Conclusion S. maltophilia is becoming an important

  7. Antibiotic resistance in the opportunistic pathogen Stenotrophomonas maltophilia.

    PubMed

    Sánchez, María B

    2015-01-01

    Stenotrophomonas maltophilia is an environmental bacterium found in the soil, associated with plants and animals, and in aquatic environments. It is also an opportunistic pathogen now causing an increasing number of nosocomial infections. The treatment of S. maltophilia is quite difficult given its intrinsic resistance to a number of antibiotics, and because it is able to acquire new resistances via horizontal gene transfer and mutations. Certainly, strains resistant to quinolones, cotrimoxale and/or cephalosporins-antibiotics commonly used to treat S. maltophilia infections-have emerged. The increasing number of available S. maltophilia genomes has allowed the identification and annotation of a large number of antimicrobial resistance genes. Most encode inactivating enzymes and efflux pumps, but information on their role in intrinsic and acquired resistance is limited. Non-typical antibiotic resistance mechanisms that also form part of the intrinsic resistome have been identified via mutant library screening. These include non-typical antibiotic resistance genes, such as bacterial metabolism genes, and non-inheritable resistant phenotypes, such as biofilm formation and persistence. Their relationships with resistance are complex and require further study.

  8. Molecular characterization of 2-chlorobiphenyl degrading Stenotrophomonas maltophilia GS-103.

    PubMed

    Somaraja, P K; Gayathri, D; Ramaiah, N

    2013-08-01

    The catabolic potential of transformer oil contaminated soil bacteria in aerobic degradation of polychlorinated biphenyls (PCB) were assessed. Transformer oil contaminated soil sample was subjected to microcosm enrichment experiments (PAS medium/biphenyl as sole carbon source). PCB-degrading activity of the enrichment cultures in PAS medium with the addition of 2-chlorobiphenyl were analysed by GC-MS indicated that, although the isolates differed in PCB-degrading capabilities, all of the enrichment cultures expressed activity toward at least some of the lower chlorinated congeners. Biphenyl-utilizing bacteria isolated from the most active PCB-degrading mixed cultures showed little taxonomic diversity and identified as Stenotrophomonas maltophilia GS-103.

  9. Dengue co-infection in a blood stream infection caused by Stenotrophomonas maltophilia: A case report.

    PubMed

    Srirangaraj, Sreenivasan; Kali, Arunava; Vijayan, Sivaranjini

    2014-01-01

    Stenotrophomonas maltophilia (S. maltophilia) is an emerging opportunistic bacterial pathogen with resistance to several commonly used antibiotics. Owing to its multidrug resistance (MDR), management of S. maltophilia blood stream infection (BSI) is challenging and requires the selection of appropriate antibiotic therapy. The presence of thrombocytopenia and shock are independent risk factors associated with increased mortality in patients with S. maltophilia BSI. We describe an unusual case of S. maltophilia BSI in a middle-age female complicated by dengue fever. We highlight the importance of early recognition of both dengue and S. maltophilia infection in management of such cases.

  10. Polymorphic Mutation Frequencies of Clinical and Environmental Stenotrophomonas maltophilia Populations▿

    PubMed Central

    Turrientes, María Carmen; Baquero, María Rosario; Sánchez, María Blanca; Valdezate, Sylvia; Escudero, Esther; Berg, Gabrielle; Cantón, Rafael; Baquero, Fernando; Galán, Juan Carlos; Martínez, José Luis

    2010-01-01

    Mutation frequencies were studied in 174 Stenotrophomonas maltophilia isolates from clinical and nonclinical environments by detecting spontaneous rifampin-resistant mutants in otherwise-susceptible populations. The distribution of mutation frequencies followed a pattern similar to that found for other bacterial species, with a modal value of 1 × 10−8. Nevertheless, the proportion of isolates showing mutation frequencies below the modal value (hypomutators) was significantly higher for S. maltophilia than those so far reported in other organisms. Low mutation frequencies were particularly frequent among environmental S. maltophilia strains (58.3%), whereas strong mutators were found only among isolates with a clinical origin. These results indicate that clinical environments might select bacterial populations with high mutation frequencies, likely by second-order selection processes. In several of the strong-mutator isolates, functional-complementation assays with a wild-type allele of the mutS gene demonstrated that the mutator phenotype was due to the impairment of MutS activity. In silico analysis of the amino acid changes present in the MutS proteins of these hypermutator strains in comparison with the normomutator isolates suggests that the cause of the defect in MutS might be a H683P amino acid change. PMID:20097818

  11. Polymorphic mutation frequencies of clinical and environmental Stenotrophomonas maltophilia populations.

    PubMed

    Turrientes, María Carmen; Baquero, María Rosario; Sánchez, María Blanca; Valdezate, Sylvia; Escudero, Esther; Berg, Gabrielle; Cantón, Rafael; Baquero, Fernando; Galán, Juan Carlos; Martínez, José Luis

    2010-03-01

    Mutation frequencies were studied in 174 Stenotrophomonas maltophilia isolates from clinical and nonclinical environments by detecting spontaneous rifampin-resistant mutants in otherwise-susceptible populations. The distribution of mutation frequencies followed a pattern similar to that found for other bacterial species, with a modal value of 1 x 10(-8). Nevertheless, the proportion of isolates showing mutation frequencies below the modal value (hypomutators) was significantly higher for S. maltophilia than those so far reported in other organisms. Low mutation frequencies were particularly frequent among environmental S. maltophilia strains (58.3%), whereas strong mutators were found only among isolates with a clinical origin. These results indicate that clinical environments might select bacterial populations with high mutation frequencies, likely by second-order selection processes. In several of the strong-mutator isolates, functional-complementation assays with a wild-type allele of the mutS gene demonstrated that the mutator phenotype was due to the impairment of MutS activity. In silico analysis of the amino acid changes present in the MutS proteins of these hypermutator strains in comparison with the normomutator isolates suggests that the cause of the defect in MutS might be a H683P amino acid change.

  12. Infective endocarditis caused by Stenotrophomonas maltophilia: A report of two cases and review of literature.

    PubMed

    Subhani, Shaik; Patnaik, Amar N; Barik, Ramachandra; Nemani, Lalita

    2016-09-01

    Stenotrophomonas maltophilia is known for nosocomial habitat. Infective endocarditis due to this organism is rare and challenging because of resistance to multiple broad-spectrum antibiotic regimens. Early detection and appropriate antibiotic based on culture sensitivity reports are the key to its management. We report the diagnosis, treatment, and outcome of two cases of infective endocarditis caused by S. maltophilia.

  13. Draft Genome Sequence of Stenotrophomonas maltophilia SBo1 Isolated from Bactrocera oleae

    PubMed Central

    Blow, Frances; Vontas, John

    2016-01-01

    Bacteria of the genus Stenotrophomonas are ubiquitous in the environment and are increasingly associated with insects. Stenotrophomonas maltophilia SBo1 was cultured from the gut of Bactrocera oleae. The draft genome sequence presented here will inform future investigations into the nature of the interaction between insects and their microbiota. PMID:27660769

  14. A molecular biological protocol to distinguish potentially human pathogenic Stenotrophomonas maltophilia from plant-associated Stenotrophomonas rhizophila.

    PubMed

    Ribbeck-Busch, Kathrin; Roder, Anja; Hasse, Dirk; de Boer, Wietse; Martínez, José Luis; Hagemann, Martin; Berg, Gabriele

    2005-11-01

    In recent years, the importance of the Gram-negative bacterium Stenotrophomonas as an opportunistic pathogen as well as in biotechnology has increased. The aim of the present study was to develop new methods for distinguishing between strains closely related to the potentially human pathogenic Stenotrophomonas maltophilia and those closely related to the plant-associated Stenotrophomonas rhizophila. To accomplish this, 58 strains were characterized by 16S rDNA sequencing and amplified ribosomal DNA restriction analysis (ARDRA), and the occurrence of specific functional genes. Based on 16S rDNA sequences, an ARDRA protocol was developed which allowed differentiation between strains of the S. maltophilia and the S. rhizophila group. As it was known that only salt-treated cells of S. rhizophila were able to synthesize the compatible solute glucosylglycerol (GG), the ggpS gene responsible for GG synthesis was used for differentiation between both species and it was confirmed that it only occurred in S. rhizophila strains. As a further genetic marker the smeD gene, which is part of the genes coding for the multidrug efflux pump SmeDEF from S. maltophilia, was used. Based on the results we propose a combination of fingerprinting techniques using the 16S rDNA and the functional genes ggpS and smeD to distinguish both Stenotrophomonas species.

  15. New strategies against Stenotrophomonas maltophilia: a serious worldwide intrinsically drug-resistant opportunistic pathogen.

    PubMed

    Brooke, Joanna S

    2014-01-01

    Stenotrophomonas maltophilia is a worldwide human opportunistic pathogen associated with serious infections in humans, and is most often recovered from respiratory tract infections. In addition to its intrinsic drug resistance, this organism may acquire resistance via multiple molecular mechanisms. New antimicrobial strategies are needed to combat S. maltophilia infections, particularly in immunocompromised patients, cystic fibrosis patients with polymicrobial infections of the lung, and in patients with chronic infections. This editorial reports on newer drugs and antimicrobial strategies and their potential for use in treatment of S. maltophilia infections, the development of new technologies to detect this organism, and identifies strategies currently in use to reduce transmission of this pathogen.

  16. Persistence and variability of Stenotrophomonas maltophilia in cystic fibrosis patients, Madrid, 1991-1998.

    PubMed Central

    Valdezate, S.; Vindel, A.; Maiz, L.; Baquero, F.; Escobar, H.; Cantón, R.

    2001-01-01

    During 1991 to 1998 at least one Stenotrophomonas maltophilia pulmonary infection was observed in 25 (24%) of 104 cystic fibrosis patients at the same unit of our hospital in Spain. Ribotyping and pulse-field gel electrophoresis (PFGE) characterization of 76 S. maltophilia isolates from these patients indicated an overall clonal incidence of 47.1%, reflecting new strains in 44% of patients with repeated positive cultures for S. maltophilia. Six patients with repeated episodes were persistently colonized (> or = 6 months) with the same strain. S. maltophilia bacterial counts were higher (geometric mean, 2.9 x 10(8) cfu/mL) in patients with repeated episodes than in those with a single episode (8.4 x 10(4) cfu/mL, p < 0.01). Single episodes of S. maltophilia occurred in patients < 10 years of age (43% [6/14]), whereas chronic colonization occurred more frequently in older patients (> 16 years of age). PMID:11266301

  17. Genome Sequence of Stenotrophomonas maltophilia Strain SmAs1, Isolated From the Asian Malaria Mosquito Anopheles stephensi

    PubMed Central

    Hughes, Grant L.; Raygoza Garay, Juan Antonio; Koundal, Vikas; Mwangi, Michael M.

    2016-01-01

    An isolate of Stenotrophomonas maltophilia was cultured from the Asian malaria vector Anopheles stephensi. Here, we present the annotated draft genome sequence of this S. maltophilia strain. This genomic resource will facilitate further characterization of bacteria associated with mosquitoes. PMID:26966198

  18. Metagenome-Assembled Draft Genome Sequence of a Novel Microbial Stenotrophomonas maltophilia Strain Isolated from Caenorhabditis remanei Tissue

    PubMed Central

    Murdock, Duncan A.; Thanthiriwatte, Chamali; Willis, John H.; Phillips, Patrick C.

    2017-01-01

    ABSTRACT Stenotrophomonas maltophilia is a Gram-negative aerobic bacterium and emerging nosocomial pathogen. Here, we present a draft genome sequence for an S. maltophilia strain assembled from a metagenomic DNA extract isolated from a laboratory stock of the nematode worm Caenorhabditis remanei. PMID:28209833

  19. Stenotrophomonas maltophilia and Vermamoeba vermiformis relationships: bacterial multiplication and protection in amoebal-derived structures.

    PubMed

    Cateau, Estelle; Maisonneuve, Elodie; Peguilhan, Samuel; Quellard, Nathalie; Hechard, Yann; Rodier, Marie-Helene

    2014-12-01

    Stenotrophomonas maltophilia, a bacteria involved in healthcare-associated infections, can be found in hospital water systems. Other microorganisms, such as Free Living amoebae (FLA), are also at times recovered in the same environment. Amongst these protozoa, many authors have reported the presence of Vermamoeba vermiformis. We show here that this amoeba enhances S. maltophilia growth and harbors the bacteria in amoebal-derived structures after 28 days in harsh conditions. These results highlight the fact that particular attention should be paid to the presence of FLA in hospital water systems, because of their potential implication in survival and growth of pathogenic bacterial species.

  20. Successful Treatment of Bloodstream Infection Due to Metallo-β-Lactamase-Producing Stenotrophomonas maltophilia in a Renal Transplant Patient

    PubMed Central

    Mojica, Maria F.; Ouellette, Christopher P.; Leber, Amy; Becknell, M. Brian; Ardura, Monica I.; Perez, Federico; Aitken, Samuel L.

    2016-01-01

    Stenotrophomonas maltophilia is an emerging multidrug-resistant (MDR) opportunistic pathogen for which new antibiotic options are urgently needed. We report our clinical experience treating a 19-year-old renal transplant recipient who developed prolonged bacteremia due to metallo-β-lactamase-producing S. maltophilia refractory to conventional treatment. The infection recurred despite a prolonged course of colistimethate sodium (colistin) but resolved with the use of a novel drug combination with clinical efficacy against the patient's S. maltophilia isolate. PMID:27551008

  1. A Stenotrophomonas maltophilia Strain Evades a Major Caenorhabditis elegans Defense Pathway

    PubMed Central

    White, Corin V.; Darby, Brian J.; Breeden, Robert J.

    2015-01-01

    Stenotrophomonas maltophilia is a ubiquitous bacterium and an emerging nosocomial pathogen. This bacterium is resistant to many antibiotics, associated with a number of infections, and a significant health risk, especially for immunocompromised patients. Given that Caenorhabditis elegans shares many conserved genetic pathways and pathway components with higher organisms, the study of its interaction with bacterial pathogens has biomedical implications. S. maltophilia has been isolated in association with nematodes from grassland soils, and it is likely that C. elegans encounters this bacterium in nature. We found that a local S. maltophilia isolate, JCMS, is more virulent than the other S. maltophilia isolates (R551-3 and K279a) tested. JCMS virulence correlates with intestinal distension and bacterial accumulation and requires the bacteria to be alive. Many of the conserved innate immune pathways that serve to protect C. elegans from various pathogenic bacteria also play a role in combating S. maltophilia JCMS. However, S. maltophilia JCMS is virulent to normally pathogen-resistant DAF-2/16 insulin-like signaling pathway mutants. Furthermore, several insulin-like signaling effector genes were not significantly differentially expressed between S. maltophilia JCMS and avirulent bacteria (Escherichia coli OP50). Taken together, these findings suggest that S. maltophilia JCMS evades the pathogen resistance conferred by the loss of DAF-2/16 pathway components. In summary, we have discovered a novel host-pathogen interaction between C. elegans and S. maltophilia and established a new animal model with which to study the mode of action of this emerging nosocomial pathogen. PMID:26644380

  2. The flavonoid galangin inhibits the L1 metallo-beta-lactamase from Stenotrophomonas maltophilia.

    PubMed

    Denny, Brian J; Lambert, Peter A; West, Patrick W J

    2002-02-19

    The flavonoid galangin inhibits the partially purified metallo-beta-lactamase from Stenotrophomonas maltophilia. The effect was not reversed by the addition of ZnCl(2) suggesting that the inhibitory effect is not related to metal chelation. The flavonoid quercetin also has some inhibitory effect against the enzyme. Using the crystal structure of the enzyme, a molecular modelling study predicts a possible orientation of galangin at the active site of the enzyme.

  3. Stenotrophomonas maltophilia in Mexico: antimicrobial resistance, biofilm formation and clonal diversity.

    PubMed

    Flores-Treviño, Samantha; Gutiérrez-Ferman, Jessica Lizzeth; Morfín-Otero, Rayo; Rodríguez-Noriega, Eduardo; Estrada-Rivadeneyra, Diego; Rivas-Morales, Catalina; Llaca-Díaz, Jorge M; Camacho-Ortíz, Adrián; Mendoza-Olazarán, Soraya; Garza-González, Elvira

    2014-11-01

    Stenotrophomonas maltophilia is an important multidrug-resistant nosocomial pathogen associated with high mortality. Our aim was to examine antimicrobial susceptibility, biofilm production and clonal relatedness of clinical isolates of S. maltophilia. S. maltophilia isolates were collected between 2006 and 2013 from two tertiary care hospitals in Mexico. Antimicrobial susceptibility was evaluated by the broth microdilution method. PCR was used to determine the presence of β-lactamase genes L1 and L2. Biofilm formation was assessed with crystal violet staining. Clonal relatedness was determined by PFGE. Among the 119 collected S. maltophilia isolates, 73 (61.3%) were from the respiratory tract. Resistance levels exceeded 75% for imipenem, meropenem, ampicillin, aztreonam, gentamicin and tobramycin. Resistance to trimethoprim-sulfamethoxazole was 32.8%. L1 and L2 genes were detected in 77.1% (91/118) and 66.9% (79/118) of isolates, respectively. All S. maltophilia strains were able to produce biofilms. Strains were classified as weak (47.9%, 57/119), moderate (38.7%, 46/119), or strong (13.4%, 16/119) biofilm producers. A total of 89 distinct PFGE types were identified and 21.6% (22/102) of the isolates were distributed in nine clusters. This is the first study in Mexico to reveal characteristics of clinical isolates of S. maltophilia. Clonal diversity data indicate low cross-transmission of S. maltophilia in a hospital setting. The high antibiotic resistance underscores the need for continuous surveillance of S. maltophilia in hospital settings in Mexico.

  4. Clinical characteristics and prognostic factors of patients with Stenotrophomonas maltophilia infections

    PubMed Central

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

    2017-01-01

    INTRODUCTION: Stenotrophomonas maltophilia earlier had limited pathogenic potential, but now with growing degree of immunosuppression in general population, it is being recognized as an important nosocomial pathogen. METHODOLOGY: A retrospective 7 years study was carried out to determine the clinical characteristics of all patients with Stenotrophomonas infections, antibiotic resistance pattern, and risk factors associated with hospital mortality. All patients with Stenotrophomonas culture positivity were identified and their medical records were reviewed. Risk factor associated with hospital mortality was analyzed. RESULTS: A total of 123 samples obtained from 88 patients were culture positive. Most patients presented with bacteremia (45, 51%) followed by pneumonia (37, 42%) and skin and soft tissue infections (6, 7%). About 23 of 88 Stenotrophomonas infected patients had co-infection. Percentage resistance to cotrimoxazole; 8 (5.4%) was lower than that for levofloxacin; 18 (12%). Twenty-eight patients died during hospital stay. Intensive Care Unit admission (P = 0.0002), mechanical ventilation (P = 0.0004), central venous catheterization (P = 0.0227), urethral catheterization (P = 0.0484), and previous antibiotic intake (P = 0.0026) were independent risk factors associated with mortality. CONCLUSION: Our findings suggest that Stenotrophomonas can cause various infections irrespective of patient's immune status and irrespective of potential source. Thus, Stenotrophomonas should be thought of as potential pathogen and its isolation should be looked with clinical suspicion. PMID:28367030

  5. Stenotrophomonas maltophilia strains from cystic fibrosis patients: genomic variability and molecular characterization of some virulence determinants.

    PubMed

    Nicoletti, Mauro; Iacobino, Angelo; Prosseda, Gianni; Fiscarelli, Ersilia; Zarrilli, Raffaele; De Carolis, Elena; Petrucca, Andrea; Nencioni, Lucia; Colonna, Bianca; Casalino, Mariassunta

    2011-01-01

    The genetic relatedness of 52 Stenotrophomonas maltophilia strains, collected from various environmental and clinical sources, including cystic fibrosis (CF) patients, as well as the presence and the expression of some virulence-associated genes were studied. Pulsed-field gel electrophoresis (PFGE) analysis identified 47 profiles and three clusters of isolates with an identical PFGE pattern considered to be indistinguishable strains. Restriction fragment length polymorphism of the gyrB gene grouped the 52 strains into nine different profiles. Most CF clinical isolates (29 out of 41) showed profile 1, while the analysis of the hypervariable regions of the 16S rRNA gene revealed five distinct allelic variations, with the majority of CF isolates (23 out of 41) belonging to sequence group 1. Furthermore, the strains were characterized for motility and expression of virulence-associated genes, including genes encoding type-1 fimbriae, proteases (StmPr1 and StmPr2) and esterase. All S. maltophilia strains exhibited a very broad range of swimming and twitching motility, while none showed swarming motility. A complete smf-1 gene was PCR-amplified only from clinically derived S. maltophilia strains. Finally, the virulence of representative S. maltophilia strains impaired in the expression of proteases and esterase activities was evaluated by infecting larvae of the wax moth Galleria mellonella. The results obtained strongly indicate that the major extracellular protease StmPr1 may be a relevant virulence factor of S. maltophilia.

  6. Update on infections caused by Stenotrophomonas maltophilia with particular attention to resistance mechanisms and therapeutic options

    PubMed Central

    Chang, Ya-Ting; Lin, Chun-Yu; Chen, Yen-Hsu; Hsueh, Po-Ren

    2015-01-01

    Stenotrophomonas maltophilia is a Gram-negative, biofilm-forming bacterium. Although generally regarded as an organism of low virulence, S. maltophilia is an emerging multi-drug resistant opportunistic pathogen in hospital and community settings, especially among immunocompromised hosts. Risk factors associated with S. maltophilia infection include underlying malignancy, cystic fibrosis, corticosteroid or immunosuppressant therapy, the presence of an indwelling central venous catheter and exposure to broad spectrum antibiotics. In this review, we provide a synthesis of information on current global trends in S. maltophilia pathogenicity as well as updated information on the molecular mechanisms contributing to its resistance to an array of antimicrobial agents. The prevalence of S. maltophilia infection in the general population increased from 0.8–1.4% during 1997–2003 to 1.3–1.68% during 2007–2012. The most important molecular mechanisms contributing to its resistance to antibiotics include β-lactamase production, the expression of Qnr genes, and the presence of class 1 integrons and efflux pumps. Trimethoprim/sulfamethoxazole (TMP/SMX) is the antimicrobial drug of choice. Although a few studies have reported increased resistance to TMP/SMX, the majority of studies worldwide show that S. maltophilia continues to be highly susceptible. Drugs with historically good susceptibility results include ceftazidime, ticarcillin-clavulanate, and fluoroquinolones; however, a number of studies show an alarming trend in resistance to those agents. Tetracyclines such as tigecycline, minocycline, and doxycycline are also effective agents and consistently display good activity against S. maltophilia in various geographic regions and across different time periods. Combination therapies, novel agents, and aerosolized forms of antimicrobial drugs are currently being tested for their ability to treat infections caused by this multi-drug resistant organism. PMID:26388847

  7. Resistance of Stenotrophomonas maltophilia to Fluoroquinolones: Prevalence in a University Hospital and Possible Mechanisms

    PubMed Central

    Jia, Wei; Wang, Jiayuan; Xu, Haotong; Li, Gang

    2015-01-01

    Objective: The purpose of this study was to investigate the clinical distribution and genotyping of Stenotrophomonas maltophilia, its resistance to antimicrobial agents, and the possible mechanisms of this drug resistance. Methods: S. maltophilia isolates were collected from clinical specimens in a university hospital in Northwestern China during the period between 2010 and 2012, and were identified to the species level with a fully automated microbiological system. Antimicrobial susceptibility testing was performed for S. maltophilia with the Kirby-Bauer disc diffusion method. The minimal inhibitory concentrations (MICs) of norfloxacin, ofloxacin, chloramphenicol, minocycline, ceftazidime, levofloxacin and ciprofloxacin against S. maltophilia were assessed using the agar dilution method, and changes in the MIC of norfloxacin, ciprofloxacin and ofloxacin were observed after the addition of reserpine, an efflux pump inhibitor. Fluoroquinolone resistance genes were detected in S. maltophilia using a polymerase chain reaction (PCR) assay, and the expression of efflux pump smeD and smeF genes was determined using a quantitative fluorescent (QF)-PCR assay. Pulsed-field gel electrophoresis (PFGE) was employed to genotype identified S. maltophilia isolates. Results: A total of 426 S. maltophilia strains were isolated from the university hospital from 2010 to 2012, consisting of 10.1% of total non-fermentative bacteria. The prevalence of norfloxacin, ciprofloxacin and ofloxacin resistance was 32.4%, 21.9% and 13.2% in the 114 S. maltophilia isolates collected from 2012, respectively. Following reserpine treatment, 19 S. maltophilia isolates positive for efflux pump were identified, and high expression of smeD and smeF genes was detected in two resistant isolates. gyrA, parC, smeD, smeE and smeF genes were detected in all 114 S. maltophilia isolates, while smqnr gene was found in 25.4% of total isolates. Glu-Lys mutation (GAA-AAA) was detected at the 151th amino acid of the

  8. Whole-genome sequencing identifies emergence of a quinolone resistance mutation in a case of Stenotrophomonas maltophilia bacteremia.

    PubMed

    Pak, Theodore R; Altman, Deena R; Attie, Oliver; Sebra, Robert; Hamula, Camille L; Lewis, Martha; Deikus, Gintaras; Newman, Leah C; Fang, Gang; Hand, Jonathan; Patel, Gopi; Wallach, Fran; Schadt, Eric E; Huprikar, Shirish; van Bakel, Harm; Kasarskis, Andrew; Bashir, Ali

    2015-11-01

    Whole-genome sequences for Stenotrophomonas maltophilia serial isolates from a bacteremic patient before and after development of levofloxacin resistance were assembled de novo and differed by one single-nucleotide variant in smeT, a repressor for multidrug efflux operon smeDEF. Along with sequenced isolates from five contemporaneous cases, they displayed considerable diversity compared against all published complete genomes. Whole-genome sequencing and complete assembly can conclusively identify resistance mechanisms emerging in S. maltophilia strains during clinical therapy.

  9. Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5.

    PubMed

    Chang, Hsiao-Chuan; Chen, Chiy-Rong; Lin, Juey-Wen; Shen, Gwan-Han; Chang, Kai-Ming; Tseng, Yi-Hsiung; Weng, Shu-Fen

    2005-03-01

    Stenotrophomonas maltophilia is one of the most prevalent opportunistic bacteria causing nosocomial infections. It has become problematic because most of the isolates are resistant to multiple antibiotics, and therefore, development of phage therapy has attracted strong attention. In this study, eight S. maltophilia phages were isolated from clinical samples including patient specimens, catheter-related devices, and wastewater. These phages can be divided into four distinct groups based on host range and digestibility of the phage DNAs with different restriction endonucleases. One of them, designated phiSMA5, was further characterized. Electron microscopy showed it resembled Myoviridae, with an isometric head (90 nm in diameter), a tail (90 nm long), a baseplate (25 nm wide), and short tail fibers. The phiSMA5 double-stranded DNA, refractory to digestion by most restriction enzymes, was tested and estimated to be 250 kb by pulsed-field gel electrophoresis. This genome size is second to that of the largest phage, phiKZ of Pseudomonas aeruginosa. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 25 virion proteins were visualized. N-terminal sequencing of four of them suggested that each of them might have had its N terminus cleaved off. Among the 87 S. maltophilia strains collected in this study, only 61 were susceptible to phiSMA5, indicating that more phages are needed toward a phage therapy strategy. Since literature search yielded no information about S. maltophilia phages, phiSMA5 appears to be the first reported.

  10. SmeOP-TolCSm Efflux Pump Contributes to the Multidrug Resistance of Stenotrophomonas maltophilia

    PubMed Central

    Lin, Cheng-Wen; Huang, Yi-Wei; Hu, Rouh-Mei

    2014-01-01

    A five-gene cluster, tolCSm-pcm-smeRo-smeO-smeP, of Stenotrophomonas maltophilia was characterized. The presence of smeOP and smeRo-pcm-tolCSm operons was verified by reverse transcription (RT)-PCR. Both operons were negatively regulated by the TetR-type transcriptional regulator SmeRo, as demonstrated by quantitative RT-PCR and a promoter-fusion assay. SmeO and SmeP were associated with TolCSm (the TolC protein of S. maltophilia) for the assembly of a resistance-nodulation-cell-division (RND)-type pump. The compounds extruded by SmeOP-TolCSm mainly included nalidixic acid, doxycycline, amikacin, gentamicin, erythromycin, leucomycin, carbonyl cyanide 3-chlorophenylhydrazone, crystal violet, sodium dodecyl sulfate, and tetrachlorosalicylanilide. PMID:24395237

  11. Genomic Potential of Stenotrophomonas maltophilia in Bioremediation with an Assessment of Its Multifaceted Role in Our Environment

    PubMed Central

    Mukherjee, Piyali; Roy, Pranab

    2016-01-01

    The gram negative bacterium Stenotrophomonas is rapidly evolving as a nosocomial pathogen in immuno-compromised patients. Treatment of Stenotrophomonas maltophilia infections is problematic because of their increasing resistance to multiple antibiotics. This article aims to review the multi-disciplinary role of Stenotrophomonas in our environment with special focus on their metabolic and genetic potential in relation to bioremediation and phytoremediation. Current and emerging treatments and diagnosis for patients infected with S. maltophilia are discussed besides their capability of production of novel bioactive compounds. The plant growth promoting characteristics of this bacterium has been considered with special reference to secondary metabolite production. Nano-particle synthesis by Stenotrophomonas has also been reviewed in addition to their applications as effective biocontrol agents in plant and animal pathogenesis. PMID:27446008

  12. Molecular characterization and ultrastructure of a new amoeba endoparasite belonging to the Stenotrophomonas maltophilia complex.

    PubMed

    Corsaro, Daniele; Müller, Karl-Dieter; Michel, Rolf

    2013-04-01

    Naegleria and Acanthamoeba spp. were recovered from biofilm of a flushing cistern in a lavatory and both were found to be infected by rod-shaped bacteria enclosed within a vacuole. These intracellular bacteria behave like parasites, causing lysis of host amoebae. The bacteria proved unculturable on bacteriological media, and but could be maintained as endocytobionts within Acanthamoeba on agar plates. A marked differential host preference was observed in co-culture assays with various strains of amoebae. Molecular phylogenetic analyses performed on almost complete 16S rDNA sequences showed that the bacteria emerged as an atypical rapidly-evolving strain within the Stenotrophomonas maltophilia complex (Gamma-Proteobacteria, Xanthomonadales).

  13. Facile biosynthesis of phosphate capped gold nanoparticles by a bacterial isolate Stenotrophomonas maltophilia

    SciTech Connect

    Nangia, Yogesh; Wangoo, Nishima; Raman Suri, C.; Sharma, Saurabh; Wu, J.-S.; Dravid, Vinayak; Shekhawat, G. S.

    2009-06-08

    We report intracellular biosynthesis of gold nanoparticles (GNPs) by a strain Stenotrophomonas maltophilia (AuRed02) isolated from the soil samples of Singhbhum gold mines, India. An aqueous solution of gold chloride was reduced to metallic gold in a suspension of disrupted cell mass of AuRed02, which progressively turns into cherry red within 8 h of incubation at 25 deg. C. The optical spectrum showed the plasmon resonance at 530 nm and analysis by transmission electron microscopy and dynamic light scattering confirmed the formation of around 40 nm GNPs. Zeta potential and Fourier transform infrared measurements confirmed GNPs are capped by negatively charged phosphate groups of NADP.

  14. Gene flow, recombination, and positive selection in Stenotrophomonas maltophilia: mechanisms underlying the diversity of the widespread opportunistic pathogen.

    PubMed

    Yu, Dong; Yin, Zhiqiu; Li, Beiping; Jin, Yuan; Ren, Hongguang; Zhou, Jing; Zhou, Wei; Liang, Long; Yue, Junjie

    2016-12-01

    Stenotrophomonas maltophilia is a global multidrug-resistant human opportunistic pathogen in clinical environments. Stenotrophomonas maltophilia is also ubiquitous in aqueous environments, soil, and plants. Various molecular typing methods have revealed that S. maltophilia exhibits high levels of phenotypic and genotypic diversity. However, information regarding the genomic diversity within S. maltophilia and the corresponding genetic mechanisms resulting in said diversity remain scarce. The genome sequences of 17 S. maltophilia strains were selected to investigate the mechanisms contributing to genetic diversity at the genome level. The core and large pan-genomes of the species were first estimated, resulting in a large, open pan-genome. A species phylogeny was also reconstructed based on 344 orthologous genes with one copy per genome, and the contribution of four evolutionary mechanisms to the species genome diversity was quantified: 15%-35% of the genes showed evidence for recombination, 0%-25% of the genes in one genome were likely gained, 0%-44% of the genes in some genomes were likely lost, and less than 0.3% of the genes in a genome were under positive selection pressures. We observed that, among the four main mechanisms, homologous recombination plays a key role in maintaining diversity in S. maltophilia. In this study, we provide an overview of evolution in S. maltophilia to provide a better understanding of its evolutionary dynamics and its relationship with genome diversity.

  15. Phenotypic Heterogeneity Affects Stenotrophomonas maltophilia K279a Colony Morphotypes and β-Lactamase Expression

    PubMed Central

    Abda, Ebrahim M.; Krysciak, Dagmar; Krohn-Molt, Ines; Mamat, Uwe; Schmeisser, Christel; Förstner, Konrad U.; Schaible, Ulrich E.; Kohl, Thomas A.; Nieman, Stefan; Streit, Wolfgang R.

    2015-01-01

    Phenotypic heterogeneity at the cellular level in response to various stresses, e.g., antibiotic treatment has been reported for a number of bacteria. In a clonal population, cell-to-cell variation may result in phenotypic heterogeneity that is a mechanism to survive changing environments including antibiotic therapy. Stenotrophomonas maltophilia has been frequently isolated from cystic fibrosis patients, can cause numerous infections in other organs and tissues, and is difficult to treat due to antibiotic resistances. S. maltophilia K279a produces the L1 and L2 β-lactamases in response to β-lactam treatment. Here we report that the patient isolate S. maltophilia K279a diverges into cellular subpopulations with distinct but reversible morphotypes of small and big colonies when challenged with ampicillin. This observation is consistent with the formation of elongated chains of bacteria during exponential growth phase and the occurrence of mainly rod-shaped cells in liquid media. RNA-seq analysis of small versus big colonies revealed differential regulation of at least seven genes among the colony morphotypes. Among those, blaL1 and blaL2 were transcriptionally the most strongly upregulated genes. Promoter fusions of blaL1 and blaL2 genes indicated that expression of both genes is also subject to high levels of phenotypic heterogeneous expression on a single cell level. Additionally, the comE homolog was found to be differentially expressed in homogenously versus heterogeneously blaL2 expressing cells as identified by RNA-seq analysis. Overexpression of comE in S. maltophilia K279a reduced the level of cells that were in a blaL2-ON mode to 1% or lower. Taken together, our data provide strong evidence that S. maltophilia K279a populations develop phenotypic heterogeneity in an ampicillin challenged model. This cellular variability is triggered by regulation networks including blaL1, blaL2, and comE. PMID:26696982

  16. Comparison of two multimetal resistant bacterial strains: Enterobacter sp. YSU and Stenotrophomonas maltophilia ORO2.

    PubMed

    Holmes, Andrew; Vinayak, Anubhav; Benton, Cherise; Esbenshade, Aaron; Heinselman, Carlisle; Frankland, Daniel; Kulkarni, Samatha; Kurtanich, Adrienne; Caguiat, Jonathan

    2009-11-01

    The Y-12 plant in Oak Ridge, TN, which manufactured nuclear weapons during World War II and the Cold War, contaminated East Fork Poplar Creek with heavy metals. The multimetal resistant bacterial strain, Stenotrophomonas maltophilia Oak Ridge strain O2 (S. maltophilia O2), was isolated from East Fork Poplar Creek. Sequence analysis of 16s rDNA suggested that our working strain of S. maltophilia O2 was a strain of Enterobacter. Phylogenetic tree analysis and biochemical tests confirmed that it belonged to an Enterobacter species. This new strain was named Enterobacter sp. YSU. Using a modified R3A growth medium, R3A-Tris, the Hg(II), Cd(II), Zn(II), Cu(II), Au(III), Cr(VI), Ag(I), As(III), and Se(IV) MICs for a confirmed strain of S. maltophilia O2 were 0.24, 0.33, 5, 5, 0.25, 7, 0.03, 14, and 40 mM, respectively, compared to 0.07, 0.24, 0.8, 3, 0.05, 0.4, 0.08, 14, and 40 mM, respectively, for Enterobacter sp. YSU. Although S. maltophilia O2 was generally more metal resistant than Enterobacter sp. YSU, in comparison to Escherichia coli strain HB101, Enterobacter sp. YSU was resistant to Hg(II), Cd(II), Zn(II), Au(III), Ag(I), As(III), and Se(IV). By studying metal resistances in these two strains, it may be possible to understand what makes one microorganism more metal resistant than another microorganism. This work also provided benchmark MICs that can be used to evaluate the metal resistance properties of other bacterial isolates from East Fork Poplar Creek and other metal contaminated sites.

  17. Effects of Fluoroquinolones and Azithromycin on Biofilm Formation of Stenotrophomonas maltophilia

    PubMed Central

    Wang, Aihua; Wang, Qinqin; Kudinha, Timothy; Xiao, Shunian; Zhuo, Chao

    2016-01-01

    Stenotrophomonas maltophilia is an opportunistic pathogen that causes respiratory and urinary tract infections, as well as wound infections in immunocompromised patients. This pathogen is difficult to treat due to increased resistance to many antimicrobial agents. We investigated the in vitro biofilm formation of S. maltophilia, including effects of fluoroquinolones (FQs) and azithromycin on biofilm formation. The organism initiated attachment to polystyrene surfaces after a 4 h incubation period, and reached maximal growth at 18–24 h. In the presence of FQs (moxifloxacin, levofloxacin or ciprofloxacin), the biofilm biomass was significantly reduced (P < 0.05). A lower concentration of moxifloxacin (10 μg/mL) exhibited a better inhibiting effect on biofilm formation than 100 μg/mL (P < 0.01), but with no difference in effect compared to the 50 μg/mL concentration (P > 0.05). However, the inhibitory effects of 10 μg/mL of levofloxacin or ciprofloxacin were slightly less pronounced than those of the higher concentrations. A combination of azithromycin and FQs significantly reduced the biofilm inhibiting effect on S. maltophilia preformed biofilms compared to azithromycin or FQs alone. We conclude that early use of clinically acceptable concentrations of FQs, especially moxifloxacin (10 μg/mL), may possibly inhibit biofilm formation by S. maltophilia. Our study provides an experimental basis for a possible optimal treatment strategy for S. maltophilia biofilm-related infections. PMID:27405358

  18. Levofloxacin Efflux and smeD in Clinical Isolates of Stenotrophomonas maltophilia.

    PubMed

    Chong, So Young; Lee, Kyungwon; Chung, Hae-Sun; Hong, Seong Geun; Suh, Younghee; Chong, Yunsop

    2017-03-01

    Trimethoprim-sulfamethoxazole is the first-line antimicrobial combination for Stenotrophomonas maltophilia infections. However, allergy or intolerance and increasing resistance limit the use of trimethoprim-sulfamethoxazole. Quinolones can be used as an alternative therapeutic option, but resistance can emerge rapidly during therapy. We analyzed the contribution of SmeABC and SmeDEF efflux pumps to levofloxacin resistance in clinical isolates of S. maltophilia. Nonduplicate clinical isolates of S. maltophilia were collected in 2010 from 11 university hospitals (n = 102). Fifty-five levofloxacin nonsusceptible (minimum inhibitory concentration [MIC] ≥4 μg/ml) and 47 susceptible (MIC ≤2 μg/ml) isolates were tested for efflux pump overexpression. Real-time reverse transcription-PCR was performed for amplification and quantification of smeB, smeC, smeD, and smeF mRNA. To determine which antimicrobials were affected by smeD overexpression, the growth rates of a levofloxacin-susceptible S. maltophilia isolate were compared by measuring absorbance of antimicrobial-supplemented Luria-Bertani broth (LB) cultures with or without triclosan. Significant relationships between sme gene overexpression and resistance were observed for smeD against levofloxacin, smeC and smeF against ceftazidime, and smeC against ticarcillin-clavulanate. The mean MICs of moxifloxacin and tigecycline did not significantly differ for isolates with or without overexpression of smeB, smeC, and smeF, but were significantly higher for isolates with smeD overexpression. The mean MICs of amikacin were significantly higher for smeC or smeF overexpressing isolates. Increased growth of a levofloxacin-susceptible isolate was observed in LB with 1/2 MIC levofloxacin in the presence of triclosan. These data suggest that the expression of smeD plays a role in levofloxacin resistance in S. maltophilia.

  19. Removal of cadmium by bioflocculant produced by Stenotrophomonas maltophilia using phenol-containing wastewater.

    PubMed

    Chen, Honggao; Zhong, Chunying; Berkhouse, Hudson; Zhang, Youlang; Lv, Yao; Lu, Wanyu; Yang, Yongbing; Zhou, Jiangang

    2016-07-01

    Bioflocculants have been applied in numerous applications including heavy metals removal. A major bottleneck for commercial application of bioflocculant is its high production cost. Phenol-containing wastewater are abundantly available. However, the toxic phenol inhibited the microbial activities in the subsequent fermentation processes. Consequently, strains that can secrete phenol-degrading enzymes and simultaneously produce bioflocculants through directly degrading the phenol are of academic and practical interests. A phenol-degrading strain, Stenotrophomonas maltophilia ZZC-06, which can produce the bioflocculant MBF-06 using phenol-containing wastewater, was isolated in this study. The effects of culture conditions including initial pH, dissolved oxygen, phenol concentration, inoculum size, and temperature on MBF-06 production were analyzed. The experimental results showed that over 90% flocculating activity was achieved when the phenol was used as a carbon source and 4.99 g/L of MBF-06 was achieved under the optimized condition: 2.0% dissolved oxygen, 800 mg/L phenol concentration, 10% inoculum size, an initial pH of 6.0, and a temperature of 30 °C. The bioflocculant MBF-06 contained 71.2% polysaccharides and 27.9% proteins. The feasibility of cadmium removal using MBF-06 was evaluated. The highest flocculating efficiency for cadmium was 81.43%. This study shows for the first time that Stenotrophomonas maltophilia ZZC-06 can directly convert phenol into a bioflocculant, which can be used to effectively remove cadmium.

  20. Outbreak of Stenotrophomonas maltophilia bacteremia among patients undergoing bone marrow transplantation: association with faulty replacement of handwashing soap.

    PubMed

    Klausner, J D; Zukerman, C; Limaye, A P; Corey, L

    1999-11-01

    Using molecular typing methods, we confirmed an outbreak of Stenotrophomonas maltophilia among bone marrow transplant patients. The likely source was a healthcare worker who may have washed with moisturizer instead of soap between patients. Hospital epidemiologists need to go beyond antibiograms when evaluating outbreaks and be vigilant about all aspects of hand washing.

  1. The DSF Quorum Sensing System Controls the Positive Influence of Stenotrophomonas maltophilia on Plants

    PubMed Central

    Alavi, Peyman; Müller, Henry; Cardinale, Massimiliano; Zachow, Christin; Sánchez, María B.; Martínez, José Luis; Berg, Gabriele

    2013-01-01

    The interaction of the Gram-negative bacterium Stenotrophomonas maltophilia with eukaryotes can improve overall plant growth and health, but can also cause opportunistic infections in humans. While the quorum sensing molecule DSF (diffusible signal factor) is responsible for the regulation of phenotypes in pathogenic Stenotrophomonas, up until now, no beneficial effects were reported to be controlled by it. Our objective was to study the function of DSF in the plant growth promoting model strain S. maltophilia R551-3 using functional and transcriptomic analyses. For this purpose, we compared the wild-type strain with a mutant deficient in the rpfF (regulation of pathogenicity factors) gene that is essential for the synthesis of DSF. Oilseed rape seeds treated with the wild-type strain showed a statistically significant increase in germination rate compared with those treated with the rpfF mutant. Similarly, the wild-type strain exhibited better plant growth promotion and a greater efficiency in colonizing oilseed rape compared to the mutant strain. Moreover, only the wild-type was capable of forming structured cell aggregates both in vitro and in the rhizosphere, a characteristic mediated by DSF. Gene transcription analyses showed that numerous genes known to play a role in plant colonization (e.g. chemotaxis, cell motility, biofilm formation, multidrug efflux pumps) are controlled by the rpf/DSF system in S. maltophilia. In addition, we detected new potential functions of spermidine, primarily for both growth promotion and stress protection. Overall, our results showed a correspondence between the regulation of DSF and the positive interaction effect with the plant host. PMID:23874407

  2. Colistin susceptibility testing: evaluation of reliability for cystic fibrosis isolates of Pseudomonas aeruginosa and Stenotrophomonas maltophilia

    PubMed Central

    Moskowitz, Samuel M.; Garber, Elizabeth; Chen, Yunhua; Clock, Sarah A.; Tabibi, Setareh; Miller, Amanda K.; Doctor, Michael; Saiman, Lisa

    2010-01-01

    Objectives Antibiotic susceptibility methods that are commonly used to test bacterial isolates from patients with cystic fibrosis are of uncertain reliability for the polymyxins. To assess the reliability of four standard testing methods, this pilot study used a challenge set that included polymyxin-resistant isolates of Pseudomonas aeruginosa and Stenotrophomonas maltophilia. Methods Twenty-five P. aeruginosa and 12 S. maltophilia isolates were tested for susceptibility to colistin (polymyxin E). Repeatability (concordance of replicates performed concurrently), reproducibility (concordance of replicates performed over time) and comparability (concordance of different methods) of agar dilution, broth microdilution, Etest and disc diffusion were assessed through the use of descriptive statistics and scatterplot analyses. Results All four methods displayed excellent repeatability (overall concordance rate of 99%). However, analysis of reproducibility revealed substantially lower rates of concordance (74% for agar dilution, 84% for broth microdilution and Etest, and 91% for disc diffusion). In addition, comparability to agar dilution of the three other methods was generally poor, with overall rates of very major error ranging from 12% for broth microdilution to 18% for Etest and disc diffusion. Conclusions Compared with agar dilution, other susceptibility testing methods give high rates of apparent false polymyxin susceptibility for cystic fibrosis isolates of P. aeruginosa and S. maltophilia. Prospective study of the correlation between in vitro susceptibility and clinical response is needed to clarify whether these discrepancies reflect oversensitivity of the agar dilution method or insensitivity of the other methods. PMID:20430789

  3. Occurrence of Stenotrophomonas maltophilia in agricultural soils and antibiotic resistance properties.

    PubMed

    Deredjian, Amélie; Alliot, Nolwenn; Blanchard, Laurine; Brothier, Elisabeth; Anane, Makram; Cambier, Philippe; Jolivet, Claudy; Khelil, Mohamed Naceur; Nazaret, Sylvie; Saby, Nicolas; Thioulouse, Jean; Favre-Bonté, Sabine

    2016-05-01

    The occurrence of Stenotrophomonas maltophilia was monitored in organic amendments and agricultural soils from various sites in France and Tunisia. S. maltophilia was detected in horse and bovine manures, and its abundance ranged from 0.294 (±0.509) × 10(3) to 880 (±33.4) × 10(3) CFU (g drywt)(-1) of sample. S. maltophilia was recovered from most tested soil samples (104/124). Its abundance varied from 0.33 (±0.52) to 414 (±50) × 10(3) CFU (g drywt)(-1) of soil and was not related to soil characteristics. Antibiotic resistance properties of a set of environmental strains were compared to a clinical set, and revealed a high diversity of antibiotic resistance profiles, given both the numbers of resistance and the phenotypes. Manure strains showed resistance phenotypes, with most of the strains resisting between 7 and 9 antibiotics. While French soil strains were sensitive to most antibiotics tested, some Tunisian strains displayed resistance phenotypes close to those of clinical French strains. Screening for metal resistance among 66 soil strains showed a positive relationship between antibiotic and metal resistance. However, the prevalence of antibiotic resistance phenotypes in the studied sites was not related to the metal content in soil samples.

  4. Interplay between intrinsic and acquired resistance to quinolones in Stenotrophomonas maltophilia.

    PubMed

    García-León, Guillermo; Salgado, Fabiola; Oliveros, Juan Carlos; Sánchez, María Blanca; Martínez, José Luis

    2014-05-01

    To analyse whether the mutation-driven resistance-acquisition potential of a given bacterium might be a function of its intrinsic resistome, quinolones were used as selective agents and Stenotrophomonas maltophilia was chosen as a bacterial model. S. maltophilia has two elements - SmQnr and SmeDEF - that are important in intrinsic resistance to quinolones. Using a battery of mutants in which either or both of these elements had been removed, the apparent mutation frequency for quinolone resistance and the phenotype of the selected mutants were found to be related to the intrinsic resistome and also depended on the concentration of the selector. Most mutants had phenotypes compatible with the overexpression of multidrug efflux pump(s); SmeDEF overexpression was the most common cause of quinolone resistance. Whole genome sequencing showed that mutations of the SmeRv regulator, which result in the overexpression of the efflux pump SmeVWX, are the cause of quinolone resistance in mutants not overexpressing SmeDEF. These results indicate that the development of mutation-driven antibiotic resistance is highly dependent on the intrinsic resistome, which, at least for synthetic antibiotics such as quinolones, did not develop as a response to the presence of antibiotics in the natural ecosystems in which S. maltophilia evolved.

  5. Infections Caused by Stenotrophomonas maltophilia in Recipients of Hematopoietic Stem Cell Transplantation

    PubMed Central

    Al-Anazi, Khalid Ahmed; Al-Jasser, Asma M.

    2014-01-01

    Stenotrophomonas maltophilia (S. maltophilia) is a globally emerging Gram-negative bacillus that is widely spread in environment and hospital equipment. Recently, the incidence of infections caused by this organism has increased, particularly in patients with hematological malignancy and in recipients of hematopoietic stem cell transplantation (HSCT) having neutropenia, mucositis, diarrhea, central venous catheters or graft versus host disease and receiving intensive cytotoxic chemotherapy, immunosuppressive therapy, or broad-spectrum antibiotics. The spectrum of infections in HSCT recipients includes pneumonia, urinary tract and surgical site infection, peritonitis, bacteremia, septic shock, and infection of indwelling medical devices. The organism exhibits intrinsic resistance to many classes of antibiotics including carbapenems, aminoglycosides, most of the third-generation cephalosporins, and other β-lactams. Despite the increasingly reported drug resistance, trimethoprim-sulfamethoxazole is still the drug of choice. However, the organism is still susceptible to ticarcillin-clavulanic acid, tigecycline, fluoroquinolones, polymyxin-B, and rifampicin. Genetic factors play a significant role not only in evolution of drug resistance but also in virulence of the organism. The outcome of patients having S. maltophilia infections can be improved by: using various combinations of novel therapeutic agents and aerosolized aminoglycosides or colistin, prompt administration of in vitro active antibiotics, removal of possible sources of infection such as infected indwelling intravascular catheters, and application of strict infection control measures. PMID:25202682

  6. The isolation and characterization of Stenotrophomonas maltophilia T4-like bacteriophage DLP6.

    PubMed

    Peters, Danielle L; Stothard, Paul; Dennis, Jonathan J

    2017-01-01

    Increasing isolation of the extremely antibiotic resistant bacterium Stenotrophomonas maltophilia has caused alarm worldwide due to the limited treatment options available. A potential treatment option for fighting this bacterium is 'phage therapy', the clinical application of bacteriophages to selectively kill bacteria. Bacteriophage DLP6 (vB_SmoM-DLP6) was isolated from a soil sample using clinical isolate S. maltophilia strain D1571 as host. Host range analysis of phage DLP6 against 27 clinical S. maltophilia isolates shows successful infection and lysis in 13 of the 27 isolates tested. Transmission electron microscopy of DLP6 indicates that it is a member of the Myoviridae family. Complete genome sequencing and analysis of DLP6 reveals its richly recombined evolutionary history, featuring a core of both T4-like and cyanophage genes, which suggests that it is a member of the T4-superfamily. Unlike other T4-superfamily phages however, DLP6 features a transposase and ends with 229 bp direct terminal repeats. The isolation of this bacteriophage is an exciting discovery due to the divergent nature of DLP6 in relation to the T4-superfamily of phages.

  7. The isolation and characterization of Stenotrophomonas maltophilia T4-like bacteriophage DLP6

    PubMed Central

    Peters, Danielle L.; Stothard, Paul

    2017-01-01

    Increasing isolation of the extremely antibiotic resistant bacterium Stenotrophomonas maltophilia has caused alarm worldwide due to the limited treatment options available. A potential treatment option for fighting this bacterium is ‘phage therapy’, the clinical application of bacteriophages to selectively kill bacteria. Bacteriophage DLP6 (vB_SmoM-DLP6) was isolated from a soil sample using clinical isolate S. maltophilia strain D1571 as host. Host range analysis of phage DLP6 against 27 clinical S. maltophilia isolates shows successful infection and lysis in 13 of the 27 isolates tested. Transmission electron microscopy of DLP6 indicates that it is a member of the Myoviridae family. Complete genome sequencing and analysis of DLP6 reveals its richly recombined evolutionary history, featuring a core of both T4-like and cyanophage genes, which suggests that it is a member of the T4-superfamily. Unlike other T4-superfamily phages however, DLP6 features a transposase and ends with 229 bp direct terminal repeats. The isolation of this bacteriophage is an exciting discovery due to the divergent nature of DLP6 in relation to the T4-superfamily of phages. PMID:28291834

  8. Iron is a signal for Stenotrophomonas maltophilia biofilm formation, oxidative stress response, OMPs expression, and virulence

    PubMed Central

    García, Carlos A.; Alcaraz, Eliana S.; Franco, Mirta A.; Passerini de Rossi, Beatriz N.

    2015-01-01

    Stenotrophomonas maltophilia is an emerging nosocomial pathogen. In many bacteria iron availability regulates, through the Fur system, not only iron homeostasis but also virulence. The aim of this work was to assess the role of iron on S. maltophilia biofilm formation, EPS production, oxidative stress response, OMPs regulation, quorum sensing (QS), and virulence. Studies were done on K279a and its isogenic fur mutant F60 cultured in the presence or absence of dipyridyl. This is the first report of spontaneous fur mutants obtained in S. maltophilia. F60 produced higher amounts of biofilms than K279a and CLSM analysis demonstrated improved adherence and biofilm organization. Under iron restricted conditions, K279a produced biofilms with more biomass and enhanced thickness. In addition, F60 produced higher amounts of EPS than K279a but with a similar composition, as revealed by ATR-FTIR spectroscopy. With respect to the oxidative stress response, MnSOD was the only SOD isoenzyme detected in K279a. F60 presented higher SOD activity than the wt strain in planktonic and biofilm cultures, and iron deprivation increased K279a SOD activity. Under iron starvation, SDS-PAGE profile from K279a presented two iron-repressed proteins. Mass spectrometry analysis revealed homology with FepA and another putative TonB-dependent siderophore receptor of K279a. In silico analysis allowed the detection of potential Fur boxes in the respective coding genes. K279a encodes the QS diffusible signal factor (DSF). Under iron restriction K279a produced higher amounts of DSF than under iron rich condition. Finally, F60 was more virulent than K279a in the Galleria mellonella killing assay. These results put in evidence that iron levels regulate, likely through the Fur system, S. maltophilia biofilm formation, oxidative stress response, OMPs expression, DSF production and virulence. PMID:26388863

  9. Nosocomial outbreak of colonization and infection with Stenotrophomonas maltophilia in preterm infants associated with contaminated tap water.

    PubMed Central

    Verweij, P. E.; Meis, J. F.; Christmann, V.; Van der Bor, M.; Melchers, W. J.; Hilderink, B. G.; Voss, A.

    1998-01-01

    Between March and May 1996 Stenotrophomonas maltophilia was cultured from endotracheal aspirate samples from five preterm infants in a neonatal intensive care unit (NICU). Four infants were superficially colonized, but a fifth died due to S. maltophilia septicaemia. S. maltophilia was cultured from tap water from three outlets in the NICU including one with a previously unnoticed defective sink drain. Water from these outlets was used to wash the preterm infants. Environmental and clinical S. maltophilia isolates yielded identical banding patterns on random arbitrary polymorphic DNA (RAPD) PCR analysis. The outbreak was controlled by reinforcement of hand disinfection, limitation of the use of tap water for hand washing and by using sterile water to wash the preterm infants. We conclude that tap water should not be used for washing preterm infants in the NICU, unless steps are taken to prevent microbial growth in the outlets. PMID:9692603

  10. Genotypic and Phenotypic Characterization of Stenotrophomonas maltophilia Strains from a Pediatric Tertiary Care Hospital in Serbia

    PubMed Central

    Madi, Haowa; Lukić, Jovanka; Vasiljević, Zorica; Biočanin, Marjan; Kojić, Milan; Jovčić, Branko; Lozo, Jelena

    2016-01-01

    Background Stenotrophomonas maltophilia is an environmental bacterium and an opportunistic pathogen usually associated with healthcare-associated infections, which has recently been recognized as a globally multi-drug resistant organism. The aim of this study was genotyping and physiological characterization of Stenotrophomonas maltophilia isolated in a large, tertiary care pediatric hospital in Belgrade, Serbia, hosting the national reference cystic fibrosis (CF) center for pediatric and adult patients. Methods We characterized 42 strains of cystic fibrosis (CF) and 46 strains of non-cystic fibrosis (non-CF) origin isolated from 2013 to 2015 in order to investigate their genetic relatedness and phenotypic traits. Genotyping was performed using sequencing of 16S rRNA gene, Pulse Field Gel Electrophoresis (PFGE) and Multi locus sequencing typing (MLST) analysis. Sensitivity to five relevant antimicrobial agents was determined, namely trimethoprim/sulfamethoxazole (TMP/SMX), chloramphenicol, ciprofloxacin, levofloxacin and tetracycline. Surface characteristics, motility, biofilm formation and adhesion to mucin were tested in all strains. Statistical approach was used to determine correlations between obtained results. Results Most of the isolates were not genetically related. Six new sequence types were determined. Strains were uniformly sensitive to all tested antimicrobial agents. The majority of isolates (89.8%) were able to form biofilm with almost equal representation in both CF and non-CF strains. Swimming motility was observed in all strains, while none of them exhibited swarming motility. Among strains able to adhere to mucin, no differences between CF and non-CF isolates were observed. Conclusions High genetic diversity among isolates implies the absence of clonal spread within the hospital. Positive correlation between motility, biofilm formation and adhesion to mucin was demonstrated. Biofilm formation and motility were more pronounced among non-CF than CF

  11. Draft Genome Sequence of Stenotrophomonas maltophilia Strain B418, a Promising Agent for Biocontrol of Plant Pathogens and Root-Knot Nematode.

    PubMed

    Wu, Yuanzheng; Wang, Yilian; Li, Jishun; Hu, Jindong; Chen, Kai; Wei, Yanli; Bazhanov, Dmitry P; Bazhanova, Alesia A; Yang, Hetong

    2015-02-19

    Stenotrophomonas maltophilia strain B418 was isolated from a barley rhizosphere in China. This bacterium exhibits broad-spectrum inhibitory activities against plant pathogens and root-knot nematode along with growth-promoting effects. Here, we present the draft genome sequence of S. maltophilia B418.

  12. In Vitro Efficacy of High-Dose Tobramycin against Burkholderia cepacia Complex and Stenotrophomonas maltophilia Isolates from Cystic Fibrosis Patients

    PubMed Central

    Ratjen, Anina; Yau, Yvonne; Wettlaufer, Jillian; Matukas, Larissa; Zlosnik, James E. A.; Speert, David P.; LiPuma, John J.; Tullis, Elizabeth

    2014-01-01

    Burkholderia cepacia complex and Stenotrophomonas maltophilia infections are associated with poor clinical outcomes in persons with cystic fibrosis (CF). The MIC50 based on planktonic growth and the biofilm concentration at which 50% of the isolates tested are inhibited (BIC50) of tobramycin were measured for 180 B. cepacia complex and 101 S. maltophilia CF isolates and were 100 μg/ml for both species. New inhalation devices that deliver high tobramycin levels to the lung may be able to exceed these MICs. PMID:25348526

  13. In vitro efficacy of high-dose tobramycin against Burkholderia cepacia complex and Stenotrophomonas maltophilia isolates from cystic fibrosis patients.

    PubMed

    Ratjen, Anina; Yau, Yvonne; Wettlaufer, Jillian; Matukas, Larissa; Zlosnik, James E A; Speert, David P; LiPuma, John J; Tullis, Elizabeth; Waters, Valerie

    2015-01-01

    Burkholderia cepacia complex and Stenotrophomonas maltophilia infections are associated with poor clinical outcomes in persons with cystic fibrosis (CF). The MIC50 based on planktonic growth and the biofilm concentration at which 50% of the isolates tested are inhibited (BIC50) of tobramycin were measured for 180 B. cepacia complex and 101 S. maltophilia CF isolates and were 100 μg/ml for both species. New inhalation devices that deliver high tobramycin levels to the lung may be able to exceed these MICs.

  14. Whole-Genome Sequencing Identifies Emergence of a Quinolone Resistance Mutation in a Case of Stenotrophomonas maltophilia Bacteremia

    PubMed Central

    Altman, Deena R.; Attie, Oliver; Sebra, Robert; Hamula, Camille L.; Lewis, Martha; Deikus, Gintaras; Newman, Leah C.; Fang, Gang; Hand, Jonathan; Patel, Gopi; Wallach, Fran; Schadt, Eric E.; Huprikar, Shirish; van Bakel, Harm; Bashir, Ali

    2015-01-01

    Whole-genome sequences for Stenotrophomonas maltophilia serial isolates from a bacteremic patient before and after development of levofloxacin resistance were assembled de novo and differed by one single-nucleotide variant in smeT, a repressor for multidrug efflux operon smeDEF. Along with sequenced isolates from five contemporaneous cases, they displayed considerable diversity compared against all published complete genomes. Whole-genome sequencing and complete assembly can conclusively identify resistance mechanisms emerging in S. maltophilia strains during clinical therapy. PMID:26324280

  15. A Polysaccharide Lyase from Stenotrophomonas maltophilia with a Unique, pH-regulated Substrate Specificity*

    PubMed Central

    MacDonald, Logan C.; Berger, Bryan W.

    2014-01-01

    Polysaccharide lyases (PLs) catalyze the depolymerization of anionic polysaccharides via a β-elimination mechanism. PLs also play important roles in microbial pathogenesis, participating in bacterial invasion and toxin spread into the host tissue via degradation of the host extracellular matrix, or in microbial biofilm formation often associated with enhanced drug resistance. Stenotrophomonas maltophilia is a Gram-negative bacterium that is among the emerging multidrug-resistant organisms associated with chronic lung infections as well as with cystic fibrosis patients. A putative alginate lyase (Smlt1473) from S. maltophilia was heterologously expressed in Escherichia coli, purified in a one-step fashion via affinity chromatography, and activity as well as specificity determined for a range of polysaccharides. Interestingly, Smlt1473 catalyzed the degradation of not only alginate, but poly-β-d-glucuronic acid and hyaluronic acid as well. Furthermore, the pH optimum for enzymatic activity is substrate-dependent, with optimal hyaluronic acid degradation at pH 5, poly-β-d-glucuronic acid degradation at pH 7, and alginate degradation at pH 9. Analysis of the degradation products revealed that each substrate was cleaved endolytically into oligomers comprised predominantly of even numbers of sugar groups, with lower accumulation of trimers and pentamers. Collectively, these results imply that Smlt1473 is a multifunctional PL that exhibits broad substrate specificity, but utilizes pH as a mechanism to achieve selectivity. PMID:24257754

  16. Effects of Green Tea Compound Epigallocatechin-3-Gallate against Stenotrophomonas maltophilia Infection and Biofilm

    PubMed Central

    Vidigal, Pedrina G.; Müsken, Mathias; Becker, Katrin A.; Häussler, Susanne; Wingender, Jost; Steinmann, Eike; Kehrmann, Jan; Gulbins, Erich; Buer, Jan; Rath, Peter Michael; Steinmann, Jörg

    2014-01-01

    We investigated the in vitro and in vivo activities of epigallocatechin-3-gallate (EGCg), a green tea component, against Stenotrophomonas maltophilia (Sm) isolates from cystic fibrosis (CF) patients. In vitro effects of EGCg and the antibiotic colistin (COL) on growth inhibition, survival, and also against young and mature biofilms of S. maltophilia were determined. Qualitative and quantitative changes on the biofilms were assessed by confocal laser scanning microscopy (CLSM). Further, in vivo effects of nebulized EGCg in C57BL/6 and Cftr mutant mice during acute Sm lung infection were evaluated. Subinhibitory concentrations of EGCg significantly reduced not only biofilm formation, but also the quantity of viable cells in young and mature biofilms. CLSM showed that EGCg-exposed biofilms exhibited either a change in total biofilm biovolume or an increase of the fraction of dead cells contained within the biofilm in a dose depended manner. Sm infected wild-type and Cftr mutant mice treated with 1,024 mg/L EGCg by inhalation exhibited significantly lower bacterial counts than those undergoing no treatment or treated with COL. EGCg displayed promising inhibitory and anti-biofilm properties against CF Sm isolates in vitro and significantly reduced Sm bacterial counts in an acute infection model with wild type and CF mice. This natural compound may represent a novel therapeutic agent against Sm infection in CF. PMID:24690894

  17. Insights into the degradation of chlorimuron-ethyl by Stenotrophomonas maltophilia D310-3.

    PubMed

    Zang, Hailian; Yu, Qi; Lv, Tongyang; Cheng, Yi; Feng, Lu; Cheng, Xiaosong; Li, Chunyan

    2016-02-01

    In this study, the effects of cultivation conditions on the degradation of chlorimuron-ethyl by Stenotrophomonas maltophilia D310-3, which exhibits a high chlorimuron-ethyl-degrading capability, were investigated. To improve the biodegradation efficiency, the cultivation conditions were optimized using response surface methodology (RSM) based on Box-Behnken design (BBD). The maximum biodegradation rate (89.9%) was obtained at the optimal conditions (culture time, 6 d; substrate concentration, 50.21 mg L(-1); pH, 5.95; temperature, 30.15 °C). The Andrews model was used to describe the dynamic change regularity of the specific degradation rate as the substrate concentration increased, and the values of the maximum specific degradation rate (q(max)), half-saturation constant (K(S)) and inhibition constant (K(i)) were 78.87 d(-1), 9180.97 mg L(-1) and 0.28 mg L(-1), respectively. Eight degradation products were captured and identified by liquid chromatography-mass spectrometry (LC-MS) and Fourier transform infrared (FTIR) spectrometry, and three possible degradation pathways are proposed based on the results of high-performance liquid chromatography (HPLC), LC-MS and FTIR analyses as well as results reported in relevant literature. To the best of our knowledge, this is the first systematic study of the degradation pathway of chlorimuron-ethyl by S. maltophilia D310-3. This study provides valuable information for further exploration of the microbial degradation of other sulfonylurea herbicides.

  18. Effects of green tea compound epigallocatechin-3-gallate against Stenotrophomonas maltophilia infection and biofilm.

    PubMed

    Vidigal, Pedrina G; Müsken, Mathias; Becker, Katrin A; Häussler, Susanne; Wingender, Jost; Steinmann, Eike; Kehrmann, Jan; Gulbins, Erich; Buer, Jan; Rath, Peter Michael; Steinmann, Jörg

    2014-01-01

    We investigated the in vitro and in vivo activities of epigallocatechin-3-gallate (EGCg), a green tea component, against Stenotrophomonas maltophilia (Sm) isolates from cystic fibrosis (CF) patients. In vitro effects of EGCg and the antibiotic colistin (COL) on growth inhibition, survival, and also against young and mature biofilms of S. maltophilia were determined. Qualitative and quantitative changes on the biofilms were assessed by confocal laser scanning microscopy (CLSM). Further, in vivo effects of nebulized EGCg in C57BL/6 and Cftr mutant mice during acute Sm lung infection were evaluated. Subinhibitory concentrations of EGCg significantly reduced not only biofilm formation, but also the quantity of viable cells in young and mature biofilms. CLSM showed that EGCg-exposed biofilms exhibited either a change in total biofilm biovolume or an increase of the fraction of dead cells contained within the biofilm in a dose depended manner. Sm infected wild-type and Cftr mutant mice treated with 1,024 mg/L EGCg by inhalation exhibited significantly lower bacterial counts than those undergoing no treatment or treated with COL. EGCg displayed promising inhibitory and anti-biofilm properties against CF Sm isolates in vitro and significantly reduced Sm bacterial counts in an acute infection model with wild type and CF mice. This natural compound may represent a novel therapeutic agent against Sm infection in CF.

  19. A new Stenotrophomonas maltophilia strain producing laccase. Use in decolorization of synthetics dyes.

    PubMed

    Galai, Said; Limam, Ferid; Marzouki, M Nejib

    2009-08-01

    Laccase activity was detected in a soil bacterium Stenotrophomonas maltophilia AAP56 identified by biochemical and molecular methods. It was produced in cells at the stationary growth phase in Luria Bertani (LB) medium added by 0.4 mM copper sulfate. The addition of CuSO(4) in culture medium improved production of laccase activity. However, one laccase enzyme was detected by native polyacrylamide gel electrophoresis. The enzyme showed syringaldazine (K (m) = 53 microM), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (K (m) = 700 microM), and pyrocatechol (K (m) = 25 microM) oxidase activity and was activated by addition of 0.1% (v/v) Triton-X-100 in the reaction mixture. Moreover, the laccase activity was increased 2.6-fold by the addition of 10 mM copper sulfate; the enzyme was totally inhibited by ethylenediaminetetraacetic acid (5 mM), suggesting that this laccase is a metal-dependant one. Decolorization activity of some synthetic dyes (methylene blue, methyl green, toluidine blue, Congo red, methyl orange, and pink) and the industrial effluent (SITEX Black) was achieved by the bacteria S. maltophilia AAP56 in the LB growth medium under shaking conditions.

  20. A Highly Thermostable Xylanase from Stenotrophomonas maltophilia: Purification and Partial Characterization.

    PubMed

    Raj, Abhay; Kumar, Sharad; Singh, Sudheer Kumar

    2013-01-01

    Seven xylanolytic bacterial strains were isolated from saw-dust dump soil. The bacterial strain X6 was selected on the basis of the highest xylanase activity with no cellulase contamination. It was identified as Stenotrophomonas maltophilia by biochemical tests and 16S rRNA gene sequencing approach. Xylanase production studies by S. maltophilia on different commercial xylans and agro-industrial residues suggested that wheat bran was the best carbon source for xylanase production (26.4 ± 0.6 IU/mL). The studies with inorganic and organic nitrogen sources suggested yeast extract as the best support for xylanase production (25 ± 0.6 IU/mL). Maximum xylanase production was observed at initial medium pH = 8.0 (23.8 ± 0.4 IU/mL) with production at pH = 7.0 and pH = 9.0 being almost comparable. Xylanase produced by S. maltophilia was purified to homogeneity using ammonium sulfate precipitation, gel filtration, and ion exchange chromatography. The final purification was 5.43-fold with recovery of 19.18%. The molecular weight of the purified xylanase protein was ~142 kDa. Both crude and purified xylanase had good stability at pH = 9.0 and 80°C with activity retention greater than 90% after 30 min incubation. The enzyme stability at high temperature and alkaline pH make it potentially effective for industrial applications.

  1. High Genetic Diversity among Stenotrophomonas maltophilia Strains Despite Their Originating at a Single Hospital

    PubMed Central

    Valdezate, Sylvia; Vindel, Ana; Martín-Dávila, Pilar; Del Saz, Begoña Sánchez; Baquero, Fernando; Cantón, Rafael

    2004-01-01

    The levels of genetic relatedness of 139 Stenotrophomonas maltophilia strains recovered from 105 hospitalized non-cystic fibrosis patients (51% from medical wards, 35% from intensive care units, and 14% from surgical wards) and 7 environmental sources in the same hospital setting during a 4-year period were typed by the pulsed-field gel electrophoresis (PFGE) technique. A total of 99 well-defined distinct XbaI PFGE patterns were identified (Simpson's discrimination index, 0.996). The dendrogram showed a Dice similarity coefficient ranging from 28 to 80%. Two major clusters (I and II), three minor clusters (III, IV, and V), and two independent branches were observed when using a 36% Dice coefficient, indicating a high diversity of genetic relatedness. It is of note that 84% of strains were grouped within two major clonal lineages. No special cluster gathering was found among strains belonging to the same sample type specimen, patients' infection or colonization status, and ward of precedence. Despite this fact, three different clones (A, B, and C) recovered from respiratory samples from six, three, and two patients, respectively, and two clones, D and E, in two bacteremic patients each, were identified. Isolation of an S. maltophilia strain belonging to the clone A profile in a bronchoscope demonstrated a common source from this clone. This study revealed a high genetic diversity of S. maltophilia isolates despite their origin from a single hospital, which may be related to the wide environmental distribution of this pathogen. However, few clones could be transmitted among different patients, yielding outbreak situations. PMID:14766838

  2. Epidemiology and outcomes of Stenotrophomonas maltophilia and Burkholderia cepacia infections among trauma patients of India: a five year experience

    PubMed Central

    Rajkumari, Nonika; Gupta, Amit K; Sharma, Kumkum; Misra, Mahesh C

    2014-01-01

    Background: Infections by uncommon non-fermenting Gram negative bacteria are on the rise, but little is known about the risk factors and drug resistance in trauma patients in India. This study explored the infections caused by Stenotrophomonas maltophilia and/or Burkholderia cepacia in trauma patients over a period of 5 years. Material and methods: Patients admitted for trauma care with S. maltophilia and/or B. cepacia isolated from clinical specimens were enrolled. Characteristics regarding the strain isolation, drug sensitivity pattern, multidrug resistance (MDR), patient, outcomes, and differentiation of true infection from colonisation were observed. Results: Of the total 233 isolates, 102 were S. maltophilia and 131 were B. cepacia; 4.3% were responsible for polymicrobial infections with other bacteria. There were more B. cepacia MDR isolates than S. maltophilia. Maximum resistance was found to tetracycline (86.7%) and tobramycin (86.7%) in B. cepacia and to co-trimoxazole (68.7%) in S. maltophilia. Of these, 21 (16.03%) had a fatal outcome and the remaining 111 (84.7%) were discharged healthy. The in-hospital mortality rate associated with B. cepacia was much (16%) higher than S. maltophilia (13%) at this centre. Conclusion: The analysis of epidemiology and outcome of these infections will help to inform their management and treatment.

  3. In Vitro Killing Effect of Moxifloxacin on Clinical Isolates of Stenotrophomonas maltophilia Resistant to Trimethoprim-Sulfamethoxazole

    PubMed Central

    Giamarellos-Bourboulis, Evangelos J.; Karnesis, Lazaros; Galani, Irene; Giamarellou, Helen

    2002-01-01

    The time-kill effect of moxifloxacin on 20 genetically distinct isolates of Stenotrophomonas maltophilia resistant to trimethoprim-sulfamethoxazole was studied. The majority (80%) were killed by a concentration equivalent to four times the MIC; the MIC induced a transient decrease in bacterial counts at 4 h, followed by regrowth. No effect was detected in four isolates. These results merit further clinical consideration. PMID:12435710

  4. Isolation and characterization of a novel strain of Stenotrophomonas maltophilia possessing various dioxygenases for monocyclic hydrocarbon degradation

    PubMed Central

    Urszula, Guzik; Izabela, Greń; Danuta, Wojcieszyńska; Sylwia, Łabużek

    2009-01-01

    A Gram-negative bacterium, designated as strain KB2, was isolated from activated sludge and was found to utilize different aromatic substrates as sole carbon and energy source. On the basis of morphological and physiochemical characteristics and 16S rRNA gene sequence analysis, the isolated strain KB2 was identified as Stenotrophomonas maltophilia. Strain KB2 is from among different Stenotrophomonas maltophilia strains the first one described as exhibiting the activities of three types of dioxygenases depending on the structure of the inducer. The cells grown on benzoate and catechol showed mainly catechol 1,2-dioxygenase activity. The activity of 2,3-dioxygenase was detected after phenol induction. Protocatechuate 3,4-dioxygenase was found in crude cell extracts of this strain after incubation with 4-hydroxybenzoic acid, protocatechuic acid and vanillic acid. Because of broad spectrum of dioxygenases’ types that Stenotrophomonas maltophilia KB2 can exhibit, this strain appears to be very powerful and useful tool in the biotreatment of wastewaters and in soil decontamination. PMID:24031359

  5. Aflatoxin B1 Degradation by Stenotrophomonas Maltophilia and Other Microbes Selected Using Coumarin Medium#

    PubMed Central

    Guan, Shu; Ji, Cheng; Zhou, Ting; Li, Junxia; Ma, Qiugang; Niu, Tiangui

    2008-01-01

    Aflatoxin B1 (AFB1) is one of the most harmful mycotoxins in animal production and food industry. A safe, effective and environmentally sound detoxification method is needed for controlling this toxin. In this study, 65 samples were screened from various sources with vast microbial populations using a newly developed medium containing coumarin as the sole carbon source. Twenty five single-colony bacterial isolates showing AFB1 reduction activity in a liquid culture medium were selected from the screen. Isolate 35-3, obtained from tapir feces and identified to be Stenotrophomonas maltophilia, reduced AFB1 by 82.5% after incubation in the liquid medium at 37 °C for 72 h. The culture supernatant of isolate 35-3 was able to degrade AFB1 effectively, whereas the viable cells and cell extracts were far less effective. Factors influencing AFB1 degradation by the culture supernatant were investigated. Activity was reduced to 60.8% and 63.5% at 20 °C and 30 °C, respectively, from 78.7% at 37 °C. The highest degradation rate was 84.8% at pH 8 and the lowest was only 14.3% at pH 4.0. Ions Mg2+ and Cu2+ were activators for AFB1 degradation, however ion Zn2+ was a strong inhibitor. Treatments with proteinase K, proteinase K plus SDS and heating significantly reduced or eradicated the degradation activity of the culture supernatant. The results indicated that the degradation of AFB1 by S. maltophilia 35-3 was enzymatic and could have a great potential in industrial applications. PMID:19325817

  6. Decoding the genetic and functional diversity of the DSF quorum-sensing system in Stenotrophomonas maltophilia

    PubMed Central

    Huedo, Pol; Yero, Daniel; Martinez-Servat, Sònia; Ruyra, Àngels; Roher, Nerea; Daura, Xavier; Gibert, Isidre

    2015-01-01

    Stenotrophomonas maltophilia uses the Diffusible Signal Factor (DSF) quorum sensing (QS) system to mediate intra- and inter-specific signaling and regulate virulence-related processes. The components of this system are encoded by the rpf cluster, with genes rpfF and rpfC encoding for the DSF synthase RpfF and sensor RpfC, respectively. Recently, we have shown that there exist two variants of the rpf cluster (rpf-1 and rpf-2), distinguishing two groups of S. maltophilia strains. Surprisingly, only rpf-1 strains produce detectable DSF, correlating with their ability to control biofilm formation, swarming motility and virulence. The evolutive advantage of acquiring two different rpf clusters, the phylogenetic time point and mechanism of this acquisition and the conditions that activate DSF production in rpf-2 strains, are however not known. Examination of this cluster in various species suggests that its variability originated most probably by genetic exchange between rhizosphere bacteria. We propose that rpf-2 variant strains make use of a strategy recently termed as “social cheating.” Analysis of cellular and extracellular fatty acids (FAs) of strains E77 (rpf-1) and M30 (rpf-2) suggests that their RpfFs have also a thioesterase activity that facilitates the release of unspecific FAs to the medium in addition to DSF. Production of DSF in rpf-1 strains appears in fact to be modulated by some of these extracellular FAs in addition to other factors such as temperature and nutrients, while in rpf-2 strains DSF biosynthesis is derepressed only upon detection of DSF itself, suggesting that they require cohabitation with DSF-producer bacteria to activate their DSF regulatory machinery. Finally, we show that the mixed rpf-1/rpf-2 population presents synergism in DSF production and virulence capacity in an in vivo infection model. Recovery and quantification of DSF from co-infected animals correlates with the observed mortality rate. PMID:26284046

  7. Antibiogram of Stenotrophomonas maltophilia Isolated From Nkonkobe Municipality, Eastern Cape Province, South Africa

    PubMed Central

    Adegoke, Anthony Ayodeji; Okoh, Anthony I.

    2014-01-01

    Background: Assessment of resistance genes is imperative, as they become disseminated to bacterial flora in plants and to the indigenous bacterial community, and thus ultimately contributes to the clinical problems of antibiotic resistant pathogens. Objectives: The research was to assess the antibiotic characteristics and incidence of sul3 genes of Stenotrophomonas maltophilia isolates recovered from rhizospheres plant in Nkonkobe Municipality. Materials and Methods: Identification and assessment of resistance genes (sul2 and sul3 genes) were carried out using polymerase chain reaction (PCR). Analytical profile index (API) was used for biochemical characterization for identification before the PCR. Antibiotic susceptibility test was carried out using the approved guidelines and standards of Clinical Laboratory Standard Institute (CLSI). Results: A total of 125 isolates were identified, composed of 120 (96%) from grass root rhizosphere and 5 (4%) from soil butternut root rhizosphere. In vitro antibiotic susceptibility tests showed varying resistances to meropenem (8.9%), cefuroxime (95.6 %), ampicillin-sulbactam (53.9%), ceftazidime (10.7%), cefepime (29.3 %), minocycline (2.2%), kanamycin (56.9%), ofloxacin (2.9%), levofloxacin (1.3%), moxifloxacin (2.8%), ciprofloxacin (24.3%), gatifloxacin (1.3%), polymyxin B (2.9 %), cotrimoxazole (26.1%), trimethoprim (98.6%) and aztreonam (58%). The isolates were susceptible to the fluoroquinolones (74.3-94.7%), polymycin (97.1%) and meropenem (88.1%). The newest sulphonamide resistance gene, sul3, was detected among the trimethoprim-sulfamethoxazole (cotrimoxazole)-resistant isolates, while the most frequent sulphonamide-resistant gene in animal source isolates, sul2, was not. Conclusions: The commensal S. maltophilia isolates in the Nkonkobe Municipality environment harbored the resistant gene sul3 as clinical counterparts, especially from the perspective of reservoirs of antibiotic resistance determinants. PMID:25789125

  8. Comparative Genomics of Environmental and Clinical Stenotrophomonas maltophilia Strains with Different Antibiotic Resistance Profiles

    PubMed Central

    Youenou, Benjamin; Favre-Bonté, Sabine; Bodilis, Josselin; Brothier, Elisabeth; Dubost, Audrey; Muller, Daniel; Nazaret, Sylvie

    2015-01-01

    Stenotrophomonas maltophilia, a ubiquitous Gram-negative γ-proteobacterium, has emerged as an important opportunistic pathogen responsible for nosocomial infections. A major characteristic of clinical isolates is their high intrinsic or acquired antibiotic resistance level. The aim of this study was to decipher the genetic determinism of antibiotic resistance among strains from different origins (i.e., natural environment and clinical origin) showing various antibiotic resistance profiles. To this purpose, we selected three strains isolated from soil collected in France or Burkina Faso that showed contrasting antibiotic resistance profiles. After whole-genome sequencing, the phylogenetic relationships of these 3 strains and 11 strains with available genome sequences were determined. Results showed that a strain’s phylogeny did not match their origin or antibiotic resistance profiles. Numerous antibiotic resistance coding genes and efflux pump operons were revealed by the genome analysis, with 57% of the identified genes not previously described. No major variation in the antibiotic resistance gene content was observed between strains irrespective of their origin and antibiotic resistance profiles. Although environmental strains generally carry as many multidrug resistant (MDR) efflux pumps as clinical strains, the absence of resistance–nodulation–division (RND) pumps (i.e., SmeABC) previously described to be specific to S. maltophilia was revealed in two environmental strains (BurA1 and PierC1). Furthermore the genome analysis of the environmental MDR strain BurA1 showed the absence of SmeABC but the presence of another putative MDR RND efflux pump, named EbyCAB on a genomic island probably acquired through horizontal gene transfer. PMID:26276674

  9. A Stenotrophomonas maltophilia Multilocus Sequence Typing Scheme for Inferring Population Structure▿ †

    PubMed Central

    Kaiser, Sabine; Biehler, Klaus; Jonas, Daniel

    2009-01-01

    Stenotrophomonas maltophilia is an opportunistic, highly resistant, and ubiquitous pathogen. Strains have been assigned to genogroups using amplified fragment length polymorphism. Hence, isolates of environmental and clinical origin predominate in different groups. A multilocus sequence typing (MLST) scheme was developed using a highly diverse selection of 70 strains of various ecological origins from seven countries on all continents including strains of the 10 previously defined genogroups. Sequence data were assigned to 54 sequence types (ST) based on seven loci. Indices of association for all isolates and clinical isolates of 2.498 and 2.562 indicated a significant linkage disequilibrium, as well as high congruence of tree topologies from different loci. Potential recombination events were detected in one-sixth of all ST. Calculation of the mean divergence between and within predicted clusters confirmed previously defined groups and revealed five additional groups. Consideration of the different ecological origins showed that 18 out of 31 respiratory tract isolates, including 12 out of 19 isolates from cystic fibrosis (CF) patients, belonged to genogroup 6. In contrast, 16 invasive strains isolated from blood cultures were distributed among nine different genogroups. Three genogroups contained isolates of strictly environmental origin that also featured high sequence distances to other genogroups, including the S. maltophilia type strain. On the basis of this MLST scheme, isolates can be assigned to the genogroups of this species in order to further scrutinize the population structure of this species and to unravel the uneven distribution of environmental and clinical isolates obtained from infected, colonized, or CF patients. PMID:19251858

  10. Stenotrophomonas maltophilia Phenotypic and Genotypic Diversity during a 10-year Colonization in the Lungs of a Cystic Fibrosis Patient

    PubMed Central

    Pompilio, Arianna; Crocetta, Valentina; Ghosh, Dipankar; Chakrabarti, Malabika; Gherardi, Giovanni; Vitali, Luca Agostino; Fiscarelli, Ersilia; Di Bonaventura, Giovanni

    2016-01-01

    The present study was carried out to understand the adaptive strategies developed by Stenotrophomonas maltophilia for chronic colonization of the cystic fibrosis (CF) lung. For this purpose, 13 temporally isolated strains from a single CF patient chronically infected over a 10-year period were systematically characterized for growth rate, biofilm formation, motility, mutation frequencies, antibiotic resistance, and pathogenicity. Pulsed-field gel electrophoresis (PFGE) showed over time the presence of two distinct groups, each consisting of two different pulsotypes. The pattern of evolution followed by S. maltophilia was dependent on pulsotype considered, with strains belonging to pulsotype 1.1 resulting to be the most adapted, being significantly changed in all traits considered. Generally, S. maltophilia adaptation to CF lung leads to increased growth rate and antibiotic resistance, whereas both in vivo and in vitro pathogenicity as well as biofilm formation were decreased. Overall, our results show for the first time that S. maltophilia can successfully adapt to a highly stressful environment such as CF lung by paying a “biological cost,” as suggested by the presence of relevant genotypic and phenotypic heterogeneity within bacterial population. S. maltophilia populations are, therefore, significantly complex and dynamic being able to fluctuate rapidly under changing selective pressures. PMID:27746770

  11. Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli.

    PubMed

    Ribitsch, D; Heumann, S; Karl, W; Gerlach, J; Leber, R; Birner-Gruenberger, R; Gruber, K; Eiteljoerg, I; Remler, P; Siegert, P; Lange, J; Maurer, K H; Berg, G; Guebitz, G M; Schwab, H

    2012-01-01

    A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45°C. Specific activity of StmPr2 determined with suc-L-Ala-L-Ala-L-Pro-l-Phe-p-nitroanilide as the substrate was 17±2U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2.

  12. Global and local selection acting on the pathogen Stenotrophomonas maltophilia in the human lung

    PubMed Central

    Chung, Hattie; Lieberman, Tami D.; Vargas, Sara O.; Flett, Kelly B.; McAdam, Alexander J.; Priebe, Gregory P.; Kishony, Roy

    2017-01-01

    Bacterial populations diversify during infection into distinct subpopulations that coexist within the human body. Yet, it is unknown to what extent subpopulations adapt to location-specific selective pressures as they migrate and evolve across space. Here we identify bacterial genes under local and global selection by testing for spatial co-occurrence of adaptive mutations. We sequence 552 genomes of the pathogen Stenotrophomonas maltophilia across 23 sites of the lungs from a patient with cystic fibrosis. We show that although genetically close isolates colocalize in space, distant lineages with distinct phenotypes separated by adaptive mutations spread throughout the lung, suggesting global selective pressures. Yet, for one gene (a distant homologue of the merC gene implicated in metal resistance), mutations arising independently in two lineages colocalize in space, providing evidence for location-specific selection. Our work presents a general framework for understanding how selection acts upon a pathogen that colonizes and evolves across the complex environment of the human body. PMID:28102223

  13. High-level quinolone resistance is associated with the overexpression of smeVWX in Stenotrophomonas maltophilia clinical isolates.

    PubMed

    García-León, G; Ruiz de Alegría Puig, C; García de la Fuente, C; Martínez-Martínez, L; Martínez, J L; Sánchez, M B

    2015-05-01

    Stenotrophomonas maltophilia is the only known bacterium in which quinolone-resistant isolates do not present mutations in the genes encoding bacterial topoisomerases. The expression of the intrinsic quinolone resistance elements smeDEF, smeVWX and Smqnr was analysed in 31 clinical S. maltophilia isolates presenting a minimum inhibitory concentration (MIC) range to ciprofloxacin between 0.5 and > 32 μg/mL; 11 (35.5%) overexpressed smeDEF, 2 (6.5%) presenting the highest quinolone MICs overexpressed smeVWX and 1 (3.2%) overexpressed Smqnr. Both strains overexpressing smeVWX presented changes at the Gly266 position of SmeRv, the repressor of smeVWX. Changes at the same position were previously observed in in vitro selected S. maltophilia quinolone-resistant mutants, indicating this amino acid is highly relevant for the activity of SmeRv in repressing smeVWX expression. For the first time SmeVWX overexpression is associated with quinolone resistance of S. maltophilia clinical isolates.

  14. Antibiotic susceptibility of sulfamethoxazole-trimethoprim resistant Stenotrophomonas maltophilia strains isolated at a tertiary care centre in Hungary.

    PubMed

    Juhász, Emese; Pongrácz, Júlia; Iván, Miklós; Kristóf, Katalin

    2015-09-01

    Sulfamethoxazole-trimethoprim (SXT) is the drug-of-choice in Stenotrophomonas maltophilia caused infections. There has been an increase in resistance to SXT of S. maltophilia over recent years. In this study 30 S. maltophilia clinical isolates resistant to SXT were investigated. Antibiotic susceptibilities for ciprofloxacin, moxifloxacin, levofloxacin, doxycycline, tigecycline, ceftazidime, colistin and chloramphenicol were determined by broth microdilution method. None of the strains were susceptible to ciprofloxacin, tigecycline, ceftazidime or colistin. Only 37% of the isolates were susceptible to levofloxacin or moxifloxacin. Two isolates resistant to all tested antibiotic agents and two others susceptible only to doxycycline were further investigated: susceptibility for combinations of antibiotics was analyzed by checkerboard technique. According to the fractional inhibitory concentration indices calculated, moxifloxacin plus ceftazidime combination was found to be synergistic in each case. Genetic testing revealed the predominance of sul1 gene. Our study concluded that the range of effective antibiotic agents is even more limited in infections caused by SXT-resistant S. maltophilia. In these cases, in vitro synergistic antibiotic combinations could be potential therapeutic options.

  15. Stenotrophomonas maltophilia Virulence and Specific Variations in Trace Elements during Acute Lung Infection: Implications in Cystic Fibrosis

    PubMed Central

    Crocetta, Valentina; Consalvo, Ada; Zappacosta, Roberta; Di Ilio, Carmine; Di Bonaventura, Giovanni

    2014-01-01

    Metal ions are necessary for the proper functioning of the immune system, and, therefore, they might have a significant influence on the interaction between bacteria and host. Ionic dyshomeostasis has been recently observed also in cystic fibrosis (CF) patients, whose respiratory tract is frequently colonized by Stenotrophomonas maltophilia. For the first time, here we used an inductively mass spectrometry method to perform a spatial and temporal analysis of the pattern of changes in a broad range of major trace elements in response to pulmonary infection by S. maltophilia. To this, DBA/2 mouse lungs were comparatively infected by a CF strain and by an environmental one. Our results showed that pulmonary ionomic profile was significantly affected during infection. Infected mice showed increased lung levels of Mg, P, S, K, Zn, Se, and Rb. To the contrary, Mn, Fe, Co, and Cu levels resulted significantly decreased. Changes of element concentrations were correlated with pulmonary bacterial load and markers of inflammation, and occurred mostly on day 3 post-exposure, when severity of infection culminated. Interestingly, CF strain – significantly more virulent than the environmental one in our murine model - provoked a more significant impact in perturbing pulmonary metal homeostasis. Particularly, exposure to CF strain exclusively increased P and K levels, while decreased Fe and Mn ones. Overall, our data clearly indicate that S. maltophilia modulates pulmonary metal balance in a concerted and virulence-dependent manner highlighting the potential role of the element dyshomeostasis during the progression of S. maltophilia infection, probably exacerbating the harmful effects of the loss of CF transmembrane conductance regulator function. Further investigations are required to understand the biological significance of these alterations and to confirm they are specifically caused by S. maltophilia. PMID:24586389

  16. Genome Sequence of a Multidrug-Resistant Strain of Stenotrophomonas maltophilia with Carbapenem Resistance, Isolated from King Abdullah Medical City, Makkah, Saudi Arabia

    PubMed Central

    Abdel-Haleem, Alyaa M.; Rchiad, Zineb; Khan, Babar K.; Abdallah, Abdallah M.; Naeem, Raeece; Nikhat Sheerin, Shalam; Solovyev, Victor; Ahmed, Abdalla

    2015-01-01

    The emergence and spread of multidrug-resistant (MDR) bacteria have been regarded as major challenges among health care-associated infections worldwide. Here, we report the draft genome sequence of an MDR Stenotrophomonas maltophilia strain isolated in 2014 from King Abdulla Medical City, Makkah, Saudi Arabia. PMID:26472828

  17. Draft Genome Sequence of Stenotrophomonas maltophilia CBF10-1, an Organophosphate-Degrading Bacterium Isolated from Ranch Soil in Fairchilds, Texas

    PubMed Central

    Damania, Ashish

    2016-01-01

    Stenotrophomonas maltophilia CBF10-1 was isolated from a ranch in Fairchilds, Texas, USA. Its genome reveals a highly adaptable microorganism with a large complement of antibiotic and heavy metal resistance genes, efflux pumps, multidrug transporters, and xenobiotic degradation pathways. PMID:27174285

  18. Honeydew honey as a potent antibacterial agent in eradication of multi-drug resistant Stenotrophomonas maltophilia isolates from cancer patients.

    PubMed

    Majtan, Juraj; Majtanova, Lubica; Bohova, Jana; Majtan, Viktor

    2011-04-01

    Multi-drug resistance in nosocomial pathogens is a continually evolving and alarming problem in health care units. Since ancient times, honey has been used successfully for the treatment of a broad spectrum of infections with no risk of resistance development. This study investigated the antibacterial activity of two natural honeys, namely honeydew and manuka, against 20 nosocomial multi-drug resistant Stenotrophomonas maltophilia (S. maltophilia) isolates from cancer patients. An antibiotic susceptibility test was carried out using the disk diffusion method with 20 antibiotic disks. The antibacterial activity of honey was determined using a broth dilution method. The concentration of honey used in the study was within the range of 3.75% to 25% (w/v). All 20 clinical isolates were multi-drug resistant against 11 to 19 antibiotics. The MICs for honeydew honey ranged from 6.25% to 17.5%, while those for active manuka honey ranged from 7.5% to 22.5%. Honeydew honey had lower MICs than manuka honey against 16 of the tested isolates. This study showed that Slovak honeydew honey has exceptional antibacterial activity against multi-drug resistant S. maltophilia isolates and was more efficient than manuka honey (UMF 15+). Honeydew honey with strong antibacterial activity could be used as a potential agent to eradicate multi-drug resistant clinical isolates.

  19. Surfactant-enhanced biodegradation of high molecular weight polycyclic aromatic hydrocarbons by stenotrophomonas maltophilia

    PubMed

    Boonchan; Britz; Stanley

    1998-08-20

    The objectives of this study were to isolate and evaluate microorganisms with the ability to degrade high molecular weight polycyclic aromatic hydrocarbons (PAHs) in the presence of synthetic surfactants. Stenotrophomonas maltophilia VUN 10,010, isolated from PAH-contaminated soil, utilized pyrene as a sole carbon and energy source and also degraded other high molecular weight PAHs containing up to seven benzene rings. Various synthetic surfactants were tested for their ability to improve the PAH degradation rate of strain VUN 10,010. Anionic and cationic surfactants were highly toxic to this strain, and the Tween series was used as a growth substrate. Five nonionic surfactants (Brij 35, Igepal CA-630, Triton X-100, Tergitol NP-10, and Tyloxapol) were not utilized by, and were less toxic to, strain VUN 10,010. MSR and log Km values were determined for fluoranthene, pyrene, and benzo[a]pyrene in the presence of these nonionic surfactants and their apparent solubility was increased by a minimum of 250-fold in the presence of 10 g L-1 of all surfactants. The rate of pyrene degradation by strain VUN 10,010 was enhanced by the addition of four of the nonionic surfactants (5-10 g L-1); however, 5 g L-1 Igepal CA-630 inhibited pyrene degradation and microbial growth. The specific growth rate of VUN 10,010 on pyrene was increased by 67% in the presence of 10 g L-1 Brij 35 or Tergitol NP-10. The addition of Brij 35 and Tergitol NP-10 to media containing a single high molecular weight PAH (four and five benzene rings) as the sole carbon source increased the maximum specific PAH degradation rate and decreased the lag period normally seen for PAH degradation. The addition of Tergitol NP-10 to VUN 10,010 cultures which contained a PAH mixture (three to seven benzene rings) substantially improved the overall degradation rate of each PAH and increased the specific growth rate of VUN 10,010 by 30%. Evaluation of the use of VUN 10,010 for degrading high molecular weight PAHs in

  20. A function of SmeDEF, the major quinolone resistance determinant of Stenotrophomonas maltophilia, is the colonization of plant roots.

    PubMed

    García-León, Guillermo; Hernández, Alvaro; Hernando-Amado, Sara; Alavi, Peyman; Berg, Gabriele; Martínez, José Luis

    2014-08-01

    Quinolones are synthetic antibiotics, and the main cause of resistance to these antimicrobials is mutation of the genes encoding their targets. However, in contrast to the case for other organisms, such mutations have not been found in quinolone-resistant Stenotrophomonas maltophilia isolates, in which overproduction of the SmeDEF efflux pump is a major cause of quinolone resistance. SmeDEF is chromosomally encoded and highly conserved in all studied S. maltophilia strains; it is an ancient element that evolved over millions of years in this species. It thus seems unlikely that its main function would be resistance to quinolones, a family of synthetic antibiotics not present in natural environments until the last few decades. Expression of SmeDEF is tightly controlled by the transcriptional repressor SmeT. Our work shows that plant-produced flavonoids can bind to SmeT, releasing it from smeDEF and smeT operators. Antibiotics extruded by SmeDEF do not impede the binding of SmeT to DNA. The fact that plant-produced flavonoids specifically induce smeDEF expression indicates that they are bona fide effectors regulating expression of this resistance determinant. Expression of efflux pumps is usually downregulated unless their activity is needed. Since smeDEF expression is triggered by plant-produced flavonoids, we reasoned that this efflux pump may have a role in the colonization of plants by S. maltophilia. Our results showed that, indeed, deletion of smeE impairs S. maltophilia colonization of plant roots. Altogether, our results indicate that quinolone resistance is a recent function of SmeDEF and that colonization of plant roots is likely one original function of this efflux pump.

  1. Clinical Characteristics of Stenotrophomonas maltophilia Bacteremia: A Regional Report and a Review of a Japanese Case Series

    PubMed Central

    Ebara, Hirotaka; Hagiya, Hideharu; Haruki, Yuto; Kondo, Eisei; Otsuka, Fumio

    2017-01-01

    Objective Stenotrophomonas maltophilia is an emerging nosocomial pathogen that causes fatal infections in critically ill or immunocompromised patients. S. maltophilia bacteremia (SMB) is a rare condition, and its clinical characteristics in Japanese settings are not well known. Methods The medical charts of patients with SMB were retrospectively reviewed at two medical facilities (Okayama University Hospital and Tsuyama Chuo Hospital) for seven years. The data were analyzed along with those previously reported from other Japanese facilities. Result A total of 181 patients (110 men and 71 women) were evaluated. The major underlying diseases included hematologic malignancy (36.5%), solid organ malignancy (25.4%), and neutropenia (31.5%). The recent use of carbapenem was seen in 56.9% of the cases in total, and more than one-third of the patients in our hospitals were treated with carbapenem at the onset of SMB. Of 28 (63.6%) of 44 cases treated for S. maltophilia, those who did not survive were more likely to have been treated with broad-spectrum antibiotics. A multivariate analysis revealed that a higher updated Charlson Comorbidity Index [odds ratio (95% confidence interval), 1.75 (1.11-2.75); p=0.015] and intubation [odds ratio (95% confidence interval), 12.6 (1.62-97.9); p=0.016] were associated with mortality in our cases. Pathogens were often resistant to ceftazidime but susceptible to minocycline, trimethoprim/sulfamethoxazole, and fluoroquinolones. The overall mortality rates within 30 and 90 days were 37.5% and 62.5%, respectively. Conclusion The clinical characteristics of SMB in Japanese cases were similar to those reported from other countries. Clinicians should be aware that breakthrough infection by S. maltophilia may occur during administration of carbapenem. PMID:28090041

  2. Characteristics of Aspergillus fumigatus in Association with Stenotrophomonas maltophilia in an In Vitro Model of Mixed Biofilm

    PubMed Central

    Melloul, Elise; Luiggi, Stéphanie; Anaïs, Leslie; Arné, Pascal; Costa, Jean-Marc; Fihman, Vincent; Briard, Benoit; Dannaoui, Eric; Guillot, Jacques; Decousser, Jean-Winoc; Beauvais, Anne; Botterel, Françoise

    2016-01-01

    Background Biofilms are communal structures of microorganisms that have long been associated with a variety of persistent infections poorly responding to conventional antibiotic or antifungal therapy. Aspergillus fumigatus fungus and Stenotrophomonas maltophilia bacteria are examples of the microorganisms that can coexist to form a biofilm especially in the respiratory tract of immunocompromised patients or cystic fibrosis patients. The aim of the present study was to develop and assess an in vitro model of a mixed biofilm associating S. maltophilia and A. fumigatus by using analytical and quantitative approaches. Materials and Methods An A. fumigatus strain (ATCC 13073) expressing a Green Fluorescent Protein (GFP) and an S. maltophilia strain (ATCC 13637) were used. Fungal and bacterial inocula (105 conidia/mL and 106 cells/mL, respectively) were simultaneously deposited to initiate the development of an in vitro mixed biofilm on polystyrene supports at 37°C for 24 h. The structure of the biofilm was analysed via qualitative microscopic techniques like scanning electron and transmission electron microscopy, and fluorescence microscopy, and by quantitative techniques including qPCR and crystal violet staining. Results Analytic methods revealed typical structures of biofilm with production of an extracellular matrix (ECM) enclosing fungal hyphae and bacteria. Quantitative methods showed a decrease of A. fumigatus growth and ECM production in the mixed biofilm with antibiosis effect of the bacteria on the fungi seen as abortive hyphae, limited hyphal growth, fewer conidia, and thicker fungal cell walls. Conclusion For the first time, a mixed A. fumigatus—S. maltophilia biofilm was validated by various analytical and quantitative approaches and the bacterial antibiosis effect on the fungus was demonstrated. The mixed biofilm model is an interesting experimentation field to evaluate efficiency of antimicrobial agents and to analyse the interactions between the

  3. Predictive Studies Suggest that the Risk for the Selection of Antibiotic Resistance by Biocides Is Likely Low in Stenotrophomonas maltophilia

    PubMed Central

    Sánchez, María Blanca; Decorosi, Francesca; Viti, Carlo; Oggioni, Marco Rinaldo; Martínez, José Luis; Hernández, Alvaro

    2015-01-01

    Biocides are used without restriction for several purposes. As a consequence, large amounts of biocides are released without any control in the environment, a situation that can challenge the microbial population dynamics, including selection of antibiotic resistant bacteria. Previous work has shown that triclosan selects Stenotrophomonas maltophilia antibiotic resistant mutants overexpressing the efflux pump SmeDEF and induces expression of this pump triggering transient low-level resistance. In the present work we analyze if two other common biocides, benzalkonium chloride and hexachlorophene, trigger antibiotic resistance in S. maltophilia. Bioinformatic and biochemical methods showed that benzalkonium chloride and hexachlorophene bind the repressor of smeDEF, SmeT. Only benzalkonium chloride triggers expression of smeD and its effect in transient antibiotic resistance is minor. None of the hexachlorophene-selected mutants was antibiotic resistant. Two benzalkonium chloride resistant mutants presented reduced susceptibility to antibiotics and were impaired in growth. Metabolic profiling showed they were more proficient than their parental strain in the use of some dipeptides. We can then conclude that although bioinformatic predictions and biochemical studies suggest that both hexachlorophene and benzalkonium chloride should induce smeDEF expression leading to transient S. maltophilia resistance to antibiotics, phenotypic assays showed this not to be true. The facts that hexachlorophene resistant mutants are not antibiotic resistant and that the benzalkonium chloride resistant mutants presenting altered susceptibility to antibiotics were impaired in growth suggests that the risk for the selection (and fixation) of S. maltophilia antibiotic resistant mutants by these biocides is likely low, at least in the absence of constant selection pressure. PMID:26201074

  4. Predictive Studies Suggest that the Risk for the Selection of Antibiotic Resistance by Biocides Is Likely Low in Stenotrophomonas maltophilia.

    PubMed

    Sánchez, María Blanca; Decorosi, Francesca; Viti, Carlo; Oggioni, Marco Rinaldo; Martínez, José Luis; Hernández, Alvaro

    2015-01-01

    Biocides are used without restriction for several purposes. As a consequence, large amounts of biocides are released without any control in the environment, a situation that can challenge the microbial population dynamics, including selection of antibiotic resistant bacteria. Previous work has shown that triclosan selects Stenotrophomonas maltophilia antibiotic resistant mutants overexpressing the efflux pump SmeDEF and induces expression of this pump triggering transient low-level resistance. In the present work we analyze if two other common biocides, benzalkonium chloride and hexachlorophene, trigger antibiotic resistance in S. maltophilia. Bioinformatic and biochemical methods showed that benzalkonium chloride and hexachlorophene bind the repressor of smeDEF, SmeT. Only benzalkonium chloride triggers expression of smeD and its effect in transient antibiotic resistance is minor. None of the hexachlorophene-selected mutants was antibiotic resistant. Two benzalkonium chloride resistant mutants presented reduced susceptibility to antibiotics and were impaired in growth. Metabolic profiling showed they were more proficient than their parental strain in the use of some dipeptides. We can then conclude that although bioinformatic predictions and biochemical studies suggest that both hexachlorophene and benzalkonium chloride should induce smeDEF expression leading to transient S. maltophilia resistance to antibiotics, phenotypic assays showed this not to be true. The facts that hexachlorophene resistant mutants are not antibiotic resistant and that the benzalkonium chloride resistant mutants presenting altered susceptibility to antibiotics were impaired in growth suggests that the risk for the selection (and fixation) of S. maltophilia antibiotic resistant mutants by these biocides is likely low, at least in the absence of constant selection pressure.

  5. Stenotrophomonas maltophilia as a part of normal oral bacterial flora in captive snakes and its susceptibility to antibiotics.

    PubMed

    Hejnar, Petr; Bardon, Jan; Sauer, Pavel; Kolár, Milan

    2007-04-15

    Only little is known about normal oral bacterial flora in captive snakes containing Stenotrophomonas maltophilia. This microbe has been reported as a causative agent of numerous infections in reptiles. Therefore, the goal of the study was to detect its presence in the mouths of a significant number of healthy captive snakes and determining its susceptibility to antibiotics at 30 and 37 degrees C. The isolates were obtained in 1999-2005 from mouth swabs of 115 snakes of 12 genera and 22 species-most often Elaphe guttata (24 individuals; 20.9%). Susceptibility to 24 antibiotics was tested by the microdilution method. The microbe was demonstrated in 34 (29.6%) individuals. Overall, 47 strains of S. maltophilia were acquired. Evaluation using PFGE profiles and antibiograms resulted in confirmation of one strain of S. maltophilia in 23 (20.0%) individuals, two strains in nine (7.8%) and three in two (1.8%) snakes. All tested antibiotics were more effective at 37 degrees C, with the partial exception of cotrimoxazole and cefoperazone/sulbactam. At a temperature of 37 degrees C, the lowest frequency of resistance to levofloxacin (no resistant strains), cotrimoxazole and ofloxacin (97.9% of susceptible strains) was recorded. At 30 degrees C, the most active agents were cotrimoxazole (97.9% of susceptible strains), levofloxacin (91.5%) and ofloxacin (85.1%). In conclusion, S. maltophilia is present in the mouths of about one third of healthy captive snakes, showing good susceptibility to cotrimoxazole, some fluoroquinolones and aminoglycosides. The antibiotics (particularly aminoglycosides) are more effective at 37 degrees C.

  6. A Function of SmeDEF, the Major Quinolone Resistance Determinant of Stenotrophomonas maltophilia, Is the Colonization of Plant Roots

    PubMed Central

    García-León, Guillermo; Hernández, Alvaro; Hernando-Amado, Sara; Alavi, Peyman; Berg, Gabriele

    2014-01-01

    Quinolones are synthetic antibiotics, and the main cause of resistance to these antimicrobials is mutation of the genes encoding their targets. However, in contrast to the case for other organisms, such mutations have not been found in quinolone-resistant Stenotrophomonas maltophilia isolates, in which overproduction of the SmeDEF efflux pump is a major cause of quinolone resistance. SmeDEF is chromosomally encoded and highly conserved in all studied S. maltophilia strains; it is an ancient element that evolved over millions of years in this species. It thus seems unlikely that its main function would be resistance to quinolones, a family of synthetic antibiotics not present in natural environments until the last few decades. Expression of SmeDEF is tightly controlled by the transcriptional repressor SmeT. Our work shows that plant-produced flavonoids can bind to SmeT, releasing it from smeDEF and smeT operators. Antibiotics extruded by SmeDEF do not impede the binding of SmeT to DNA. The fact that plant-produced flavonoids specifically induce smeDEF expression indicates that they are bona fide effectors regulating expression of this resistance determinant. Expression of efflux pumps is usually downregulated unless their activity is needed. Since smeDEF expression is triggered by plant-produced flavonoids, we reasoned that this efflux pump may have a role in the colonization of plants by S. maltophilia. Our results showed that, indeed, deletion of smeE impairs S. maltophilia colonization of plant roots. Altogether, our results indicate that quinolone resistance is a recent function of SmeDEF and that colonization of plant roots is likely one original function of this efflux pump. PMID:24837376

  7. Microbial degradation and detoxification of high molecular weight polycyclic aromatic hydrocarbons by Stenotrophomonas maltophilia strain VUN 10,003.

    PubMed

    Juhasz, A L; Stanley, G A; Britz, M L

    2000-05-01

    The ability of Stenotrophomonas maltophilia strain VUN 10,003 to degrade and detoxify high molecular weight polycyclic aromatic hydrocarbons (PAHs) was evaluated in a basal liquid medium. Using high cell density inocula of strain VUN 10,003, the concentration of pyrene, fluoranthene, benz[a]anthracene, benzo[a]pyrene, dibenz[a, h]anthracene and coronene decreased by 98, 45, 26, 22, 22 and 55% over periods ranging from 5 to 42 d. When a PAH mixture containing three- to seven-ring compounds was used, degradation of both low and high molecular weight compounds occurred concurrently. Mutagenicity assays (Ames Test) demonstrated a decrease in the mutagenic potential of dichloromethane culture extracts from all cultures containing single PAH over the incubation period, corresponding to the decrease in the concentration of the PAH. These observations indicate that strain VUN 10,003 could be used for the detoxification of PAH-contaminated wastes.

  8. [Methods for extraction of exopolymeric complex in plankton and biofilm growth mode of Stenotrophomonas maltophilia 22M].

    PubMed

    Boretskaia, M A; Suslova, O S

    2013-01-01

    The optimal methods for the extraction of exopolymeric complex (EPS) of Stenotrophomonas maltophilia 22M was determined. That EPS was synthesized in plankton and biofilm growth mode on the mild steel surface. It is desirable to use different physical and chemical methods for studying the EPS composition (carbohydrates and proteins) depending on the bacteria growth mode. In this way the interaction with ion exchange resin was the most effective for plankton growth mode to determine the maximum amount of carbohydrates (9.5 microg/ml), and the impact of heating to determine protein (3.9 microg/ml). For EPS biofilm in order to obtain maximum amount of carbohydrate it is desirable to use heating (30 microg/ml) and centrifugation (35 microg/ml). It is recommended to determine protein in the biofilm EPS after treatment with heating (3.75 microg/ml) and centrifugation (3.75 microg/ml).

  9. Purification and characterization of novel organic solvent tolerant 98kDa alkaline protease from isolated Stenotrophomonas maltophilia strain SK.

    PubMed

    Waghmare, Shailesh R; Gurav, Aparna A; Mali, Sonal A; Nadaf, Naiem H; Jadhav, Deepak B; Sonawane, Kailas D

    2015-03-01

    Ability of microorganisms to grow at alkaline pH makes them an attractive target for several industrial applications. Thus, search for new extremozyme producing microorganisms must be a continuous exercise. Hence, we isolated a potent alkaline protease producing bacteria from slaughter house soil. The morphological, biochemical and 16S rDNA gene sequencing studies revealed that the isolated bacteria is Stenotrophomonas maltophilia strain SK. Alkaline protease from S. maltophilia strain SK was purified by using ammonium sulphate precipitation and DEAE-cellulose ion exchange column chromatography. The purified enzyme was optimally active at pH 9.0 and temperature 40°C with broad substrate specificity. It was observed that the metal ions such as Ca(++), Mg(++) and Fe(+++) completely repressed the enzyme activity. The enzyme was stable in presence of various water miscible solvents like ethanol, methanol, isopropanol at 25% (v/v) concentration and less stable at 37.5% (v/v) concentration. These robust properties of enzyme might be applicable for various applications in detergent and pharmaceutical industries.

  10. Genome-Wide Identification of Genes Necessary for Biofilm Formation by Nosocomial Pathogen Stenotrophomonas maltophilia Reveals that Orphan Response Regulator FsnR Is a Critical Modulator

    PubMed Central

    Kang, Xiu-Min; Wang, Fang-Fang; Zhang, Huan

    2014-01-01

    Stenotrophomonas maltophilia is a Gram-negative bacterial pathogen of increasing concern to human health. Most clinical isolates of S. maltophilia efficiently form biofilms on biotic and abiotic surfaces, making this bacterium resistant to a number of antibiotic treatments and therefore difficult to eliminate. To date, very few studies have investigated the molecular and regulatory mechanisms responsible for S. maltophilia biofilm formation. Here we constructed a random transposon insertion mutant library of S. maltophilia ATCC 13637 and screened 14,028 clones. A total of 46 nonredundant genes were identified. Mutants of these genes exhibited marked changes in biofilm formation, suggesting that multiple physiological pathways, including extracellular polysaccharide production, purine synthesis, transportation, and peptide and lipid synthesis, are involved in bacterial cell aggregation. Of these genes, 20 putatively contributed to flagellar biosynthesis, indicating a critical role for cell motility in S. maltophilia biofilm formation. Genetic and biochemical evidence demonstrated that an orphan response regulator, FsnR, activated transcription of at least two flagellum-associated operons by directly binding to their promoters. This regulatory protein plays a fundamental role in controlling flagellar assembly, cell motility, and biofilm formation. These results provide a genetic basis to systematically study biofilm formation of S. maltophilia. PMID:25480754

  11. Synergistic activities of macrolide antibiotics against Pseudomonas aeruginosa, Burkholderia cepacia, Stenotrophomonas maltophilia, and Alcaligenes xylosoxidans isolated from patients with cystic fibrosis.

    PubMed

    Saiman, Lisa; Chen, Yunhua; Gabriel, Pablo San; Knirsch, Charles

    2002-04-01

    Azithromycin and clarithromycin were paired with other antibiotics to test synergistic activity against 300 multidrug-resistant pathogens isolated from cystic fibrosis (CF) patients. Clarithromycin-tobramycin was most active against Pseudomonas aeruginosa and inhibited 58% of strains. Azithromycin-trimethoprim-sulfamethoxazole, azithromycin-ceftazidime, and azithromycin-doxycycline or azithromycin-trimethoprim-sulfamethoxazole inhibited 40, 20, and 22% of Stenotrophomonas maltophilia, Burkholderia cepacia complex, and Achromobacter (Alcaligenes) xylosoxidans strains, respectively.

  12. Stenotrophomonas maltophilia responds to exogenous AHL signals through the LuxR solo SmoR (Smlt1839)

    PubMed Central

    Martínez, Paula; Huedo, Pol; Martinez-Servat, Sònia; Planell, Raquel; Ferrer-Navarro, Mario; Daura, Xavier; Yero, Daniel; Gibert, Isidre

    2015-01-01

    Quorum Sensing (QS) mediated by Acyl Homoserine Lactone (AHL) molecules are probably the most widespread and studied among Gram-negative bacteria. Canonical AHL systems are composed by a synthase (LuxI family) and a regulator element (LuxR family), whose genes are usually adjacent in the genome. However, incomplete AHL-QS machinery lacking the synthase LuxI is frequently observed in Proteobacteria, and the regulator element is then referred as LuxR solo. It has been shown that certain LuxR solos participate in interspecific communication by detecting signals produced by different organisms. In the case of Stenotrophomonas maltophilia, a preliminary genome sequence analysis revealed numerous putative luxR genes, none of them associated to a luxI gene. From these, the hypothetical LuxR solo Smlt1839, here designated SmoR, presents a conserved AHL binding domain and a helix-turn-helix DNA binding motif. Its genomic organization—adjacent to hchA gene—indicate that SmoR belongs to the new family “LuxR regulator chaperone HchA-associated.” AHL-binding assays revealed that SmoR binds to AHLs in-vitro, at least to oxo-C8-homoserine lactone, and it regulates operon transcription, likely by recognizing a conserved palindromic regulatory box in the hchA upstream region. Supplementation with concentrated supernatants from Pseudomonas aeruginosa, which contain significant amounts of AHLs, promoted swarming motility in S. maltophilia. Contrarily, no swarming stimulation was observed when the P. aeruginosa supernatant was treated with the lactonase AiiA from Bacillus subtilis, confirming that AHL contributes to enhance the swarming ability of S. maltophilia. Finally, mutation of smoR resulted in a swarming alteration and an apparent insensitivity to the exogenous AHLs provided by P. aeruginosa. In conclusion, our results demonstrate that S. maltophilia senses AHLs produced by neighboring bacteria through the LuxR solo SmoR, regulating population behaviors such as swarming

  13. Stenotrophomonas maltophilia responds to exogenous AHL signals through the LuxR solo SmoR (Smlt1839).

    PubMed

    Martínez, Paula; Huedo, Pol; Martinez-Servat, Sònia; Planell, Raquel; Ferrer-Navarro, Mario; Daura, Xavier; Yero, Daniel; Gibert, Isidre

    2015-01-01

    Quorum Sensing (QS) mediated by Acyl Homoserine Lactone (AHL) molecules are probably the most widespread and studied among Gram-negative bacteria. Canonical AHL systems are composed by a synthase (LuxI family) and a regulator element (LuxR family), whose genes are usually adjacent in the genome. However, incomplete AHL-QS machinery lacking the synthase LuxI is frequently observed in Proteobacteria, and the regulator element is then referred as LuxR solo. It has been shown that certain LuxR solos participate in interspecific communication by detecting signals produced by different organisms. In the case of Stenotrophomonas maltophilia, a preliminary genome sequence analysis revealed numerous putative luxR genes, none of them associated to a luxI gene. From these, the hypothetical LuxR solo Smlt1839, here designated SmoR, presents a conserved AHL binding domain and a helix-turn-helix DNA binding motif. Its genomic organization-adjacent to hchA gene-indicate that SmoR belongs to the new family "LuxR regulator chaperone HchA-associated." AHL-binding assays revealed that SmoR binds to AHLs in-vitro, at least to oxo-C8-homoserine lactone, and it regulates operon transcription, likely by recognizing a conserved palindromic regulatory box in the hchA upstream region. Supplementation with concentrated supernatants from Pseudomonas aeruginosa, which contain significant amounts of AHLs, promoted swarming motility in S. maltophilia. Contrarily, no swarming stimulation was observed when the P. aeruginosa supernatant was treated with the lactonase AiiA from Bacillus subtilis, confirming that AHL contributes to enhance the swarming ability of S. maltophilia. Finally, mutation of smoR resulted in a swarming alteration and an apparent insensitivity to the exogenous AHLs provided by P. aeruginosa. In conclusion, our results demonstrate that S. maltophilia senses AHLs produced by neighboring bacteria through the LuxR solo SmoR, regulating population behaviors such as swarming

  14. Surveillance of Dihydropteroate Synthase Genes in Stenotrophomonas maltophilia by LAMP: Implications for Infection Control and Initial Therapy

    PubMed Central

    Zhao, Jin; Xing, Yubin; Liu, Wei; Ni, Wentao; Wei, Chuanqi; Wang, Rui; Liu, Yunxi; Liu, Youning

    2016-01-01

    Stenotrophomonas maltophilia is a common nosocomial pathogen that causes high morbidity and mortality. Because of its inherent extended antibiotic resistance, therapeutic options for S. maltophilia are limited, and sulfamethoxazole/trimethoprim (SXT) is the only first-line antimicrobial recommended. However, with the spread of dihydropteroate synthase (sul1 and sul2) genes, global emergence of SXT resistance has been reported. There is an urgent need to develop a rapid and sensitive but cost-efficient method to monitor the dissemination of sul genes. In this study, we developed loop-mediated isothermal amplification (LAMP) assays for sul1 and sul2 using real-time turbidity and hydroxy naphthol blue coloration methods. The assays could quickly detect sul genes with high sensitivity and specificity. The LAMP detection limit was 0.74 pg/reaction of extracted genomic DNA for sul1 and 2.6 pg/reaction for sul2, which were both 10-fold more sensitive than the corresponding traditional PCR assays. Additionally, the LAMP assays could positively amplify DNA from sul1-producing strains, but not from the negative controls. We then used the LAMP assays to investigate the dissemination of sul genes among S. maltophilia isolates from patients in three hospitals in Beijing, China. Among 450 non-duplicated samples collected during 2012–2014, 56 (12.4%) strains were SXT-resistant. All these SXT-resistant strains were positive for sul genes, with 35 (62.5%) carrying sul1, 17 (30.4%) carrying sul2, and 4 (7.1%) carrying both sul1 and sul2, which indicated that sul genes were the predominant resistance mechanism. Of 394 SXT-susceptible strains, 16 were also sul-positive. To provide epidemiological data for the appropriate choice of antimicrobials for treatment of sul-positive S. maltophilia, we further tested the susceptibility to 18 antimicrobials. Among these, sul-positive strains showed the highest susceptibility to tetracycline derivatives, especially minocycline (MIC50/MIC90, 0

  15. Changes in fatty acid composition of Stenotrophomonas maltophilia KB2 during co-metabolic degradation of monochlorophenols.

    PubMed

    Nowak, Agnieszka; Greń, Izabela; Mrozik, Agnieszka

    2016-12-01

    The changes in the cellular fatty acid composition of Stenotrophomonas maltophilia KB2 during co-metabolic degradation of monochlorophenols in the presence of phenol as well as its adaptive mechanisms to these compounds were studied. It was found that bacteria were capable of degrading 4-chlorophenol (4-CP) completely in the presence of phenol, while 2-chlorophenol (2-CP) and 3-chlorophenol (3-CP) they degraded partially. The analysis of the fatty acid profiles indicated that adaptive mechanisms of bacteria depended on earlier exposure to phenol, which isomer they degraded, and on incubation time. In bacteria unexposed to phenol the permeability and structure of their membranes could be modified through the increase of hydroxylated and cyclopropane fatty acids, and straight-chain and hydroxylated fatty acids under 2-CP, 3-CP and 4-CP exposure, respectively. In the exposed cells, regardless of the isomer they degraded, the most important changes were connected with the increase of the contribution of branched fatty acid on day 4 and the content of hydroxylated fatty acids on day 7. The changes, particularly in the proportion of branched fatty acids, could be a good indicator for assessing the progress of the degradation of monochlorophenols by S. maltophilia KB2. In comparison, in phenol-degrading cells the increase of cyclopropane and straight-chain fatty acid content was established. These findings indicated the degradative potential of the tested strain towards the co-metabolic degradation of persistent chlorophenols, and extended the current knowledge about the adaptive mechanisms of these bacteria to such chemicals.

  16. A linkage between SmeIJK efflux pump, cell envelope integrity, and σE-mediated envelope stress response in Stenotrophomonas maltophilia.

    PubMed

    Huang, Yi-Wei; Liou, Rung-Shiuan; Lin, Yi-Tsung; Huang, Hsin-Hui; Yang, Tsuey-Ching

    2014-01-01

    Resistance nodulation division (RND) efflux pumps, such as the SmeIJK pump of Stenotrophomonas maltophilia, are known to contribute to the multidrug resistance in Gram-negative bacteria. However, some RND pumps are constitutively expressed even though no antimicrobial stresses occur, implying that there should be some physical implications for these RND pumps. In this study, the role of SmeIJK in antimicrobials resistance, envelope integrity, and σE-mediated envelope stress response (ESR) of S. maltophilia was assessed. SmeIJK was involved in the intrinsic resistance of S. maltophilia KJ to aminoglycosides and leucomycin. Compared with the wild-type KJ, the smeIJK deletion mutant exhibited growth retardation in the MH medium, an increased sensitivity to membrane-damaging agents (MDAs), as well as activation of an σE-mediated ESR. Moreover, the expression of smeIJK was further induced by sub-lethal concentrations of MDAs or surfactants in an σE-dependent manner. These data collectively suggested an alternative physiological role of smeIJK in cell envelope integrity maintenance and σE-mediated ESR beyond the efflux of antibiotics. Because of the necessity of the physiological role of SmeIJK in protecting S. maltophilia from the envelope stress, smeIJK is constitutively expressed, which, in turn, contributes the intrinsic resistance to aminoglycoside and leucomycin. This is the first demonstration of the linkage among RND-type efflux pump, cell envelope integrity, and σE-mediated ESR in S. maltophilia.

  17. Degradation of abamectin by newly isolated Stenotrophomonas maltophilia ZJB-14120 and characterization of its abamectin-tolerance mechanism.

    PubMed

    Wang, Yuan-Shan; Zheng, Xing-Chang; Hu, Qi-Wei; Zheng, Yu-Guo

    2015-06-01

    An abamectin (ABM)-degrading bacterium, Stenotrophomonas maltophilia ZJB-14120, was isolated and identified. This strain is capable of degrading 84.82% of ABM at an initial concentration of 200 mg/L over a 48 h incubation period. This strain showed efficient biodegradation ability (7.81 mg/L/h) to ABM and high tolerance (1000 mg/L) to all macrolides tested. In addition to ABM, emamectin, erythromycin and spiramycin can also be degraded by this strain. Modifications involving either reduction of the double bond between C22-C23 or replacement of the C25-group of ABM with a cyclohexyl group can completely inhibit biodegradation of ABM. The ABM-degrading capability of strain ZJB-14120 is likely to be intrinsic to its metabolism and could be inhibited by incubating with erythromycin, azithromycin, spiramycin or rifampicin. A new and successive degradation pathway was proposed based on metabolite analysis. Although there is evidence for metabolite inhibition, this strain has high ABM degradation activity and reusability. Further investigation showed that activated macrolide efflux pump(s) and an undetermined mechanism for regulating the intracellular ABM concentration are responsible for normal uptake of essential metabolites while pumping out excess harmful compounds. Strain ZJB-14120 may provide efficient treatment of water and soil contaminated by toxic levels of abamectin and emamectin.

  18. A Patient Presenting with Cholangitis due to Stenotrophomonas Maltophilia and Pseudomonas Aeruginosa Successfully Treated with Intrabiliary Colistine

    PubMed Central

    Pérez, Pablo N.; Ramírez, María A.; Fernández, José A.; de Guevara, Laura Ladrón

    2014-01-01

    Anatomical barriers for antibiotic penetration can pose a particular challenge in the clinical setting. Stenotrophomonas maltophilia (SM) and Pseudomonas aeruginosa (PA) are two pathogens capable of developing multiple drug-resistance (MDR) mechanisms. We report the case of a 56-year-old female patient with a permanent percutaneous transhepatic biliary drainage (PTBD), who was admitted to our hospital with a cholangitis due to a MDR Escherichia coli strain. Upon admission, culture-guided antimicrobial therapy was conducted and the biliary catheter was replaced, with poor clinical response. Subsequently, SM and PA were detected. Treatment with fosfomycin and colistine was initiated, again without adequate response. Systemic colistine and tigecycline along with an intrabiliary infusion of colistine for 5 days was then used, followed by parenteral fosfomycin and tigecycline for 7 days. The patient was then successfully discharged. This is the first case report we are aware of on the use of intrabiliary colistine. It describes a new approach to treating cholangitis by MDR bacteria in patients with a PTBD. PMID:25002957

  19. A Patient Presenting with Cholangitis due to Stenotrophomonas Maltophilia and Pseudomonas Aeruginosa Successfully Treated with Intrabiliary Colistine.

    PubMed

    Pérez, Pablo N; Ramírez, María A; Fernández, José A; de Guevara, Laura Ladrón

    2014-05-13

    Anatomical barriers for antibiotic penetration can pose a particular challenge in the clinical setting. Stenotrophomonas maltophilia (SM) and Pseudomonas aeruginosa (PA) are two pathogens capable of developing multiple drug-resistance (MDR) mechanisms. We report the case of a 56-year-old female patient with a permanent percutaneous transhepatic biliary drainage (PTBD), who was admitted to our hospital with a cholangitis due to a MDR Escherichia coli strain. Upon admission, culture-guided antimicrobial therapy was conducted and the biliary catheter was replaced, with poor clinical response. Subsequently, SM and PA were detected. Treatment with fosfomycin and colistine was initiated, again without adequate response. Systemic colistine and tigecycline along with an intrabiliary infusion of colistine for 5 days was then used, followed by parenteral fosfomycin and tigecycline for 7 days. The patient was then successfully discharged. This is the first case report we are aware of on the use of intrabiliary colistine. It describes a new approach to treating cholangitis by MDR bacteria in patients with a PTBD.

  20. In vitro efficacy of copper and silver ions in eradicating Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii: implications for on-site disinfection for hospital infection control.

    PubMed

    Huang, Hsin-I; Shih, Hsiu-Yun; Lee, Chien-Ming; Yang, Thomas C; Lay, Jiunn-Jyi; Lin, Yusen E

    2008-01-01

    Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter baumannii are major opportunistic waterborne pathogens causing hospital-acquired infections. Copper-silver ionization has been shown to be effective in controlling Legionella colonization in hospital water systems. The objective was to determine the efficacy of copper and silver ions alone and in combination in eradicating P. aeruginosa, S. maltophilia and A. baumannii at the concentration applied to Legionella control. Kill curve experiments and mathematical modeling were conducted at copper and silver ion concentrations of 0.1, 0.2, 0.4, 0.8 and 0.01, 0.02, 0.04, 0.08 mg/L, respectively. The combinations of copper and silver ions were tested at concentrations of 0.2/0.02 and 0.4/0.04 mg/L, respectively. Initial organism concentration was ca. of 3 x 10(6)cfu/mL, and viability of the test organisms was assessed at predetermined time intervals. Samples (0.1 mL) withdrawn were mixed with 10 microL neutralizer solution immediately, serially diluted and plated in duplicate onto blood agar plates. The culture plates were incubated for 48 h at 37 degrees C and enumerated for the cfu (detection limit 10 cfu/mL). The results showed all copper ion concentrations tested (0.1-0.8 mg/L) achieved more than 99.999% reduction of P. aeruginosa which appears to be more susceptible to copper ions than S. maltophilia and A. baumannii. Silver ions concentration of 0.08 mg/L achieved more than 99.999% reduction of P. aeruginosa, S. maltophilia and A. baumannii in 6, 12 and 96 h, respectively. Combination of copper and silver ions exhibited a synergistic effect against P. aeruginosa and A. baumannii while the combination exhibited an antagonistic effect against S. maltophilia. Ionization may have a potential to eradicate P. aeruginosa, S. maltophilia and A. baumannii from hospital water systems.

  1. Current Situation of Antimicrobial Resistance and Genetic Differences in Stenotrophomonas maltophilia Complex Isolates by Multilocus Variable Number of Tandem Repeat Analysis

    PubMed Central

    Song, Jae-Hoon

    2016-01-01

    Background Stenotrophomonas maltophilia is one of several opportunistic pathogens of growing significance. Several studies on the molecular epidemiology of S. maltophilia have shown clinical isolates to be genetically diverse. Materials and Methods A total of 121 clinical isolates tentatively identified as S. malophilia from seven tertiary-care hospitals in Korea from 2007 to 2011 were included. Species and groups were identified using partial gyrB gene sequences and antimicrobial susceptibility testing was performed using a broth microdilution method. Multi locus variable number of tandem repeat analysis (MLVA) surveys are used for subtyping. Results Based on partial gyrB gene sequences, 118 isolates were identified as belonging to the S. maltophilia complex. For all S. maltophilia isolates, the resistance rates to trimethoprime-sulfamethoxazole (TMP/SMX) and levofloxacin were the highest (both, 30.5%). Resistance rate to ceftazidime was 28.0%. 11.0% and 11.9% of 118 S. maltophilia isolates displayed resistance to piperacillin/tazobactam and tigecycline, respectively. Clade 1 and Clade 2 were definitely distinguished from the data of MLVA with amplification of loci. All 118 isolates were classified into several clusters as its identification. Conclusion Because of high resistance rates to TMP/SMX and levofloxacin, the clinical laboratory department should consider providing the data about other antimicrobial agents and treatment of S. maltophilia infections with a combination of antimicrobials can be considered in the current practice. The MLVA evaluated in this study provides a fast, portable, relatively low cost genotyping method that can be employed in genotypic linkage or transmission networks comparing to analysis of the gyrB gene. PMID:28032486

  2. Structural and Functional Analysis of SmeT, the Repressor of the Stenotrophomonas maltophilia Multidrug Efflux Pump SmeDEF*

    PubMed Central

    Hernández, Alvaro; Maté, María J.; Sánchez-Díaz, Patricia C.; Romero, Antonio; Rojo, Fernando; Martínez, José L.

    2009-01-01

    Stenotrophomonas maltophilia is an opportunistic pathogen characterized for its intrinsic low susceptibility to several antibiotics. Part of this low susceptibility relies on the expression of chromosomally encoded multidrug efflux pumps, with SmeDEF being the most relevant antibiotic resistance efflux pump so far studied in this bacterial species. Expression of smeDEF is down-regulated by the SmeT repressor, encoded upstream smeDEF, in its complementary DNA strand. In the present article we present the crystal structure of SmeT and analyze its interactions with its cognate operator. Like other members of the TetR family of transcriptional repressors, SmeT behaves as a dimer and presents some common structural features with other TetR proteins like TtgR, QacR, and TetR. Differing from other TetR proteins for which the structure is available, SmeT turned out to have two extensions at the N and C termini that might be relevant for its function. Besides, SmeT presents the smallest binding pocket so far described in the TetR family of transcriptional repressors, which may correlate with a specific type and range of effectors. In vitro studies revealed that SmeT binds to a 28-bp pseudopalindromic region, forming two complexes. This operator region was found to overlap the promoters of smeT and smeDEF. This finding is consistent with a role for SmeT simultaneously down-regulating smeT and smeDEF transcription, likely by steric hindrance on RNA polymerase binding to DNA. PMID:19324881

  3. Identification of Electrode Respiring, Hydrocarbonoclastic Bacterial Strain Stenotrophomonas maltophilia MK2 Highlights the Untapped Potential for Environmental Bioremediation

    PubMed Central

    Venkidusamy, Krishnaveni; Megharaj, Mallavarapu

    2016-01-01

    Electrode respiring bacteria (ERB) possess a great potential for many biotechnological applications such as microbial electrochemical remediation systems (MERS) because of their exoelectrogenic capabilities to degrade xenobiotic pollutants. Very few ERB have been isolated from MERS, those exhibited a bioremediation potential toward organic contaminants. Here we report once such bacterial strain, Stenotrophomonas maltophilia MK2, a facultative anaerobic bacterium isolated from a hydrocarbon fed MERS, showed a potent hydrocarbonoclastic behavior under aerobic and anaerobic environments. Distinct properties of the strain MK2 were anaerobic fermentation of the amino acids, electrode respiration, anaerobic nitrate reduction and the ability to metabolize n-alkane components (C8–C36) of petroleum hydrocarbons (PH) including the biomarkers, pristine and phytane. The characteristic of diazoic dye decolorization was used as a criterion for pre-screening the possible electrochemically active microbial candidates. Bioelectricity generation with concomitant dye decolorization in MERS showed that the strain is electrochemically active. In acetate fed microbial fuel cells (MFCs), maximum current density of 273 ± 8 mA/m2 (1000 Ω) was produced (power density 113 ± 7 mW/m2) by strain MK2 with a coulombic efficiency of 34.8%. Further, the presence of possible alkane hydroxylase genes (alkB and rubA) in the strain MK2 indicated that the genes involved in hydrocarbon degradation are of diverse origin. Such observations demonstrated the potential of facultative hydrocarbon degradation in contaminated environments. Identification of such a novel petrochemical hydrocarbon degrading ERB is likely to offer a new route to the sustainable bioremedial process of source zone contamination with simultaneous energy generation through MERS. PMID:28018304

  4. Antibacterial and Cytotoxic Efficacy of Extracellular Silver Nanoparticles Biofabricated from Chromium Reducing Novel OS4 Strain of Stenotrophomonas maltophilia

    PubMed Central

    Oves, Mohammad; Khan, Mohammad Saghir; Zaidi, Almas; Ahmed, Arham S.; Ahmed, Faheem; Ahmad, Ejaz; Sherwani, Asif; Owais, Mohammad; Azam, Ameer

    2013-01-01

    Biofabricated metal nanoparticles are generally biocompatible, inexpensive, and ecofriendly, therefore, are used preferably in industries, medical and material science research. Considering the importance of biofabricated materials, we isolated, characterized and identified a novel bacterial strain OS4 of Stenotrophomonas maltophilia (GenBank: JN247637.1). At neutral pH, this Gram negative bacterial strain significantly reduced hexavalent chromium, an important heavy metal contaminant found in the tannery effluents and minings. Subsequently, even at room temperature the supernatant of log phase grown culture of strain OS4 also reduced silver nitrate (AgNO3) to generate nanoparticles (AgNPs). These AgNPs were further characterized by UV–visible, Nanophox particle size analyzer, XRD, SEM and FTIR. As evident from the FTIR data, plausibly the protein components of supernatant caused the reduction of AgNO3. The cuboid and homogenous AgNPs showed a characteristic UV-visible peak at 428 nm with average size of ∼93 nm. The XRD spectra exhibited the characteristic Bragg peaks of 111, 200, 220 and 311 facets of the face centred cubic symmetry of nanoparticles suggesting that these nanoparticles were crystalline in nature. From the nanoparticle release kinetics data, the rapid release of AgNPs was correlated with the particle size and increasing surface area of the nanoparticles. A highly significant antimicrobial activity against medically important bacteria by the biofabricated AgNPs was also revealed as decline in growth of Staphylococcus aureus (91%), Escherichia coli (69%) and Serratia marcescens (66%) substantially. Additionally, different cytotoxic assays showed no toxicity of AgNPs to liver function, RBCs, splenocytes and HeLa cells, hence these particles were safe to use. Therefore, this novel bacterial strain OS4 is likely to provide broad spectrum benefits for curing chromium polluted sites, for biofabrication of AgNPs and ultimately in the nanoparticle based

  5. Systematic Mutational Analysis of Histidine Kinase Genes in the Nosocomial Pathogen Stenotrophomonas maltophilia Identifies BfmAK System Control of Biofilm Development

    PubMed Central

    Zheng, Liu; Wang, Fang-Fang; Ren, Bao-Zhen; Liu, Wei

    2016-01-01

    The Gram-negative bacterium Stenotrophomonas maltophilia lives in diverse ecological niches. As a result of its formidable capabilities of forming biofilm and its resistance to multiple antibiotic agents, the bacterium is also a nosocomial pathogen of serious threat to the health of patients whose immune systems are suppressed or compromised. Besides the histidine kinase RpfC, the two-component signal transduction system (TCS), which is the canonical regulatory machinery used by most bacterial pathogens, has never been experimentally investigated in S. maltophilia. Here, we annotated 62 putative histidine kinase genes in the S. maltophilia genome and successfully obtained 51 mutants by systematical insertional inactivation. Phenotypic characterization identified a series of mutants with deficiencies in bacterial growth, swimming motility, and biofilm development. A TCS, named here BfmA-BfmK (Smlt4209-Smlt4208), was genetically confirmed to regulate biofilm formation in S. maltophilia. Together with interacting partner prediction and chromatin immunoprecipitation screens, six candidate promoter regions bound by BfmA in vivo were identified. We demonstrated that, among them, BfmA acts as a transcription factor that binds directly to the promoter regions of bfmA-bfmK and Smlt0800 (acoT), a gene encoding an acyl coenzyme A thioesterase that is associated with biofilm development, and positively controls their transcription. Genome-scale mutational analyses of histidine kinase genes and functional dissection of BfmK-BfmA regulation in biofilm provide genetic information to support more in-depth studies on cellular signaling in S. maltophilia, in the context of developing novel approaches to fight this important bacterial pathogen. PMID:26873318

  6. Modification of surface and enzymatic properties of Achromobacter denitrificans and Stenotrophomonas maltophilia in association with diesel oil biodegradation enhanced with alkyl polyglucosides.

    PubMed

    Sałek, Karina; Zgoła-Grześkowiak, Agnieszka; Kaczorek, Ewa

    2013-11-01

    The article concerns the influence of selected alkyl polyglucosides on biodegradation, cell surface and enzymatic properties of Stenotrophomonas maltophilia and Achromobacter denitrificans. The biodegradation of diesel oil depends on several factors including type and the amount of surfactant as well as bacterial genera used in the process. Nevertheless, a careful selection of these variables must be made as some bacterial strains prefer to use surfactants as their carbon source. This leads to the lowered biodegradation of diesel oil as can be observed for the tested S. maltophilia strain. Alkyl polyglucosides influenced the cell surface properties of both of the tested strains in slightly different ways. Especially for A. denitrificans, for which the hydrophobicity increased with concentration of both--Lutensol GD 70 and Glucopon 215 in diesel oil-surfactant systems. Moreover, judging by the efficiency of biodegradation, the most effective process was observed in the presence of Lutensol GD 70 (240 and 360 mg L(-1)) with biodegradation rising from 32% (without surfactant) to 68%. No such relation was observed for S. maltophilia.

  7. Antibiotic susceptibility of Stenotrophomonas (Xanthomonas) maltophilia: comparative (NCCLS criteria) evaluation of antimicrobial drugs with the agar dilution and the agar disk diffusion (Bauer-Kirby) tests.

    PubMed

    Traub, W H; Leonhard, B; Bauer, D

    1998-01-01

    Ninety-six clinical isolates of Stenotrophomonas maltophilia were examined with the agar dilution method for susceptibility to 19 antimicrobial drugs. Doxycycline, cotrimoxazole, timentin, ofloxacin, fosfomycin, and piperacillin + tazobactam, in that order, inhibited the majority of strains. All isolates were resistant to nitrofurantoin. Concurrent disk susceptibility (Bauer-Kirby method) testing, using currently valid NCCLS interpretative criteria for Pseudomonas aeruginosa, uncovered a significant incidence of very major (category I), major (category II), and minor (categories III and IV) discrepancies for aminoglycosides, cephalosporins, chloramphenicol, and piperacillin + tazobactam and ticarcillin + clavulanic acid. Therefore, new interpretative criteria indicative of intermediate (I) susceptibility of S. maltophilia to these various antibiotics were proposed. In addition, new intermediate susceptibility criteria were proposed for the two beta-lactam-beta-lactamase inhibitor combinations. It was recommended to exclude ciprofloxacin from test batteries against this microorganism due to the wide scatter of minimal inhibitory concentration values and diameters of inhibition zones; the same was true for polymyxin B. It is hoped that the proposed modified, species-specific criteria will improve the clinical utility of laboratory-generated disk antibiograms with respect to the inherently multiple antibiotic-resistant, opportunistic pathogen S. maltophilia.

  8. Polymerase chain reaction for the detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Burkholderia cepacia in sputum of patients with cystic fibrosis.

    PubMed

    Karpati, F; Jonasson, J

    1996-12-01

    Occurrence of Pseudomonas aeruginosa, Stenotrophomonas (Xanthomonas) maltophilia and Burkholderia (Pseudomonas) cepacia in sputum of cystic fibrosis (CF) patients was demonstrated with a simple and rapid polymerase chain reaction (PCR) technique. The PCR was performed with a set of three primer pairs based on 16S rRNA sequences after sputum preparation with dithiothreitol and NaOH lysis. All three pathogens could be individually detected by the use of this technique. To prevent carry-over contamination, dUTP and uracil-N-glycosylase were included in the reaction. The amplicons were visualized by agarose gel electrophoresis. Sputum culture was performed on all samples. Ninety specimens from CF patients were analysed. The sensitivity for the detection of P. aeruginosa was 37/40 (93%) compared to culture. Bacterial growth of P. aeruginosa was found in three cases, where PCR amplicons were not detected, while PCR was positive in five cases, where culture did not reveal the presence of this bacterium. For this reason, the specificity was 45/50 (90%). For S. maltophilia, the PCR was less sensitive than culture (positive in three of six cases). In our series, B. cepacia was detected by culture in one case and this was also detected by PCR. There were no false-positive PCR results regarding S. maltophilia or B. cepacia. Thus, combined PCR-based detection of these three clinically relevant bacteria in sputum samples from CF patients can be performed by a reliable technique in a relatively simple manner. The present data indicate a high sensitivity and specificity for P. aeruginosa. The lower sensitivity observed for the detection of S. maltophilia in sputum and B. cepacia, as estimated from laboratory strains, may depend on PCR conditions and genetic heterogeneity, respectively. The greatest gains with this method can be made when it is used for the early detection of P. aeruginosa in sputum-producing CF patients.

  9. High activity catechol 1,2-dioxygenase from Stenotrophomonas maltophilia strain KB2 as a useful tool in cis,cis-muconic acid production.

    PubMed

    Guzik, Urszula; Hupert-Kocurek, Katarzyna; Sitnik, Małgorzata; Wojcieszyńska, Danuta

    2013-06-01

    This is the first report of a catechol 1,2-dioxygenase from Stenotrophomonas maltophilia strain KB2 with high activity against catechol and its methyl derivatives. This enzyme was maximally active at pH 8.0 and 40 °C and the half-life of the enzyme at this temperature was 3 h. Kinetic studies showed that the value of K m and V max was 12.8 μM and 1,218.8 U/mg of protein, respectively. During our studies on kinetic properties of the catechol 1,2-dioxygenase we observed substrate inhibition at >80 μM. The nucleotide sequence of the gene encoding the S. maltophilia strain KB2 catechol 1,2-dioxygenase has high identity with other catA genes from members of the genus Pseudomonas. The deduced 314-residue sequence of the enzyme corresponds to a protein of molecular mass 34.5 kDa. This enzyme was inhibited by competitive inhibitors (phenol derivatives) only by ca. 30 %. High tolerance against condition changes is desirable in industrial processes. Our data suggest that this enzyme could be of use as a tool in production of cis,cis-muconic acid and its derivatives.

  10. Minocycline activity tested against Acinetobacter baumannii complex, Stenotrophomonas maltophilia, and Burkholderia cepacia species complex isolates from a global surveillance program (2013).

    PubMed

    Flamm, Robert K; Castanheira, Mariana; Streit, Jennifer M; Jones, Ronald N

    2016-07-01

    Clinical isolates of Acinetobacter baumannii complex (1312), Stenotrophomonas maltophilia (464), and Burkholderia cepacia species complex (30) were selected from medical centers in the United States (USA), Europe and the Mediterranean (EU-M) region, Latin America, and Asia Pacific. Only one isolate per infected patient episode was included and local identifications were confirmed by the monitoring laboratory. Susceptibility testing was performed at the monitoring laboratory using the reference broth microdilution method as described by Clinical and Laboratory Standards Institute (CLSI). A. baumannii complex were classified as MDR (multi-drug resistant [MDR]; nonsusceptible to ≥1 agent in ≥3 antimicrobial classes) and extensively drug-resistant (XDR; nonsusceptible to ≥1 agent in all but ≤2 antimicrobial classes). A total of 81.6% of A. baumannii complex were MDR. Colistin was the most active agent against MDR A. baumannii complex. Minocycline was the most active "tetracycline" against these organisms based on susceptibility. Against B. cepacia, trimethoprim-sulfamethoxazole (MIC90, 2 μg/mL; 100.0% susceptible) was the most active agent tested. Overall, minocycline was the most active tetracycline tested against A. baumannii complex and S. maltophilia isolates collected from patients throughout EU-M, USA, Latin America, and the Asia-Pacific. Minocycline, particularly the intravenous formulation, has activity against several ESKAPE pathogens and merits consideration in seriously ill patients where treatment options may be limited due to the presence of MDR bacteria.

  11. Biodegradation of polycyclic aromatic hydrocarbons by an acidophilic Stenotrophomonas maltophilia strain AJH1 isolated from a mineral mining site in Saudi Arabia.

    PubMed

    Arulazhagan, P; Al-Shekri, K; Huda, Q; Godon, J J; Basahi, J M; Jeyakumar, D

    2017-01-01

    The present study aims at analyzing the degradation of polycyclic aromatic hydrocarbons (PAHs) at acidic conditions (pH = 2) by acidophilic Stenotrophomonas maltophilia strain AJH1 (KU664513). The strain AJH1 was obtained from an enrichment culture obtained from soil samples of mining area in the presence of PAH as sole sources of carbon and energy. Strain AJH1was able to degrade low (anthracene, phenanthrene, naphthalene, fluorene) and high (pyrene, benzo(e)pyrene and benzo(k)fluoranthene) molecular weight PAHs in acidophilic mineral salt medium at pH 2, with removal rates of up to 95% (LMW PAH) and 80% (HMW PAH), respectively. In addition, strain AJH1 treated petroleum wastewater with 89 ± 1.1% COD removal under acidic condition (pH 2) in a continuously stirred reactor. Acidophilic S. maltophilia strain AJH1, hence holds the promise as an effective degrader for biological treatment of PAHs contaminated wastewater at acidic pH.

  12. Draft Genome Sequences of Stenotrophomonas maltophilia Strains Sm32COP, Sm41DVV, Sm46PAILV, SmF3, SmF22, SmSOFb1, and SmCVFa1, Isolated from Different Manures in France

    PubMed Central

    Bodilis, Josselin; Youenou, Benjamin; Briolay, Jérome; Brothier, Elisabeth; Favre-Bonté, Sabine

    2016-01-01

    Stenotrophomonas maltophilia is a major opportunistic human pathogen responsible for nosocomial infections. Here, we report the draft genome sequences of Sm32COP, Sm41DVV, Sm46PAILV, SmF3, SmF22, SmSOFb1, and SmCVFa1, isolated from different manures in France, which provide insights into the genetic determinism of intrinsic or acquired antibiotic resistance in this species. PMID:27540065

  13. Determination of crystal structures of proteins of unknown identity using a marathon molecular replacement procedure: structure of Stenotrophomonas maltophilia phosphate-binding protein.

    PubMed

    Hatti, Kaushik; Gulati, Ashutosh; Srinivasan, Narayanaswamy; Murthy, M R N

    2016-10-01

    During the past decade, the authors have collected a few X-ray diffraction data sets from protein crystals that appeared to be easy cases of molecular replacement but failed to yield structures even after extensive trials. Here, the use of a large-scale molecular replacement method that explores all structurally characterized domains as phasing models to determine the structure corresponding to two data sets collected at 1.9 and 2.3 Å resolution is reported. These two structures were of the same protein independently crystallized in 2007 and 2011. The structures derived are virtually identical and were found to consist of two compact globular domains connected by a hinge. The high resolution of one of these data sets enabled inference of the amino-acid sequence from the electron-density map. The deduced sequence is nearly identical to that of a protein from the multidrug-resistant bacterium Stenotrophomonas maltophilia. Although the structure of this protein has not been determined previously, it is homologous to the well studied DING proteins which mediate the cellular uptake of phosphate ions. The final electron-density maps from both of the data sets revealed a large density at the interface of the two globular domains that is likely to represent a phosphate ion. Thus, the structure is likely to be that of a phosphate-binding protein encoded by the S. maltophilia genome (SmPBP; PDB entry 5j1d). The nature of the phosphate-binding site of SmPBP closely resembles that of Pseudomonas fluorescens DING (PfluDING), which displays remarkable discrimination between the closely similar phosphate and arsenate ions. The results presented here illustrate that routine crystallization trials may occasionally lead to the serendipitous crystallization of a protein of unknown identity and brute-force molecular replacement through `fold space' might allow the identification of the unknown protein.

  14. Delineation of Stenotrophomonas maltophilia isolates from cystic fibrosis patients by fatty acid methyl ester profiles and matrix-assisted laser desorption/ionization time-of-flight mass spectra using hierarchical cluster analysis and principal component analysis.

    PubMed

    Vidigal, Pedrina Gonçalves; Mosel, Frank; Koehling, Hedda Luise; Mueller, Karl Dieter; Buer, Jan; Rath, Peter Michael; Steinmann, Joerg

    2014-12-01

    Stenotrophomonas maltophilia is an opportunist multidrug-resistant pathogen that causes a wide range of nosocomial infections. Various cystic fibrosis (CF) centres have reported an increasing prevalence of S. maltophilia colonization/infection among patients with this disease. The purpose of this study was to assess specific fingerprints of S. maltophilia isolates from CF patients (n = 71) by investigating fatty acid methyl esters (FAMEs) through gas chromatography (GC) and highly abundant proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and to compare them with isolates obtained from intensive care unit (ICU) patients (n = 20) and the environment (n = 11). Principal component analysis (PCA) of GC-FAME patterns did not reveal a clustering corresponding to distinct CF, ICU or environmental types. Based on the peak area index, it was observed that S. maltophilia isolates from CF patients produced significantly higher amounts of fatty acids in comparison with ICU patients and the environmental isolates. Hierarchical cluster analysis (HCA) based on the MALDI-TOF MS peak profiles of S. maltophilia revealed the presence of five large clusters, suggesting a high phenotypic diversity. Although HCA of MALDI-TOF mass spectra did not result in distinct clusters predominantly composed of CF isolates, PCA revealed the presence of a distinct cluster composed of S. maltophilia isolates from CF patients. Our data suggest that S. maltophilia colonizing CF patients tend to modify not only their fatty acid patterns but also their protein patterns as a response to adaptation in the unfavourable environment of the CF lung.

  15. Protocatechuate 3,4-dioxygenase: a wide substrate specificity enzyme isolated from Stenotrophomonas maltophilia KB2 as a useful tool in aromatic acid biodegradation.

    PubMed

    Guzik, Urszula; Hupert-Kocurek, Katarzyna; Sitnik, Małgorzata; Wojcieszyńska, Danuta

    2014-01-01

    Protocatechuate 3,4-dioxygenases (P34Os) catalyze the reaction of the ring cleavage of aromatic acid derivatives. It is a key reaction in many xenobiotic metabolic pathways. P34Os characterize narrow substrate specificity. This property is an unfavorable feature in the biodegradation process because one type of pollution is rarely present in the environment. Thus, the following study aimed at the characterization of a P34O from Stenotrophomonas maltophilia KB2, being able to utilize a wide spectrum of aromatic carboxylic acids. A total of 3 mM vanillic acid and 4-hydroxybenzoate were completely degraded during 8 and 4.5 h, respectively. When cells of strain KB2 were grown on 9 mM 4-hydroxybenzoate, P34O was induced. Biochemical analysis revealed that the examined enzyme was similar to other known P34Os, but showed untypical wide substrate specificity. A high activity of P34O against 2,4- and 3,5-dihydroxybenzoate was observed. As these substrates do not possess ortho configuration hydroxyl groups, it is postulated that their cleavage could be connected with their monodentate binding of substrate to the active site. Since this enzyme characterizes untypical wide substrate specificity it makes it a useful tool in applications for environmental clean-up purposes.

  16. Differentiation of pulmonary bacterial pathogens in cystic fibrosis by volatile metabolites emitted by their in vitro cultures: Pseudomonas aeruginosa, Staphylococcus aureus, Stenotrophomonas maltophilia and the Burkholderia cepacia complex.

    PubMed

    Dryahina, Kseniya; Sovová, Kristýna; Nemec, Alexandr; Španěl, Patrik

    2016-08-10

    As a contribution to the continuing search for breath biomarkers of lung and airways infection in patients with cystic fibrosis, CF, we have analysed the volatile metabolites released in vitro by Pseudomonas aeruginosa and other bacteria involved in respiratory infections in these patients, i.e. those belonging to the Burkholderia cepacia complex, Staphylococcus aureus or Stenotrophomonas maltophilia. These opportunistic pathogens are generally harmless to healthy people but they may cause serious infections in patients with severe underlying disease or impaired immunity such as CF patients. Volatile organic compounds emitted from the cultures of strains belonging to the above-mentioned four taxa were analysed by selected ion flow tube mass spectrometry. In order to minimize the effect of differences in media composition all strains were cultured in three different liquid media. Multivariate statistical analysis reveals that the four taxa can be well discriminated by the differences in the headspace VOC concentration profiles. The compounds that should be targeted in breath as potential biomarkers of airway infection were identified for each of these taxa of CF pathogens.

  17. Copper Enhanced Monooxygenase Activity and FT-IR Spectroscopic Characterisation of Biotransformation Products in Trichloroethylene Degrading Bacterium: Stenotrophomonas maltophilia PM102

    PubMed Central

    Mukherjee, Piyali; Roy, Pranab

    2013-01-01

    Stenotrophomonas maltophilia PM102 (NCBI GenBank Acc. no. JQ797560) is capable of growth on trichloroethylene as the sole carbon source. In this paper, we report the purification and characterisation of oxygenase present in the PM102 isolate. Enzyme activity was found to be induced 10.3-fold in presence of 0.7 mM copper with a further increment to 14.96-fold in presence of 0.05 mM NADH. Optimum temperature for oxygenase activity was recorded at 36°C. The reported enzyme was found to have enhanced activity at pH 5 and pH 8, indicating presence of two isoforms. Maximum activity was seen on incubation with benzene compared to other substrates like TCE, chloroform, toluene, hexane, and petroleum benzene. Km and Vmax for benzene were 3.8 mM and 340 U/mg/min and those for TCE were 2.1 mM and 170 U/mg/min. The crude enzyme was partially purified by ammonium sulphate precipitation followed by dialysis. Zymogram analysis revealed two isoforms in the 70% purified enzyme fraction. The activity stain was more prominent when the native gel was incubated in benzene as substrate in comparison to TCE. Crude enzyme and purified enzyme fractions were assayed for TCE degradation by the Fujiwara test. TCE biotransformation products were analysed by FT-IR spectroscopy. PMID:24083236

  18. Effects of Endobacterium (Stenotrophomonas maltophilia) on Pathogenesis-Related Gene Expression of Pine Wood Nematode (Bursaphelenchus xylophilus) and Pine Wilt Disease

    PubMed Central

    He, Long-Xi; Wu, Xiao-Qin; Xue, Qi; Qiu, Xiu-Wen

    2016-01-01

    Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, is responsible for devastating epidemics in pine trees in Asia and Europe. Recent studies showed that bacteria carried by the PWN might be involved in PWD. However, the molecular mechanism of the interaction between bacteria and the PWN remained unclear. Now that the whole genome of B. xylophilus (Bursaphelenchus xylophilus) is published, transcriptome analysis is a unique method to study the role played by bacteria in PWN. In this study, the transcriptome of aseptic B. xylophilus, B. xylophilus treated with endobacterium (Stenotrophomonas maltophilia NSPmBx03) and fungus B. xylophilus were sequenced. We found that 61 genes were up-regulated and 830 were down-regulated in B. xylophilus after treatment with the endobacterium; 178 genes were up-regulated and 1122 were down-regulated in fungus B. xylophilus compared with aseptic B. xylophilus. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to study the significantly changed biological functions and pathways for these differentially expressed genes. Many pathogenesis-related genes, including glutathinone S-transferase, pectate lyase, ATP-binding cassette transporter and cytochrome P450, were up-regulated after B. xylophilus were treated with the endobacterium. In addition, we found that bacteria enhanced the virulence of PWN. These findings indicate that endobacteria might play an important role in the development and virulence of PWN and will improve our understanding of the regulatory mechanisms involved in the interaction between bacteria and the PWN. PMID:27231904

  19. Inactivation of SmeSyRy Two-Component Regulatory System Inversely Regulates the Expression of SmeYZ and SmeDEF Efflux Pumps in Stenotrophomonas maltophilia

    PubMed Central

    Lin, Yi-Tsung; Ning, Hsiao-Chen; Yang, Tsuey-Ching

    2016-01-01

    SmeYZ efflux pump is a critical pump responsible for aminoglycosides resistance, virulence-related characteristics (oxidative stress susceptibility, motility, and secreted protease activity), and virulence in Stenotrophomonas maltophilia. However, the regulatory circuit involved in SmeYZ expression is little known. A two-component regulatory system (TCS), smeRySy, transcribed divergently from the smeYZ operon is the first candidate to be considered. To assess the role of SmeRySy in smeYZ expression, the smeRySy isogenic deletion mutant, KJΔRSy, was constructed by gene replacement strategy. Inactivation of smeSyRy correlated with a higher susceptibility to aminoglycosides concomitant with an increased resistance to chloramphenicol, ciprofloxacin, tetracycline, and macrolides. To elucidate the underlying mechanism responsible for the antimicrobials susceptibility profiles, the SmeRySy regulon was firstly revealed by transcriptome analysis and further confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and promoter transcription fusion constructs assay. The results demonstrate that inactivation of smeRySy decreased the expression of SmeYZ pump and increased the expression of SmeDEF pump, which underlies the ΔsmeSyRy-mediated antimicrobials susceptibility profile. To elucidate the cognate relationship between SmeSy and SmeRy, a single mutant, KJΔRy, was constructed and the complementation assay of KJΔRSy with smeRy were performed. The results support that SmeSy-SmeRy TCS is responsible for the regulation of smeYZ operon; whereas SmeSy may be cognate with another unidentified response regulator for the regulation of smeDEF operon. The impact of inverse expression of SmeYZ and SmeDEF pumps on physiological functions was evaluated by mutants construction, H2O2 susceptibility test, swimming, and secreted protease activity assay. The increased expression of SmeDEF pump in KJΔRSy may compensate, to some extents, the SmeYZ downexpression

  20. Potential novel therapeutic strategies in cystic fibrosis: antimicrobial and anti-biofilm activity of natural and designed α-helical peptides against Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia

    PubMed Central

    2012-01-01

    Background Treatment of cystic fibrosis-associated lung infections is hampered by the presence of multi-drug resistant pathogens, many of which are also strong biofilm producers. Antimicrobial peptides, essential components of innate immunity in humans and animals, exhibit relevant in vitro antimicrobial activity although they tend not to select for resistant strains. Results Three α-helical antimicrobial peptides, BMAP-27 and BMAP-28 of bovine origin, and the artificial P19(9/B) peptide were tested, comparatively to Tobramycin, for their in vitro antibacterial and anti-biofilm activity against 15 Staphylococcus aureus, 25 Pseudomonas aeruginosa, and 27 Stenotrophomonas maltophilia strains from cystic fibrosis patients. All assays were carried out in physical-chemical experimental conditions simulating a cystic fibrosis lung. All peptides showed a potent and rapid bactericidal activity against most P. aeruginosa, S. maltophilia and S. aureus strains tested, at levels generally higher than those exhibited by Tobramycin and significantly reduced biofilm formation of all the bacterial species tested, although less effectively than Tobramycin did. On the contrary, the viability-reducing activity of antimicrobial peptides against preformed P. aeruginosa biofilms was comparable to and, in some cases, higher than that showed by Tobramycin. Conclusions The activity shown by α-helical peptides against planktonic and biofilm cells makes them promising “lead compounds” for future development of novel drugs for therapeutic treatment of cystic fibrosis lung disease. PMID:22823964

  1. In vitro activity of levofloxacin against planktonic and biofilm Stenotrophomonas maltophilia lifestyles under conditions relevant to pulmonary infection in cystic fibrosis, and relationship with SmeDEF multidrug efflux pump expression.

    PubMed

    Pompilio, Arianna; Crocetta, Valentina; Verginelli, Fabio; Bonaventura, Giovanni Di

    2016-07-01

    The activity of levofloxacin against planktonic and biofilm Stenotrophomonas maltophilia cells and the role played by the multidrug efflux pump SmeDEF were evaluated under conditions relevant to the cystic fibrosis (CF) lung. MIC, MBC and MBEC of levofloxacin were assessed, against five CF strains, under 'standard' (CLSI-recommended) and 'CF-like' (pH 6.8, 5% CO2, in a synthetic CF sputum) conditions. Levofloxacin was tested against biofilms at concentrations (10, 50 and 100 μg mL(-1)) corresponding to achievable serum levels and sputum levels by aerosolisation. smeD expression was evaluated, under both conditions, in planktonic and biofilm cells by RT-PCR. The bactericidal effect of levofloxacin was decreased, in three out of five strains tested, under 'CF-like' conditions (MBC: 2-4 vs 8-16 μg mL(-1), under 'standard' and 'CF-like' conditions, respectively). Biofilm was intrinsically resistant to levofloxacin, regardless of conditions tested (MBECs ≥ 100 μg mL(-1) for all strains). Only under 'CF-like' conditions, smeD expression increased during planktonic-to-biofilm transition, and in biofilm cells compared to stationary planktonic cells. Our findings confirmed that S. maltophilia biofilm is intrinsically resistant to therapeutic concentrations of levofloxacin. Under conditions relevant to CF, smeD overexpression could contribute to levofloxacin resistance. Further studies are warranted to define the clinical relevance of our findings.

  2. Genome Sequence of Type Strains of Genus Stenotrophomonas

    PubMed Central

    Patil, Prashant P.; Midha, Samriti; Kumar, Sanjeet; Patil, Prabhu B.

    2016-01-01

    Genomic resource of type strains and historically important strains of genus Stenotrophomonas allowed us to reveal the existence of 18 distinct species by applying modern phylogenomic criterions. Apart from Stenotrophomonas maltophilia, S. africana represents another species of clinical importance. Interestingly, Pseudomonas hibsicola, P. beteli, and S. pavani that are of plant origin are closer to S. maltophilia than the majority of the environmental isolates. The genus has an open pan-genome. By providing the case study on genes encoding metallo-β-lactamase and Clustered Regularly Interspaced Short Palindrome Repeats (CRISPR) regions, we have tried to show the importance of this genomic dataset in understanding its ecology. PMID:27014232

  3. Stenotrophomonas, Mycobacterium, and Streptomyces in home dust and air: associations with moldiness and other home/family characteristics

    EPA Science Inventory

    Abstract Aims: (1) To investigate the dustborne and airborne bacterial concentrations of three emerging moisture-related bacteria: Stenotrophomonas maltophilia, Streptomyces, and Mycobacterium. (2) To study the association between these bacteria concentrations and Environmenta...

  4. The versatility and adaptation of bacteria from the genus Stenotrophomonas

    SciTech Connect

    Ryan, R.P.; van der Lelie, D.; Monchy, S.; Cardinale, M.; Taghavi, S.; Crossman, L.; Avison, M. B.; Berg, G.; Dow, J. M.

    2009-07-01

    The genus Stenotrophomonas comprises at least eight species. These bacteria are found throughout the environment, particularly in close association with plants. Strains of the most predominant species, Stenotrophomonas maltophilia, have an extraordinary range of activities that include beneficial effects for plant growth and health, the breakdown of natural and man-made pollutants that are central to bioremediation and phytoremediation strategies and the production of biomolecules of economic value, as well as detrimental effects, such as multidrug resistance, in human pathogenic strains. Here, we discuss the versatility of the bacteria in the genus Stenotrophomonas and the insight that comparative genomic analysis of clinical and endophytic isolates of S. maltophilia has brought to our understanding of the adaptation of this genus to various niches.

  5. Aph(3′)-IIc, an Aminoglycoside Resistance Determinant from Stenotrophomonas maltophilia▿

    PubMed Central

    Okazaki, Aki; Avison, Matthew B.

    2007-01-01

    We report the characterization of an intrinsic, chromosomally carried aph(3′)-IIc gene from Stenotrophomonas maltophilia clinical isolate K279a, encoding an aminoglycoside phosphotransferase enzyme that significantly increases MICs of kanamycin, neomycin, butirosin, and paromomycin when expressed in Escherichia coli. Disruption of aph(3′)-IIc in K279a results in decreased MICs of these drugs. PMID:17088477

  6. Phylogenetic Analysis of Stenotrophomonas spp. Isolates Contributes to the Identification of Nosocomial and Community-Acquired Infections

    PubMed Central

    Cerezer, Vinicius Godoy; Pasternak, Jacyr; Franzolin, Marcia Regina; Moreira-Filho, Carlos Alberto

    2014-01-01

    Stenotrophomonas ssp. has a wide environmental distribution and is also found as an opportunistic pathogen, causing nosocomial or community-acquired infections. One species, S. maltophilia, presents multidrug resistance and has been associated with serious infections in pediatric and immunocompromised patients. Therefore, it is relevant to conduct resistance profile and phylogenetic studies in clinical isolates for identifying infection origins and isolates with augmented pathogenic potential. Here, multilocus sequence typing was performed for phylogenetic analysis of nosocomial isolates of Stenotrophomonas spp. and, environmental and clinical strains of S. maltophilia. Biochemical and multidrug resistance profiles of nosocomial and clinical strains were determined. The inferred phylogenetic profile showed high clonal variability, what correlates with the adaptability process of Stenotrophomonas to different habitats. Two clinical isolates subgroups of S. maltophilia sharing high phylogenetic homogeneity presented intergroup recombination, thus indicating the high permittivity to horizontal gene transfer, a mechanism involved in the acquisition of antibiotic resistance and expression of virulence factors. For most of the clinical strains, phylogenetic inference was made using only partial ppsA gene sequence. Therefore, the sequencing of just one specific fragment of this gene would allow, in many cases, determining whether the infection with S. maltophilia was nosocomial or community-acquired. PMID:24818127

  7. Screening for endophytic nitrogen-fixing bacteria in Brazilian sugar cane varieties used in organic farming and description of Stenotrophomonas pavanii sp. nov.

    PubMed

    Ramos, Patrícia L; Van Trappen, Stefanie; Thompson, Fabiano L; Rocha, Rafael C S; Barbosa, Heloiza R; De Vos, Paul; Moreira-Filho, Carlos A

    2011-04-01

    A Gram-negative, rod-shaped, non-spore-forming and nitrogen-fixing bacterium, designated ICB 89(T), was isolated from stems of a Brazilian sugar cane variety widely used in organic farming. 16S rRNA gene sequence analysis revealed that strain ICB 89(T) belonged to the genus Stenotrophomonas and was most closely related to Stenotrophomonas maltophilia LMG 958(T), Stenotrophomonas rhizophila LMG 22075(T), Stenotrophomonas nitritireducens L2(T), [Pseudomonas] geniculata ATCC 19374(T), [Pseudomonas] hibiscicola ATCC 19867(T) and [Pseudomonas] beteli ATCC 19861(T). DNA-DNA hybridization together with chemotaxonomic data and biochemical characteristics allowed the differentiation of strain ICB 89(T) from its nearest phylogenetic neighbours. Therefore, strain ICB 89(T) represents a novel species, for which the name Stenotrophomonas pavanii sp. nov. is proposed. The type strain is ICB 89(T) ( = CBMAI 564(T)  = LMG 25348(T)).

  8. Virulence genes in clinical and environmental Stenotrophomas maltophilia isolates: a genome sequencing and gene expression approach.

    PubMed

    Adamek, Martina; Linke, Burkhard; Schwartz, Thomas

    2014-01-01

    The rate of nosocomial infections with the opportunistic pathogen Stenotrophomonas maltophilia has remarkably increased in the last decade. To determine S. maltophilia virulence genes, the complete genome sequences of two S. maltophilia isolates were compared. The clinical strain SKK35 was proved virulent in an amoeba host-pathogen model, and wastewater strain RA8 was determined as non-virulent in the amoeba model. The genome sequences of three additional S. maltophilia strains, K279a (clinical, non-virulent against amoeba), R511-3 and SKA14 (both environmental, non-virulent against amoeba) were taken into account as reference strains. We were able to show that all clinical and environmental S. maltophilia strains presented comparable distribution of so far identified potential virulence genes, regardless to their virulence potential against amoebae. Aside from that, strain SKK35 was found harboring a putative, strain specific pathogenicity island, encoding two proteins from the RTX (repeats-in-toxin) family. The actual expression of the RTX genes was verified in growth experiments in different culture media containing blood or blood components and in co-cultures with amoeba.

  9. Complete Genome Sequence of Lactobacillus plantarum CGMCC 8198

    PubMed Central

    Dong, Qing-Qing; Hu, Hai-Jie; Wang, Qiu-Tong; Gu, Xiang-Chao; Zhou, Hao; Zhou, Wen-Juan; Ni, Xiao-Meng

    2017-01-01

    ABSTRACT We report the complete genome sequence of Lactobacillus plantarum CGMCC 8198, a novel probiotic strain isolated from fermented herbage. We have determined the complete genome sequence of strain L. plantarum CGMCC 8198, which consists of genes that are likely to be involved in dairy fermentation and that have probiotic qualities. PMID:28183756

  10. [Environmental factors affecting the succinic acid production by Actinobacillus succinogenes CGMCC 1593].

    PubMed

    Zheng, Pu; Zhou, Wei; Ni, Ye; Jiang, Min; Wei, Ping; Sun, Zhihao

    2008-06-01

    Actinobacillus succinogenes is a promising candidate for the production of bio-based succinic acid. Previously, we isolated a succinic acid-producing strain Actinobacillus succinogenes CGMCC 1593 from bovine rumen. In this paper, the influence of the environmental factors such as gas phase, pH, ORP, on succinic acid production by A. succinogenes CGMCC 1593 was studied. The results showed that CO2 was the optimum gas phase for anaerobic fermentation ofA. succinogenes CGMCC 1593 as well as one of the substrate for the succinic acid synthesis. Using MgCO3 as a pH regulator, the pH was maintained within 7.1-6.2 during the anaerobic fermentation for the cell growth and acid production of A. succinogenes CGMCC 1593. Our results showed that low initial ORP was disadvantageous for the growth of A. succinogenes CGMCC 1593 and an ORP of -270 mV was demonstrated to be beneficial to the succinic acid production. By adding Na2S.9H2O to decrease ORP to -270 mV at the end of exponential growth phase in batch culture of A. succinogenes CGMCC 1593, the succinic acid concentration reached 37 g/L and the yield of succinic acid was 129% at 48 h. This work might provide valuable information for further optimization of succinic acid fermentation by A. succinogenes CGMCC 1593.

  11. Characterization of Bacterial Cellulose by Gluconacetobacter hansenii CGMCC 3917.

    PubMed

    Feng, Xianchao; Ullah, Niamat; Wang, Xuejiao; Sun, Xuchun; Li, Chenyi; Bai, Yun; Chen, Lin; Li, Zhixi

    2015-10-01

    In this study, comprehensive characterization and drying methods on properties of bacterial cellulose were analyzed. Bacterial cellulose was prepared by Gluconacetobacter hansenii CGMCC 3917, which was mutated by high hydrostatic pressure (HHP) treatment. Bacterial cellulose is mainly comprised of cellulose Iα with high crystallinity and purity. High-water holding and absorption capacity were examined by reticulated structure. Thermogravimetric analysis showed high thermal stability. High tensile strength and Young's modulus indicated its mechanical properties. The rheological analysis showed that bacterial cellulose had good consistency and viscosity. These results indicated that bacterial cellulose is a potential food additive and also could be used for a food packaging material. The high textural stability during freeze-thaw cycles makes bacterial cellulose an effective additive for frozen food products. In addition, the properties of bacterial cellulose can be affected by drying methods. Our results suggest that the bacterial cellulose produced from HHP-mutant strain has an effective characterization, which can be used for a wide range of applications in food industry.

  12. Stenotrophomonas, Mycobacterium, and Streptomyces in home dust and air: Associations with moldiness and other home/family characteristics

    PubMed Central

    Kettleson, Eric; Kumar, Sudhir; Reponen, Tiina; Vesper, Stephen; Méheust, Delphine; Grinshpun, Sergey A.; Adhikari, Atin

    2013-01-01

    Respiratory illnesses have been linked to children’s exposures to water-damaged homes. Therefore, understanding the microbiome in water-damaged homes is critical to preventing these illnesses. Few studies have quantified bacterial contamination, especially specific species, in water-damaged homes. We collected air and dust samples in twenty-one low-mold homes and twenty-one high-mold homes. The concentrations of three bacteria/genera, Stenotrophomonas maltophilia, Streptomyces sp. and Mycobacterium sp., were measured in air and dust samples using quantitative PCR (QPCR). The concentrations of the bacteria measured in the air samples were not associated with any specific home characteristic based on multiple regression models. However, higher concentrations of S. maltophilia in the dust samples were associated with water damage, i.e. with higher floor surface moisture and higher concentrations of moisture-related mold species. The concentrations of Streptomyces and Mycobacterium sp. had similar patterns and may be partially determined by human and animal occupants and outdoor sources of these bacteria. PMID:23397905

  13. De-novo synthesis of 2-phenylethanol by Enterobacter sp. CGMCC 5087

    PubMed Central

    2014-01-01

    Background 2-phenylethanl (2-PE) and its derivatives are important chemicals, which are widely used in food materials and fine chemical industries and polymers and it’s also a potentially valuable alcohol for next-generation biofuel. However, the biosynthesis of 2-PE are mainly biotransformed from phenylalanine, the price of which barred the production. Therefore, it is necessary to seek more sustainable technologies for 2-PE production. Results A new strain which produces 2-PE through the phenylpyruvate pathway was isolated and identified as Enterobacter sp. CGMCC 5087. The strain is able to use renewable monosaccharide as the carbon source and NH4Cl as the nitrogen source to produce 2-PE. Two genes of rate-limiting enzymes, chorismate mutase p-prephenate dehydratase (PheA) and 3-deoxy-d-arabino-heptulosonic acid 7-phosphate synthase (DAHP), were cloned from Escherichia coli and overexpressed in E. sp. CGMCC 5087. The engineered E. sp. CGMCC 5087 produces 334.9 mg L-1 2-PE in 12 h, which is 3.26 times as high as the wild strain. Conclusions The phenylpyruvate pathway and the substrate specificity of 2-keto-acid decarboxylase towards phenylpyruvate were found in E. sp. CGMCC 5087. Combined with the low-cost monosaccharide as the substrate, the finding provides a novel and potential way for 2-PE production. PMID:24766677

  14. Synthetic Peptides Derived from Bovine Lactoferricin Exhibit Antimicrobial Activity against E. coli ATCC 11775, S. maltophilia ATCC 13636 and S. enteritidis ATCC 13076.

    PubMed

    Huertas Méndez, Nataly De Jesús; Vargas Casanova, Yerly; Gómez Chimbi, Anyelith Katherine; Hernández, Edith; Leal Castro, Aura Lucia; Melo Diaz, Javier Mauricio; Rivera Monroy, Zuly Jenny; García Castañeda, Javier Eduardo

    2017-03-12

    Linear, dimeric, tetrameric, and cyclic peptides derived from lactoferricin B-containing non-natural amino acids and the RWQWR motif were synthesized, purified, and characterized using RP-HPLC, MALDI-TOF mass spectrometry, and circular dichroism. The antibacterial activity of peptides against Escherichia coli ATCC 11775, Stenotrophomonas maltophilia ATCC 13636, and Salmonella enteritidis ATCC 13076 was evaluated. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. The synthetic bovine lactoferricin exhibited antibacterial activity against E. coli ATCC 11775 and S. enteritidis ATCC 13076. The dimeric peptide (RRWQWR)₂K-Ahx exhibited the highest antibacterial activity against the tested bacterial strain. The monomeric, cyclic, tetrameric, and palindromic peptides containing the RWQWR motif exhibited high and specific activity against E. coli ATCC 11775. The results suggest that short peptides derived from lactoferricin B could be considered as potential candidates for the development of antibacterial agents against infections caused by E. coli.

  15. Specific gonadotropin binding to Pseudomonas maltophilia.

    PubMed

    Richert, N D; Ryan, R J

    1977-03-01

    Binding of 125I-labeled human chorionic gonadotropin to Pseudomonas maltophilia is dependent on time, temperature, and pH and the binding to this procaryotic species is hormone-specific and saturable. The equilibrium dissociation constant is 2.3 X 10(-9) M. There are no cooperative interactions between binding sites (Hill coefficient, 1.05). The number of sites is estimaated as 240 fmol/100 mug of protein. NaCl and KCl, at concentrations from 1 to 10 mM, have no effect on binding. Divalent cations (Mg2+ and Ca2+) and 1 mM EDTA inhibit hormone binding. Binding is destroyed by heat or by treatment with Pronase of alpha-chymotrypsin and is increased by phospholipase C. Binding of the labeled gonadotropin is not observed with other gram-negative organisms--e.g., Escherichia coli, Pseudomonas testosteroni, Pseudomonas aeruginosa, Enterobacter aerogenes, or Enterobacter cloacae.

  16. Central metabolic pathways of Aureobasidium pullulans CGMCC1234 for pullulan production.

    PubMed

    Sheng, Long; Liu, Chang; Tong, Qunyi; Ma, Meihu

    2015-12-10

    With the purpose of understanding the metabolic network of Aureobasidium pullulans, the central metabolic pathways were confirmed by the activities of the key enzymes involved in different pathways. The effect of different iodoacetic acid concentrations on pullulan fermentation was also investigated in this paper. The activities of phosphofructokinases and glucose-6-phosphate dehydrogenase existed in A. pullulans CGMCC1234, whereas 2-keto-3-deoxy-6-phosphogluconate aldolase activity was not detected. We proposed that the central metabolic pathways of A. pullulans CGMCC1234 included EMP and PPP, but no ED. Pullulan production declined fast as the iodoacetic acid increased, while cell growth offered upgrade firstly than descending latter tendency. Compared to the control group, the ratio of ATP/ADP of 0.60 mM iodoacetic acid group was lower at different stages of pullulan fermentation. The findings revealed that low concentration of iodoacetic acid might impel carbon flux flow toward the PPP, but reduce the flux of the EMP.

  17. Complete genome sequence of a benzo[a]pyrene-degrading bacterium Altererythrobacter epoxidivorans CGMCC 1.7731(T).

    PubMed

    Li, Zheng-Yang; Wu, Yue-Hong; Huo, Ying-Yi; Cheng, Hong; Wang, Chun-Sheng; Xu, Xue-Wei

    2016-02-01

    Altererythrobacter epoxidivorans CGMCC 1.7731(T) is a Gram-negative bacterium isolated from marine sediments. It is able to utilize benzo[a]pyrene as sole carbon and energy source. Here, we describe the complete genome sequence and annotation of A. epoxidivorans CGMCC 1.7731(T). The genome has a size of 2,786,256 bp (61.50 mol% G+C content), which consists of 2773 coding genes, 43 tRNA genes and 3 rRNA genes. According to the genome information, strain A. epoxidivorans CGMCC 1.7731(T) encodes 22 genes related to degradation of benzo[a]pyrene. These genes may have potential in bioremediation of PAH-polluted environments.

  18. Biodegradation and dissolution of polyaromatic hydrocarbons by Stenotrophomonas sp.

    PubMed

    Tiwari, Bhagyashree; Manickam, N; Kumari, Smita; Tiwari, Akhilesh

    2016-09-01

    The aim of this work was to study the biodegradation capabilities of a locally isolated bacterium, Stenotrophomonas sp. strain IITR87 to degrade the polycyclic aromatic hydrocarbons and also check the preferential biodegradation of polycyclic aromatic hydrocarbons (PAHs). From preferential substrate degradation studies, it was found that Stenotrophomonas sp. strain IITR87 first utilized phenanthrene (three membered ring), followed by pyrene (four membered ring), then benzo[α]pyrene (five membered ring). Dissolution study of PAHs with surfactants, rhamnolipid and tritonX-100 showed that the dissolution of PAHs increased in the presence of surfactants.

  19. Potential probiotic attributes of a new strain of Bacillus coagulans CGMCC 9951 isolated from healthy piglet feces.

    PubMed

    Gu, Shao-Bin; Zhao, Li-Na; Wu, Ying; Li, Shi-Chang; Sun, Jian-Rui; Huang, Jing-Fang; Li, Dan-Dan

    2015-06-01

    A new strain of Bacillus coagulans CGMCC 9551, which has a broad range of antibacterial activities against six main pathogenic bacteria including Escherichia coli O8, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar enteritidis, Streptococcus suis, Listeria monocytogenes and Pasteurella multocida, was isolated from healthy piglet feces. In adhesion assay, the isolate exhibited a stronger adhesion to pig intestinal mucus than that of B. subtilis JT143 and L. acidophilus LY24 respectively isolated from BioPlus(®)2B and FloraFIT(®) Probiotics (P < 0.05). The adhesion activity reached 44.5 ± 3.2, 48.9 ± 2.6, 42.6 ± 3.3 and 37.6 ± 2.4% to jejunum, ileum, transverse colon and sigmoid colon, separately. The survival rate of B. coagulans CGMCC 9551 was reduced by only 20% at 4 h exposure under 0.9% w/v bile salt. The strain was fully resistant to pH 2 for 2 h with 90.1 ± 3.5% survival and susceptible to 15 antibiotics commonly used in veterinary medicine. Additionally, the bacteria showed amylase, protease and cellulase activities. The safety assessment demonstrated the lack of toxicity potential in B. coagulans CGMCC 9551 by ligated rabbit ileal loop assay, acute and subchronic toxicity test. These results implied that that the new strain of B. coagulans CGMCC 9951 isolated from healthy piglet feces has promising probiotic characteristics and offers desirable opportunities for its successful commercialization as one excellent candidate probiotic.

  20. Statistical experimental design optimization of rhamsan gum production by Sphingomonas sp. CGMCC 6833.

    PubMed

    Xu, Xiao-Ying; Dong, Shu-Hao; Li, Sha; Chen, Xiao-Ye; Wu, Ding; Xu, Hong

    2015-04-01

    Rhamsan gum is a type of water-soluble exopolysaccharide produced by species of Sphingomonas bacteria. The optimal fermentation medium for rhamsan gum production by Sphingomonas sp. CGMCC 6833 was explored definition. Single-factor experiments indicate that glucose, soybean meal, K(2)HPO(4) and MnSO(4) compose the optimal medium along with and initial pH 7.5. To discover ideal cultural conditions for rhamsan gum production in a shake flask culture, response surface methodology was employed, from which the following optimal ratio was derived: 5.38 g/L soybean meal, 5.71 g/L K(2)HPO(4) and 0.32 g/L MnSO(4). Under ideal fermentation rhamsan gum yield reached 19.58 g/L ± 1.23 g/L, 42.09% higher than that of the initial medium (13.78 g/L ± 1.38 g/L). Optimizing the fermentation medium results in enhanced rhamsan gum production.

  1. Purification and characterisation of a bifunctional alginate lyase from novel Isoptericola halotolerans CGMCC 5336.

    PubMed

    Dou, Wenfang; Wei, Dan; Li, Hui; Li, Heng; Rahman, Muhammad Masfiqur; Shi, Jinsong; Xu, Zhenghong; Ma, Yanhe

    2013-11-06

    A novel halophilic alginate-degrading microorganism was isolated from rotten seaweed and identified as Isoptericola halotolerans CGMCC5336. The lyase from the strain was purified to homogeneity by combining of ammonium sulfate fractionation and anion-exchange chromatography with a specific activity of 8409.19 U/ml and a recovery of 25.07%. This enzyme was a monomer with a molecular mass of approximately 28 kDa. The optimal temperature and pH were 50 °C and pH 7.0, respectively. The lyase maintained stability at neutral pH (7.0-8.0) and temperatures below 50 °C. Metal ions including Na(+), Mg(2+), Mn(2+), and Ca(2+) notably increased the activity of the enzyme. With sodium alginate as the substrate, the Km and Vmax were 0.26 mg/ml and 1.31 mg/ml min, respectively. The alginate lyase had substrate specificity for polyguluronate and polymannuronate units in alginate molecules, indicating its bifunctionality. These excellent characteristics demonstrated the potential applications in alginate oligosaccharides production with low polymerisation degrees.

  2. Production of D-tagatose, a functional sweetener, utilizing alginate immobilized Lactobacillus fermentum CGMCC2921 cells.

    PubMed

    Xu, Zheng; Li, Sha; Fu, Fenggen; Li, Guixiang; Feng, Xiaohai; Xu, Hong; Ouyang, Pingkai

    2012-02-01

    D-tagatose is a ketohexose that can be used as a novel functional sweetener in foods, beverages, and dietary supplements. This study was aimed at developing a high-yielding D-tagatose production process using alginate immobilized Lactobacillus fermentum CGMCC2921 cells. For the isomerization from D-galactose into D-tagatose, the immobilized cells showed optimum temperature and pH at 65 °C and 6.5, respectively. The alginate beads exhibited a good stability after glutaraldehyde treatment and retained 90% of the enzyme activity after eight cycles (192 h at 65 °C) of batch conversion. The addition of borate with a molar ratio of 1.0 to D-galactose led to a significant enhancement in the D-tagatose yield. Using commercial β-galactosidase and immobilized L. fermentum cells, D-tagatose was successfully obtained from lactose after a two-step biotransformation. The relatively high conversion rate and productivity from D-galactose to D-tagatose of 60% and 11.1 g l⁻¹ h⁻¹ were achieved in a packed-bed bioreactor. Moreover, lactobacilli have been approved as generally recognized as safe organisms, which makes this L. fermentum strain an attracting substitute for recombinant Escherichia coli cells among D-tagatose production progresses.

  3. Protective effect of Bifidobacterium infantis CGMCC313-2 on ovalbumin-induced airway asthma and β-lactoglobulin-induced intestinal food allergy mouse models

    PubMed Central

    Liu, Meng-Yun; Yang, Zhen-Yu; Dai, Wen-Kui; Huang, Jian-Qiong; Li, Yin-Hu; Zhang, Juan; Qiu, Chuang-Zhao; Wei, Chun; Zhou, Qian; Sun, Xin; Feng, Xin; Li, Dong-Fang; Wang, He-Ping; Zheng, Yue-Jie

    2017-01-01

    AIM To determine whether oral administration of Bifidobacterium infantis CGMCC313-2 (B. infantis CGMCC313-2) inhibits allergen-induced airway inflammation and food allergies in a mouse model. METHODS Ovalbumin (OVA)-induced allergic asthma and β-lactoglobulin-induced food allergy mouse models were used in this study. Following oral administration of B. infantis CGMCC313-2 during or after allergen sensitization, histopathologic changes in the lung and intestine were evaluated by hematoxylin and eosin (HE) staining. In the allergic asthma mouse model, we evaluated the proportion of lung-infiltrating inflammatory cells. OVA-specific IgE and IgG1 levels in serum and cytokine levels in bronchoalveolar lavage fluid (BALF) were also assessed. In the food allergy mouse model, the levels of total IgE and cytokines in serum were measured. RESULTS Oral administration of B. infantis CGMCC313-2 during or after allergen sensitization suppressed allergic inflammation in lung and intestinal tissues, while the proportion of infiltrating inflammatory cells was significantly decreased in the BALF of allergic asthma mice. Moreover, B. infantis CGMCC313-2 decreased the serum levels of total IgE in food allergy mice, and reductions in IgE and IgG1 were also observed in OVA-induced allergic asthma mice. The expression of interleukin-4 (IL-4) and IL-13 in both serum and BALF was suppressed following the administration of B. infantis CGMCC313-2, while an effect on serum IL-10 levels was not observed. CONCLUSION B. infantis CGMCC313-2 inhibits the secretion of allergen-induced IgE, IL-4 and IL-13, and attenuates allergic inflammation.

  4. Cloning, purification, crystallization and preliminary X-ray diffraction of the OleC protein from Stenotrophomonas maltophilia involved in head-to-head hydrocarbon biosynthesis

    SciTech Connect

    Frias, JA; Goblirsch, BR; Wackett, LP; Wilmot, CM

    2010-08-28

    OleC, a biosynthetic enzyme involved in microbial hydrocarbon biosynthesis, has been crystallized. Synchrotron X-ray diffraction data have been collected to 3.4 A resolution. The crystals belonged to space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 98.8, c = 141.0 A.

  5. Degradation potential of protocatechuate 3,4-dioxygenase from crude extract of Stenotrophomonas maltophilia strain KB2 immobilized in calcium alginate hydrogels and on glyoxyl agarose.

    PubMed

    Guzik, Urszula; Hupert-Kocurek, Katarzyna; Krysiak, Marta; Wojcieszyńska, Danuta

    2014-01-01

    Microbial intradiol dioxygenases have been shown to have a great potential for bioremediation; however, their structure is sensitive to various environmental and chemical agents. Immobilization techniques allow for the improvement of enzyme properties. This is the first report on use of glyoxyl agarose and calcium alginate as matrixes for the immobilization of protocatechuate 3,4-dioxygenase. Multipoint attachment of the enzyme to the carrier caused maintenance of its initial activity during the 21 days. Immobilization of dioxygenase in calcium alginate or on glyoxyl agarose resulted in decrease in the optimum temperature by 5 °C and 10 °C, respectively. Entrapment of the enzyme in alginate gel shifted its optimum pH towards high-alkaline pH while immobilization of the enzyme on glyoxyl agarose did not influence pH profile of the enzyme. Protocatechuate 3,4-dioygenase immobilized in calcium alginate showed increased activity towards 2,5-dihydroxybenzoate, caffeic acid, 2,3-dihydroxybenzoate, and 3,5-dihydroxybenzoate. Slightly lower activity of the enzyme was observed after its immobilization on glyoxyl agarose. Entrapment of the enzyme in alginate gel protected it against chelators and aliphatic alcohols while its immobilization on glyoxyl agarose enhanced enzyme resistance to inactivation by metal ions.

  6. Associated mortality and clinical characteristics of nosocomial Pseudomonas maltophilia in a university hospital.

    PubMed Central

    Morrison, A J; Hoffmann, K K; Wenzel, R P

    1986-01-01

    We studied the spectrum of clinical disease in 99 patients with nosocomial Pseudomonas maltophilia isolates at the University of Virginia Hospital from 1981 through 1984. The annual rate of isolation increased from 7.1 to 14.1 per 10,000 patient discharges. A crude mortality rate of 43% was documented in all patients from whom the organism was cultured, and the data include 12 patients with nosocomial bacteremia (four deaths). Risk factors associated with death for patients having a P. maltophilia isolate included the following: requirement for care in any intensive care unit during hospitalization (P = 0.0001), patient age over 40 years (P = 0.002), and a pulmonary source for the P. maltophilia isolate (P = 0.003). All P. maltophilia isolates were susceptible to trimethoprim-sulfamethoxazole, 60% of the isolates were resistant to all aminoglycosides (amikacin, tobramycin, and gentamicin), and more than 75% of the isolates were resistant to all beta-lactam antibiotics. The antibiotic susceptibility pattern allows for a niche exploitable in the hospital microbial environment by an organism with a marked associated mortality. PMID:3487553

  7. Rapid biodegradation of organophosphorus pesticides by Stenotrophomonas sp. G1.

    PubMed

    Deng, Shuyan; Chen, Yao; Wang, Daosheng; Shi, Taozhong; Wu, Xiangwei; Ma, Xin; Li, Xiangqiong; Hua, Rimao; Tang, Xinyun; Li, Qing X

    2015-10-30

    Organophosphorus insecticides have been widely used, which are highly poisonous and cause serious concerns over food safety and environmental pollution. A bacterial strain being capable of degrading O,O-dialkyl phosphorothioate and O,O-dialkyl phosphate insecticides, designated as G1, was isolated from sludge collected at the drain outlet of a chlorpyrifos manufacture plant. Physiological and biochemical characteristics and 16S rDNA gene sequence analysis suggested that strain G1 belongs to the genus Stenotrophomonas. At an initial concentration of 50 mg/L, strain G1 degraded 100% of methyl parathion, methyl paraoxon, diazinon, and phoxim, 95% of parathion, 63% of chlorpyrifos, 38% of profenofos, and 34% of triazophos in 24 h. Orthogonal experiments showed that the optimum conditions were an inoculum volume of 20% (v/v), a substrate concentration of 50 mg/L, and an incubation temperature in 40 °C. p-Nitrophenol was detected as the metabolite of methyl parathion, for which intracellular methyl parathion hydrolase was responsible. Strain G1 can efficiently degrade eight organophosphorus pesticides (OPs) and is a very excellent candidate for applications in OP pollution remediation.

  8. Poly-γ-glutamic acid produced from Bacillus licheniformis CGMCC 2876 as a potential substitute for polyacrylamide in the sugarcane industry.

    PubMed

    Yan, Shan; Yao, Haosheng; Chen, Zhen; Zeng, Shengquan; Xi, Xi; Wang, Yuanpeng; He, Ning; Li, Qingbiao

    2015-01-01

    As an environmentally friendly and industrially useful biopolymer, poly-γ-glutamic acid (γ-PGA) from Bacillus licheniformis CGMCC 2876 was characterized by the high-resolution mass spectrometry and (1)H NMR. A flocculating activity of 11,474.47 U mL(-1) obtained with γ-PGA, and the effects of carbon sources, ions, and chemical properties (D-/L-composition and molecular weight) on the production and flocculating activity of γ-PGA were discussed. Being a bioflocculant in the sugar refinery process, the color and turbidity of the sugarcane juice was IU 1,877.36 and IU 341.41 with 0.8 ppm of γ-PGA, respectively, which was as good as the most widely used chemically synthesized flocculant in the sugarcane industry--polyacrylamide with 1 ppm. The γ-PGA produced from B. licheniformis CGMCC 2876 could be a promising alternate of chemically synthesized flocculants in the sugarcane industry.

  9. A novel glutamate transport system in poly(γ-glutamic acid)-producing strain Bacillus subtilis CGMCC 0833.

    PubMed

    Wu, Qun; Xu, Hong; Zhang, Dan; Ouyang, Pingkai

    2011-08-01

    Bacillus subtilis CGMCC 0833 is a poly(γ-glutamic acid) (γ-PGA)-producing strain. It has the capacity to tolerate high concentration of extracellular glutamate and to utilize glutamate actively. Such a high uptake capacity was owing to an active transport system for glutamate. Therefore, a specific transport system for L-glutamate has been observed in this strain. It was a novel transport process in which glutamate was symported with at least two protons, and an inward-directed sodium gradient had no stimulatory effect on it. K(m) and V(m) for glutamate transport were estimated to be 67 μM and 152 nmol⁻¹ min⁻¹ mg⁻¹ of protein, respectively. The transport system showed structural specificity and stereospecificity and was strongly dependent on extracellular pH. Moreover, it could be stimulated by Mg²⁺, NH₄⁺, and Ca²⁺. In addition, the glutamate transporter in this strain was studied at the molecular level. As there was no important mutation of the transporter protein, it appeared that the differences of glutamate transporter properties between this strain and other B. subtilis strains were not due to the differences of the amino acid sequence and the structure of transporter protein. This is the first extensive report on the properties of glutamate transport system in γ-PGA-producing strain.

  10. Production of bacterial cellulose by Gluconacetobacter hansenii CGMCC 3917 using only waste beer yeast as nutrient source.

    PubMed

    Lin, Dehui; Lopez-Sanchez, Patricia; Li, Rui; Li, Zhixi

    2014-01-01

    In order to improve the use of waste beer yeast (WBY) for bacterial cellulose production by Gluconacetobacter hansenii CGMCC 3917, a two-step pre-treatment was designed. First WBY was treated by 4 methods: 0.1M NaOH treatment, high speed homogenizer, ultrasonication and microwave treatment followed by hydrolysis (121°C, 20 min) under mild acid condition (pH 2). The optimal pre-treatment conditions were evaluated by the reducing sugar yield after hydrolysis. 15% WBY treated by ultrasonication for 40 min had the highest reducing sugar yield (29.19%), followed by NaOH treatment (28.98%), high speed homogenizer (13.33%) and microwaves (13.01%). Treated WBY hydrolysates were directly supplied as only nutrient source for BC production. A sugar concentration of 3% WBY hydrolysates treated by ultrasonication gave the highest BC yield (7.02 g/L), almost 6 times as that from untreated WBY (1.21 g/L). Furthermore, the properties of the BC were as good as those obtained from the conventional chemical media.

  11. Sophorolipids production from rice straw via SO3 micro-thermal explosion by Wickerhamiella domercqiae var. sophorolipid CGMCC 1576.

    PubMed

    Liu, Xin-Ge; Ma, Xiao-Jing; Yao, Ri-Sheng; Pan, Chun-Yu; He, Hua-Bing

    2016-12-01

    A novel lignocellulose material, holocellulose from rice straw via the pretreatment of SO3 micro-thermal explosion, was developed to produce sophorolipids (SLs) with Wickerhamiella domercqiae var. sophorolipid CGMCC 1576. The influence factors of inoculum dose, yeast extract concentration and pH regulators (chemical regents used for adjusting/influencing pH) was investigated and discussed. Results showed that W. domercqiae can grow in the rice straw holocellulose hydrolysate, and acquire relative high SL yield of 53.70 ± 2.61 g/L in shake flask culture. Inoculum dose, yeast extract concentration and pH regulator made obvious influence on fermentation parameters, especially on final broth pH and SLs production. Furthermore, there is a strong negative linear correlation existing between final broth pH and lactonic SL or ratio of lac SL/tot SL. Additionally, comparison between SL production and non-glucose carbon sources, culture methods, microbes in previous reports was carried out. These results will be benefit for acquiring SL mixture with suitable lac SL/tot SL ratio for specific purpose and scope economically.

  12. Indirect Manganese Removal by Stenotrophomonas sp. and Lysinibacillus sp. Isolated from Brazilian Mine Water

    PubMed Central

    Barboza, Natália Rocha; Amorim, Soraya Sander; Santos, Pricila Almeida; Reis, Flávia Donária; Cordeiro, Mônica Mendes; Guerra-Sá, Renata; Leão, Versiane Albis

    2015-01-01

    Manganese is a contaminant in the wastewaters produced by Brazilian mining operations, and the removal of the metal is notoriously difficult because of the high stability of the Mn(II) ion in aqueous solutions. To explore a biological approach for removing excessive amounts of aqueous Mn(II), we investigated the potential of Mn(II) oxidation by both consortium and bacterial isolates from a Brazilian manganese mine. A bacterial consortium was able to remove 99.7% of the Mn(II). A phylogenetic analysis of isolates demonstrated that the predominant microorganisms were members of Stenotrophomonas, Bacillus, and Lysinibacillus genera. Mn(II) removal rates between 58.5% and 70.9% were observed for Bacillus sp. and Stenotrophomonas sp. while the Lysinibacillus isolate 13P removes 82.7%. The catalytic oxidation of Mn(II) mediated by multicopper oxidase was not properly detected; however, in all of the experiments, a significant increase in the pH of the culture medium was detected. No aggregates inside the cells grown for a week were found by electronic microscopy. Nevertheless, an energy-dispersive X-ray spectroscopy of the isolates revealed the presence of manganese in Stenotrophomonas sp. and Lysinibacillus sp. grown in K medium. These results suggest that members of Stenotrophomonas and Lysinibacillus genera were able to remove Mn(II) by a nonenzymatic pathway. PMID:26697496

  13. Revealing Differences in Metabolic Flux Distributions between a Mutant Strain and Its Parent Strain Gluconacetobacter xylinus CGMCC 2955

    PubMed Central

    Liu, Miao; Yang, Xiao-Ning; Zhu, Hui-Xia; Jia, Yuan-Yuan; Jia, Shi-Ru; Piergiovanni, Luciano

    2014-01-01

    A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955) using DEC (diethyl sulfate) and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct) concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA) cycle was obtained in mutant strain (57.0%) compared with parent strain (17.0%). It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP) and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH), which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53–6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain. PMID:24901455

  14. Clostridium Butyricum CGMCC0313.1 Modulates Lipid Profile, Insulin Resistance and Colon Homeostasis in Obese Mice

    PubMed Central

    Shang, Haixiao

    2016-01-01

    Obesity is associated with a cluster of metabolic disorders and systemic low-grade inflammation involving multiple organs. Recent findings have suggested that intestine is a key organ altered in response to high fat diet (HFD) feeding. Probiotics mainly lactobacillus strains have earlier been implicated in alleviating metabolic disorders. Here we aimed to examine the effects of a naturally occurring butyrate-producing probiotic clostridium butyricum CGMCC0313.1 (CB0313.1) in limiting the development of HFD-induced obesity. Mice treated with CB0313.1 exhibited reduced lipid accumulation in liver and serum, lower circulating insulin levels and improved glucose tolerance and insulin sensitivity. Furthermore, CB0313.1 administration reversed the HFD-induced colonic inflammation as evidenced by reduced tumor necrosis factor (TNF)-α level and increases the interleukin (IL)-10 and IL-22 levels in colon tissue. Additionally to colonic inflammation, CB0313.1 also reduced the colon permeability by upregulating the tight junction (TJ) proteins (claudin-1 and occludin) and contributed to a decreased circulating endotoxin level. In colon content, CB0313.1 administration restored the reduced production of butyrate and other short chain fatty acids (SCFAs) caused by HFD feeding. In adipose tissue, lower transcriptional levels of pro-inflammatory TNF-α, IL-6, IL-1β and monocyte chemotactic protein (MCP)-1 in adipose tissue were observed in CB0313.1-treated mice. Collectively, our data demonstrated that CB0313.1, targeting colon inflammation and permeability, ameliorated HFD-induced obesity, insulin resistance as well as adipose inflammation. PMID:27123997

  15. High efficiency transformation of stevioside into a single mono-glycosylated product using a cyclodextrin glucanotransferase from Paenibacillus sp. CGMCC 5316.

    PubMed

    Yu, Xuejian; Yang, Jinshui; Li, Baozhen; Yuan, Hongli

    2015-12-01

    Stevioside is a non-caloric, natural, high-intensity sweetener. However, the bitter aftertaste of stevioside restricts its utilization for human consumption and limits its application in the food industry. In this study, a high efficiency enzymatic modification system was investigated to improve stevioside taste quality. A cyclodextrin glucanotransferase (CGTase) producing strain Paenibacillus sp. CGMCC 5316 was isolated from Stevia planting soil. With starch as glycosyl donor, this CGTase can transform stevioside into a single specific product which is an isomer of rebaudioside A and identified as mono-glycosylated stevioside. The taste of stevioside is improved noticeably by generating mono-glycosylated stevioside, which possesses a sucrose-like taste and has sweetness increased significantly by 35.4%. Next, the parameters influencing CGTase production were optimized. Compared to initial conditions, CGTase activity increased by 214.7% under optimum conditions of 3.9 g/L starch, 17.9 g/L tryptone, and 67.6 h of culture time, and the transglycosylation rate of stevioside was remarkably increased by 284.8%, reaching 85.6%. This CGTase modification system provides a promising solution for improving the sweetness and taste quality of stevioside. The efficiency of CGTase transformation can be greatly increased by optimizing the culture conditions of Paenibacillus sp. CGMCC 5316.

  16. Complete Genome Sequence of a Copper-Resistant Bacterium from the Citrus Phyllosphere, Stenotrophomonas sp. Strain LM091, Obtained Using Long-Read Technology

    PubMed Central

    Richard, Damien; Boyer, Claudine; Lefeuvre, Pierre

    2016-01-01

    The Stenotrophomonas genus shows great adaptive potential including resistance to multiple antimicrobials, opportunistic pathogenicity, and production of numerous secondary metabolites. Using long-read technology, we report the sequence of a plant-associated Stenotrophomonas strain originating from the citrus phyllosphere that displays a copper resistance phenotype. PMID:27979933

  17. The production and molecular properties of the zinc beta-lactamase of Pseudomonas maltophilia IID 1275.

    PubMed Central

    Bicknell, R; Emanuel, E L; Gagnon, J; Waley, S G

    1985-01-01

    The production and purification of a tetrameric zinc beta-lactamase from Pseudomonas maltophilia IID 1275 were greatly improved. Three charge variants were isolated by chromatofocusing. The subunits each contain two atomic proportions of zinc and (in two of the variants) one residue of cysteine. The thiol group is not required for activity, nor does it appear to bind to the metal. Replacement of zinc by cobalt, cadmium or nickel takes place at a measurable rate, and gives enzymes that are less active than the zinc enzyme. The properties of this enzyme differ from those of the other known zinc beta-lactamase, beta-lactamase II from Bacillus cereus. The amino acid sequence of the N-terminal 32 residues was determined; there is no similarity to the N-terminal sequences of other beta-lactamases. PMID:3931629

  18. Engineering chlorpyrifos-degrading Stenotrophomonas sp. YC-1 for heavy metal accumulation and enhanced chlorpyrifos degradation.

    PubMed

    Liu, Ruihua; Jiang, Hong; Xu, Ping; Qiao, Chuanling; Zhou, Qixing; Yang, Chao

    2014-11-01

    Many ecosystems are currently co-contaminated with pesticides and heavy metals, such as chlorpyrifos and cadmium. A promising strategy to remediate mixed chlorpyrifos-cadmium-contaminated sites is the use of chlorpyrifos-degrading bacteria endowed with cadmium removal capabilities. In this work, a gene coding for synthetic phytochelatins (EC20) with high cadmium-binding capacity was introduced into a chlorpyrifos-degrading bacterium, Stenotrophomonas sp. YC-1, resulting in an engineered strain with both cadmium accumulation and chlorpyrifos degradation capabilities. To improve the cadmium-binding efficiency of whole cells, EC20 was displayed on the cell surface of Stenotrophomonas sp. YC-1 using the truncated ice nucleation protein (INPNC) anchor. The surface localization of the INPNC-EC20 fusion protein was demonstrated by cell fractionation, Western blot analysis, and immunofluorescence microscopy. Expression of EC20 on the cell surface not only improved cadmium binding, but also alleviated the cellular toxicity of cadmium. As expected, the chlorpyrifos degradation rate was reduced in the presence of cadmium for cells without EC20 expression. However, expression of EC20 (higher cadmium accumulation) completely restored the level of chlorpyrifos degradation. These results demonstrated that EC20 expression not only enhanced cadmium accumulation, but also reduced the toxic effect of cadmium on chlorpyrifos degradation.

  19. Characterization of three antifungal calcite-forming bacteria, Arthrobacter nicotianae KNUC2100, Bacillus thuringiensis KNUC2103, and Stenotrophomonas maltophilia KNUC2106, derived from the Korean islands, Dokdo and their application on mortar.

    PubMed

    Park, Jong-Myong; Park, Sung-Jin; Ghim, Sa-Youl

    2013-09-28

    Crack remediation on the surface of cement mortar using microbiological calcium carbonate (CaCO3) precipitation (MICP) has been investigated as a microbial sealing agent on construction materials. However, MICP research has never acknowledged the antifungal properties of calcite-forming bacteria (CFB). Since fungal colonization on concrete surfaces can trigger biodeterioration processes, fungi on concrete buildings have to be prevented. Therefore, to develop a microbial sealing agent that has antifungal properties to remediate cement cracks without deteriorative fungal colonization, we introduced an antifungal CFB isolated from oceanic islands (Dokdo islands, territory of South Korea, located at the edge of the East Sea in Korea.). The isolation of CFB was done using B4 or urea-CaCl2 media. Furthermore, antifungal assays were done using the pairing culture and disk diffusion methods. Five isolated CFB showed CaCO3 precipitation and antifungal activities against deteriorative fungal strains. Subsequently, five candidate bacteria were identified using 16S rDNA sequence analysis. Crack remediation, fungi growth inhibition, and water permeability reduction of antifungal CFB-treated cement surfaces were tested. All antifungal CFB showed crack remediation abilities, but only three strains (KNUC2100, 2103, and 2106) reduced the water permeability. Furthermore, these three strains showed fungi growth inhibition. This paper is the first application research of CFB that have antifungal activity, for an eco-friendly improvement of construction materials.

  20. Isolation of new Stenotrophomonas bacteriophages and genomic characterization of temperate phage S1.

    PubMed

    García, Pilar; Monjardín, Cristina; Martín, Rebeca; Madera, Carmen; Soberón, Nora; Garcia, Eva; Meana, Alvaro; Suárez, Juan E

    2008-12-01

    Twenty-two phages that infect Stenotrophomonas species were isolated through sewage enrichment and prophage induction. Of them, S1, S3, and S4 were selected due to their wide host ranges compared to those of the other phages. S1 and S4 are temperate siphoviruses, while S3 is a virulent myovirus. The genomes of S3 and S4, about 33 and 200 kb, were resistant to restriction digestion. The lytic cycles lasted 30 min for S3 and about 75 min for S1 and S4. The burst size for S3 was 100 virions/cell, while S1 and S4 produced about 75 virus particles/cell. The frequency of bacteriophage-insensitive host mutants, calculated by dividing the number of surviving colonies by the bacterial titer of a parallel, uninfected culture, ranged between 10(-5) and 10(-6) for S3 and 10(-3) and 10(-4) for S1 and S4. The 40,287-bp genome of S1 contains 48 open reading frames (ORFs) and 12-bp 5' protruding cohesive ends. By using a combination of bioinformatics and experimental evidence, functions were ascribed to 21 ORFs. The morphogenetic and lysis modules are well-conserved, but no lysis-lysogeny switch or DNA replication gene clusters were recognized. Two major clusters of genes with respect to transcriptional orientation were observed. Interspersed among them were lysogenic conversion genes encoding phosphoadenosine phosphosulfate reductase and GspM, a protein involved in the general secretion system II. The attP site of S1 may be located within a gene that presents over 75% homology to a Stenotrophomonas chromosomal determinant.

  1. Metabolic flux analysis of Arthrobacter sp. CGMCC 3584 for cAMP production based on 13C tracer experiments and gas chromatography-mass spectrometry.

    PubMed

    Niu, Huanqing; Chen, Yong; Yao, Shiwei; Liu, Lixia; Yang, Chen; Li, Bingbing; Liu, Dong; Xie, Jingjing; Chen, Xiaochun; Wu, Jinglan; Ying, Hanjie

    2013-12-01

    Arthrobacter sp. CGMCC 3584 are able to produce cAMP from glucose by the purine synthesis pathway via de novo or salvage biosynthesis. In order to gain an improved understanding of its metabolism, (13)C-labeling experiment and gas chromatography-mass spectrometry (GC-MS) analysis were employed to determine the metabolic network structure and estimate the intracellular fluxes. GC-MS analysis helps to reflect the activity of the intracellular pathways and reactions. The metabolic network mainly contains glycolytic and pentose phosphate pathways, the tricarboxylic acid cycle, and the inactive glyoxylate shunt. Hypoxanthine as a precursor of cAMP and sodium fluoride as an inhibitor of glycolysis were found to increase the cAMP production, as well as the flux through the PP pathway. The effects of adding hypoxanthine and sodium fluoride are discussed based on the enzyme assays and metabolic flux analysis. In conclusion, our results provide quantitative insights into how cells manipulate the metabolic network under different culture conditions and this may be of value in metabolic regulation for desirable production.

  2. Stenotrophomonas sp. RZS 7, a novel PHB degrader isolated from plastic contaminated soil in Shahada, Maharashtra, Western India.

    PubMed

    Wani, S J; Shaikh, S S; Tabassum, B; Thakur, R; Gulati, A; Sayyed, R Z

    2016-12-01

    This paper reports an isolation and identification of novel poly-β-hydroxybutyrate (PHB) degrading bacterium Stenotrophomonas sp. RZS 7 and studies on its extracellular PHB degrading depolymerase enzyme. The bacterium isolated from soil samples of plastic contaminated sites of municipal area in Shahada, Maharashtra, Western India. It was identified as Stenotrophomonas sp. RZS 7 based on polyphasic approach. The bacterium grew well in minimal salt medium (MSM) and produced a zone (4.2 mm) of PHB hydrolysis on MSM containing PHB as the only source of nutrient. An optimum yield of enzyme was obtained on the fifth day of incubation at 37 °C and at pH 6.0. Further increase in enzyme production was recorded with Ca(2+) ions, while other metal ions like Fe(2+) (1 mM) and chemical viz. mercaptoethanol severally affected the production of enzyme.

  3. Biosynthesis of gold and silver nanoparticles using a novel marine strain of Stenotrophomonas.

    PubMed

    Malhotra, Ankit; Dolma, Kunzes; Kaur, Navjot; Rathore, Y S; Ashish; Mayilraj, S; Choudhury, Anirban Roy

    2013-08-01

    The present study aims at exploiting marine microbial diversity for biosynthesis of metal nanoparticles and also investigates role of microbial proteins in the process of bio-mineralization of gold and silver. This is the first report for concurrent production of gold and silver nanoparticles (AuNPs and AgNPs) by extracellular secretion of a novel strain of Stenotrophomonas, isolated from Indian marine origin. This novel strain has faster rate kinetics for AgNPs synthesis than any other organism reported earlier. The nanoparticles were further characterized using UV-vis spectrophotometer, TEM, DLS and EDAX confirming their size ranging from 10-50 nm and 40-60 nm in dimensions for AuNPs and AgNPs, respectively. TEM analysis indicated formation of multi-shaped nanoparticles with heterogeneous size distribution in both the cases. Finally, the SDS-PAGE analysis of extracellular media supernatant suggested a potential involvement of certain low molecular weight secretory proteins in AuNPs and AgNPs biosynthesis.

  4. Biodegradation of DDT by Stenotrophomonas sp. DDT-1: Characterization and genome functional analysis

    NASA Astrophysics Data System (ADS)

    Pan, Xiong; Lin, Dunli; Zheng, Yuan; Zhang, Qian; Yin, Yuanming; Cai, Lin; Fang, Hua; Yu, Yunlong

    2016-02-01

    A novel bacterium capable of utilizing 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) as the sole carbon and energy source was isolated from a contaminated soil which was identified as Stenotrophomonas sp. DDT-1 based on morphological characteristics, BIOLOG GN2 microplate profile, and 16S rDNA phylogeny. Genome sequencing and functional annotation of the isolate DDT-1 showed a 4,514,569 bp genome size, 66.92% GC content, 4,033 protein-coding genes, and 76 RNA genes including 8 rRNA genes. Totally, 2,807 protein-coding genes were assigned to Clusters of Orthologous Groups (COGs), and 1,601 protein-coding genes were mapped to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The degradation half-lives of DDT increased with substrate concentration from 0.1 to 10.0 mg/l, whereas decreased with temperature from 15 °C to 35 °C. Neutral condition was the most favorable for DDT biodegradation. Based on genome annotation of DDT degradation genes and the metabolites detected by GC-MS, a mineralization pathway was proposed for DDT biodegradation in which it was orderly converted into DDE/DDD, DDMU, DDOH, and DDA via dechlorination, hydroxylation, and carboxylation, and ultimately mineralized to carbon dioxide. The results indicate that the isolate DDT-1 is a promising bacterial resource for the removal or detoxification of DDT residues in the environment.

  5. Astaxanthin preparation by fermentation of esters from Haematococcus pluvialis algal extracts with Stenotrophomonas species.

    PubMed

    Dong, Hao; Li, Xuemin; Xue, Changhu; Mao, Xiangzhao

    2016-05-01

    Natural astaxanthin (Ax) is an additive that is widely used because of its beneficial biochemical functions. However, the methods used to produce free Ax have drawbacks. Chemical saponification methods produce several by-products, and lipase-catalyzed hydrolysis methods are not cost effective. In this study, a bacterial strain of Stenotrophomonas sp. was selected to enzymatically catalyze the saponification of Ax esters to produce free all-trans-Ax. Through single-factor experiments and a Box-Behnken design, the optimal fermentation conditions were determined as follows: a seed culture age of 37.79 h, an inoculum concentration of 5.92%, and an initial broth pH of 6.80. Under these conditions, a fermentation curve was drawn, and the optimal fermentation time was shown to be 60 h. At 60 h, the degradation rate of the Ax esters was 98.08%, and the yield of free all-trans-Ax was 50.130 μg/mL. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:649-656, 2016.

  6. Biodegradation of DDT by Stenotrophomonas sp. DDT-1: Characterization and genome functional analysis.

    PubMed

    Pan, Xiong; Lin, Dunli; Zheng, Yuan; Zhang, Qian; Yin, Yuanming; Cai, Lin; Fang, Hua; Yu, Yunlong

    2016-02-18

    A novel bacterium capable of utilizing 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) as the sole carbon and energy source was isolated from a contaminated soil which was identified as Stenotrophomonas sp. DDT-1 based on morphological characteristics, BIOLOG GN2 microplate profile, and 16S rDNA phylogeny. Genome sequencing and functional annotation of the isolate DDT-1 showed a 4,514,569 bp genome size, 66.92% GC content, 4,033 protein-coding genes, and 76 RNA genes including 8 rRNA genes. Totally, 2,807 protein-coding genes were assigned to Clusters of Orthologous Groups (COGs), and 1,601 protein-coding genes were mapped to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. The degradation half-lives of DDT increased with substrate concentration from 0.1 to 10.0 mg/l, whereas decreased with temperature from 15 °C to 35 °C. Neutral condition was the most favorable for DDT biodegradation. Based on genome annotation of DDT degradation genes and the metabolites detected by GC-MS, a mineralization pathway was proposed for DDT biodegradation in which it was orderly converted into DDE/DDD, DDMU, DDOH, and DDA via dechlorination, hydroxylation, and carboxylation, and ultimately mineralized to carbon dioxide. The results indicate that the isolate DDT-1 is a promising bacterial resource for the removal or detoxification of DDT residues in the environment.

  7. Once-Daily Amikacin Dosing in Burn Patients Treated with Continuous Venovenous Hemofiltration

    DTIC Science & Technology

    2011-10-01

    Stenotrophomonas maltophilia (11 isolates), Enterobacter aerogenes (9 isolates), Aeromonas hydro- philia (6 isolates), Serratia marcescens (4 isolates), Escherichia...64 Acinetobacter baumannii (35) 39.1% 23 64/64 Other Enterobacteriaciae (43) Stenotrophomonas maltophilia (11), Enterobacter aerogenes (9...Patient and infection characteristics . Sixty patients received amikacin and had sufficient dosing and postinfusion data to calculate pharmacokinetic

  8. Microcystin-degrading activity of an indigenous bacterial strain Stenotrophomonas acidaminiphila MC-LTH2 isolated from Lake Taihu.

    PubMed

    Yang, Fei; Zhou, Yuanlong; Yin, Lihong; Zhu, Guangcan; Liang, Geyu; Pu, Yuepu

    2014-01-01

    Microcystin-LR (MC-LR) and microcystin-RR (MC-RR) produced by harmful cyanobacterial blooms (HCBs) pose substantial threats to the ecosystem and public health due to their potential hepatotoxicity. Degradation of microcystins (MCs) by indigenous bacteria represents a promising method for removing MCs from fresh water without harming the aquatic environment, but only a few microcystin (MC)-degrading bacteria have been isolated and had their mechanisms reported. This study aimed to isolate indigenous bacteria from Lake Taihu, and investigate the capability and mechanism of MC degradation by these bacteria. During a Microcystis bloom, an indigenous MC-degrading bacterium designated MC-LTH2 was successfully isolated from Lake Taihu, and identified as Stenotrophomonas acidaminiphila based on phylogenetic analysis. In the presence of MC-LR together with MC-RR, the strain MC-LTH2 was capable of totally degrading both simultaneously in 8 days, at rates of 3.0 mg/(L⋅d) and 5.6 mg/(L⋅d), respectively. The degradation rates of MCs were dependent on temperature, pH, and initial MC concentration. Adda (3-amino-9-methoxy-2, 6, 8-trimethyl-10-phenyldeca-4, 6-dienoic acid) was detected as an intermediate degradation product of MCs using high performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS). To the best of our knowledge, this is the first report of Stenotrophomonas acidaminiphila capable of degrading two MC analogues and other compounds containing Adda residue completely under various conditions, although the mlrA gene in the strain was not detected. These results indicate the Stenotrophomonas acidaminiphila strain MC-LTH2 possesses a significant potential to be used in bioremediation of water bodies contaminated by MC-LR and MC-RR, and is potentially involved in the degradation of MCs during the disappearance of the HCBs in Lake Taihu.

  9. Root-microbe systems: the effect and mode of interaction of Stress Protecting Agent (SPA) Stenotrophomonas rhizophila DSM14405T

    PubMed Central

    Alavi, Peyman; Starcher, Margaret R.; Zachow, Christin; Müller, Henry; Berg, Gabriele

    2013-01-01

    Stenotrophomonas rhizophila has great potential for applications in biotechnology and biological control due to its ability to both promote plant growth and protect roots against biotic and a-biotic stresses, yet little is known about the mode of interactions in the root-environment system. We studied mechanisms associated with osmotic stress using transcriptomic and microscopic approaches. In response to salt or root extracts, the transcriptome of S. rhizophila DSM14405T changed drastically. We found a notably similar response for several functional gene groups responsible for general stress protection, energy production, and cell motility. However, unique changes in the transcriptome were also observed: the negative regulation of flagella-coding genes together with the up-regulation of the genes responsible for biofilm formation and alginate biosynthesis were identified as a single mechanism of S. rhizophila DSM14405T against salt shock. However, production and excretion of glucosylglycerol (GG) were found as a remarkable mechanism for the stress protection of this Stenotrophomonas strain. For S. rhizophila treated with root exudates, the shift from the planktonic lifestyle to a sessile one was measured as expressed in the down-regulation of flagellar-driven motility. These findings fit well with the observed positive regulation of host colonization genes and microscopic images that show different colonization patterns of oilseed rape roots. Spermidine, described as a plant growth regulator, was also newly identified as a protector against stress. Overall, we identified mechanisms of Stenotrophomonas to protect roots against osmotic stress in the environment. In addition to both the changes in life style and energy metabolism, phytohormons, and osmoprotectants were also found to play a key role in stress protection. PMID:23717321

  10. Thrombus Degradation by Fibrinolytic Enzyme of Stenotrophomonas sp. Originated from Indonesian Soybean-Based Fermented Food on Wistar Rats

    PubMed Central

    Tjandrawinata, Raymond R.

    2016-01-01

    Objective. To evaluate thrombus degrading effect of a fibrinolytic enzyme from food origin Stenotrophomonas sp. of Indonesia. Methods. Prior to animal study, the enzyme safety was tested using cell culture. The effect on expression of tissue plasminogen activator was also analysed in the cell culture. For in vivo studies, 25 Wistar rats were used: normal control, negative control, treatment groups with crude and semipurified enzyme given orally at 25 mg/kg, and positive control group which received Lumbrokinase at 25 mg/kg. Blood clot in the tail was induced by kappa carrageenan injection at 1 mg/kg BW. Results. Experiment with cell culture confirmed the enzyme safety at the concentration used and increased expression of tPA. Decreasing of thrombus was observed in the positive group down to 70.35 ± 23.11% of the negative control animals (100%). The thrombus observed in the crude enzyme treatment was down to 56.99 ± 15.95% and 71.5 ± 15.7% for semipurified enzyme. Scanning electron microscopy showed clearly that bood clots were found in the animals injected with kappa carrageenan; however, in the treatment and positive groups, the clot was much reduced. Conclusions. Oral treatment of enzyme from Stenotrophomonas sp. of Indonesian fermented food was capable of degrading thrombus induced in Wistar rats. PMID:27635131

  11. Degradation of Microcystin-LR and RR by a Stenotrophomonas sp. Strain EMS Isolated from Lake Taihu, China

    PubMed Central

    Chen, Jian; Hu, Liang Bin; Zhou, Wei; Yan, Shao Hua; Yang, Jing Dong; Xue, Yan Feng; Shi, Zhi Qi

    2010-01-01

    A bacterial strain EMS with the capability of degrading microcystins (MCs) was isolated from Lake Taihu, China. The bacterium was tentatively identified as a Stenotrophomonas sp. The bacterium could completely consume MC-LR and MC-RR within 24 hours at a concentration of 0.7 μg/mL and 1.7 μg/mL, respectively. The degradation of MC-LR and MC-RR by EMS occurred preferentially in an alkaline environment. In addition, mlrA gene involved in the degradation of MC-LR and MC-RR was detected in EMS. Due to the limited literature this gene has rare homologues. Sequencing analysis of the translated protein from mlrA suggested that MlrA might be a transmembrane protein, which suggests a possible new protease family having unique function. PMID:20479990

  12. Biosurfactant production by the crude oil degrading Stenotrophomonas sp. B-2: chemical characterization, biological activities and environmental applications.

    PubMed

    Gargouri, Boutheina; Contreras, María Del Mar; Ammar, Sonda; Segura-Carretero, Antonio; Bouaziz, Mohamed

    2016-11-26

    In this work, biosurfactant-producing microorganisms were isolated from hydrocarbon-contaminated water collected from Tunisian oilfield. After enrichment and isolation, different bacterial strains were preliminary studied for their biosurfactant/bioemulsifier properties when using crude oil as the unique carbon source. In particular, the isolate strain B-2, a Gram-negative, rod-shaped bacterium, efficiently emulsified crude oil. The extracellular biosurfactant product from this strain presented an emulsification activity above 70% and a hydrophobicity of 71%. In addition, a diameter of 6 cm was observed in the oil displacement test. The characterization of B-2 strain using 16S rDNA sequencing enables us to find a high degree of similarity with various members of the genus Stenotrophomonas (with a percentage of similarity of 99%). The emulsification activity of Stenotrophomonas biosurfactant B-2 was maintained in a wide range of pH (2 to 6), temperature (4 to 55 °C), and salinity (0 to 50 g L(-1)) conditions. It also enhanced the solubility of phenanthrene in water and could be used in the re-mobilization of hydrocarbon-contaminated environment. In addition, this biosurfactant exhibited antimicrobial and antioxidant properties. Infrared spectroscopy suggested potential lipidic and peptidic moieties, and mass spectrometry-based analyses showed that the biosurfactant contains mainly cyclic peptidic structures belonging to the class of diketopiperazines. Therefore, the B-2 strain is a promising biosurfactant-producing microorganism and its derived biosurfactant presents a wide range of industrial applications.

  13. Potential of biosynthesized silver nanoparticles using Stenotrophomonas sp. BHU-S7 (MTCC 5978) for management of soil-borne and foliar phytopathogens

    PubMed Central

    Mishra, Sandhya; Singh, Braj Raj; Naqvi, Alim H.; Singh, H. B.

    2017-01-01

    Stenotrophomonas sp. is emerging as a popular microbe of global concern with various potential ecological roles. Biosynthesis of gold and silver nanoparticles (AgNPs) using this bacterial strain has shown promising applications in life sciences. However, there is no report on efficient agricultural applications of biosynthesized AgNPs using Stenotrophomonas sp. In this regard, successful biosynthesis of AgNPs using Stenotrophomonas sp. BHU-S7 (MTCC 5978) was monitored by Uv-visible spectrum showing surface plasmon resonance (SPR) peak at 440 nm. The biosynthesized AgNPs were spherical with an average mean size of ~12 nm. The antifungal efficacy of biosynthesized AgNPs against foliar and soil-borne phytopathogens was observed. The inhibitory impact of AgNPs (2, 4, 10 μg/ml) on conidial germination was recorded under in vitro conditions. Interestingly, sclerotia of Sclerotium rolfsii exposed to AgNPs failed to germinate on PDA medium and in soil system. Moreover, AgNPs treatment successfully managed collar rot of chickpea caused by S. rolfsii under greenhouse conditions. The reduced sclerotia germination, phenolic acids induction, altered lignification and H2O2 production was observed to be the probable mechanisms providing protection to chickpea against S. rolfsii. Our data revealed that AgNPs treated plants are better equipped to cope with pathogen challenge pointing towards their robust applications in plant disease management. PMID:28345581

  14. Optimization of medium components and physicochemical parameters to simultaneously enhance microbial growth and production of lypolitic enzymes by Stenotrophomonas sp.

    PubMed

    Mazzucotelli, Cintia Anabela; Agüero, María Victoria; Del Rosario Moreira, María; Ansorena, María Roberta

    2016-05-01

    The optimization of lipase and esterase production (LP and EP) and bacterial growth (BG) of a Stenotrophomonas sp. strain was developed. For this purpose, the effect of five different medium components and three physicochemical parameters were evaluated using a Plackett-Burman statistical design. Among eight variables, stirring speed, pH, and peptone concentration were found to be the most effective factors on the three responses under evaluation. An optimization study applying Box-Behnken response surface methodology was used to study the interactive effects of the three selected variables on LP/EP and microorganism growth. Predicted models were found to be significant with high regression coefficients (90%-99%). By using the desirability function approach, the optimum condition applying simultaneous optimization of the three responses under study resulted to be: stirring speed of 100 rpm, pH of 7.5, and a peptone concentration of 10 g/L, with a desirability value of 0.977. Under these optimal conditions, it is possible to achieve in the optimized medium a 15-fold increase in esterase productivity, a 117-fold increase in lipase production, and a 9-log CFU/mL increase in BG, compared with the basal medium without agitation.

  15. Whole-genome sequence assembly of Pediococcus pentosaceus LI05 (CGMCC 7049) from the human gastrointestinal tract and comparative analysis with representative sequences from three food-borne strains

    PubMed Central

    2014-01-01

    Background Strains of Pediococcus pentosaceus from food and the human gastrointestinal tract have been widely identified, and some have been reported to reduce inflammation, encephalopathy, obesity and fatty liver in animals. In this study, we sequenced the whole genome of P. pentosaceus LI05 (CGMCC 7049), which was isolated from the fecal samples of healthy volunteers, and determined its ability to reduce acute liver injury. No other genomic information for gut-borne P. pentosaceus is currently available in the public domain. Results We obtained the draft genome of P. pentosaceus LI05, which was 1,751,578 bp in size and possessed a mean G + C content of 37.3%. This genome encoded an abundance of proteins that were protective against acids, bile salts, heat, oxidative stresses, enterocin A, arsenate and universal stresses. Important adhesion proteins were also encoded by the genome. Additionally, P. pentosaceus LI05 genes encoded proteins associated with the biosynthesis of not only three antimicrobials, including prebacteriocin, lysin and colicin V, but also vitamins and functional amino acids, such as riboflavin, folate, biotin, thiamine and gamma-aminobutyrate. A comparison of P. pentosaceus LI05 with all known genomes of food-borne P. pentosaceus strains (ATCC 25745, SL4 and IE-3) revealed that it possessed four novel exopolysaccharide biosynthesis proteins, additional putative environmental stress tolerance proteins and phage-related proteins. Conclusions This work demonstrated the probiotic properties of P. pentosaceus LI05 from the gut and the three other food-borne P. pentosaceus strains through genomic analyses. We have revealed the major genomic differences between these strains, providing a framework for understanding the probiotic effects of strain LI05, which exhibits unique physiological and metabolic properties. PMID:25349631

  16. Detection of Bacillus and Stenotrophomonas species growing in an organic acid and endocrine-disrupting chemical-rich environment of distillery spent wash and its phytotoxicity.

    PubMed

    Chandra, Ram; Kumar, Vineet

    2017-01-01

    Sugarcane molasses-based distillery spent wash (DSW) is well known for its toxicity and complex mixture of various recalcitrant organic pollutants with acidic pH, but the chemical nature of these pollutants is unknown. This study revealed the presence of toxic organic acids (butanedioic acid bis(TMS)ester; 2-hydroxysocaproic acid; benzenepropanoic acid, α-[(TMS)oxy], TMS ester; vanillylpropionic acid, bis(TMS)), and other recalcitrant organic pollutants (2-furancarboxylic acid, 5-[[(TMS)oxy] methyl], TMS ester; benzoic acid 3-methoxy-4-[(TMS)oxy], TMS ester; and tricarballylic acid 3TMS), which are listed as endocrine-disrupting chemicals. In addition, several major heavy metals were detected, including Fe (163.947), Mn (4.556), Zn (2.487), and Ni (1.175 mg l(-1)). Bacterial community analysis by restriction fragment length polymorphism revealed that Bacillus and Stenotrophomonas were dominant autochthonous bacterial communities belonging to the phylum Firmicutes and γ-Proteobacteria, respectively. The presence of Bacillus and Stenotrophomonas species in highly acidic environments indicated its broad range adaptation. These findings indicated that these autochthonous bacterial communities were pioneer taxa for in situ remediation of this hazardous waste during ecological succession. Further, phytotoxicity assay of DSW with Phaseolus mungo L. and Triticum aestivum revealed that T. aestivum was more sensitive than P. mungo L. in the seed germination test. The results of this study may be useful for monitoring and toxicity assessment of sugarcane molasses-based distillery waste at disposal sites.

  17. Biodegradation of fenvalerate and 3-phenoxybenzoic acid by a novel Stenotrophomonas sp. strain ZS-S-01 and its use in bioremediation of contaminated soils.

    PubMed

    Chen, Shaohua; Yang, Liu; Hu, Meiying; Liu, Jingjing

    2011-04-01

    A bacterial strain ZS-S-01, newly isolated from activated sludge, could effectively degrade fenvalerate and its hydrolysis product 3-phenoxybenzoic acid (3-PBA). Based on the morphology, physiological biochemical characteristics, and 16 S rDNA sequence, strain ZS-S-01 was identified as Stenotrophomonas sp. Strain ZS-S-01 could also degrade and utilize deltamethrin, beta-cypermethrin, beta-cyfluthrin, and cyhalothrin as substrates for growth. Strain ZS-S-01 was capable of degrading fenvalerate rapidly without a lag phase over a wide range of pH and temperature, even in the presence of other carbon sources, and metabolized it to yield 3-PBA, then completely degraded it. No persistent accumulative product was detected by HPLC and GC/MS analysis. Studies on biodegradation in various soils showed that strain ZS-S-01 demonstrated efficient degradation of fenvalerate and 3-PBA (both 50 mg·kg(-1)) with a rate constant of 0.1418-0.3073 d(-1), and half-lives ranged from 2.3 to 4.9 days. Compared with the controls, the half-lives for fenvalerate and 3-PBA reduced by 16.9-156.3 days. These results highlight strain ZS-S-01 may have potential for use in bioremediation of pyrethroid-contaminated environment.

  18. Optimization of Crude Oil and PAHs Degradation by Stenotrophomonas rhizophila KX082814 Strain through Response Surface Methodology Using Box-Behnken Design

    PubMed Central

    Virupakshappa, Praveen Kumar Siddalingappa; Mishra, Gaurav; Mehkri, Mohammed Ameenuddin

    2016-01-01

    The present paper describes the process optimization study for crude oil degradation which is a continuation of our earlier work on hydrocarbon degradation study of the isolate Stenotrophomonas rhizophila (PM-1) with GenBank accession number KX082814. Response Surface Methodology with Box-Behnken Design was used to optimize the process wherein temperature, pH, salinity, and inoculum size (at three levels) were used as independent variables and Total Petroleum Hydrocarbon, Biological Oxygen Demand, and Chemical Oxygen Demand of crude oil and PAHs as dependent variables (response). The statistical analysis, via ANOVA, showed coefficient of determination R2 as 0.7678 with statistically significant P value 0.0163 fitting in second-order quadratic regression model for crude oil removal. The predicted optimum parameters, namely, temperature, pH, salinity, and inoculum size, were found to be 32.5°C, 9, 12.5, and 12.5 mL, respectively. At this optimum condition, the observed and predicted PAHs and crude oil removal were found to be 71.82% and 79.53% in validation experiments, respectively. The % TPH results correlate with GC/MS studies, BOD, COD, and TPC. The validation of numerical optimization was done through GC/MS studies and % removal of crude oil. PMID:28116165

  19. γ-Dodecelactone production from safflower oil via 10-hydroxy-12(Z)-octadecenoic acid intermediate by whole cells of Candida boidinii and Stenotrophomonas nitritireducens.

    PubMed

    Jo, Ye-Seul; An, Jung-Ung; Oh, Deok-Kun

    2014-07-16

    Candida boidinii was selected as a γ-dodecelactone producer because of the highest production of γ-dodecelactone from 10-hydroxy-12(Z)-octadecenoic acid among the 11 yeast strains tested. Under the reaction conditions of pH 5.5 and 25 °C with 5 g/L 10-hydroxy-12(Z)-octadecenoic acid and 30 g/L cells, whole C. boidinii cells produced 2.1 g/L γ-dodecelactone from 5 g/L 10-hydroxy-12(Z)-octadecenoic acid after 6 h, with a conversion yield of 64% (mol/mol) and a volumetric productivity of 350 mg/L/h. The production of γ-dodecelactone from safflower oil was performed by lipase hydrolysis reaction and two-step whole-cell biotransformation using Stenotrophomonas nitritireducens and C. boidinii. γ-Dodecelactone at 1.88 g/L was produced from 7.5 g/L safflower oil via 5 g/L 10-hydroxy-12(Z)-octadecenoic acid intermediate by these reactions after 8 h of reaction time, with a volumetric productivity of 235 mg/L/h and a conversion yield of 25% (w/w). To the best of the authors' knowledge, this is the highest volumetric productivity and conversion yield reported to date for the production of γ-lactone from natural oils.

  20. Optimization of Crude Oil and PAHs Degradation by Stenotrophomonas rhizophila KX082814 Strain through Response Surface Methodology Using Box-Behnken Design.

    PubMed

    Virupakshappa, Praveen Kumar Siddalingappa; Krishnaswamy, Manjunatha Bukkambudhi; Mishra, Gaurav; Mehkri, Mohammed Ameenuddin

    2016-01-01

    The present paper describes the process optimization study for crude oil degradation which is a continuation of our earlier work on hydrocarbon degradation study of the isolate Stenotrophomonas rhizophila (PM-1) with GenBank accession number KX082814. Response Surface Methodology with Box-Behnken Design was used to optimize the process wherein temperature, pH, salinity, and inoculum size (at three levels) were used as independent variables and Total Petroleum Hydrocarbon, Biological Oxygen Demand, and Chemical Oxygen Demand of crude oil and PAHs as dependent variables (response). The statistical analysis, via ANOVA, showed coefficient of determination R(2) as 0.7678 with statistically significant P value 0.0163 fitting in second-order quadratic regression model for crude oil removal. The predicted optimum parameters, namely, temperature, pH, salinity, and inoculum size, were found to be 32.5°C, 9, 12.5, and 12.5 mL, respectively. At this optimum condition, the observed and predicted PAHs and crude oil removal were found to be 71.82% and 79.53% in validation experiments, respectively. The % TPH results correlate with GC/MS studies, BOD, COD, and TPC. The validation of numerical optimization was done through GC/MS studies and % removal of crude oil.

  1. Gene cloning of an efficiency oleate hydratase from Stenotrophomonas nitritireducens for polyunsaturated fatty acids and its application in the conversion of plant oils to 10-hydroxy fatty acids.

    PubMed

    Kang, Woo-Ri; Seo, Min-Ju; Shin, Kyung-Chul; Park, Jin-Byung; Oh, Deok-Kun

    2017-01-01

    Hydroxy fatty acids are used as precursors of lactones and dicarboxylic acids, as starting materials of polymers, and as additives in coatings and paintings. Stenotrophomonas nitritireducens efficiently converts cis-9 polyunsaturated fatty acids (PUFAs) to 10-hydroxy fatty acids. However, gene encoding enzyme involved in this conversion has not been identified to date. We purified a putative fatty acid double-bond hydratase from S. nitritireducens by ultrafiltration and HiPrep DEAE FF and Resource Q ion exchange chromatographies. Peptide sequences of the purified enzyme were obtained by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis. Sequence of the partial gene encoding this putative fatty acid double-bond hydratase was determined by degenerate polymerase chain reaction (PCR) based on the peptide sequences. The remaining gene sequence was identified by rapid amplification of cDNA ends using cDNA of S. nitritireducens as a template, and the full-length gene was cloned subsequently. The expressed enzyme was identified as an oleate hydratase by determining its kinetic parameters toward unsaturated fatty acids. S. nitritireducens oleate hydratase showed higher activity toward PUFAs compared with other available oleate hydratases. This suggested that the enzyme could be used effectively to convert plant oils to 10-hydroxy fatty acids because these oils contained unsaturated fatty acids such as oleic acid (OA) and linoleic acid (LA) and PUFAs such as α-linolenic acid and/or γ-linolenic acid. The enzyme converted soybean oil and perilla seed oil hydrolyzates containing 10 mM total unsaturated fatty acids, including OA, LA, and ALA, to 8.87 and 8.70 mM total 10-hydroxy fatty acids, respectively, in 240 min. To our knowledge, this is the first study on the biotechnological conversion of PUFA-containing oils to hydroxy fatty acids. Biotechnol. Bioeng. 2017;114: 74-82. © 2016 Wiley Periodicals, Inc.

  2. Complete Genome Sequencing of Stenotrophomonas acidaminiphila ZAC14D2_NAIMI4_2, a Multidrug-Resistant Strain Isolated from Sediments of a Polluted River in Mexico, Uncovers New Antibiotic Resistance Genes and a Novel Class-II Lasso Peptide Biosynthesis Gene Cluster.

    PubMed

    Vinuesa, Pablo; Ochoa-Sánchez, Luz Edith

    2015-12-10

    Here, we report the first complete genome sequence of a Stenotrophomonas acidaminiphila strain, generated with PacBio RS II single-molecule real-time technology, consisting of a single circular chromosome of 4.13 Mb. We annotated mobile genetic elements and natural product biosynthesis clusters, including a novel class-II lasso peptide with a 7-residue macrolactam ring.

  3. Treatment of Polymicrobial Osteomyelitis with Ceftolozane-Tazobactam: Case Report and Sensitivity Testing of Isolates

    PubMed Central

    Jolliff, Jeffrey C.; Joson, Jeremiah; Heidari, Arash; Johnson, Royce

    2016-01-01

    Stenotrophomonas maltophilia is an inherently multidrug resistant (MDR) opportunistic pathogen with many mechanisms of resistance. SENTRY studies reveal decreasing sensitivities of S. maltophilia to trimethoprim-sulfamethoxazole and fluoroquinolones. Ceftolozane-tazobactam (Zerbaxa, Merck & Co., Inc.) a novel intravenous combination agent of a third-generation cephalosporin and β-lactamase inhibitor was demonstrated to have in vitro activity against many Gram-positive, Gram-negative, and MDR organisms. Data for ceftolozane-tazobactam's use outside of Food and Drug Administration (FDA) approved indications has been limited thus far to two case reports which demonstrated its efficacy in pan-resistant Pseudomonas aeruginosa pneumonia. Herein, we describe the first published case of treatment of MDR S. maltophilia in polymicrobial osteomyelitis with long-term (>14 days) ceftolozane-tazobactam and metronidazole. Ceftolozane-tazobactam may offer a possible alternative for clinicians faced with limited options in the treatment of resistant pathogens including MDR S. maltophilia. PMID:27437155

  4. Infectious Complications of Open Type III Tibial Fractures among Combat Casualties

    DTIC Science & Technology

    2007-08-15

    Escherichia coli 2 1 Stenotrophomonas maltophilia 1 0 Unknown 1 0 Gram-positive Coagulase-negative staphylococci 3 7 Enterococcus species 3 0 MSSA 2b 5...aeruginosa (6) Amputation (9)c 8 25 (M, IIIb) CoNS ( Escherichia coli and En- terobacter species recovered from broth) (30) Imipenem-cilastatin (2...nonunion wound; this organisim was a methicillin- resistant Staphylococcus aureus strain that was inadvertently not treated. Five of 35 patients

  5. A Case of Sinusitis Caused by Schizophyllum Commune and Bacteria in Acute Myelocytic Leukemia.

    PubMed

    Yin, Xiuyun; Liang, Yuying; Zeng, Lijun; Chen, Shuiping

    2015-01-01

    Schizophyllum commune infections have been rarely reported. Here we reported a rare case of sinusitis in an acute myelocytic leukemia patient, who was co-infected by Escherichia coli, Stenotrophomonas maltophilia, and basidiomycetous fungi (Schizophyllum commune) in sinuses. Considering the in vitro and in vivo anti-fungal activity of voriconazole, it might be a good option to treat Schizophyllum commune infections when antifungal susceptibility testing is not available. When severe side effects occur, amphotericin B or itraconazole might be subsequent choice.

  6. Nosocomial Infections with IMP-19−Producing Pseudomonas aeruginosa Linked to Contaminated Sinks, France

    PubMed Central

    Amoureux, Lucie; Riedweg, Karena; Chapuis, Angélique; Bador, Julien; Siebor, Eliane; Péchinot, André; Chrétien, Marie-Lorraine; de Curraize, Claire

    2017-01-01

    We isolated IMP-19–producing Pseudomonas aeruginosa from 7 patients with nosocomial infections linked to contaminated sinks in France. We showed that blaIMP-19 was located on various class 1 integrons among 8 species of gram-negative bacilli detected in sinks: P. aeruginosa, Achromobacter xylosoxidans, A. aegrifaciens, P. putida, Stenotrophomonas maltophilia, P. mendocina, Comamonas testosteroni, and Sphingomonas sp. PMID:28098548

  7. Transcriptome profiling of heat-resistant strain Bacillus licheniformis CGMCC3962 producing Maotai flavor.

    PubMed

    Wu, Qun; Xu, Yan

    2012-02-29

    Although Maotai flavor liquor is exclusive due to its soy sauce flavor, knowledge of its key compound and production mechanism is still scarce until now. To gain insight into the production mechanism of soy sauce flavor, a soy sauce flavor producing strain with high efficiency and heat-resistant capability was obtained, and the metabolic mechanism of the strain was investigated with the technique of microarray profiling. Because high temperature was a key factor for soy sauce flavor production, the global gene expression of this heat-resistant strain fermented at 55 °C was analyzed. Except for the responsive increase of heat shock proteins, which maintained cell survival during heat stress, biosynthesis of cysteine was also up-regulated. In addition, some metabolites were significantly increased when cysteine was added to the fermentation medium, such as 2,3-butanediol, 3-hydroxy-2-butanone, and tetramethylpyrazine, which were important flavor compounds in soy sauce flavor liquor and might be related with soy sauce flavor. The results indicated that cysteine might play an important role in the formation of soy sauce flavor compound, and it might act as an indirect precursor or stimulator of soy sauce flavor formation. This was the first use of the microarray profiling tool to investigate the fermentative strains for Chinese traditional liquor, which would allow a deeper insight into the mechanism of the formation of soy sauce flavor compound.

  8. [Purification and characterization of a novel alpha-galactosidase from penicillium sp. F63 CGMCC1669].

    PubMed

    Mi, Shi-jun; Bai, Ying-guo; Meng, Kun; Wang, Ya-ru; Yao, Bin; Shi, Xiu-yun; Huang, Huo-qing; Zhang, Yu-hong; Shi, Peng-jun

    2007-02-01

    An a-galactosidase-producing fungus was screened out of 26 filamentous fungi isolated from soil by us. Phylogenetic analysis based on the alignment of 18S rDNA sequences, combined with the morphological identification, indicated that the strain F63 was a member of the genus Penicillium. The a-galactosidase from Penicillium sp. F63 was purified to apparent homogeneity by ammonium sulfate precipitation, ion-exchange and gel filtration chromatography. The molecular size of the purified enzyme is approximately 82kDa estimated by SDS-PAGE. The a-galactosidase has an optimum pH of 5.0 and an optimum temperature of 45 degrees C. The enzyme is stable between pH5.0 and 6.0 below 40 degrees C. The a-galactosidase activity is slightly inhibited by Ag+ , which is dissimilar to other a-galactosidases. Kinetic studies of the a-galactosidase showed that the Km and the Vmax for pNPG are 1.4mmol/L and 1.556mmol/L. min(-1) x mg- 1, respectively. The enzyme is able to degrade natural substrates such as melibiose, raffinose and stachyose but not galactose-containing polysaccharides. The alpha-galactosidase was identified by MALDI-TOF-MS and its inner peptides were sequenced by ESI-MS/MS. The results show that the a-galactosidase is a novel one.

  9. Understanding the influence of Tween 80 on pullulan fermentation by Aureobasidium pullulans CGMCC1234.

    PubMed

    Sheng, Long; Tang, Guiyue; Su, Peng; Zhang, Jinling; Xiao, Qian; Tong, Qunyi; Ma, Meihu

    2016-01-20

    In this paper, several new perspectives concerned with the effect of Tween 80 promoting pullulan production were presented. With the presence of Tween 80, the maximum pullulan yield increased by 41% (53.04 g/L). Meanwhile, the carbon source was consumed faster and the broth viscosity was higher. The lower final pH suggested that Tween 80 could protect the integrity of the mycelia. The dispersed filaments were not easily entangled with each other and less pellets were formed in the Tween 80 culture broth. FT-IR spectrum analysis indicated that the evaluated sample structure was coincided with commercial pullulan. The molecular weight of sample significantly dropped comparing with the control. The above findings indicated that Tween 80 facilitated the uptake of nutrient from surroundings to the microorganism and the release of pullulan into the extracellular fluid. These results were useful in better understanding the regulation and optimization of efficient pullulan fermentation.

  10. Isolation and Characterization of α-Endosulfan Degrading Bacteria from the Microflora of Cockroaches.

    PubMed

    Ozdal, Murat; Ozdal, Ozlem Gur; Alguri, Omer Faruk

    2016-01-01

    Extensive applications of organochlorine pesticides like endosulfan have led to the contamination of soil and environments. Five different bacteria were isolated from cockroaches living in pesticide contaminated environments. According to morphological, physiological, biochemical properties, and total cellular fatty acid profile by Fatty Acid Methyl Esters (FAMEs), the isolates were identified as Pseudomonas aeruginosa G1, Stenotrophomonas maltophilia G2, Bacillus atrophaeus G3, Citrobacter amolonaticus G4 and Acinetobacter lwoffii G5. This is the first study on the bacterial flora of Blatta orientalis evaluated for the biodegradation of α-endosulfan. After 10 days of incubation, the biodegradation yields obtained from P. aeruginosa G1, S. maltophilia G2, B. atrophaeus G3, C. amolonaticus G4 and A. lwoffii G5 were 88.5% , 85.5%, 64.4%, 56.7% and 80.2%, respectively. As a result, these bacterial strains may be utilized for biodegradation of endosulfan polluted soil and environments.

  11. [Pathogenic bacteria in cystic fibrosis].

    PubMed

    Mariani-Kurkdjian, P; Bingen, E

    2003-09-01

    Since the CF gene identification in 1989 and despite the improvement of our knowledge in the physiopathology of the disease, bronchopulmonary infection determines the vital prognosis. Following Staphylococcus aureus infection, patients are colonized or colonized by Pseudomonas aeruginosa, greatly involved in the pulmonary deterioration. Other bacteria may be involved Burkholderia cepacia, Stenotrophomonas maltophilia, Alcaligenes sp. Intensive antibiotic treatment of primocolonisation helps to prevent or delay chronic colonisation. Chronic colonization needs a rational long term antibiotic strategy to prevent the occurrence of multiresistant germs; antibiotic cures are performed every 3 or 4 months before pulmonary exacerbation symptoms.

  12. Mutualistic growth of the sulfate-reducer Desulfovibrio vulgaris Hildenborough with different carbohydrates.

    PubMed

    Santana, M M; Portillo, M C; Gonzalez, J M

    2012-01-01

    Desulfovibrio vulgaris Hildenborough genome presents a phosphotransferase system putatively involved in the transport of carbohydrates. However, utilization of sugars by this sulfate-reducing bacterium has never been reported. Herein, we have observed proliferation of D. vulgaris Hildenborough with some carbohydrates, in mutualism with Stenotrophomonas maltophilia, a non-fermentative, gram-negative gammaproteobacterium, or Microbacterium, a gram-positive actinobacterium. These results suggest the importance of feedback interactions between different heterotrophic bacterial species including the alternative for D. vulgaris of exploiting additional organic resources and novel habitats. Thus, D. vulgaris strongly participates in the mineralization of carbohydrates both in complex natural and artificial systems.

  13. Q-PCR based bioburden assessment of drinking water throughout treatment and delivery to the International Space Station

    NASA Technical Reports Server (NTRS)

    Newcombe, David; Stuecker, Tara; La Duc, Myron; Venkateswaran, Kasthuri

    2005-01-01

    Previous studies indicated evidence of opportunistic pathogens samples obtained during missions to the International Space Station (ISS). This study utilized TaqMan quantitative PCR to determine specific gene abundance in potable and non-potable ISS waters. Probe and primer sets specific to the small subunit rRNA genes were used to elucidate overall bacterial rRNA gene numbers. while those specific for Burkholderia cepacia and Stenotrophomonas maltophilia were optimized and used to probe for the presence of these two opportunistic pathogens. This research builds upon previous microbial diversity studies of ISS water and demonstrates the utility of Q-PCR tool to examine water quality.

  14. Bacterial strains isolated from PCB-contaminated sediments and their use for bioaugmentation strategy in microcosms.

    PubMed

    Dudášová, Hana; Lukáčová, Lucia; Murínová, Slavomíra; Puškárová, Andrea; Pangallo, Domenico; Dercová, Katarína

    2014-04-01

    This study was focused on the characterization of 15 bacterial strains isolated from long-term PCB-contaminated sediment located at the Strážsky canal in eastern part of Slovakia, in the surroundings of a former PCB producer. PCB-degrading strains were isolated and identified as Microbacterium oleivorans, Stenotrophomonas maltophilia, Brevibacterium sp., Ochrobactrum anthropi, Pseudomonas mandelii, Rhodococcus sp., Achromobacter xylosoxidans, Stenotrophomonas sp., Ochrobactrum sp., Pseudomonas aeruginosa, and Starkeya novella by the 16S rRNA gene sequence phylogenetic analysis. This study presents a newly isolated bacterial strain S. novella with PCB-degrading ability in liquid medium as well as in sediment. For A. xylosoxidans, the bphA gene was identified. The best growth ability in the presence of all sole carbon sources (biphenyl and PCBs vapor) was obtained for Ochrobactrum sp. and Rhodococcus sp. Uncultured Achromobacter sp. showed the highest potential for bioaugmentation of PCB-contaminated sediment.

  15. Isolation and characterization of alkane degrading bacteria from petroleum reservoir waste water in Iran (Kerman and Tehran provenances).

    PubMed

    Hassanshahian, Mehdi; Ahmadinejad, Mohammad; Tebyanian, Hamid; Kariminik, Ashraf

    2013-08-15

    Petroleum products spill and leakage have become two major environmental challenges in Iran. Sampling was performed in the petroleum reservoir waste water of Tehran and Kerman Provinces of Iran. Alkane degrading bacteria were isolated by enrichment in a Bushnel-Hass medium, with hexadecane as sole source of carbon and energy. The isolated strains were identified by amplification of 16S rDNA gene and sequencing. Specific primers were used for identification of alkane hydroxylase gene. Fifteen alkane degrading bacteria were isolated and 8 strains were selected as powerful degradative bacteria. These 8 strains relate to Rhodococcus jostii, Stenotrophomonas maltophilia, Achromobacter piechaudii, Tsukamurella tyrosinosolvens, Pseudomonas fluorescens, Rhodococcus erythropolis, Stenotrophomonas maltophilia, Pseudomonas aeruginosa genera. The optimum concentration of hexadecane that allowed high growth was 2.5%. Gas chromatography results show that all strains can degrade approximately half of hexadecane in one week of incubation. All of the strains have alkane hydroxylase gene which are important for biodegradation. As a result, this study indicates that there is a high diversity of degradative bacteria in petroleum reservoir waste water in Iran.

  16. Evaluation of colistin susceptibility in multidrug-resistant clinical isolates from cystic fibrosis, France.

    PubMed

    Biswas, S; Dubus, J-C; Reynaud-Gaubert, M; Stremler, N; Rolain, J-M

    2013-11-01

    The emergence of multidrug-resistant (MDR) bacteria in cystic fibrosis (CF) patients has led to the use of colistin drug and the emergence of colistin-resistant Gram-negative bacteria. The aim of this study was to compare the disk diffusion and Etest methods for colistin susceptibility testing on MDR bacteria associated with CF from Marseille, France. Forty-nine MDR clinical isolates (27 Stenotrophomonas maltophilia, 22 Achromobacter xylosoxidans) were used in this study. Disk diffusion and Etest assays were used to assess the reliability of these two techniques. For S. maltophilia, 25 out of 27 isolates had low minimum inhibitory concentrations (MICs, 0.125-0.75 mg/L), whereas two isolates displayed high MICs (32 mg/L). Similarly, 19 out of 22 A. xylosoxidans isolates had low MICs (0.75-3.0 mg/L), whereas three isolates had high MICs (32-256 mg/L). The diameters of zone inhibition with a 50-μg colistin disk displayed a good correlation with the MICs obtained by the Etest. Susceptible and resistant strains were eventually separated using a disk diffusion assay at a cut-off of ≤ 12 mm for a 50-μg disk. Colistin displayed excellent activity against S. maltophilia and A. xylosoxidans and the disk diffusion assay could be confidently used to determine the susceptibility to colistin for MDR Gram-negative bacteria in the context of CF.

  17. Insights into Cystic Fibrosis Polymicrobial Consortia: The Role of Species Interactions in Biofilm Development, Phenotype, and Response to In-Use Antibiotics.

    PubMed

    Magalhães, Andreia P; Lopes, Susana P; Pereira, Maria O

    2016-01-01

    Cystic Fibrosis (CF) airways disease involves complex polymicrobial infections where different bacterial species can interact and influence each other and/or even interfere with the whole community. To gain insights into the role that interactions between Pseudomonas aeruginosa in co-culture with Staphylococcus aureus, Inquilinus limosus, and Stenotrophomonas maltophilia may play in infection, the reciprocal effect during biofilm formation and the response of dual biofilms toward ciprofloxacin under in vitro atmospheres with different oxygen availabilities were evaluated. Biofilm formation kinetics showed that the growth of S. aureus, I. limosus, and S. maltophilia was disturbed in the presence of P. aeruginosa, under both aerobic and anaerobic environments. On the other hand, under aerobic conditions, I. limosus led to a decrease in biofilm mass production by P. aeruginosa, although biofilm-cells viability remains unaltered. The interaction between S. maltophilia and P. aeruginosa positively influenced dual biofilm development by increasing its biomass. Compared with monocultures, biomass of P. aeruginosa+ S. aureus biofilms was significantly reduced by reciprocal interference. When grown in dual biofilms with P. aeruginosa, ciprofloxacin was less effective against S. aureus, I. limosus, and S. maltophilia, with increasing antibiotic doses leading to drastic inhibitions of P. aeruginosa cultivability. Therefore, P. aeruginosa might be responsible for the protection of the whole dual consortia against ciprofloxacin activity. Based on the overall data, it can be speculated that reciprocal interferences occur between the different bacterial species in CF lung, regardless the level of oxygen. The findings also suggest that alterations of bacterial behavior due to species interplay may be important for disease progression in CF infection.

  18. Nontuberculous Mycobacteria, Fungi, and Opportunistic Pathogens in Unchlorinated Drinking Water in the Netherlands

    PubMed Central

    van der Kooij, Dick

    2013-01-01

    The multiplication of opportunistic pathogens in drinking water supplies might pose a threat to public health. In this study, distributed unchlorinated drinking water from eight treatment plants in the Netherlands was sampled and analyzed for fungi, nontuberculous mycobacteria (NTM), and several opportunistic pathogens by using selective quantitative PCR methods. Fungi and NTM were detected in all drinking water samples, whereas Legionella pneumophila, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Aspergillus fumigatus were sporadically observed. Mycobacterium avium complex and Acanthamoeba spp. were not detected. Season had no influence on the occurrence of these organisms, except for NTM and S. maltophilia, which were present in higher numbers in the summer. Opportunistic pathogens were more often observed in premise plumbing water samples than in samples from the distribution system. The lowest number of these organisms was observed in the finished water at the plant. Thus, fungi, NTM, and some of the studied opportunistic pathogens can multiply in the distribution and premise plumbing systems. Assimilable organic carbon (AOC) and/or total organic carbon (TOC) had no clear effects on fungal and NTM numbers or on P. aeruginosa- and S. maltophilia-positive samples. However, L. pneumophila was detected more often in water with AOC concentrations above 10 μg C liter−1 than in water with AOC levels below 5 μg C liter−1. Finally, samples that contained L. pneumophila, P. aeruginosa, or S. maltophilia were more frequently positive for a second opportunistic pathogen, which shows that certain drinking water types and/or sampling locations promote the growth of multiple opportunistic pathogens. PMID:23160134

  19. Nontuberculous mycobacteria, fungi, and opportunistic pathogens in unchlorinated drinking water in The Netherlands.

    PubMed

    van der Wielen, Paul W J J; van der Kooij, Dick

    2013-02-01

    The multiplication of opportunistic pathogens in drinking water supplies might pose a threat to public health. In this study, distributed unchlorinated drinking water from eight treatment plants in the Netherlands was sampled and analyzed for fungi, nontuberculous mycobacteria (NTM), and several opportunistic pathogens by using selective quantitative PCR methods. Fungi and NTM were detected in all drinking water samples, whereas Legionella pneumophila, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Aspergillus fumigatus were sporadically observed. Mycobacterium avium complex and Acanthamoeba spp. were not detected. Season had no influence on the occurrence of these organisms, except for NTM and S. maltophilia, which were present in higher numbers in the summer. Opportunistic pathogens were more often observed in premise plumbing water samples than in samples from the distribution system. The lowest number of these organisms was observed in the finished water at the plant. Thus, fungi, NTM, and some of the studied opportunistic pathogens can multiply in the distribution and premise plumbing systems. Assimilable organic carbon (AOC) and/or total organic carbon (TOC) had no clear effects on fungal and NTM numbers or on P. aeruginosa- and S. maltophilia-positive samples. However, L. pneumophila was detected more often in water with AOC concentrations above 10 μg C liter(-1) than in water with AOC levels below 5 μg C liter(-1). Finally, samples that contained L. pneumophila, P. aeruginosa, or S. maltophilia were more frequently positive for a second opportunistic pathogen, which shows that certain drinking water types and/or sampling locations promote the growth of multiple opportunistic pathogens.

  20. Insights into Cystic Fibrosis Polymicrobial Consortia: The Role of Species Interactions in Biofilm Development, Phenotype, and Response to In-Use Antibiotics

    PubMed Central

    Magalhães, Andreia P.; Lopes, Susana P.; Pereira, Maria O.

    2017-01-01

    Cystic Fibrosis (CF) airways disease involves complex polymicrobial infections where different bacterial species can interact and influence each other and/or even interfere with the whole community. To gain insights into the role that interactions between Pseudomonas aeruginosa in co-culture with Staphylococcus aureus, Inquilinus limosus, and Stenotrophomonas maltophilia may play in infection, the reciprocal effect during biofilm formation and the response of dual biofilms toward ciprofloxacin under in vitro atmospheres with different oxygen availabilities were evaluated. Biofilm formation kinetics showed that the growth of S. aureus, I. limosus, and S. maltophilia was disturbed in the presence of P. aeruginosa, under both aerobic and anaerobic environments. On the other hand, under aerobic conditions, I. limosus led to a decrease in biofilm mass production by P. aeruginosa, although biofilm-cells viability remains unaltered. The interaction between S. maltophilia and P. aeruginosa positively influenced dual biofilm development by increasing its biomass. Compared with monocultures, biomass of P. aeruginosa+ S. aureus biofilms was significantly reduced by reciprocal interference. When grown in dual biofilms with P. aeruginosa, ciprofloxacin was less effective against S. aureus, I. limosus, and S. maltophilia, with increasing antibiotic doses leading to drastic inhibitions of P. aeruginosa cultivability. Therefore, P. aeruginosa might be responsible for the protection of the whole dual consortia against ciprofloxacin activity. Based on the overall data, it can be speculated that reciprocal interferences occur between the different bacterial species in CF lung, regardless the level of oxygen. The findings also suggest that alterations of bacterial behavior due to species interplay may be important for disease progression in CF infection. PMID:28133457

  1. Effect of bacteria on survival and growth of Acanthamoeba castellanii.

    PubMed

    Wang, X; Ahearn, D G

    1997-04-01

    The growth and survival of Acanthamoeba castellanii in the presence of Gram-negative bacteria such as Pseudomonas aeruginosa, Escherichia coli, Serratia marcescens, and Stenotrophomonas maltophilia varied with the densities and species of bacteria. All species of bacteria suspended in a buffered saline at densities of 10(5) to 10(6)/ml supported the growth and survival of 10(6)/ml trophozoites of Acanthamoeba castellanii in a buffered saline solution. At densities of bacteria to amoebae of 100:1 or greater, growth and survival of A. castellanii were suppressed, particularly by P. aeruginosa. In an enrichment medium, the rapid growth of most co-inoculated bacteria inhibited the growth and survival of the amoeba.

  2. Capillary isoelectric focusing and fluorometric detection of proteins and microorganisms dynamically modified by poly(ethylene glycol) pyrenebutanoate.

    PubMed

    Horka, Marie; Ruzicka, Filip; Horký, Jaroslav; Holá, Veronika; Slais, Karel

    2006-12-15

    The nonionogenic pyrene-based tenside, poly(ethylene glycol) pyrenebutanoate, was prepared and applied in capillary isoelectric focusing with fluorometric detection. This dye was used here as a buffer additive in capillary isoelectric focusing for a dynamic modification of the sample of proteins and microorganisms. The values of the isoelectric points of the labeled bioanalytes were calculated with use of the fluorescent pI markers and were found comparable with pI of the native compounds. The mixed cultures of proteins and microorganisms, Escherichia coli CCM 3954, Staphylococcus epidermidis CCM 4418, Proteus vulgaris, Enterococcus faecalis CCM 4224, and Stenotrophomonas maltophilia, the strains of the yeast cells, Candida albicans CCM 8180, Candida krusei, Candida parapsilosis, Candida glabrata, Candida tropicalis, and Saccharomyces cerevisiae were reproducibly focused and separated by the suggested technique. Using UV excitation for the on-column fluorometric detection, the minimum detectable amount was down to 10 cells injected on the separation capillary.

  3. Microorganisms Isolated from Blood Cultures of Febrile Neutropenic Patients in ‹bn-i Sina Hospital.

    PubMed

    Arıkan Akan, Özay

    2003-12-05

    Patients with profound neutropenia have increased risk of septicemia associated with significant morbidity. To provide the appropriate broad-spectrum antimicrobial cover, documentation of causative agents and their antimicrobial susceptibilities should be established in each hospital. During 2001 in Ibn-i Sina Hospital Hematology unit, among 125 isolates from blood cultures of febrile neutropenic patients, gram-negative bacteria was prevalent (56.8%). Among the gram-positives (34.4% of isolates) coagulase-negative staphylococci (CNS) were the predominant bacteria (15/43) followed by Staphylococcus aureus (12/43). Escherichia coli (23/71) and Klebsiella spp. (15/71) were the most common species among 71 gram-negative bacteria. Nonfermentative gram-negative bacilli were 21.6% of the isolates. Increase in the isolation rate of Acinetobacter baumannii (7 strains) and Stenotrophomonas maltophilia (6 strains) was noticed.

  4. Inhibitory Activity and Chemical Characterization of Daucus carota subsp. maximus Essential Oils.

    PubMed

    Gaglio, Raimondo; Barbera, Marcella; Aleo, Aurora; Lommatzsch, Ines; Mantia, Tommaso La; Settanni, Luca

    2017-02-07

    The essential oils (EOs) of green seeds from Daucus carota subsp. maximus growing wild in Pantelleria island (Sicily, Italy) were characterised. EOs were extracted by steam distillation, examined for their inhibitory properties against food-borne Gram positive and Gram negative bacteria and analysed for the chemical composition by gas chromatography (GC) and mass spectrometry (MS). Undiluted EOs showed a large inhibition spectrum against Gram positive strains and also vs Acinetobacter spp. and Stenotrophomonas maltophilia. The minimum inhibition concentration (MIC) was in the range 1.25 - 2.50 μl/ml for the most sensitive strains. The chemical analysis indicated that D. carota subsp. maximus EOs included 34 compounds (5 monoterpene hydrocarbons, 6 oxygenated monoterpenes, 14 sesquiterpene hydrocarbons, 4 oxygenated sesquiterpenes, camphorene and 4 other compounds), accounting for 95.48% of the total oil, and that the major chemicals were carotol, β-bisabolene and isoelemicin. This article is protected by copyright. All rights reserved.

  5. An Evaluation of Microbial and Chemical Contamination Sources Related to the Deterioration of Tap Water Quality in the Household Water Supply System

    PubMed Central

    Lee, Yoonjin

    2013-01-01

    The predominant microorganisms in samples taken from shower heads in residences in the Korean city “N” were Stenotrophomonas maltophilia, Sphingomonas paucimobilis, Acidovorax temperans, and Microbacterium lacticum. Legionella was not detected in this case. The volatile organic compounds (VOCs) vinylacetate, NN-DMA, cis-1,2-dichloroethylene, epichlorohydrin, and styrene were measured in five types of plastic pipes: PVC, PB, PP, PE, and cPVC. The rate of multiplication of the heterotrophic plate count (HPC) attached on the copper pipe in contact with hot tap water was higher than the rate for the copper pipe in contact with cold tap water. Biofilm accumulation on stainless steel pipes with added acetate (3 mg/L) was 2.56 times higher than the non-supplemented condition. Therefore, the growth of HPC in the pipe system was affected by the type and availability of nutrients and depended on variables such as heating during the hot water supply. PMID:24018837

  6. Dynamics of Seed-Borne Rice Endophytes on Early Plant Growth Stages

    PubMed Central

    Hardoim, Pablo R.; Hardoim, Cristiane C. P.; van Overbeek, Leonard S.; van Elsas, Jan Dirk

    2012-01-01

    Bacterial endophytes are ubiquitous to virtually all terrestrial plants. With the increasing appreciation of studies that unravel the mutualistic interactions between plant and microbes, we increasingly value the beneficial functions of endophytes that improve plant growth and development. However, still little is known on the source of established endophytes as well as on how plants select specific microbial communities to establish associations. Here, we used cultivation-dependent and -independent approaches to assess the endophytic bacterrial community of surface-sterilized rice seeds, encompassing two consecutive rice generations. We isolated members of nine bacterial genera. In particular, organisms affiliated with Stenotrophomonas maltophilia and Ochrobactrum spp. were isolated from both seed generations. PCR-based denaturing gradient gel electrophoresis (PCR-DGGE) of seed-extracted DNA revealed that approximately 45% of the bacterial community from the first seed generation was found in the second generation as well. In addition, we set up a greenhouse experiment to investigate abiotic and biotic factors influencing the endophytic bacterial community structure. PCR-DGGE profiles performed with DNA extracted from different plant parts showed that soil type is a major effector of the bacterial endophytes. Rice plants cultivated in neutral-pH soil favoured the growth of seed-borne Pseudomonas oryzihabitans and Rhizobium radiobacter, whereas Enterobacter-like and Dyella ginsengisoli were dominant in plants cultivated in low-pH soil. The seed-borne Stenotrophomonas maltophilia was the only conspicuous bacterial endophyte found in plants cultivated in both soils. Several members of the endophytic community originating from seeds were observed in the rhizosphere and surrounding soils. Their impact on the soil community is further discussed. PMID:22363438

  7. Transformation and Precipitation of Toxic Metals by ’Pseudomonas maltophilia

    DTIC Science & Technology

    1991-05-31

    Jackson has used plasmid DNA derived from strain 0-2 to transform various metal-resistance phenotypes into a recipient strain of E. coli. He has...PAGE 1 Of AUTRACT’ UNCLASSTJT UNCLASSIFIED UN CLAS SIF IED UL I4N7d.l~0SO Stanbato @Form 29S ( Rev . 24E91 A. RESEARCH OBJECTIVES The aims of this project...Barton and Thomas Zocco at Iew Mexico State University and Los Alamos National Laboratory, respectively. Examination by SEM of cultures of 0-2 grown in the

  8. Microbial contamination of embryos and semen during long term banking in liquid nitrogen.

    PubMed

    Bielanski, A; Bergeron, H; Lau, P C K; Devenish, J

    2003-04-01

    We report on microbial contamination of embryos and semen cryopreserved in sealed plastic straws and stored for 6-35 years in liquid nitrogen. There were 32 bacterial and 1 fungal species identified from randomly drawn liquid nitrogen, frozen semen, and embryos samples stored in 8 commercial and 8 research facility liquid nitrogen (LN) tanks. The identified bacteria represented commensal or environmental microorganisms and some, such as Escherichia coli, were potential or opportunistic pathogens for humans and animals. Stenotrophomonas maltophilia was the most common contaminant identified from the samples and was further shown to significantly suppress fertilization and embryonic development in vitro. Analysis of the strains by pulsed field gel electrophoresis revealed restriction patterns with no relatedness indicating that there was no apparent cross-contamination of S. maltophilia strains between the germplasm and liquid nitrogen samples. In addition, no transmission of bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1) from infected semen and embryos straws to clean germplasm stored in the same LN tanks or LN was detected.

  9. Complementary treatment of contact lens-induced corneal ulcer using honey: a case report.

    PubMed

    Majtanova, Nora; Vodrazkova, Erika; Kurilova, Veronika; Horniackova, Miroslava; Cernak, Martin; Cernak, Andrej; Majtan, Juraj

    2015-02-01

    The aim of this study was to report the complementary use of honey for treatment of a contact lens-induced corneal ulcer. A 23-year-old contact lens user presented with a corneal ulcer in her left eye. She had visual acuity reduced to hand movement. There was a history of wearing contact lenses while swimming in a lake seven days before presentation. The cultures from corneal scrapings and contact lenses were positive for Klebsiella oxytoca, Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Pseudomonas spp. The treatment with topical levofloxacin and 25% (w/v) γ-irradiated honeydew honey solution was effective and the patient achieved final best corrected visual acuity of affected eye. In addition to positive clinical outcome, honeydew honey was shown to be highly effective in vitro against ocular isolates, in particular S. maltophilia. The minimal inhibitory concentrations for honeydew honey ranged from 5% to 10%. These results demonstrate that honey is a promising antibacterial agent in management of corneal ulcers. Moreover, honey exhibits anti-biofilm and anti-inflammatory properties, and thus becomes an interesting ophthalmologic agent.

  10. Effects of pathology dyes on Raman bone spectra

    NASA Astrophysics Data System (ADS)

    Esmonde-White, Karen A.; Esmonde-White, Francis W. L.; Morris, Michael D.; Roessler, Blake J.

    2013-05-01

    We report an overlooked source of artifacts for clinical specimens, where unexpected and normally negligible contaminants can skew the interpretation of results. During an ongoing study of bone fragments from diabetic osteomyelitis, strong Raman signatures were found, which did not correspond with normal bone mineral or matrix. In a bone biopsy from the calcaneus of a patient affected by diabetic osteomyelitis, Raman microspectroscopic analysis revealed regions with both abnormal mineral and degraded collagen in addition to normal bone. Additional bands indicated a pathological material. Stenotrophomonas maltophilia was identified in the wound culture by independent microbiologic examination. We initially assigned the unusual bands to xanthomonadin, a bacterial pigment from S. maltophilia. However, the same bands were also found more than a year later on a second specimen that had been noticeably contaminated with pathology marking dye. Drop deposition/Raman spectroscopy of commonly used pathology dyes revealed that a blue tissue-marking dye was responsible for the unusual bands in both specimens, even in the first specimen where there was no visible evidence of contamination.

  11. Microbial contamination of suction tubes attached to suction instruments and preventive methods.

    PubMed

    Yorioka, Katsuhiro; Oie, Shigeharu; Kamiya, Akira

    2010-03-01

    We investigated the microbial contamination of suction tubes attached to wall-type suction instruments. Microbial contamination of suction tubes used for endoscopy or sputum suction in hospital wards was examined before and after their disinfection. In addition, disinfection and washing methods for suction tubes were evaluated. Suction tubes (n=33) before disinfection were contaminated with 10(2)-10(8) colony-forming units (cfu)/tube. The main contaminants were Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia. The suction tubes were disinfected with sodium hypochlorite (n=11) or hot water (n=11), or by an automatic tube cleaner (n=11). After 2-h immersion in 0.1% (1,000 ppm) sodium hypochlorite, 10(3)-10(7) cfu/tube of bacteria were detected in all 11 tubes examined. After washing in hot running water (65 degrees C), 10(3)-10(7) cfu/tube were detected in 3 of the 11 examined tubes. The bacteria detected in the suction tubes after disinfection with sodium hypochlorite or hot water were P. aeruginosa, A. baumannii, and S. maltophilia. On the other hand, after washing with warm water (40 degrees C) using the automatic tube cleaner, contamination was found to be <20 cfu/tube (lower detection limit, 20 cfu/tube) in all 11 tubes examined. These results suggest the usefulness of washing with automatic tube cleaners.

  12. Bacteria associated with Amblyomma cajennense tick eggs

    PubMed Central

    Machado-Ferreira, Erik; Vizzoni, Vinicius Figueiredo; Piesman, Joseph; Gazeta, Gilberto Salles; Soares, Carlos Augusto Gomes

    2015-01-01

    Abstract Ticks represent a large group of pathogen vectors that blood feed on a diversity of hosts. In the Americas, the Ixodidae ticks Amblyomma cajennense are responsible for severe impact on livestock and public health. In the present work, we present the isolation and molecular identification of a group of culturable bacteria associated with A. cajennense eggs from females sampled in distinct geographical sites in southeastern Brazil. Additional comparative analysis of the culturable bacteria from Anocentor nitens, Rhipicephalus sanguineus and Ixodes scapularis tick eggs were also performed. 16S rRNA gene sequence analyses identified 17 different bacterial types identified as Serratia marcescens, Stenotrophomonas maltophilia, Pseudomonas fluorescens, Enterobacter spp., Micrococcus luteus, Ochrobactrum anthropi, Bacillus cereus and Staphylococcus spp., distributed in 12 phylogroups. Staphylococcus spp., especially S. sciuri, was the most prevalent bacteria associated with A. cajennense eggs, occurring in 65% of the samples and also frequently observed infecting A. nitens eggs. S. maltophilia, S. marcescens and B. cereus occurred infecting eggs derived from specific sampling sites, but in all cases rising almost as pure cultures from infected A. cajennense eggs. The potential role of these bacterial associations is discussed and they possibly represent new targets for biological control strategies of ticks and tick borne diseases. PMID:26537602

  13. Factors Associated with Worse Lung Function in Cystic Fibrosis Patients with Persistent Staphylococcus aureus

    PubMed Central

    den Reijer, Martijn; Wiedemann, Bärbel; Tümmler, Burkhard; Ellemunter, Helmut; Dübbers, Angelika; Küster, Peter; Ballmann, Manfred; Koerner-Rettberg, Cordula; Große-Onnebrink, Jörg; Heuer, Eberhardt; Sextro, Wolfgang; Mainz, Jochen G.; Hammermann, Jutta; Riethmüller, Joachim; Graepler-Mainka, Ute; Staab, Doris; Wollschläger, Bettina; Szczepanski, Rüdiger; Schuster, Antje; Tegtmeyer, Friedrich-Karl; Sutharsan, Sivagurunathan; Wald, Alexandra; Nofer, Jerzy-Roch; van Wamel, Willem; Becker, Karsten; Peters, Georg

    2016-01-01

    Background Staphylococcus aureus is an important pathogen in cystic fibrosis (CF). However, it is not clear which factors are associated with worse lung function in patients with persistent S. aureus airway cultures. Our main hypothesis was that patients with high S. aureus density in their respiratory specimens would more likely experience worsening of their lung disease than patients with low bacterial loads. Methods Therefore, we conducted an observational prospective longitudinal multi-center study and assessed the association between lung function and S. aureus bacterial density in respiratory samples, co-infection with other CF-pathogens, nasal S. aureus carriage, clinical status, antibiotic therapy, IL-6- and IgG-levels against S. aureus virulence factors. Results 195 patients from 17 centers were followed; each patient had an average of 7 visits. Data were analyzed using descriptive statistics and generalized linear mixed models. Our main hypothesis was only supported for patients providing throat specimens indicating that patients with higher density experienced a steeper lung function decline (p<0.001). Patients with exacerbations (n = 60), S. aureus small-colony variants (SCVs, n = 84) and co-infection with Stenotrophomonas maltophilia (n = 44) had worse lung function (p = 0.0068; p = 0.0011; p = 0.0103). Patients with SCVs were older (p = 0.0066) and more often treated with trimethoprim/sulfamethoxazole (p = 0.0078). IL-6 levels positively correlated with decreased lung function (p<0.001), S. aureus density in sputa (p = 0.0016), SCVs (p = 0.0209), exacerbations (p = 0.0041) and co-infections with S. maltophilia (p = 0.0195) or A. fumigatus (p = 0.0496). Conclusions In CF-patients with chronic S. aureus cultures, independent risk factors for worse lung function are high bacterial density in throat cultures, exacerbations, elevated IL-6 levels, presence of S. aureus SCVs and co-infection with S. maltophilia. Trial Registration ClinicalTrials.gov NCT

  14. Biodiesel Production: Utilization of Loofah Sponge to Immobilize Rhizopus chinensis CGMCC #3.0232 Cells as a Whole-Cell Biocatalyst.

    PubMed

    He, Qiyang; Xia, Qianjun; Wang, Yuejiao; Li, Xun; Zhang, Yu; Hu, Bo; Wang, Fei

    2016-07-28

    Rhizopus chinensis cells immobilized on loofah (Luffa cylindrica) sponges were used to produce biodiesel via the transesterification of soybean oil. In whole-cell immobilization, loofah sponge is considered to be a superior alternative to conventional biomass carriers because of its biodegradable and renewable properties. During cell cultivation, Rhizopus chinensis mycelia can spontaneously and firmly adhere to the surface of loofah sponge particles. The optimal conditions for processing 9.65 g soybean oil at 40°C and 180 rpm using a 3:1 methanol-to-oil molar ratio were found to be 8% cell addition and 3-10% water content (depending on the oil's weight). Under optimal conditions, an over 90% methyl ester yield was achieved after the first reaction batch. The operational stability of immobilized Rhizopus chinensis cells was assayed utilizing a 1:1 methanol-to-oil molar ratio, thus resulting in a 16.5-fold increase in half-life when compared with immobilized cells of the widely studied Rhizopus oryzae. These results suggest that transesterification of vegetable oil using Rhizopus chinensis whole cells immobilized onto loofah sponge is an effective approach for biodiesel production.

  15. Longitudinal Metagenomic Analysis of Hospital Air Identifies Clinically Relevant Microbes

    PubMed Central

    King, Paula; Pham, Long K.; Waltz, Shannon; Sphar, Dan; Yamamoto, Robert T.; Conrad, Douglas; Taplitz, Randy; Torriani, Francesca

    2016-01-01

    We describe the sampling of sixty-three uncultured hospital air samples collected over a six-month period and analysis using shotgun metagenomic sequencing. Our primary goals were to determine the longitudinal metagenomic variability of this environment, identify and characterize genomes of potential pathogens and determine whether they are atypical to the hospital airborne metagenome. Air samples were collected from eight locations which included patient wards, the main lobby and outside. The resulting DNA libraries produced 972 million sequences representing 51 gigabases. Hierarchical clustering of samples by the most abundant 50 microbial orders generated three major nodes which primarily clustered by type of location. Because the indoor locations were longitudinally consistent, episodic relative increases in microbial genomic signatures related to the opportunistic pathogens Aspergillus, Penicillium and Stenotrophomonas were identified as outliers at specific locations. Further analysis of microbial reads specific for Stenotrophomonas maltophilia indicated homology to a sequenced multi-drug resistant clinical strain and we observed broad sequence coverage of resistance genes. We demonstrate that a shotgun metagenomic sequencing approach can be used to characterize the resistance determinants of pathogen genomes that are uncharacteristic for an otherwise consistent hospital air microbial metagenomic profile. PMID:27482891

  16. Longitudinal Metagenomic Analysis of Hospital Air Identifies Clinically Relevant Microbes.

    PubMed

    King, Paula; Pham, Long K; Waltz, Shannon; Sphar, Dan; Yamamoto, Robert T; Conrad, Douglas; Taplitz, Randy; Torriani, Francesca; Forsyth, R Allyn

    2016-01-01

    We describe the sampling of sixty-three uncultured hospital air samples collected over a six-month period and analysis using shotgun metagenomic sequencing. Our primary goals were to determine the longitudinal metagenomic variability of this environment, identify and characterize genomes of potential pathogens and determine whether they are atypical to the hospital airborne metagenome. Air samples were collected from eight locations which included patient wards, the main lobby and outside. The resulting DNA libraries produced 972 million sequences representing 51 gigabases. Hierarchical clustering of samples by the most abundant 50 microbial orders generated three major nodes which primarily clustered by type of location. Because the indoor locations were longitudinally consistent, episodic relative increases in microbial genomic signatures related to the opportunistic pathogens Aspergillus, Penicillium and Stenotrophomonas were identified as outliers at specific locations. Further analysis of microbial reads specific for Stenotrophomonas maltophilia indicated homology to a sequenced multi-drug resistant clinical strain and we observed broad sequence coverage of resistance genes. We demonstrate that a shotgun metagenomic sequencing approach can be used to characterize the resistance determinants of pathogen genomes that are uncharacteristic for an otherwise consistent hospital air microbial metagenomic profile.

  17. Correlation between circuital current, Cu(II) reduction and cellular electron transfer in EAB isolated from Cu(II)-reduced biocathodes of microbial fuel cells.

    PubMed

    Shen, Jingya; Huang, Liping; Zhou, Peng; Quan, Xie; Puma, Gianluca Li

    2017-04-01

    The performance of four indigenous electrochemically active bacteria (EAB) (Stenotrophomonas maltophilia JY1, Citrobacter sp. JY3, Pseudomonas aeruginosa JY5 and Stenotrophomonas sp. JY6) was evaluated for Cu(II) reduction on the cathodes of microbial fuel cells (MFCs). These EAB were isolated from well adapted mixed cultures on the MFC cathodes operated for Cu(II) reduction. The relationship between circuital current, Cu(II) reduction rate, and cellular electron transfer processes was investigated from a mechanistic point of view using X-ray photoelectron spectroscopy, scanning electronic microscopy coupled with energy dispersive X-ray spectrometry, linear sweep voltammetry and cyclic voltammetry. JY1 and JY5 exhibited a weak correlation between circuital current and Cu(II) reduction. A much stronger correlation was observed for JY3 followed by JY6, demonstrating the relationship between circuital current and Cu(II) reduction for these species. In the presence of electron transfer inhibitors (2,4-dinitrophenol or rotenone), significant inhibition on JY6 activity and a weak effect on JY1, JY3 and JY5 was observed, confirming a strong correlation between cellular electron transfer processes and either Cu(II) reduction or circuital current. This study provides evidence of the diverse functions played by these EAB, and adds to a deeper understanding of the capabilities exerted by diverse EAB associated with Cu(II) reduction.

  18. Fluorescent probe based subcellular distribution of Cu(II) ions in living electrotrophs isolated from Cu(II)-reduced biocathodes of microbial fuel cells.

    PubMed

    Tao, Ye; Xue, Hua; Huang, Liping; Zhou, Peng; Yang, Wei; Quan, Xie; Yuan, Jinxiu

    2017-02-01

    Based on the four indigenous electrotrophs (Stenotrophomonas maltophilia JY1, Citrobacter sp. JY3, Pseudomonas aeruginosa JY5 and Stenotrophomonas sp. JY6) isolated from well adapted Cu(II)-reduced biocathodes of microbial fuel cells (MFCs), a rhodamine based Cu(II) fluorescent probe was used to imaginably and quantitatively track subcellular Cu(II) ions in these electrotrophs. Cathodic electrons led to more Cu(II) ions (14.3-30.1%) in the intracellular sites at operation time of 2-3h with Cu(II) removal rates of 2.90-3.64mg/Lh whereas the absence of cathodic electrons prolonged the appearance of more Cu(II) ions (16.6-22.5%) to 5h with Cu(II) removal rates of 1.96-2.28mg/Lh. This study illustrates that cathodic electrons directed more Cu(II) ions for quicker entrance into the electrotrophic cytoplasm, and gives an alternative approach for developing imaging and functionally tracking Cu(II) ions in the electrotrophs of MFCs.

  19. Emergence of Carbapenem Resistant Non-Fermenting Gram-Negative Bacilli Isolated in an ICU of a Tertiary Care Hospital

    PubMed Central

    Agarwal, Sonika; Khanduri, Sushant; Gupta, Shalini

    2017-01-01

    Introduction The emergence and spread of Multi-Drug Resistant (MDR) Non-Fermenting Gram-Negative Bacilli (NFGNB) in Intensive Care Units (ICU) and their genetic potential to transmit diverse antibiotic resistance regardless of their ability to ferment glucose poses a major threat in hospitals. The complex interplay of clonal spread, persistence, transmission of resistance elements and cell-cell interaction leads to the difficulty in controlling infections caused by these multi drug-resistant strains. Among non-fermenting Gram-negative rods, the most clinically significant species Pseudomonas aeruginosa, Acinetobacter baumannii and Stenotrophomonas maltophilia are increasingly acquiring resistant to carbapenems. Carbapenems once considered as a backbone of treatment of life threatening infections appears to be broken as the resistance to carbapenems is on rise. Aim To document the prevalence of carbapenem resistance in non-fermenting Gram-negative bacilli isolated from patients with respiratory tract infections in the ICU of Himalayan Institute of Medical Sciences, Dehradun. Materials and Methods This is a cross-sectional study conducted in ICU patients between October 2015 to March 2016. A total of 366 lower respiratory tract samples were collected from 356 patients with clinical evidence of lower respiratory tract infections in form of Endotracheal (ET) aspirate, Tracheal Tube (TT) aspirate and Bronchoalveolar Lavage (BAL) specimen. Organism identification and the susceptibility testing was done by using an automated system VITEK 2. Results Out of 366 samples received 99 NFGNB were isolated and most common sample was ET aspirate sample 256 (64.5%). Acinetobacter baumannii was the most common NFGNB isolated 63 (63.63%) followed by Pseudomonas aeruginosa 25 (25.25%), Elizabethkingia meningoseptica seven (7.07%) and Strenotrophomonas maltophilia four (4.04%). We observed that 90.5% Acinetobacter baumannii were resistant to imipenem and 95.2% resistant to meropenem

  20. Occurrence and Multiple Antibiotic Resistance Profiles of Non-fermentative Gram-Negative Microflora in Five Brands of Non-carbonated French Bottled Spring Water.

    PubMed

    Mary; Defives; Hornez

    2000-05-01

    Five brands of French bottled mineral water were analyzed by heterotrophic plate counts (HPC) and for the presence of multiple antibiotic resistant bacteria. HPC at 22 degrees C were around 10(4) colony forming units ml(-1) on R2A medium. Enumeration on PCA/10, MH, and especially PCA and King B media was less efficient. At 37 degrees C, HPC were two to three orders of magnitude less than at 22 degrees C. Moreover, phenotypic diversity (7 to 15 phenotypes) was optimal on R2A incubated at 22 degrees C. All isolates were identified as non-fermentative Gram-negative rods and 75% were non-identifiable with the API 20NE system. Stenotrophomonas maltophilia and fluorescent Pseudomonas were isolated on VIA and CFC selective agar media, respectively. Burkholderia cepacia strains were not isolated on BCSA medium. The species S. maltophilia was found in 33%, 28%, and 11% of sample from springs A, D, and E, respectively. Independent of brand, isolates from HPC media were less efficient to achieve confluent growth in 18 h on MH at 30 or 37 degrees C (0 to 40%) than isolates from selective media (28 to 63%). Seventy percent of the total isolates from dominant microflora (1-5 x 10(3) CFU ml(-1) on HPC media) were resistant against two or four antibiotics. The antibiotics concerned were principally aztreonam, ampicillin, and nalidixic acid. The remaining dominant bacteria showed a 6-9 multiple antibiotic resistant (MAR) pattern. All isolates were susceptible to newer antimicrobial agents. Owing to their low nutrient and temperature requirements, these isolates are unlikely to cause concern to public heath. Fifty percent of strains isolated from selective media (non-dominant microflora, 4-40 CFU l(-1)) showed a 10-18 MAR pattern and 33%, identified as S. maltophilia, a 20-27 MAR pattern. However, minocycline was effective against all isolates. Owing to its low concentration, colonization of human intestine by MAR S. maltophilia is unlikely.

  1. Degradation and mineralization of high-molecular-weight polycyclic aromatic hydrocarbons by defined fungal-bacterial cocultures.

    PubMed

    Boonchan, S; Britz, M L; Stanley, G A

    2000-03-01

    This study investigated the biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons (PAHs) in liquid media and soil by bacteria (Stenotrophomonas maltophilia VUN 10,010 and bacterial consortium VUN 10,009) and a fungus (Penicillium janthinellum VUO 10, 201) that were isolated from separate creosote- and manufactured-gas plant-contaminated soils. The bacteria could use pyrene as their sole carbon and energy source in a basal salts medium (BSM) and mineralized significant amounts of benzo[a]pyrene cometabolically when pyrene was also present in BSM. P. janthinellum VUO 10,201 could not utilize any high-molecular-weight PAH as sole carbon and energy source but could partially degrade these if cultured in a nutrient broth. Although small amounts of chrysene, benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene were degraded by axenic cultures of these isolates in BSM containing a single PAH, such conditions did not support significant microbial growth or PAH mineralization. However, significant degradation of, and microbial growth on, pyrene, chrysene, benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene, each as a single PAH in BSM, occurred when P. janthinellum VUO 10,201 and either bacterial consortium VUN 10,009 or S. maltophilia VUN 10,010 were combined in the one culture, i.e., fungal-bacterial cocultures: 25% of the benzo[a]pyrene was mineralized to CO(2) by these cocultures over 49 days, accompanied by transient accumulation and disappearance of intermediates detected by high-pressure liquid chromatography. Inoculation of fungal-bacterial cocultures into PAH-contaminated soil resulted in significantly improved degradation of high-molecular-weight PAHs, benzo[a]pyrene mineralization (53% of added [(14)C]benzo[a]pyrene was recovered as (14)CO(2) in 100 days), and reduction in the mutagenicity of organic soil extracts, compared with the indigenous microbes and soil amended with only axenic inocula.

  2. Metallo-β-Lactamase Producers in Environmental Microbiota: New Molecular Class B Enzyme in Janthinobacterium lividum

    PubMed Central

    Rossolini, Gian Maria; Condemi, Maria Adelaide; Pantanella, Fabrizio; Docquier, Jean-Denis; Amicosante, Gianfranco; Thaller, Maria Cristina

    2001-01-01

    Eleven environmental samples from different sources were screened for the presence of metallo-β-lactamase-producing bacteria by using a selective enrichment medium containing a carbapenem antibiotic and subsequently testing each isolate for production of EDTA-inhibitable carbapenemase activity. A total of 15 metallo-β-lactamase-producing isolates, including 10 Stenotrophomonas maltophilia isolates, 3 Chryseobacterium spp., one Aeromonas hydrophila isolate, and one Janthinobacterium lividum isolate (a species in which production of metallo-β-lactamase activity was not previously reported), were obtained from 8 samples. In the J. lividum isolate, named JAC1, production of metallo-β-lactamase activity was elicited upon exposure to β-lactams. Screening of a JAC1 genomic library for clones showing a reduced imipenem susceptibility led to the isolation of a metallo-β-lactamase determinant encoding a new member (named THIN-B) of the highly divergent subclass B3 lineage of metallo-β-lactamases. THIN-B is most closely related (35.6% identical residues) to the L1 enzyme of S. maltophilia and more distantly related to the FEZ-1 enzyme of Legionella gormanii (27.8% identity) and to the GOB-1 enzyme of Chryseobacterium meningosepticum (24.2% identity). Sequences related to blaTHIN-B, and inducible production of metallo-β-lactamase activity, were also detected in the J. lividum type strain DSM1522. Expression of the blaTHIN-B gene in Escherichia coli resulted in decreased susceptibility to several β-lactams, including penicillins, cephalosporins (including cephamycins and oxyimino cephalosporins), and carbapenems, revealing a broad substrate specificity of the enzyme. The results of this study indicated that metallo-β-lactamase-producing bacteria are widespread in the environment and identified a new molecular class B enzyme in the environmental species J. lividum. PMID:11181369

  3. Specific and functional diversity of endophytic bacteria from pine wood nematode Bursaphelenchus xylophilus with different virulence.

    PubMed

    Wu, Xiao-Qin; Yuan, Wei-Min; Tian, Xiao-Jing; Fan, Ben; Fang, Xin; Ye, Jian-Ren; Ding, Xiao-Lei

    2013-01-01

    Pine wilt disease (PWD) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, is one of the most devastating diseases of Pinus spp. The PWN was therefore listed as one of the most dangerous forest pests in China meriting quarantine. Virulence of the PWN is closely linked with the spread of PWD. However, main factors responsible for the virulence of PWNs are still unclear. Recently epiphytic bacteria carried by PWNs have drawn much attention. But little is known about the relationship between endophytic bacteria and virulence of B. xylophilus. In this research, virulence of ten strains of B. xylophilus from different geographical areas in six provinces of China and four pine species were tested with 2-year-old seedlings of Pinus thunbergii. Endophytic bacteria were isolated from PWNs with different virulence to investigate the relationship between the bacteria and PWN virulence. Meanwhile, the carbon metabolism of endophytic bacteria from highly and low virulent B. xylophilus was analyzed using Biolog plates (ECO). The results indicated that ten strains of PWNs showed a wide range of virulence. Simultaneously, endophytic bacteria were isolated from 90% of the B. xylophilus strains. The dominant endophytic bacteria in the nematodes were identified as species of Stenotrophomonas, Achromobacter, Ewingella, Leifsonia, Rhizobium, and Pseudomonas using molecular and biochemical methods. Moreover, S. maltophilia, and A. xylosoxidans subsp. xylosoxidans were the predominant strains. Most of the strains (80%) from P. massoniana contained either S. maltophilia, A. xylosoxidans, or both species. There was a difference between the abilities of the endophytic bacteria to utilize carbon sources. Endophytic bacteria from highly virulent B. xylophilus had a relatively high utilization rate of carbohydrate and carboxylic acids, while bacteria from low virulent B. xylophilus made better use of amino acids. In conclusion, endophytic bacteria widely exist in B. xylophilus

  4. Ventilator-associated pneumonia and the importance of education of ICU nurses on prevention – Preliminary results

    PubMed Central

    Mogyoródi, Bence; Dunai, Erzsébet; Gál, János; Iványi, Zsolt

    2016-01-01

    Background and aims Ventilator-associated pneumonia (VAP) increases intensive care unit (ICU) length of stay, ICU mortality, the number of ventilator days, and costs. We implemented a VAP bundle and investigated its efficacy on prevention. Materials and methods A prospective observational study was conducted between January 1, 2015 and December 31, 2015 in a 12-bed multidisciplinary ICU. The bundle was implemented on July 02, 2015. Comparative analysis was performed before and after the implementation of the bundle. The compliance of the nurses was also studied. Results The incidence of VAP was 21.5/1,000 ventilator days (95% CI: 14.17–31.10) in the first phase and 12.0/1,000 ventilator days (95% CI: 7.2–19.49) in the second phase. Relative risk reduction was 44% (95% CI: −0.5 to 0.98). Most common bacteria identified during the first phase were Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Staphylococcus aureus; and in the second phase P. aeruginosa, Acinetobacter baumannii, and S. maltophilia were identified. Significant improvement was achieved in the head-of-bed elevation (p = 0.004), oral care (p = 0.01), hand hygiene (p < 0.001), endotracheal suctioning (p = 0.004), and removal of condensate (p = 0.043). Discussion The incidence of VAP showed tendency for reduction. The prevalence of nursing-dependent bacteria decreased and compliance in following prevention methods increased. Conclusion These results underline the importance of education of prevention methods. PMID:28180003

  5. Prevalence of Multidrug-Resistant Bacteria on Fresh Vegetables Collected from Farmers' Markets in Connecticut.

    PubMed

    Karumathil, Deepti Prasad; Yin, Hsin-Bai; Kollanoor-Johny, Anup; Venkitanarayanan, Kumar

    2016-08-01

    This study determined the prevalence of multidrug-resistant (MDR) Acinetobacter baumannii on fresh vegetables collected from farmers' markets in Connecticut. One hundred samples each of fresh carrots, potatoes, and lettuce were sampled and streaked on selective media, namely Leeds Acinetobacter and MDR Acinetobacter agars. All morphologically different colonies from MDR Acinetobacter agar were identified by using Gram staining, biochemical tests, and PCR. In addition, susceptibility of the isolates to 10 antibiotics commonly used in humans, namely imipenem, ceftriaxone, cefepime, minocycline, erythromycin, colistin-sulfate, streptomycin, neomycin, doxycycline, and rifampin was determined by using an antibiotic disk diffusion assay. The results revealed that only two samples of potato and one sample of lettuce yielded A. baumannii. In addition, all carrot samples were found to be negative for the organism. However, several other opportunistic, MDR human pathogens, such as Burkholderia cepacia (1% potatoes, 5% carrots, and none in lettuce), Stenotrophomonas maltophilia (6% potatoes, 2% lettuce, and none in carrots), and Pseudomonas luteola (9% potatoes, 3% carrots, and none in lettuce) were recovered from the vegetables. Antibiotic susceptibility screening of the isolates revealed high resistance rates for the following: ceftriaxone (6 of 6), colistin-sulfate (5 of 6), erythromycin (5 of 6), and streptomycin (4 of 6) in B. cepacia; colistin-sulfate (11 of 11) and imipenem (10 of 11) in P. luteola; colistin-sulfate (8 of 8), ceftriaxone (8 of 8), cefepime (7 of 8), erythromycin (5 of 8), and imipenem (4 of 8) in S. maltophilia; and imipenem (3 of 3), ceftriaxone (3 of 3), erythromycin (3 of 3), and streptomycin (3 of 3) in A. baumannii. The results revealed the presence of MDR bacteria, including human pathogens on fresh produce, thereby highlighting the potential health risk in consumers, especially those with a compromised immune system.

  6. Degradation and mineralization of high-molecular-weight polycyclic aromatic hydrocarbons by defined fungal-bacterial cocultures

    SciTech Connect

    Boonchan, S.; Britz, M.L.; Stanley, G.A.

    2000-03-01

    This study investigated the biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons (PAHs) in liquid media and soil by bacteria (Stenotrophomonas maltophilia VUN 10,010 and bacterial consortium VUN 10,009) and a fungus (Penicillium janthinellum VUO 10,201) that were isolated from separate creosote- and manufactured-gas plant-contaminated soils. The bacteria could use pyrene as their sole carbon and energy source in a basal salts medium (BSM) and mineralized significant amounts of benzo[a]pyrene cometabolically when pyrene was also present in BSM. P. janthinellum VUO 10,201 could not utilize any high-molecular-weight PAH as sole carbon and energy source but could partially degrade these if cultured in a nutrient broth. Although small amounts of chrysene, benz[a]pyrene, and dibenz[a,h]anthracene were degraded by axenic cultures of these isolates in BSM containing a single PAH, such conditions did not support significant microbial growth or PAH mineralization. However, significant degradation of, and microbial growth on, pyrene, chrysene, benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene, each as a single PAH in BSM, occurred when P. janthinellum VUO 10,201 and either bacterial consortium VUN 10,009 or S. maltophilia VUN 10,010 were combined in the one culture, i.e., fungal-bacterial cocultures: 25% of the benzo[a]pyrene was mineralized to CO{sub 2} by these cocultures over 49 days, accompanied by transient accumulation and disappearance of intermediates detected by high-pressure liquid chromatography. Inoculation of fungal-bacterial cocultures into PAH-contaminated soil resulted in significantly improved degradation of high-molecular-weight PAHs, benzo[a]pyrene mineralization, and reduction in the mutagenicity of organic soil extracts, compared with the indigenous microbes and soil amended with only axenic inocula.

  7. Impact of untreated urban waste on the prevalence and antibiotic resistance profiles of human opportunistic pathogens in agricultural soils from Burkina Faso.

    PubMed

    Youenou, Benjamin; Hien, Edmond; Deredjian, Amélie; Brothier, Elisabeth; Favre-Bonté, Sabine; Nazaret, Sylvie

    2016-12-01

    This study examined the long-term effects of the landfill disposal of untreated urban waste for soil fertilization on the prevalence and antibiotic resistance profiles of various human opportunistic pathogens in soils from Burkina Faso. Samples were collected at three sites in the periphery of Ouagadougou during two campaigns in 2008 and 2011. At each site, amendment led to changes in physico-chemical characteristics as shown by the increase in pH, CEC, total C, total N, and metal contents. Similarly, the numbers of total heterotrophic bacteria were higher in the amended fields than in the control ones. No sanitation indicators, i.e., coliforms, Staphylococci, and Enterococci, were detected. Pseudomonas aeruginosa and Burkholderia cepacia complex (Bcc) were detected at a low level in one amended field. Stenotrophomonas maltophilia was detected from both campaigns at the three sites in the amended fields and only once in an unamended field. Diversity analysis showed some opportunistic pathogen isolates to be closely related to reference clinical strains responsible for nosocomial- or community-acquired infections in Northern countries. Antibiotic resistance tests showed that P. aeruginosa and Bcc isolates had a wild-type phenotype and that most S. maltophilia isolates had a multi-drug resistance profile with resistance to 7 to 15 antibiotics. Then we were able to show that amendment led to an increase of some human opportunistic pathogens including multi-drug resistant isolates. Although the application of untreated urban waste increases both soil organic matter content and therefore soil fertility, the consequences of this practice on human health should be considered.

  8. Low Prevalence of Carbapenem-Resistant Bacteria in River Water: Resistance Is Mostly Related to Intrinsic Mechanisms.

    PubMed

    Tacão, Marta; Correia, António; Henriques, Isabel S

    2015-10-01

    Carbapenems are last-resort antibiotics to handle serious infections caused by multiresistant bacteria. The incidence of resistance to these antibiotics has been increasing and new resistance mechanisms have emerged. The dissemination of carbapenem resistance in the environment has been overlooked. The main goal of this research was to assess the prevalence and diversity of carbapenem-resistant bacteria in riverine ecosystems. The presence of frequently reported carbapenemase-encoding genes was inspected. The proportion of imipenem-resistant bacteria was on average 2.24 CFU/ml. Imipenem-resistant strains (n=110) were identified as Pseudomonas spp., Stenotrophomonas maltophilia, Aeromonas spp., Chromobacterium haemolyticum, Shewanella xiamenensis, and members of Enterobacteriaceae. Carbapenem-resistant bacteria were highly resistant to other beta-lactams such as quinolones, aminoglycosides, chloramphenicol, tetracyclines, and sulfamethoxazole/trimethoprim. Carbapenem resistance was mostly associated with intrinsically resistant bacteria. As intrinsic resistance mechanisms, we have identified the blaCphA gene in 77.3% of Aeromonas spp., blaL1 in all S. maltophilia, and blaOXA-48-like in all S. xiamenensis. As acquired resistance mechanisms, we have detected the blaVIM-2 gene in six Pseudomonas spp. (5.45%). Integrons with gene cassettes encoding resistance to aminoglycosides (aacA and aacC genes), trimethoprim (dfrB1b), and carbapenems (blaVIM-2) were found in Pseudomonas spp. Results suggest that carbapenem resistance dissemination in riverine ecosystems is still at an early stage. Nevertheless, monitoring these aquatic compartments for the presence of resistance genes and its host organisms is essential to outline strategies to minimize resistance dissemination.

  9. EPS production and bioremoval of heavy metals by mixed and pure bacterial cultures isolated from Ankara Stream.

    PubMed

    Kiliç, Nur Koçberber; Kürkçü, Güliz; Kumruoğlu, Durna; Dönmez, Gönül

    2015-01-01

    This study is focused on isolation of Ni(II), Cu(II) and Cr(VI) resistant bacteria to assess their exopolysaccharide (EPS) production and related bioremoval capacities. Mixed cultures had higher heavy metal removal capacity in media with molasses (MAS) than the control cultures lacking this carbon (AS) containing 50 mg/l of heavy metal. The yields were 32%, 75.7%, and 51.1% in MAS, while the corresponding values were 29%, 55.1%, and 34.5% in AS, respectively. Purification of the strains 1, 5 and 6 present in the mixed cultures decreased the bioremoval capacities of the mixed culture samples, although these strains produced higher EPS amounts in MAS agar. Strain 5 had the highest Cu(II) (69.1%) and Cr(VI) (43.1%) removal rates at 25 mg/l initial concentration of each pollutant with EPS amounts of 0.74 g/l and 1.05 g/l, respectively. This strain was identified as Stenotrophomonas maltophilia. The presented data show that especially mixed and also pure cultures of bacterial strains isolated from Ankara Stream could be assessed as potential bioremoval agents in the treatment of Cu(II) or Cr(VI) containing wastewaters.

  10. Role of a moderately halophilic bacterial consortium in the biodegradation of polyaromatic hydrocarbons.

    PubMed

    Arulazhagan, P; Vasudevan, N

    2009-02-01

    Polycyclic aromatic hydrocarbons are ubiquitous pollutants in the environment, and most high molecular weight PAHs cause mutagenic, teratogenic and potentially carcinogenic effects. While several strains have been identified that degrade PAHs, the present study is focused on the degradation of PAHs in a marine environment by a moderately halophilic bacterial consortium. The bacterial consortium was isolated from a mixture of marine water samples collected from seven different sites in Chennai, India. The low molecular weight (LMW) PAHs phenanthrene and fluorine, and the high molecular weight (HMW) PAHs pyrene and benzo(e)pyrene were selected for the degradation study. The consortium metabolized both LMW and HMW PAHs. The consortium was also able to degrade PAHs present in crude oil-contaminated saline wastewater. The bacterial consortium was able to degrade 80% of HMW PAHs and 100% of LMW PAHs in the saline wastewater. The strains present in the consortium were identified as Ochrobactrum sp., Enterobacter cloacae and Stenotrophomonas maltophilia. This study reveals that these bacteria have the potential to degrade different PAHs in saline wastewater.

  11. Microbial Surveillance of Potable Water Sources of the International Space Station

    NASA Technical Reports Server (NTRS)

    Bruce, Rebekah J.; Ott, C. Mark; Skuratov, Vladimir M.; Pierson, Duane L.

    2005-01-01

    To mitigate risk to the crew, the microbial surveillance of the quality of potable water sources of the International Space Station (ISS) has been ongoing since before the arrival of the first permanent crew. These water sources have included stored ground-supplied water, water produced by the shuttle fuel cells during flight, and ISS humidity condensate that is reclaimed and processed. Monitoring was accomplished using a self-contained filter designed to allow bacterial growth and enumeration during flight. Upon return to earth, microbial isolates were identified using 16S ribosomal gene sequencing. While the predominant isolates were common Gramnegative bacteria including Ralstonia eutropha, Methylobacterium fujisawaense, and Spingomonas paucimobilis, opportunistic pathogens such as Stenotrophomonas maltophilia and Pseudomonas aeruginosa were also isolated. Results of in-flight enumeration have indicated a fluctuation of bacterial counts above system design specifications. Additional in-flight monitoring capability for the specific detection of coliforms was added in 2004; no coliforms have been detected from any potable water source. Neither the bacterial concentrations nor the identification of the isolates recovered from these samples has suggested a threat to crew health.

  12. Laboratory diagnosis, clinical management and infection control of the infections caused by extensively drug-resistant Gram-negative bacilli: a Chinese consensus statement.

    PubMed

    Guan, X; He, L; Hu, B; Hu, J; Huang, X; Lai, G; Li, Y; Liu, Y; Ni, Y; Qiu, H; Shao, Z; Shi, Y; Wang, M; Wang, R; Wu, D; Xie, C; Xu, Y; Yang, F; Yu, K; Yu, Y; Zhang, J; Zhuo, C

    2016-03-01

    Extensively drug-resistant (XDR) Gram-negative bacilli (GNB) are defined as bacterial isolates susceptible to two or fewer antimicrobial categories. XDR-GNB mainly occur in Enterobacteriaceae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. The prevalence of XDR-GNB is on the rise in China and in other countries, and it poses a major public health threat as a result of the lack of adequate therapeutic options. A group of Chinese clinical experts, microbiologists and pharmacologists came together to discuss and draft a consensus on the laboratory diagnosis, clinical management and infection control of XDR-GNB infections. Lists of antimicrobial categories proposed for antimicrobial susceptibility testing were created according to documents from the Clinical Laboratory Standards Institute (CLSI), the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and the United States Food and Drug Administration (FDA). Multiple risk factors of XDR-GNB infections are analyzed, with long-term exposure to extended-spectrum antimicrobials being the most important one. Combination therapeutic regimens are summarized for treatment of XDR-GNB infections caused by different bacteria based on limited clinical studies and/or laboratory data. Most frequently used antimicrobials used for the combination therapies include aminoglycosides, carbapenems, colistin, fosfomycin and tigecycline. Strict infection control measures including hand hygiene, contact isolation, active screening, environmental surface disinfections, decolonization and restrictive antibiotic stewardship are recommended to curb the XDR-GNB spread.

  13. Diversity and Antimicrobial Properties of Lactic Acid Bacteria Isolated from Rhizosphere of Olive Trees and Desert Truffles of Tunisia

    PubMed Central

    Najjari, Afef; Turki, Yousra; Jaballah, Sana; Boudabous, Abdelatif; Ouzari, Hadda

    2013-01-01

    A total of 119 lactic acid bacteria (LAB) were isolated, by culture-dependant method, from rhizosphere samples of olive trees and desert truffles and evaluated for different biotechnological properties. Using the variability of the intergenic spacer 16S-23S and 16S rRNA gene sequences, the isolates were identified as the genera Lactococcus, Pediococcus, Lactobacillus, Weissella, and Enterococcus. All the strains showed proteolytic activity with variable rates 42% were EPS producers, while only 10% showed the ability to grow in 9% NaCl. In addition, a low rate of antibiotic resistance was detected among rhizospheric enterococci. Furthermore, a strong antibacterial activity against plant and/or pathogenic bacteria of Stenotrophomonas maltophilia, Pantoea agglomerans, Pseudomonas savastanoi, the food-borne Staphylococcus aureus, and Listeria monocytogenes was recorded. Antifungal activity evaluation showed that Botrytis cinerea was the most inhibited fungus followed by Penicillium expansum, Verticillium dahliae, and Aspergillus niger. Most of the active strains belonged to the genera Enterococcus and Weissella. This study led to suggest that environmental-derived LAB strains could be selected for technological application to control pathogenic bacteria and to protect food safety from postharvest deleterious microbiota. PMID:24151598

  14. Histamine, cadaverine, and putrescine produced in vitro by enterobacteriaceae and pseudomonadaceae isolated from spinach.

    PubMed

    Lavizzari, T; Breccia, M; Bover-Cid, S; Vidal-Carou, M C; Veciana-Nogués, M T

    2010-02-01

    A total of 364 bacterial isolates, obtained from spinach leaves, were assayed in a decarboxylase broth containing histidine, lysine, and ornithine to check their ability to produce biogenic amines, and then quantified by high-performance liquid chromatography. Among these isolates, 240 formed cadaverine, 208 formed putrescine, and 196 formed histamine, in widely varying amounts. They frequently produced more than one biogenic amine. Klebsiella pneumoniae subsp. pneumoniae and Morganella morganii were the main histamine producers, with mean values of 1,600 and 2,440 mg/liter, respectively, followed by Pantoea spp. 3 (1,710 mg/liter) and Hafnia alvei (2,500 mg/liter). Enterobacter amnigenus and Enterobacter cloacae produced particularly high amounts of putrescine, with mean values of 2,340 and 2,890 mg/liter, respectively. The strongest cadaverine formation was shown by Serratia liquefaciens (3,300 mg/liter), Serratia marcescens (3,280 mg/liter), and Stenotrophomonas maltophilia (1,000 mg/liter).

  15. Preventing microbial colonisation of catheters: antimicrobial and antibiofilm activities of cellobiose dehydrogenase.

    PubMed

    Thallinger, Barbara; Argirova, Maya; Lesseva, Magdalena; Ludwig, Roland; Sygmund, Christoph; Schlick, Angelika; Nyanhongo, Gibson S; Guebitz, Georg M

    2014-11-01

    The ability of cellobiose dehydrogenase (CDH) to produce hydrogen peroxide (H(2)O(2)) for antimicrobial and antibiofilm functionalisation of urinary catheters was investigated. A recombinantly produced CDH from Myriococcum thermophilum was shown to completely inhibit the growth of Escherichia coli and Staphylococcus aureus both in liquid and solid media when supplemented with either 0.8 mM or 2 mM cellobiose as substrate. Biofilm formation on silicone films was prevented by CDH when supplemented with 1mM cellobiose. The CDH/cellobiose system also successfully inhibited many common urinary catheter-colonising micro-organisms, including multidrug-resistant S. aureus, Staphylococcus epidermidis, Proteus mirabilis, Stenotrophomonas maltophilia, Acinetobacter baumannii and Pseudomonas aeruginosa. Interestingly, CDH was also able to produce H(2)O(2) during oxidation of extracellular polysaccharides (exPS) formed by micro-organisms in the absence of cellobiose. The H(2)O(2) production and consequently antimicrobial and antibiofilm activities on these exPS were enhanced by incorporation of glycoside hydrolases such as amylases. Hydrolysis of polysaccharides by these enzymes increases the number of terminal reducing sugars as substrates for CDH as well as destabilises the biofilm. Furthermore, CDH suspended in catheter lubricants killed bacteria in biofilms colonising catheters. Incorporation of the CDH/cellobiose system in the lubricant therefore makes it an easy strategy for preventing microbial colonisation of catheters.

  16. Evaluation of the effectiveness of manual and automated dialyzers reprocessing after multiple reuses.

    PubMed

    Toniolo, Alexandra do Rosário; Ribeiro, Maíra Marques; Ishii, Marina; da Silva, Cely Barreto; Jenné Mimica, Lycia Mara; Graziano, Kazuko Uchikawa

    2016-06-01

    A cross-sectional study was conducted to evaluate the effectiveness of manual and automated dialyzer reprocessing. Dialyzers were filled with fluid thioglycollate medium from blood and dialysate chambers after being reprocessed and chemically sterilized with 0.2% peracetic acid. They were incubated for 14 days at 35°C ± 2°C, and microbiologic analysis was performed. Microorganisms were identified in 3 of the 11 samples (27.3%) from the blood chambers: Sphingomonas paucimobilis (2/3) and Penicillium spp (1/3) and in 11 of the 11 samples (100%) from the dialysate chambers: S paucimobilis (7/11), Stenotrophomonas maltophilia (4/11), Pseudomonas aeruginosa (3/11), Candida spp (1/11), and Acinetobacter baumannii (1/11). Of the 4 manually reprocessed dialyzers, gram-positive bacillus were identified in 1 sample (25%) from the blood chamber, and Bacillus spp and Burkholderia spp were identified in 1 sample (25%) from the dialysate chamber. The dialyzers reprocessing can pose risks safety because of exposure patient to microorganisms.

  17. Rhizobacterial characterization for quality control of eucalyptus biogrowth promoter products.

    PubMed

    Zarpelon, Talyta Galafassi; Guimarães, Lúcio Mauro da Silva; Alfenas-Zerbini, Poliane; Lopes, Eli Sidney; Mafia, Reginaldo Gonçalves; Alfenas, Acelino Couto

    Plant growth-promoting rhizobacteria strains from special formulations have been used to optimize eucalyptus cutting production. To undertake quality control for the formulated products, the rhizobacterial strains should be characterized to assess their purity and authentication. In the present study, we characterized nine strains of rhizobacteria, including three Bacillus subtilis (S1, S2 and 3918), two Pseudomonas sp. (MF4 and FL2), P. putida (MF2), P. fulva (Ca), Frateuria aurantia (R1), and Stenotrophomonas maltophilia (CIIb). The strains were differentiated by colony morphology after 24h of incubation in three different solid state culture media (glucose-nutritive agar, 523 medium and yeast extract-mannitol agar), sensitivity to a panel of 28 antibiotics (expressed according to the formation of inhibition halos of bacterial growth in the presence of antibiotics), and PCR-RFLP profiles of the 16S rDNA gene produced using nine restriction enzymes. It was possible to differentiate all nine strains of rhizobacteria using their morphological characteristics and sensitivity to antibiotics. The molecular analysis allowed us to separate the strains CIIb, FL2 and R1 from the strains belonging to the genera Bacillus and Pseudomonas. By using these three methods concomitantly, we were able to determine strain purity and perform the authentication.

  18. Assessing the xylanolytic bacterial diversity during the malting process.

    PubMed

    Malfliet, Sofie; Justé, Annelies; Crauwels, Sam; Willems, Kris; De Cooman, Luc; Lievens, Bart; Aerts, Guido

    2013-12-01

    The presence of microorganisms producing cell wall hydrolyzing enzymes such as xylanases during malting can improve mash filtration behavior and consequently have potential for more efficient wort production. In this study, the xylanolytic bacterial community during malting was assessed by isolation and cultivation on growth media containing arabinoxylan, and identification by 16S rRNA gene sequencing. A total of 33 species-level operational taxonomic units (OTUs) were found, taking into account a 3% sequence dissimilarity cut-off, belonging to four phyla (Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria) and 25 genera. Predominant OTUs represented xylanolytic bacteria identified as Sphingobacterium multivorum, Stenotrophomonas maltophilia, Aeromonas hydrophila and Pseudomonas fulva. DNA fingerprinting of all xylanolytic isolates belonging to S. multivorum obtained in this study revealed shifts in S. multivorum populations during the process. Xylanase activity was determined for a selection of isolates, with Cellulomonas flavigena showing the highest activity. The xylanase of this species was isolated and purified 23.2-fold by ultrafiltration, 40% ammonium sulfate precipitation and DEAE-FF ion-exchange chromatography and appeared relatively thermostable. This study will enhance our understanding of the role of microorganisms in the barley germination process. In addition, this study may provide a basis for microflora management during malting.

  19. Diaryl-substituted azolylthioacetamides: Inhibitor discovery of New Delhi metallo-β-lactamase-1 (NDM-1).

    PubMed

    Zhang, Yi-Lin; Yang, Ke-Wu; Zhou, Ya-Jun; LaCuran, Alecander E; Oelschlaeger, Peter; Crowder, Michael W

    2014-11-01

    The emergence and spread of antibiotic-resistant pathogens is a global public health problem. Metallo-β-lactamases (MβLs) such as New Delhi MβL-1 (NDM-1) are principle contributors to the emergence of resistance because of their ability to hydrolyze almost all known β-lactam antibiotics including penicillins, cephalosporins, and carbapenems. A clinical inhibitor of MBLs has not yet been found. In this study we developed eighteen new diaryl-substituted azolylthioacetamides and found all of them to be inhibitors of the MβL L1 from Stenotrophomonas maltophilia (Ki < 2 μM), thirteen to be mixed inhibitors of NDM-1 (Ki < 7 μM), and four to be broad-spectrum inhibitors of all four tested MβLs CcrA from Bacteroides fragilis, NDM-1 and ImiS from Aeromonas veronii, and L1 (Ki < 52 μM), which are representative of the B1a, B1b, B2, and B3 subclasses, respectively. Docking studies revealed that the azolylthioacetamides, which have the broadest inhibitory activity, coordinate to the Zn(II) ion(s) preferentially via the triazole moiety, while other moieties interact mostly with the conserved active site residues Lys224 (CcrA, NDM-1, and ImiS) or Ser221 (L1).

  20. Capillary isoelectric focusing of proteins and microorganisms in dynamically modified fused silica with UV detection.

    PubMed

    Horká, Marie; Růzicka, Filip; Horký, Jaroslav; Holá, Veronika; Slais, Karel

    2006-09-01

    We suggest a method for the reproducible and efficient capillary isoelectric focusing of proteins and microorganisms in the pH gradient 3-10. The method involves the segmental injection of the simple ampholytes, the solution of the selected electrolytes, and the sample mixture of bioanalytes and carrier ampholytes to the fused silica capillaries dynamically modified by poly(ethylene glycol), PEG 4000, which is added to the catholyte, the anolyte and injected solutions. In order to receive the reproducible results, the capillaries were rinsed by the mixture of acetone/ethanol between analyses. For the tracing of the pH gradients the low-molecular-mass pI markers were used. The simple proteins and the mixed cultures of microorganisms, Saccharomyces cerevisiae CCM 8191, Escherichia coli CCM 3954, Candida albicans CCM 8180, Candida parapsilosis, Candida krusei, Staphylococcus aureus, Streptococcus agalactiae CCM 6187, Enterococcus faecalis CCM 4224, Staphylococcus epidermidis CCM 4418 and Stenotrophomonas maltophilia, were focused and separated by the method suggested. The minimum detectable number of microbial cells was 5x10(2) to 1x10(3) with on-column UV detection at 280 nm.

  1. Metagenomic analysis of medicinal Cannabis samples; pathogenic bacteria, toxigenic fungi, and beneficial microbes grow in culture-based yeast and mold tests.

    PubMed

    McKernan, Kevin; Spangler, Jessica; Helbert, Yvonne; Lynch, Ryan C; Devitt-Lee, Adrian; Zhang, Lei; Orphe, Wendell; Warner, Jason; Foss, Theodore; Hudalla, Christopher J; Silva, Matthew; Smith, Douglas R

    2016-01-01

    Background: The presence of bacteria and fungi in medicinal or recreational Cannabis poses a potential threat to consumers if those microbes include pathogenic or toxigenic species. This study evaluated two widely used culture-based platforms for total yeast and mold (TYM) testing marketed by 3M Corporation and Biomérieux, in comparison with a quantitative PCR (qPCR) approach marketed by Medicinal Genomics Corporation. Methods: A set of 15 medicinal Cannabis samples were analyzed using 3M and Biomérieux culture-based platforms and by qPCR to quantify microbial DNA. All samples were then subjected to next-generation sequencing and metagenomics analysis to enumerate the bacteria and fungi present before and after growth on culture-based media. Results: Several pathogenic or toxigenic bacterial and fungal species were identified in proportions of >5% of classified reads on the samples, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Ralstonia pickettii, Salmonella enterica, Stenotrophomonas maltophilia, Aspergillus ostianus, Aspergillus sydowii, Penicillium citrinum and Penicillium steckii. Samples subjected to culture showed substantial shifts in the number and diversity of species present, including the failure of Aspergillus species to grow well on either platform. Substantial growth of Clostridium botulinum and other bacteria were frequently observed on one or both of the culture-based TYM platforms. The presence of plant growth promoting (beneficial) fungal species further influenced the differential growth of species in the microbiome of each sample. Conclusions: These findings have important implications for the Cannabis and food safety testing industries.

  2. Prevalence of antibiotic-resistant bacteria in herbal products.

    PubMed

    Brown, Joseph C; Jiang, Xiuping

    2008-07-01

    The objective of this study was to determine the prevalence of antibiotic-resistant bacteria in various herbal products. Twenty-nine herbal supplements (18 traditional and 11 organic products) were purchased from stores and analyzed microbiologically. Total bacterial counts were determined by pour plate and surface spreading on tryptic soy agar (TSA). Antibiotic-resistant bacteria were enumerated on TSA supplemented with ceftriaxone (64 microg/ml) or tetracycline (16 microg/ml). Total bacterial counts ranged from <5 to 2.9 x 10(5) CFU/g. Ceftriaxone- and tetracycline-resistant bacteria were detected in ground garlic samples at 1.1 x 10(2) CFU/g and 3.0 x 102 CFU/g, respectively. Traditional and organic onion powder samples contained tetracycline-resistant bacteria at 17 and 28 CFU/g and ceftriaxone-resistant bacteria at 35 and 2.0 x 10(3) CFU/g, respectively. Other products such as ginger, rosemary, mustard, and goldenseal contained low levels of resistant bacteria. Fifty-two isolates were further evaluated against nine antibiotics, and the prevalence of antibiotic resistance was in the following order: ampicillin, nalidixic acid, trimethoprim, ceftriaxone, and streptomycin. Resistant bacteria were identified as Bacillus spp., Erwinia spp., and Ewingella americana. Staphylococcus spp., Enterobacter cloacae, and Stenotrophomonas maltophilia also were isolated. The presence of antibiotic-resistant bacteria and pathogens in these herbal products suggests that production and use of these products may need further evaluation.

  3. Antibiotic resistance and extended-spectrum β-lactamases in isolated bacteria from seawater of Algiers beaches (Algeria).

    PubMed

    Alouache, Souhila; Kada, Mohamed; Messai, Yamina; Estepa, Vanesa; Torres, Carmen; Bakour, Rabah

    2012-01-01

    The aim of the study was to evaluate bacterial antibiotic resistance in seawater from four beaches in Algiers. The most significant resistance rates were observed for amoxicillin and ticarcillin, whereas they were relatively low for ceftazidime, cefotaxime and imipenem. According to sampling sites, the highest resistance rates were recorded for 2 sites subjected to chemical and microbiological inputs (amoxicillin, 43% and 52%; ticarcillin, 19.6% and 47.7%), and for 2 sites relatively preserved from anthropogenic influence, resistance rates were lowest (amoxicillin, 1.5% and 16%; ticarcillin, 0.8% and 2.6%). Thirty-four bacteria resistant to imipenem (n=14) or cefotaxime (n=20) were identified as Pseudomonas aeruginosa (n=15), Pseudomonas fluorescens (7), Stenotrophomonas maltophilia (4), Burkholderia cepacia (2), Bordetella sp. (1), Pantoea sp. (1), Acinetobacter baumannii (1), Chryseomonas luteola (1), Ochrobactrum anthropi (1) and Escherichia coli (1). Screening for extended spectrum β-lactamase showed the presence of CTX-M-15 β-lactamase in the E. coli isolate, and the encoding gene was transferable in association with the IncI1 plasmid of about 50 kbp. Insertion sequence ISEcp1B was located upstream of the CTX-M-15 gene. This work showed a significant level of resistance to antibiotics, mainly among environmental saprophytic bacteria. Transmissible CTX-M-15 was detected in E. coli; this may mean that contamination of the environment by resistant bacteria may cause the spread of resistance genes.

  4. Antibiotic Resistance and Extended-Spectrum β-Lactamases in Isolated Bacteria from Seawater of Algiers Beaches (Algeria)

    PubMed Central

    Alouache, Souhila; Kada, Mohamed; Messai, Yamina; Estepa, Vanesa; Torres, Carmen; Bakour, Rabah

    2012-01-01

    The aim of the study was to evaluate bacterial antibiotic resistance in seawater from four beaches in Algiers. The most significant resistance rates were observed for amoxicillin and ticarcillin, whereas they were relatively low for ceftazidime, cefotaxime and imipenem. According to sampling sites, the highest resistance rates were recorded for 2 sites subjected to chemical and microbiological inputs (amoxicillin, 43% and 52%; ticarcillin, 19.6% and 47.7%), and for 2 sites relatively preserved from anthropogenic influence, resistance rates were lowest (amoxicillin, 1.5% and 16%; ticarcillin, 0.8% and 2.6%). Thirty-four bacteria resistant to imipenem (n=14) or cefotaxime (n=20) were identified as Pseudomonas aeruginosa (n=15), Pseudomonas fluorescens(7), Stenotrophomonas maltophilia(4), Burkholderia cepacia(2), Bordetella sp. (1), Pantoea sp. (1), Acinetobacter baumannii(1), Chryseomonas luteola(1), Ochrobactrum anthropi(1) and Escherichia coli(1). Screening for extended spectrum β-lactamase showed the presence of CTX-M-15 β-lactamase in the E. coli isolate, and the encoding gene was transferable in association with the IncI1 plasmid of about 50 kbp. Insertion sequence ISEcp1B was located upstream of the CTX-M-15 gene. This work showed a significant level of resistance to antibiotics, mainly among environmental saprophytic bacteria. Transmissible CTX-M-15 was detected in E. coli; this may mean that contamination of the environment by resistant bacteria may cause the spread of resistance genes. PMID:22095134

  5. Physiological traits of the symbiotic bacterium Teredinibacter turnerae isolated from the mangrove shipworm Neoteredo reynei.

    PubMed

    Trindade-Silva, Amaro E; Machado-Ferreira, Erik; Senra, Marcus V X; Vizzoni, Vinicius F; Yparraguirre, Luciana A; Leoncini, Orilio; Soares, Carlos A G

    2009-07-01

    Nutrition in the Teredinidae family of wood-boring mollusks is sustained by cellulolytic/nitrogen fixing symbiotic bacteria of the Teredinibacter clade. The mangrove Teredinidae Neoteredo reynei is popularly used in the treatment of infectious diseases in the north of Brazil. In the present work, the symbionts of N. reynei, which are strictly confined to the host's gills, were conclusively identified as Teredinibacter turnerae. Symbiont variants obtained in vitro were able to grow using casein as the sole carbon/nitrogen source and under reduced concentrations of NaCl. Furthermore, cellulose consumption in T. turnerae was clearly reduced under low salt concentrations. As a point of interest, we hereby report first hand that T. turnerae in fact exerts antibiotic activity. Furthermore, this activity was also affected by NaCl concentration. Finally, T. turnerae was able to inhibit the growth of Gram-negative and Gram-positive bacteria, this including strains of Sphingomonas sp., Stenotrophomonas maltophilia, Bacillus cereus and Staphylococcus sciuri. Our findings introduce new points of view on the ecology of T. turnerae, and suggest new biotechnological applications for this marine bacterium.

  6. Effect of bacterial interference on biofilm development by Legionella pneumophila.

    PubMed

    Guerrieri, Elisa; Bondi, Moreno; Sabia, Carla; de Niederhäusern, Simona; Borella, Paola; Messi, Patrizia

    2008-12-01

    In the ecology of Legionella pneumophila a crucial role may be played by its relationship with the natural flora; thus we investigated the interactions between Legionella and other aquatic bacteria, particularly within biofilms. Among 80 aquatic bacteria screened for the production of bacteriocin-like substances (BLSs), 66.2% of them were active against L. pneumophila. The possible effect of some of these aquatic bacteria on the development and stability of L. pneumophila biofilms was studied. Pseudomonas fluorescens, the best BLS producer, showed the greatest negative effect on biofilm formation and strongly enhanced the detachment of Legionella. Pseudomonas aeruginosa, Burkholderia cepacia, Pseudomonas putida, Aeromonas hydrophila, and Stenotrophomonas maltophilia, although producing BLSs at different levels, were less active in the biofilm experiments. Acinetobacter lwoffii did not produce any antagonistic compound and was the only one able to strongly enhance L. pneumophila biofilm. Our results highlight that BLS production may contribute to determining the fate of L. pneumophila within ecological niches. The interactions observed in this study are important features of L. pneumophila ecology, which knowledge may lead to more effective measures to control the persistence of the germ in the environment.

  7. Biodegradation of phenanthrene in bioaugmented microcosm by consortium ASP developed from coastal sediment of Alang-Sosiya ship breaking yard.

    PubMed

    Patel, Vilas; Patel, Janki; Madamwar, Datta

    2013-09-15

    A phenanthrene-degrading bacterial consortium (ASP) was developed using sediment from the Alang-Sosiya shipbreaking yard at Gujarat, India. 16S rRNA gene-based molecular analyses revealed that the bacterial consortium consisted of six bacterial strains: Bacillus sp. ASP1, Pseudomonas sp. ASP2, Stenotrophomonas maltophilia strain ASP3, Staphylococcus sp. ASP4, Geobacillus sp. ASP5 and Alcaligenes sp. ASP6. The consortium was able to degrade 300 ppm of phenanthrene and 1000 ppm of naphthalene within 120 h and 48 h, respectively. Tween 80 showed a positive effect on phenanthrene degradation. The consortium was able to consume maximum phenanthrene at the rate of 46 mg/h/l and degrade phenanthrene in the presence of other petroleum hydrocarbons. A microcosm study was conducted to test the consortium's bioremediation potential. Phenanthrene degradation increased from 61% to 94% in sediment bioaugmented with the consortium. Simultaneously, bacterial counts and dehydrogenase activities also increased in the bioaugmented sediment. These results suggest that microbial consortium bioaugmentation may be a promising technology for bioremediation.

  8. Decontamination effects of low-temperature plasma generated by corona discharge. Part II: new insights.

    PubMed

    Scholtz, V; Julák, J; Kríha, V; Mosinger, J; Kopecká, S

    2007-01-01

    The second part of our paper presents the results of experiments with the decontamination of surfaces by low-temperature plasma generated by corona discharge in air at atmospheric pressure. A simple device is described and the effects of the corona discharge on model microorganisms, viz. the yeast Candida albicans, Gram-negative bacteria Escherichia coli, Enterobacter aerogenes, Neisseria sicca, Stenotrophomonas maltophilia, Gram-positive bacteria Deinococcus radiodurans, Enterococcus faecium, Staphylococcus epidermidis, Streptococcus sanguinis, and vegetative and spore forms of Geobacillus stearothermophilus are discussed. A similar microbicidal effect after about one-minute exposure was observed in all vegetative forms of the microorganisms. Measurement in growth inhibition zones on a semisolid medium was used to determine the dependence of the microbicidal effect on exposure time and the distance between electrodes. Counting of colonies served to assess the microbicidal effect of the discharge on contaminated inert surfaces observable after more than 1 min exposure. Geobacillus stearothermophilus spores were found to have several times lower susceptibility to the action of the discharge and the microbicidal effect was observed only after an 8 min exposure. Reaction with the iodide reagent did not unambiguously demonstrate the difference between ozone and singlet oxygen as presumed active components of the corona. The area distribution of reactive oxygen species was determined; it was found to differ from the Wartburg law depending on exposure time. Qualitative evidence was obtained on the penetration of the reactive oxygen species into the semisolid medium.

  9. Multi-channel microfluidic biosensor platform applied for online monitoring and screening of biofilm formation and activity.

    PubMed

    Bruchmann, Julia; Sachsenheimer, Kai; Rapp, Bastian E; Schwartz, Thomas

    2015-01-01

    Bacterial colonization of surfaces and interfaces has a major impact on various areas including biotechnology, medicine, food industries, and water technologies. In most of these areas biofilm development has a strong impact on hygiene situations, product quality, and process efficacies. In consequence, biofilm manipulation and prevention is a fundamental issue to avoid adverse impacts. For such scenario online, non-destructive biofilm monitoring systems become important in many technical and industrial applications. This study reports such a system in form of a microfluidic sensor platform based on the combination of electrical impedance spectroscopy and amperometric current measurement, which allows sensitive online measurement of biofilm formation and activity. A total number of 12 parallel fluidic channels enable real-time online screening of various biofilms formed by different Pseudomonas aeruginosa and Stenotrophomonas maltophilia strains and complex mixed population biofilms. Experiments using disinfectant and antibiofilm reagents demonstrate that the biofilm sensor is able to discriminate between inactivation/killing of bacteria and destabilization of biofilm structures. The impedance and amperometric sensor data demonstrated the high dynamics of biofilms as a consequence of distinct responses to chemical treatment strategies. Gene expression of flagellar and fimbrial genes of biofilms grown inside the microfluidic system supported the detected biofilm growth kinetics. Thus, the presented biosensor platform is a qualified tool for assessing biofilm formation in specific environments and for evaluating the effectiveness of antibiofilm treatment strategies.

  10. Characterization of Contaminants from a Sanitized Milk Processing Plant

    PubMed Central

    Cleto, Sara; Matos, Sónia; Kluskens, Leon; Vieira, Maria João

    2012-01-01

    Milk processing lines offer a wide variety of microenvironments where a diversity of microorganisms can proliferate. We sampled crevices and junctions where, due to deficient reach by typical sanitizing procedures, bacteria can survive and establish biofilms. The sampling sites were the holding cell, cold storage tank, pasteurizer and storage tank - transfer pump junction. The culturable bacteria that were isolated after the sanitation procedure were predominantly Pseudomonas spp., Serratia spp, Staphylococcus sciuri and Stenotrophomonas maltophilia. We assayed several phenotypic characteristics such as the ability to secrete enzymes and siderophores, as well as the capacity of the strains to form biofilms that might contribute to their survival in a mixed species environment. The Pseudomonas spp. isolates were found to either produce proteases or lecithinases at high levels. Interestingly, protease production showed an inverse correlation with siderophore production. Furthermore, all of the Serratia spp. isolates were strong biofilm formers and spoilage enzymes producers. The organisms identified were not mere contaminants, but also producers of proteins with the potential to lower the quality and shelf-life of milk. In addition, we found that a considerable number of the Serratia and Pseudomonas spp. isolated from the pasteurizer were capable of secreting compounds with antimicrobial properties. PMID:22761957

  11. Improved method for effective screening of ACC (1-aminocyclopropane-1-carboxylate) deaminase producing microorganisms.

    PubMed

    Patil, Chandrashekhar; Suryawanshi, Rahul; Koli, Sunil; Patil, Satish

    2016-12-01

    Aminocyclopropane-1-carboxylate deaminase (ACCD) producing microorganisms support plant growth under a variety of biotic and abiotic stress conditions such as drought, soil salinity, flooding, heavy metal pollution and phyto-pathogen attack. Available screening methods for ACCD give idea only about its primary microbial ACCD activity than the actual potential. In the present investigation, we have simply improved screening method by incorporating pH indicator dyes (phenol red and bromothymol blue) in ACC containing medium. This modification is based on the basic principle that ACCD action releases ammonia which can be detected by color change and zone around the bacterial colony. High color intensity and zone around the colony indicates most potent producer, colony showing only a color change indicates moderate potential and no change in colony color indicates least efficiency. Enzymatic bioassays as well as root elongation studies revealed that ACC-deaminase activity of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Bacillus subtilis clearly corresponds to their growth on dye incorporated ACC medium. This method could be used to complement the existing screening methods and to speed up the targeted isolation of agriculturally important microorganisms.

  12. Potency and Spectrum of Activity of AN3365, a Novel Boron-Containing Protein Synthesis Inhibitor, Tested against Clinical Isolates of Enterobacteriaceae and Nonfermentative Gram-Negative Bacilli

    PubMed Central

    Alley, M. R. K.; Sader, Helio S.; Biedenbach, Douglas J.; Jones, Ronald N.

    2013-01-01

    AN3365 (MIC50/90, 0.5/1 μg/ml) was active against Enterobacteriaceae, including a subset of Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae strains (MIC50/90, 1/2 μg/ml). AN3365 inhibited 98.0 and 92.2% of wild-type (MIC50/90, 2/8 μg/ml) and carbapenem-resistant (MIC50/90, 4/8 μg/ml) Pseudomonas aeruginosa strains, respectively, at ≤8 μg/ml. AN3365 also demonstrated activity against wild-type Acinetobacter baumannii (MIC50/90, 2/8 μg/ml) and Stenotrophomonas maltophilia (MIC50/90, 2/4 μg/ml), while it was less active against multidrug-resistant A. baumannii (MIC50/90, 8/16 μg/ml) and Burkholderia cepacia (MIC50/90, 8/32 μg/ml). PMID:23507283

  13. Physicochemical and Antibacterial Properties of Chitosan Extracted from Waste Shrimp Shells

    PubMed Central

    Vilar Junior, José Carlos; Ribeaux, Daylin Rubio; Alves da Silva, Carlos Alberto

    2016-01-01

    This research aims to study the production of chitosan from shrimp shell (Litopenaeus vannamei) of waste origin using two chemical methodologies involving demineralization, deproteinization, and the degree of deacetylation. The evaluation of the quality of chitosan from waste shrimp shells includes parameters for the yield, physical chemistry characteristics by infrared spectroscopy (FT-IR), the degree of deacetylation, and antibacterial activity. The results showed (by Method 1) extraction yields for chitin of 33% and for chitosan of 49% and a 76% degree of deacetylation. Chitosan obtained by Method 2 was more efficient: chitin (36%) and chitosan (63%), with a high degree of deacetylation (81.7%). The antibacterial activity was tested against Gram-negative bacteria (Stenotrophomonas maltophilia and Enterobacter cloacae) and Gram-positive Bacillus subtilis and the Minimum Inhibitory Concentrations (MIC) and the Minimum Bactericidal Concentration (MBC) were determined. Method 2 showed that extracted chitosan has good antimicrobial potential against Gram-positive and Gram-negative bacteria and that the process is viable. PMID:27478443

  14. Application of a continuously stirred tank bioreactor (CSTR) for bioremediation of hydrocarbon-rich industrial wastewater effluents.

    PubMed

    Gargouri, Boutheina; Karray, Fatma; Mhiri, Najla; Aloui, Fathi; Sayadi, Sami

    2011-05-15

    A continuously stirred tank bioreactor (CSTR) was used to optimize feasible and reliable bioprocess system in order to treat hydrocarbon-rich industrial wastewaters. A successful bioremediation was developed by an efficient acclimatized microbial consortium. After an experimental period of 225 days, the process was shown to be highly efficient in decontaminating the wastewater. The performance of the bioaugmented reactor was demonstrated by the reduction of COD rates up to 95%. The residual total petroleum hydrocarbon (TPH) decreased from 320 mg TPH l(-1) to 8 mg TPH l(-1). Analysis using gas chromatography-mass spectrometry (GC-MS) identified 26 hydrocarbons. The use of the mixed cultures demonstrated high degradation performance for hydrocarbons range n-alkanes (C10-C35). Six microbial isolates from the CSTR were characterized and species identification was confirmed by sequencing the 16S rRNA genes. The partial 16S rRNA gene sequences demonstrated that 5 strains were closely related to Aeromonas punctata (Aeromonas caviae), Bacillus cereus, Ochrobactrum intermedium, Stenotrophomonas maltophilia and Rhodococcus sp. The 6th isolate was affiliated to genera Achromobacter. Besides, the treated wastewater could be considered as non toxic according to the phytotoxicity test since the germination index of Lepidium sativum ranged between 57 and 95%. The treatment provided satisfactory results and presents a feasible technology for the treatment of hydrocarbon-rich wastewater from petrochemical industries and petroleum refineries.

  15. Investigation of bacterial pathogens on 70 frequently used environmental surfaces in a large urban U.S. university.

    PubMed

    Brooke, Joanna S; Annand, John W; Hammer, Angela; Dembkowski, Karen; Shulman, Stanford T

    2009-01-01

    After reports of increased severity of bacterial infections from community institutions, a broad spectrum of 70 surfaces was sampled for potential bacterial pathogens in the morning and afternoon of one day per week over three consecutive weeks in a large U.S. university. Surfaces included public telephone mouthpieces, water fountain drains, student computer keyboards and desks, and buttons on elevators, vending machines, and photocopiers. A total of 420 samples was obtained. Bacterial counts were high on telephone mouthpieces, up to 168.8 colony-forming units (CFUs).cm(-2) of surface area. Stenotrophomonas maltophilia was isolated from 60% of fountain drains. Ninety percent of the keyboards showed positive bacterial cultures in the afternoon sampling. Staphylococcus aureus was identified on keyboards, telephone mouthpieces, and an elevator button. No S. aureus were methicillin-resistant. The swab sampling method reduced bacterial counts to less than or equal to 2.0 CFU.cm(-2) on keyboards and telephone mouthpieces. Disinfectants for possible use in cleaning of telephones, water fountain drains, and keyboards are discussed.

  16. Characterizing Novel Thermophilic Amylase Producing Bacteria From Taptapani Hot Spring, Odisha, India

    PubMed Central

    Sen, Sudip Kumar; Raut, Sangeeta; Satpathy, Soumya; Rout, Prangya Ranjan; Bandyopadhyay, Bidyut; Das Mohapatra, Pradeep Kumar

    2014-01-01

    Background: Amylases play a vital role in biotechnological studies and rank an important position in the world enzyme market (25% to 33%). Bioprocess method of amylase production is more effective than the other sources, since the technique is easy, cost effective, fast, and the enzymes of required properties can be procured. Objectives: The current study aimed to report the characteristics of novel amylase producing bacterial strains isolated from Taptapani hot spring, Odisha, India. Materials and Methods: Bacterial strains were isolated by dilution plating method from the water samples collected from Taptapani Hot Spring, Odisha and screened for amylase production through starch hydrolysis. The bacterial isolates were identified morphologically, biochemically, and finally by 16S rDNA profiling. Results: Based on the morphological, physiological, biochemical characteristics and the molecular characterization, the isolates SS1, SS2, and SS3 were identified as Bacillus barbaricus, Aeromonas veroni, and Stenotrophomonas maltophilia, respectively. The approximate molecular weight of enzymes from SS1, SS2, and SS3 strains were 19 kDa, 56 kDa and 49 kDa, respectively. Conclusions: The current report isolates, characterizes, and demonstrates the novel heat-adapted amylase-producing bacteria SS1, SS2 and SS3 from Taptapani hot spring, indicating its potentiality and stability under acidic conditions. PMID:25741425

  17. In Vitro Assessment of the Antimicrobial Efficacy of Optimized Nitroglycerin-Citrate-Ethanol as a Nonantibiotic, Antimicrobial Catheter Lock Solution for Prevention of Central Line-Associated Bloodstream Infections

    PubMed Central

    Reitzel, Ruth A.; Hirsh-Ginsberg, Cheryl; Murray, Kimberly; Chaftari, Anne-Marie; Hachem, Ray; Raad, Issam

    2016-01-01

    The rapid, broad-spectrum, biofilm-eradicating activity of the combination of 0.01% nitroglycerin, 7% citrate, and 20% ethanol and its potential as a nonantibiotic, antimicrobial catheter lock solution (ACLS) were previously reported. Here, a nitroglycerin-citrate-ethanol (NiCE) ACLS optimized for clinical assessment was developed by reducing the nitroglycerin and citrate concentrations and increasing the ethanol concentration. Biofilm-eradicating activity was sustained when the ethanol concentration was increased from 20 to 22% which fully compensated for reducing the citrate concentration from 7% to 4% as well as the nitroglycerin concentration from 0.01% to 0.0015% or 0.003%. The optimized formulations demonstrated complete and rapid (2 h) eradication of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-intermediate Staphylococcus aureus (VISA), methicillin-resistant Staphylococcus epidermidis (MRSE), vancomycin-resistant enterococci (VRE), multidrug-resistant (MDR) Pseudomonas aeruginosa, MDR Klebsiella pneumoniae, MDR Enterobacter cloacae, MDR Acinetobacter baumannii, MDR Escherichia coli, MDR Stenotrophomonas maltophilia, Candida albicans, and Candida glabrata biofilms. The optimized NiCE lock solutions demonstrated anticoagulant activities comparable to those of heparin lock solutions. NiCE lock solution was significantly more effective than taurolidine-citrate-heparin lock solution in eradicating biofilms of Staphylococcus aureus and Candida glabrata. The optimized, nonantibiotic, heparin-free NiCE lock solution demonstrates rapid broad-spectrum biofilm eradication as well as effective anticoagulant activity, making NiCE a high-quality ACLS candidate for clinical assessment. PMID:27297475

  18. Isolation of Vermamoeba vermiformis and associated bacteria in hospital water.

    PubMed

    Pagnier, Isabelle; Valles, Camille; Raoult, Didier; La Scola, Bernard

    2015-03-01

    To detect new potential pathogens in hospital water, we isolated free-living amoebae in water samples taken from three different hospitals in Marseille (France). The samples were inoculated in media containing saline buffer and various bacteria as nutrient sources. The isolated amoebae were identified by gene sequencing. Among the 105 water samples, taken from 19 sites, we isolated 14 amoebae, of which 9 Vermamoeba vermiformis and 5 Acanthamoeba sp. None of the amoebae showed the presence of obligate bacterial endosymbionts. Because V. vermiformis was most commonly isolated, we used an axenic collection strain to isolate amoeba-resistant bacteria from the same sites. The isolated bacterial species included Stenotrophomonas maltophilia and Legionella sp. Legionella taurinensis was isolated for the first time in association with amoebae. A strict intracellular bacterium was isolated, that may represent a new genus among the Chlamydiales. We propose that it be named "Candidatus Rubidus massiliensis". Our study shows that the isolation and identification of new pathogens associated with amoebae, which were previously performed using Acanthamoeba sp., should instead use V. vermiformis because this organism is more commonly associated with humans and is an essential complement of Acanthamoeba sp. co-culture to study the ecology of hospital water supplies.

  19. Assessment of chitin decomposer diversity within an upland grassland.

    PubMed

    Krsek, M; Wellington, E M

    2001-09-01

    The breakdown of chitin within an acidic upland grassland was studied. The aim was to provide a molecular characterisation of microorganisms involved in chitin degradation in the soil using soil microcosms and buried litter bags containing chitin. The investigation involved an examination of the effects of liming on the microbial communities within the soil and their chitinolytic activity. Microcosm experiments were designed to study the influence of lime and chitin enrichment on the grassland soil bacterial community ex situ under controlled environmental conditions. Bacterial and actinomycete counts were determined and total community DNA was extracted from the microcosms and from chitin bags buried at the experimental site. PCR based on specific 16S rRNA target sequences provided products for DGGE analysis to determine the structure of bacterial and actinomycete communities. Chitinase activity was assessed spectrophotometrically using chitin labelled with remazol brilliant violet. Both liming and chitin amendment increased bacterial and actinomycete viable counts and the chitinase activity. DGGE band patterns confirmed changes in bacterial populations under the influence of both treatments. PCR products amplified from DNA isolated from chitin bags were cloned and sequenced. Only a few matched known species but a prominent coloniser of chitin proved to be Stenotrophomonas maltophilia.

  20. The effect of different growth regimes on the endophytic bacterial communities of the fern, Dicksonia sellowiana hook (Dicksoniaceae)

    PubMed Central

    de Araújo Barros, Irene; Luiz Araújo, Welington; Lúcio Azevedo, João

    2010-01-01

    Endophytic bacteria associated with the fern Dicksonia sellowiana were investigated. The bacterial communities from the surface-sterilized pinnae and rachis segments of the plants from the Brazilian Atlantic Rainforest that grew in native field conditions were compared with the bacterial communities from plants grown in greenhouses and plants that were initially grown in greenhouses and then transferred to the forest. From 540 pinnae and 540 rachis segments, 163 (30.2%) and 346 (64.2%) were colonized by bacteria, respectively. The main bacterial genera and species that were isolated included Bacillus spp. ( B. cereus, B. megaterium, B. pumilus and B. subtilis ) , Paenibacillus sp. , Amphibacillus sp. , Gracilibacillus sp. , Micrococcus sp. and Stenotrophomonas spp. ( S. maltophilia and S. nitroreducens ). B. pumilus was the most frequently isolated bacterial species . Amphibacillus and Gracilibacillus were reported as endophytes for the first time. Other commonly found bacterial genera were not observed in D. sellowiana , which may reflect preferences of specific bacterial communities inside this fern or detection limitations due to the isolation procedures. Plants that were grown in greenhouses and plants that were reintroduced into the forest displayed more bacterial genera and species diversity than native field plants, suggesting that reintroduction shifts the bacterial diversity. Endophytic bacteria that displayed antagonistic properties against different microorganisms were detected, but no obvious correlation was found between their frequencies with plant tissues or with plants from different growth regimes. This paper reports the first isolation of endophytic bacteria from a fern. PMID:24031575

  1. Antibacterial activities of grepafloxacin, ciprofloxacin, ofloxacin and fleroxacin.

    PubMed

    Barry, A L; Fuchs, P C

    1997-02-01

    Grepafloxacin (OPC-17116) and three other fluoroquinolones were tested against 461 bacterial isolates representing 44 species. Grepafloxacin was superior to ciprofloxacin, ofloxacin and fleroxacin in its activity against the Gram-positive cocci, including staphylococci, pneumococci and hemolytic streptococci. Attempts to select grepafloxacin-resistant mutants from a methicillin-resistant Staphylococcus aureus strain were unsuccessful, although ciprofloxacin readily selected quinolone-resistant mutants of that strain. Among Gram-negative bacilli other than Escherichia coli, mutants were readily selected by in vitro exposure to either grepafloxacin or ciprofloxacin. All such mutants showed complete cross resistance to six other quinolones. The four study drugs were active against nalidixic acid-susceptible strains belonging to 13 species of the Enterobacteriaceae, but ciprofloxacin was the most potent drug studied. Against Acinetobacter spp. and Stenotrophomonas maltophilia, grepafloxacin was two- to four-times more active than ciprofloxacin. However, against Pseudomonas aeruginosa, ciprofloxacin was two- to four-times more potent than grepafloxacin. Grepafloxacin and ciprofloxacin were both found to be bactericidal agents.

  2. The effect of different growth regimes on the endophytic bacterial communities of the fern, Dicksonia sellowiana hook (Dicksoniaceae).

    PubMed

    de Araújo Barros, Irene; Luiz Araújo, Welington; Lúcio Azevedo, João

    2010-10-01

    Endophytic bacteria associated with the fern Dicksonia sellowiana were investigated. The bacterial communities from the surface-sterilized pinnae and rachis segments of the plants from the Brazilian Atlantic Rainforest that grew in native field conditions were compared with the bacterial communities from plants grown in greenhouses and plants that were initially grown in greenhouses and then transferred to the forest. From 540 pinnae and 540 rachis segments, 163 (30.2%) and 346 (64.2%) were colonized by bacteria, respectively. The main bacterial genera and species that were isolated included Bacillus spp. ( B. cereus, B. megaterium, B. pumilus and B. subtilis ) , Paenibacillus sp. , Amphibacillus sp. , Gracilibacillus sp. , Micrococcus sp. and Stenotrophomonas spp. ( S. maltophilia and S. nitroreducens ). B. pumilus was the most frequently isolated bacterial species . Amphibacillus and Gracilibacillus were reported as endophytes for the first time. Other commonly found bacterial genera were not observed in D. sellowiana , which may reflect preferences of specific bacterial communities inside this fern or detection limitations due to the isolation procedures. Plants that were grown in greenhouses and plants that were reintroduced into the forest displayed more bacterial genera and species diversity than native field plants, suggesting that reintroduction shifts the bacterial diversity. Endophytic bacteria that displayed antagonistic properties against different microorganisms were detected, but no obvious correlation was found between their frequencies with plant tissues or with plants from different growth regimes. This paper reports the first isolation of endophytic bacteria from a fern.

  3. Frequency of fungi in respiratory samples from Turkish cystic fibrosis patients.

    PubMed

    Güngör, Ozge; Tamay, Zeynep; Güler, Nermin; Erturan, Zayre

    2013-03-01

    An increased isolation of fungi from the respiratory tract of patients with cystic fibrosis (CF) has been reported. The prevalence of different fungi in CF patients from Turkey is not known. Our aim was to determine the frequency of fungi in the respiratory tract of Turkish CF patients. We investigated a total of 184 samples from 48 patients. Samples were inoculated on Medium B+ and CHROMagar Candida. Candida albicans was the predominant yeast isolated [30 patients (62.5%)], followed by C. parapsilosis [6 (12.5%)] and C. dubliniensis 5 (10.4%). Aspergillus fumigatus was the most common filamentous fungus [5 (10.4%)] and non-fumigatus Aspergillus species were isolated from four (8.3%) patients. Staphylococcus aureus was the most frequently detected bacterium in C. albicans positive samples (53.57%). A. fumigatus and Pseudomonas aeruginosa or S. aureus were detected together in 75% of A. fumigatus positive samples each. No statistically significant relationship was detected between growth of yeast and moulds and age, gender, the use of inhaled corticosteroids or tobramycin. No significant correlation was found between the isolation of C. albicans, A. fumigatus and P. aeruginosa, Stenotrophomonas maltophilia or S. aureus, and the isolation of C. albicans and Haemophilus influenzae. Other factors which may be responsible for the increased isolation of fungi in CF need to be investigated.

  4. [Antibacterial activity of sulopenem, a new parenteral penem antibiotic].

    PubMed

    Inoue, E; Komoto, E; Taniyama, Y; Mitsuhashi, S

    1996-04-01

    Sulopenem, a new penem antibiotic, was compared with other antibiotics with regard to in vitro antibacterial and bactericidal activities, stabilization against beta-lactamases, and effect on the release of lipopolysaccharide from Gram-negative bacteria. The results are summarized as follows. 1. Sulopenem showed more potent activities than other antibiotics against both Gram-positive and Gram-negative bacteria except Pseudomonas aeruginosa. 2. Sulopenem showed potent bactericidal activities (MIC/MBC) against both Gram-positive and Gram-negative bacteria. Time kill studies against Staphylococcus aureus, Escherichia coli, Enterobacter cloacae and Citrobacter freundii showed potent bactericidal activities of sulopenem. 3. Sulopenem was found to possess a stronger activity than other antibiotics against beta-lactamase-producing strains except P. aeruginosa and Stenotrophomonas maltophilia. 4. In particular, sulopenem was found to be more stable to the hydrolysis by various beta-lactamases produced by Gram-negative bacteria than any other antibiotics tested. Vmax/Km values of sulopenem were smaller than those of cefotiam for all tested beta-lactamases, which reflected a broad antibacterial spectrum of sulopenem. 5. E. coli ML4707 exposed to sulopenem and imipenem released less endotoxin than did controls at all concentration ranges tested. In contrast, the strain exposed to ceftazidime at bacteriostatic concentrations released a large amount of endotoxin.

  5. A relatively small change in sodium chloride concentration has a strong effect on adhesion of ocular bacteria to contact lenses.

    PubMed

    Cowell, B A; Willcox, M D; Schneider, R P

    1998-06-01

    Adhesion of bacteria to hydrogel lenses is thought to be an initial step of ocular colonization allowing evasion of normal host defences. The salt concentration of media is an important parameter controlling microbial adhesion. Salinity varies from 0.97% NaCl equivalents in the open eye to 0.89% in the closed eye state. In this study, the effect of sodium chloride in the concentration range of 0.8-1.0% (w/v) NaCl on adhesion of ocular bacteria to soft contact lenses was investigated using a static adhesion assay. Pseudomonas aeruginosa was found to adhere to lenses in significantly greater amounts than Serratia marcescens, Flavobacterium meningosepticum, Stenotrophomonas maltophilia and Staphylococcus intermedius. Increasing NaCl from 0.8% to 1.0% (w/v) increased adhesion of all bacteria tested. This adhesion was strong since the organisms could not be removed by washing in low ionic buffer. Adhesion of these organisms did not correlate with their cell surface properties as determined by bacterial adhesion to hydrocarbons (BATH) and retention on sepharose columns.

  6. Effect of carbon on whole-biofilm metabolic response to high doses of streptomycin

    PubMed Central

    Jackson, Lindsay M. D.; Kroukamp, Otini; Wolfaardt, Gideon M.

    2015-01-01

    Biofilms typically exist as complex communities comprising multiple species with the ability to adapt to a variety of harsh conditions. In clinical settings, antibiotic treatments based on planktonic susceptibility tests are often ineffective against biofilm infections. Using a CO2 evolution measurement system we delineated the real-time metabolic response in continuous flow biofilms to streptomycin doses much greater than their planktonic susceptibilities. Stable biofilms from a multispecies culture (containing mainly Pseudomonas aeruginosa and Stenotrophomonas maltophilia), Gram-negative environmental isolates, and biofilms formed by pure culture P. aeruginosa strains PAO1 and PAO1 ΔMexXY (minimum planktonic inhibitory concentrations between 1.5 and 3.5 mg/l), were exposed in separate experiments to 4000 mg/l streptomycin for 4 h after which growth medium resumed. In complex medium, early steady state multispecies biofilms were susceptible to streptomycin exposure, inferred by a cessation of CO2 production. However, multispecies biofilms survived high dose exposures when there was extra carbon in the antibiotic medium, or when they were grown in defined citrate medium. The environmental isolates and PAO1 biofilms showed similar metabolic profiles in response to streptomycin; ceasing CO2 production after initial exposure, with CO2 levels dropping toward baseline levels prior to recovery back to steady state levels, while subsequent antibiotic exposure elicited increased CO2 output. Monitoring biofilm metabolic response in real-time allowed exploration of conditions resulting in vulnerability after antibiotic exposure compared to the resistance displayed following subsequent exposures. PMID:26441887

  7. PME-1, an extended-spectrum β-lactamase identified in Pseudomonas aeruginosa.

    PubMed

    Tian, Guo-Bao; Adams-Haduch, Jennifer M; Bogdanovich, Tatiana; Wang, Hong-Ning; Doi, Yohei

    2011-06-01

    A novel extended-spectrum β-lactamase (ESBL) was identified in a Pseudomonas aeruginosa clinical isolate obtained from a patient admitted to a hospital in Pennsylvania in 2008. The patient had a prolonged hospitalization in a hospital in Dubai, United Arab Emirates, before being transferred to the United States. The novel ESBL, designated PME-1 (Pseudomonas aeruginosa ESBL 1), is a molecular class A, Bush-Jacoby-Medeiros group 2be enzyme and shared 50, 43, and 41% amino acid identity with the L2 β-lactamase of Stenotrophomonas maltophilia, CTX-M-9, and KPC-2, respectively. PME-1 conferred clinically relevant resistance to ceftazidime, cefotaxime, cefepime, and aztreonam in P. aeruginosa PAO1 but not to carbapenems. Purified PME-1 showed good hydrolytic activity against ceftazidime, cefotaxime, and aztreonam, while activity against carbapenems and cefepime could not be measured. PME-1 was inhibited well by β-lactamase inhibitors, including clavulanic acid, sulbactam, and tazobactam. The bla(PME-1) gene was carried by an approximately 9-kb plasmid and flanked by tandem ISCR24 elements.

  8. High-rate biological denitrification in the cyclic rotating-bed biological reactor: Effect of COD/NO3(-), nitrate concentration and salinity and the phylogenetic analysis of denitrifiers.

    PubMed

    Jafari, Seyed Javad; Moussavi, Gholamreza; Yaghmaeian, Kamyar

    2015-12-01

    The effects of COD/NO3(-) ratio, nitrate concentration and salinity was tested on the performance of the CRBR in denitrification with catechol as carbon source. The maximum nitrate reduction attained at COD/NO3(-) ratio of 1. The CRBR operated at optimum COD/NO3(-) ratio could completely denitrify the nitrate at inlet concentration up to 1250mg/L without nitrite accumulation. The maximum denitrification rate in the CRBR was 3.56kgNO3(-)/m(3)d with a nitrate reduction efficiency of 99% when the bioreactor was operated at inlet nitrate loading rate of 3.6kgNO3(-)/m(3)d. The denitrification performance of the CRBR was not affected significantly by NaCl concentrations up to 20g/L. 16S rRNA fragment and phylogenetic analysis identified Pseudomonas resinovorans, Stenotrophomonas maltophilia and Bacillus cereus as the most abundant denitrifiers in biomass. Accordingly, the CRBR is a high-rate bioreactor and appropriate technology for treatment of nitrate-laden industrial wastewaters containing phenolic compounds and salinity.

  9. Isolation and characterization of phenol utilizing bacteria from industrial effluent-contaminated soil and kinetic evaluation of their biodegradation potential.

    PubMed

    Pal Basak, Sreela; Sarkar, Priyabrata; Pal, Priyabrata

    2014-01-01

    Microbial degradation of phenol by pure bacterial species is a well-known approach towards alleviation of environmental pollution. In this study, five phenol-degrading bacterial species designated as CUPS-1 to CUPS-5 were isolated from the oil-effluent dumped sites of Haldia Industrial area of West Bengal, India. Detailed morphological, biochemical and molecular characterization identified CUPS-3 as a novel strain- Stenotrophomonas maltophilia (GU358076), while the others could be identified as Pseudomonas (CUPS-2, 5), Delftia (CUPS-1) and Micrococcus (CUPS-4) genera, respectively. Although all of these strains utilized phenol as their sole carbon source supporting growth, three among them, CUPS-2, CUPS-3 and CUPS-5 proved potential phenol degraders and hence used for further biodegradation studies. Degradation experiments were carried out for several initial phenol concentrations of 500 mg/L, 750 mg/L, 1000 mg/L, 1250 mg/L and 1500 mg/L. The novel strain, CUPS-3 could completely degrade 500 mg/L phenol within 48 h, with 0.0937/h substrate degradation rate and 16.34 mg/L/h substrate consumption rate. The strains degraded phenol via meta-cleavage pathway. Prediction of kinetic parameters of the biodegradation was accomplished Haldane model using the experimental data of degradation rate and phenol concentration as function of time.

  10. Severe Bloodstream Infection due to KPC-Producer E coli in a Renal Transplant Recipient Treated With the Double-Carbapenem Regimen and Analysis of In Vitro Synergy Testing: A Case Report.

    PubMed

    Oliva, Alessandra; Cipolla, Alessia; Gizzi, Francesca; D'Abramo, Alessandra; Favaro, Marco; De Angelis, Massimiliano; Ferretti, Giancarlo; Russo, Gianluca; Iannetta, Marco; Mastroianni, Claudio M; Mascellino, Maria T; Vullo, Vincenzo

    2016-02-01

    Transplant recipients are at high risk of infections caused by multidrug resistant microorganisms. Due to the limited therapeutic options, innovative antimicrobial combinations against carbapenem-resistant Enterobacteriaceae causing severe infections are necessary.A 61-year-old woman with a history of congenital solitary kidney underwent renal transplantation. The postoperative course was complicated by nosocomial pneumonia due to Stenotrophomonas maltophilia and pan-sensitive Escherichia coli, successfully treated with antimicrobial therapy. On postoperative day 22, diagnosis of surgical site infection and nosocomial pneumonia with concomitant bacteremia due to a Klebisella pneumoniae carbapenemase-producer E coli was made. The patient was treated with the double-carbapenem regimen (high dose of meropenem plus ertapenem) and a potent synergistic and bactericidal activity of this un-conventional therapeutic strategy was observed in vitro. Despite a microbiological response with prompt negativity of blood cultures, the patient faced a worse outcome because of severe hemorrhagic shock.The double-carbapenem regimen might be considered as a rescue therapy in those subjects, including transplant recipients, in whom previous antimicrobial combinations failed or when colistin use might be discouraged. Performing in vitro synergy testing should be strongly encouraged in cases of infections caused by pan-drug resistant strains, especially in high-risk patients.

  11. Severe Bloodstream Infection due to KPC-Producer E coli in a Renal Transplant Recipient Treated With the Double-Carbapenem Regimen and Analysis of In Vitro Synergy Testing

    PubMed Central

    Oliva, Alessandra; Cipolla, Alessia; Gizzi, Francesca; D’Abramo, Alessandra; Favaro, Marco; De Angelis, Massimiliano; Ferretti, Giancarlo; Russo, Gianluca; Iannetta, Marco; Mastroianni, Claudio M.; Mascellino, Maria T.; Vullo, Vincenzo

    2016-01-01

    Abstract Transplant recipients are at high risk of infections caused by multidrug resistant microorganisms. Due to the limited therapeutic options, innovative antimicrobial combinations against carbapenem-resistant Enterobacteriaceae causing severe infections are necessary. A 61-year-old woman with a history of congenital solitary kidney underwent renal transplantation. The postoperative course was complicated by nosocomial pneumonia due to Stenotrophomonas maltophilia and pan-sensitive Escherichia coli, successfully treated with antimicrobial therapy. On postoperative day 22, diagnosis of surgical site infection and nosocomial pneumonia with concomitant bacteremia due to a Klebisella pneumoniae carbapenemase-producer E coli was made. The patient was treated with the double-carbapenem regimen (high dose of meropenem plus ertapenem) and a potent synergistic and bactericidal activity of this un-conventional therapeutic strategy was observed in vitro. Despite a microbiological response with prompt negativity of blood cultures, the patient faced a worse outcome because of severe hemorrhagic shock. The double-carbapenem regimen might be considered as a rescue therapy in those subjects, including transplant recipients, in whom previous antimicrobial combinations failed or when colistin use might be discouraged. Performing in vitro synergy testing should be strongly encouraged in cases of infections caused by pan-drug resistant strains, especially in high-risk patients. PMID:26886594

  12. Identification and antimicrobial susceptibility of Alcaligenes xylosoxidans isolated from patients with cystic fibrosis.

    PubMed

    Saiman, L; Chen, Y; Tabibi, S; San Gabriel, P; Zhou, J; Liu, Z; Lai, L; Whittier, S

    2001-11-01

    In the past decade, potential pathogens, including Alcaligenes species, have been increasingly recovered from cystic fibrosis (CF) patients. Accurate identification of multiply antibiotic-resistant gram-negative bacilli is critical to understanding the epidemiology and clinical implications of emerging pathogens in CF. We examined the frequency of correct identification of Alcaligenes spp. by microbiology laboratories affiliated with American CF patient care centers. Selective media, an exotoxin A probe for Pseudomonas aeruginosa, and a commercial identification assay, API 20 NE, were used for identification. The activity of antimicrobial agents against these clinical isolates was determined. A total of 106 strains from 78 patients from 49 CF centers in 22 states were studied. Most (89%) were correctly identified by the referring laboratories as Alcaligenes xylosoxidans. However, 12 (11%) strains were misidentified; these were found to be P. aeruginosa (n = 10), Stenotrophomonas maltophilia (n = 1), and Burkholderia cepacia (n = 1). Minocycline, imipenem, meropenem, piperacillin, and piperacillin-tazobactam were the most active since 51, 59, 51, 50, and 55% of strains, respectively, were inhibited. High concentrations of colistin (100 and 200 microg/ml) inhibited 92% of strains. Chloramphenicol paired with minocycline and ciprofloxacin paired with either imipenem or meropenem were the most active combinations and inhibited 40 and 32%, respectively, of strains. Selective media and biochemical identification proved to be useful strategies for distinguishing A. xylosoxidans from other CF pathogens. Standards for processing CF specimens should be developed, and the optimal method for antimicrobial susceptibility testing of A. xylosoxidans should be determined.

  13. Bloodstream infection after umbilical cord blood transplantation using reduced-intensity stem cell transplantation for adult patients.

    PubMed

    Narimatsu, Hiroto; Matsumura, Tomoko; Kami, Masahiro; Miyakoshi, Shigesaburo; Kusumi, Eiji; Takagi, Shinsuke; Miura, Yuji; Kato, Daisuke; Inokuchi, Chiho; Myojo, Tomohiro; Kishi, Yukiko; Murashige, Naoko; Yuji, Koichiro; Masuoka, Kazuhiro; Yoneyama, Akiko; Wake, Atsushi; Morinaga, Shinichi; Kanda, Yoshinobu; Taniguchi, Shuichi

    2005-06-01

    Bloodstream infection (BSI) is a significant problem after cord blood transplantation (CBT). However, little information has been reported on BSI after reduced-intensity CBT (RI-CBT). We retrospectively reviewed the medical records of 102 patients. The median age of the patients was 55 years (range, 17-79 years). Preparative regimens comprised fludarabine 125 to 150 mg/m 2 , melphalan 80 to 140 mg/m 2 , or busulfan 8 mg/kg and total body irradiation 2 to 8 Gy. Prophylaxis against graft-versus-host disease comprised cyclosporin or tacrolimus. BSI developed within 100 days of RI-CBT in 32 patients. The cumulative incidence of BSI was 25% at day 30 and 32% at day 100. The median onset was day 15 (range, 1-98 days). Causative organisms included Pseudomonas aeruginosa (n = 12), Staphylococcus epidermidis (n = 11), Staphylococcus aureus (n = 6), Enterococcus faecium (n = 4), Enterococcus faecalis (n = 4), Stenotrophomonas maltophilia (n = 4), and others (n = 7). Of the 32 patients with BSI, 25 (84%) died within 100 days after RI-CBT. BSI was the direct cause of death in 8 patients (25%). Univariate analysis failed to identify any significant risk factors. BSI clearly represents a significant and fatal complication after RI-CBT. Further studies are warranted to determine clinical characteristics, identify patients at high risk of BSI, and establish therapeutic strategies.

  14. Xenopus laevis oocytes infected with multi-drug-resistant bacteria: implications for electrical recordings.

    PubMed

    O'Connell, Denice; Mruk, Karen; Rocheleau, Jessica M; Kobertz, William R

    2011-08-01

    The Xenopus laevis oocyte has been the workhorse for the investigation of ion transport proteins. These large cells have spawned a multitude of novel techniques that are unfathomable in mammalian cells, yet the fickleness of the oocyte has driven many researchers to use other membrane protein expression systems. Here, we show that some colonies of Xenopus laevis are infected with three multi-drug-resistant bacteria: Pseudomonas fluorescens, Pseudomonas putida, and Stenotrophomonas maltophilia. Oocytes extracted from infected frogs quickly (3-4 d) develop multiple black foci on the animal pole, similar to microinjection scars, which render the extracted eggs useless for electrical recordings. Although multi-drug resistant, the bacteria were susceptible to amikacin and ciprofloxacin in growth assays. Supplementing the oocyte storage media with these two antibiotics prevented the appearance of the black foci and afforded oocytes suitable for whole-cell recordings. Given that P. fluorescens associated with X. laevis has become rapidly drug resistant, it is imperative that researchers store the extracted oocytes in the antibiotic cocktail and not treat the animals harboring the multi-drug-resistant bacteria.

  15. Inoculation of paperboard mill sludge versus mixed culture bacteria for hydrogen production from paperboard mill wastewater.

    PubMed

    Farghaly, Ahmed; Tawfik, Ahmed; Danial, Amal

    2016-02-01

    A comparative evaluation of paperboard mill sludge (PMS) versus mixed culture bacteria (MCB) as inoculum for hydrogen production from paperboard mill wastewater (PMW) was investigated. The experiments were conducted at different initial cultivation pHs, inoculums to substrate ratios (ISRs gVS/gCOD), and hydraulic retention times (HRTs). The peak hydrogen yield (HY) of 5.29 ± 0.16 and 1.22 ± 0.11 mmol/gCODinitial was occurred at pH = 5 for MCB and PMS, respectively. At pH of 5, the HY and COD removal achieved the highest values of 2.26 ± 0.14 mmol/gCODinitial and 86 ± 1.6% at ISR = 6 for MCB, and 2.38 ± 0.25 mmol/gCODinitial and 60.4 ± 2.5% at ISRs = 3 for PMS. The maximum hydrogen production rate was 93.75 ± 8.9 mmol/day at HRT = 9.6 h from continuous upflow anaerobic reactor inoculated with MCB. Meanwhile, the 16S ribosomal RNA (rRNA) gene fragments indicated a dominance of a novel hydrogen-producing bacterium of Stenotrophomonas maltophilia for PMS microbial community. On the other hand, Escherichia fergusonii and Enterobacter hormaechei were the predominant species for MCB.

  16. Multi-Channel Microfluidic Biosensor Platform Applied for Online Monitoring and Screening of Biofilm Formation and Activity

    PubMed Central

    Bruchmann, Julia; Sachsenheimer, Kai; Rapp, Bastian E.; Schwartz, Thomas

    2015-01-01

    Bacterial colonization of surfaces and interfaces has a major impact on various areas including biotechnology, medicine, food industries, and water technologies. In most of these areas biofilm development has a strong impact on hygiene situations, product quality, and process efficacies. In consequence, biofilm manipulation and prevention is a fundamental issue to avoid adverse impacts. For such scenario online, non-destructive biofilm monitoring systems become important in many technical and industrial applications. This study reports such a system in form of a microfluidic sensor platform based on the combination of electrical impedance spectroscopy and amperometric current measurement, which allows sensitive online measurement of biofilm formation and activity. A total number of 12 parallel fluidic channels enable real-time online screening of various biofilms formed by different Pseudomonas aeruginosa and Stenotrophomonas maltophilia strains and complex mixed population biofilms. Experiments using disinfectant and antibiofilm reagents demonstrate that the biofilm sensor is able to discriminate between inactivation/killing of bacteria and destabilization of biofilm structures. The impedance and amperometric sensor data demonstrated the high dynamics of biofilms as a consequence of distinct responses to chemical treatment strategies. Gene expression of flagellar and fimbrial genes of biofilms grown inside the microfluidic system supported the detected biofilm growth kinetics. Thus, the presented biosensor platform is a qualified tool for assessing biofilm formation in specific environments and for evaluating the effectiveness of antibiofilm treatment strategies. PMID:25706987

  17. Susceptibility to levofloxacin of clinical isolates of bacteria from intensive care and haematology/oncology patients in Switzerland: a multicentre study.

    PubMed

    Siegrist, H H; Nepa, M C; Jacquet, A

    1999-06-01

    The objective of this study was to examine the susceptibility of clinical isolates to levofloxacin, a fluoroquinolone with extended activity against Gram-positive bacteria, and other antibiotics in 12 Swiss clinical microbiology laboratories using the NCCLS disc diffusion technique. Isolates were prospectively collected from intensive care units (ICUs (59%), oncology wards (7%) and other units with haematology/oncology patients (34%) from June 1995 to March 1996. The levofloxacin breakpoints used were as recommended by the manufacturer. A total of 310 Gram-positive and 580 Gram-negative isolates from the respiratory tract (36%), skin/wounds (12%), blood (16%), urine (17%) and other sources (19%) were tested. The percentage of isolates susceptible to levofloxacin was 100% for Enterococcus spp. (38 strains), Streptococcus agalactiae (13), Streptococcus pneumoniae (65), Acinetobacter spp. (11), Citrobacter diversus (6), Citrobacter freundii (17), Klebsiella oxytoca (39), Morganella morganii (16), Proteus mirabilis (20), Proteus vulgaris (23), Serratia spp. (19), Stenotrophomonas maltophilia (10) and Haemophilus influenzae (41). The percentage of isolates susceptible to levofloxacin for Staphylococcus aureus (95 strains, including 2% MRSA) was 94%, coagulase-negative staphylococci (85) 65%, Enterobacter spp. (75) 99%, Escherichia coli (111) 97%, Klebsiella pneumoniae (45) 98% and Pseudomonas aeruginosa (124) 87%. In conclusion, levofloxacin is a new fluoroquinolone to which the most common clinical isolates in Switzerland are susceptible. The susceptibility of Enterococcus spp. and S. pneumoniae to levofloxacin was particularly remarkable. This compound appears to be a promising therapeutic alternative for the treatment of Gram-positive infections.

  18. Comparative in vitro potency of amoxycillin-clavulanic acid and four oral agents against recent North American clinical isolates from a global surveillance study.

    PubMed

    Hoban, D J; Bouchillon, S K; Johnson, J L; Zhanel, G G; Butler, D L; Saunders, K A; Miller, L A; Poupard, J A

    2003-05-01

    The in vitro activity of amoxycillin-clavulanic acid was compared with four comparator oral antimicrobial agents; ampicillin, azithromycin, cefuroxime and trimethoprim-sulphamethoxazole against 4536 recent clinical isolates covering 29 species isolated in the US and Canada between 1997 and 1999. Based upon Minimum inhibitory concentrations (MICs), amoxycillin-clavulanic acid was the most active agent against many Gram-positive species and phenotypes including methicillin susceptible Staphylococcus aureus (MSSA) Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pyogenes, Streptococcus pneumoniae including penicillin intermediate and macrolide resistant strains and was as active as ampicillin against Streptococcus agalactiae, penicillin resistant S. pneumoniae and viridans streptococci. Against Enterobacteriaceae amoxycillin-clavulanic acid in general, displayed weak activity with only Proteus mirabilis and Proteus vulgaris displaying levels of susceptibility above the 90th percentile. Amoxycillin-clavulanic acid had significant activity against many species of Gram-negative non-Enterobacteriaceae including Haemophilus influenzae, Haemophilus parainfluenzae and Moraxella catarrhalis but negligible activity against Burkholderia cepacia, Pseudomonas aeruginosa and Stenotrophomonas maltophilia. Amoxycillin-clavulanic acid continues to retain excellent activity against the majority of targeted pathogens despite 20 years of clinical use.

  19. Comparative in vitro activity of gemifloxacin, ciprofloxacin, levofloxacin and ofloxacin in a North American surveillance study.

    PubMed

    Hoban, D J; Bouchillon, S K; Johnson, J L; Zhanel, G G; Butler, D L; Miller, L A; Poupard, J A

    2001-01-01

    The in vitro activity of gemifloxacin, a new fluoroquinolone, was compared to three marketed fluoroquinolones; ciprofloxacin, levofloxacin and ofloxacin against over 4,000 recent clinical isolates covering 29 species isolated in the United States and Canada between 1997-1999. Based on MIC(90)s, gemifloxacin was the most potent fluoroquinolone tested against a majority of Gram-positive isolates: Streptococcus pneumoniae, penicillin resistant S. pneumoniae, macrolide resistant S. pneumoniae, ciprofloxacin non-susceptible (MIC > or = 4 microg/mL) S. pneumoniae, S. pyogenes, S. agalactiae, viridans streptococci, Enterococcus faecalis, methicillin susceptible Staphylococcus aureus, methicillin resistant S. aureus, S. epidermidis, S. hemolyticus, and S. saprophyticus. Against Enterobacteriaceae and aerobic non-Enterobacteriaceae Gram-negatives, gemifloxacin was usually comparable to ciprofloxacin and levofloxacin and more potent than ofloxacin for the following species: Citrobacter freundii, Enterobacter aerogenes, E. cloacae, Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Morganella morganii, Proteus mirabilis, P. vulgaris, Providencia stuartii, Serratia marcescens, Acinetobacter lwoffii, A. baumannii, Burkholderia cepacia, Haemophilus influenzae, H. parainfluenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. Gemifloxacin was generally 16-64 fold more potent than the other fluoroquinolones tested against Gram-positive organisms and retains excellent activity comparable with ciprofloxacin and levofloxacin against a majority of Gram-negative pathogens.

  20. Improved catalytic efficiency, thermophilicity, anti-salt and detergent tolerance of keratinase KerSMD by partially truncation of PPC domain

    PubMed Central

    Fang, Zhen; Zhang, Juan; Du, Guocheng; Chen, Jian

    2016-01-01

    The keratinase from Stenotrophomonas maltophilia (KerSMD) is known for its high activity and pH stability in keratin degradation. However, catalytic efficiency and detergent tolerability need to be improved in order to be used for industrial application. In this work, we obtained several keratinase variants with enhanced catalytic efficiency, thermophilicity, and anti-salt and detergent tolerability by partially truncating the PPC domain of KerSMD. The variants all showed improved catalytic efficiency to synthetic substrate AAPF, with the V355 variant having the highest kcat /Km value of 143.6 s−1 mM−1. The truncation of keratinase had little effect on alkaline stability but obviously decreased collagenase activity, developing its potential application in leather treatment. The variants V380, V370, and V355 were thermophilic, with a 1.7-fold enhancement of keratinlytic activity at 60 °C when compared to the wild type. The entire truncation of PPC domain obtained the variant V355 with improved tolerance to alkalinity, salt, chaotropic agents, and detergents. The V355 variant showed more than a 40% improvement in activity under 15% (w/v) NaCl or 4% (w/v) SDS solution, showing excellent stability under harsh washing and unhairing conditions. Our work investigated how protein engineering affects the function of PPC domain of KerSMD. PMID:27298079

  1. Ciprofloxacin-resistant gram-negative bacilli in the fecal microflora of children.

    PubMed

    Qin, Xuan; Razia, Yasmin; Johnson, James R; Stapp, Jennifer R; Boster, Daniel R; Tsosie, Treva; Smith, Donna L; Braden, Christopher R; Gay, Kathryn; Angulo, Frederick J; Tarr, Phillip I

    2006-10-01

    The extent to which antibiotic-resistant bacteria are excreted by humans who have not been exposed to antibiotics is not known. Children, who rarely receive fluoroquinolones, provide opportunities to assess the frequency of fecal excretion by fluoroquinolone-naïve hosts of fluoroquinolone-resistant gram-negative bacilli. Fresh nondiarrheal stools from children were processed by screening them on agar containing ciprofloxacin to recover ciprofloxacin-resistant gram-negative bacilli. Resistant isolates were identified, and ciprofloxacin MICs were determined. Resistant Escherichia coli isolates were also analyzed for urovirulence-associated loci. Thirteen (2.9%) of 455 stools yielded ciprofloxacin-resistant E. coli (seven children), Stenotrophomonas maltophilia (four children), and Achromobacter xylosoxidans and Enterobacter aerogenes (one child each). Neither the subjects themselves nor members of their households used fluoroquinolones in the 4 weeks preceding collection. Six of the seven resistant E. coli isolates belonged to phylogenetic groups B2 and D, in which extraintestinal pathogenic E. coli bacteria are frequently found. All resistant E. coli isolates contained at least three putative E. coli virulence loci. Most ciprofloxacin-resistant bacteria were resistant to additional antibiotics. Potentially pathogenic bacteria that are resistant to therapeutically important antimicrobial agents are excreted by some humans, despite these persons' lack of exposure to the particular drugs. The sources of these resistant organisms are unknown. This underrecognized reservoir of drug-resistant potential pathogens poses public health challenges.

  2. Host-Defense Peptides with Therapeutic Potential from Skin Secretions of Frogs from the Family Pipidae

    PubMed Central

    Conlon, J. Michael; Mechkarska, Milena

    2014-01-01

    Skin secretions from frogs belonging to the genera Xenopus, Silurana, Hymenochirus, and Pseudhymenochirus in the family Pipidae are a rich source of host-defense peptides with varying degrees of antimicrobial activities and cytotoxicities to mammalian cells. Magainin, peptide glycine-leucine-amide (PGLa), caerulein-precursor fragment (CPF), and xenopsin-precursor fragment (XPF) peptides have been isolated from norepinephrine-stimulated skin secretions from several species of Xenopus and Silurana. Hymenochirins and pseudhymenochirins have been isolated from Hymenochirus boettgeri and Pseudhymenochirus merlini. A major obstacle to the development of these peptides as anti-infective agents is their hemolytic activities against human erythrocytes. Analogs of the magainins, CPF peptides and hymenochirin-1B with increased antimicrobial potencies and low cytotoxicities have been developed that are active (MIC < 5 μM) against multidrug-resistant clinical isolates of Staphylococcus aureus, Escherichia coli, Acinetobacter baumannii, Stenotrophomonas maltophilia and Klebsiella pneumoniae. Despite this, the therapeutic potential of frog skin peptides as anti-infective agents has not been realized so that alternative clinical applications as anti-cancer, anti-viral, anti-diabetic, or immunomodulatory drugs are being explored. PMID:24434793

  3. Time-kill studies of tea tree oils on clinical isolates.

    PubMed

    May, J; Chan, C H; King, A; Williams, L; French, G L

    2000-05-01

    Tea tree oil has recently emerged as an effective topical antimicrobial agent active against a wide range of organisms. Tea tree oil may have a clinical application in both the hospital and community, especially for clearance of methicillin-resistant Staphylococcus aureus (MRSA) carriage or as a hand disinfectant to prevent cross-infection with Gram-positive and Gramnegative epidemic organisms. Our study, based on the time-kill approach, determined the kill rate of tea tree oil against several multidrug-resistant organisms, including MRSA, glycopeptide-resistant enterococci, aminoglycoside-resistant klebsiellae, Pseudomonas aeruginosa and Stenotrophomonas maltophilia, and also against sensitive microorganisms. The study was performed with two chemically different tea tree oils. One was a standard oil and the other was Clone 88 extracted from a specially bred tree, which has been selected and bred for increased activity and decreased skin irritation. Our results confirm that the cloned oil had increased antimicrobial activity when compared with the standard oil. Most results indicated that the susceptibility pattern and Gram reaction of the organism did not influence the kill rate. A rapid killing time (less than 60 min) was achieved with both tea tree oils with most isolates, but MRSA was killed more slowly than other organisms.

  4. [Bacterial isolates from respiratory samples of pediatric patients with cystic fibrosis and their distribution by ages].

    PubMed

    Busquets, Natalia P; Baroni, María R; Ochoteco, María C; Zurbriggen, María L; Virgolini, Stella; Meneghetti, Fernando G

    2013-01-01

    The bacterial isolates from respiratory samples of 50 pediatric patients with cystic fibrosis, their distribution by ages and antimicrobial resistance pattern as well as the intermittence of isolations and coinfections, were investigated. Staphylococcus aureus was isolated in 72 % of patients, followed by Pseudomonas aeruginosa (58 %), Haemophilus. influenzae (56 %), and the Burkholderia cepacia complex (12 %). The frequency of resistance of P. aeruginosa isolates to β-lactam antibiotics was low (13.8 %). Fifty percent of S. aureus isolates was methicillin-resistant, and 57.1 % of H. influenza was ampicillin resistant due to β-lactamase production. In children under 4 years-old, S. aureus was predominant, followed by P. aeruginosa and H. influenzae. This order of predominance was observed in all the groups studied, except in that of children between 10 and 14 years-old. Stenotrophomonas maltophilia and Achromobacter xylosoxidans isolates were intermittent and accompanied by other microorganisms. Finally, we observed a great variety of bacterial species, which imposes stringent performance requirements for microbiological studies in all respiratory samples of these patients.

  5. Genome survey and characterization of endophytic bacteria exhibiting a beneficial effect on growth and development of poplar trees.

    PubMed

    Taghavi, Safiyh; Garafola, Craig; Monchy, Sébastien; Newman, Lee; Hoffman, Adam; Weyens, Nele; Barac, Tanja; Vangronsveld, Jaco; van der Lelie, Daniel

    2009-02-01

    The association of endophytic bacteria with their plant hosts has a beneficial effect for many different plant species. Our goal is to identify endophytic bacteria that improve the biomass production and the carbon sequestration potential of poplar trees (Populus spp.) when grown in marginal soil and to gain an insight in the mechanisms underlying plant growth promotion. Members of the Gammaproteobacteria dominated a collection of 78 bacterial endophytes isolated from poplar and willow trees. As representatives for the dominant genera of endophytic gammaproteobacteria, we selected Enterobacter sp. strain 638, Stenotrophomonas maltophilia R551-3, Pseudomonas putida W619, and Serratia proteamaculans 568 for genome sequencing and analysis of their plant growth-promoting effects, including root development. Derivatives of these endophytes, labeled with gfp, were also used to study the colonization of their poplar hosts. In greenhouse studies, poplar cuttings (Populus deltoides x Populus nigra DN-34) inoculated with Enterobacter sp. strain 638 repeatedly showed the highest increase in biomass production compared to cuttings of noninoculated control plants. Sequence data combined with the analysis of their metabolic properties resulted in the identification of many putative mechanisms, including carbon source utilization, that help these endophytes to thrive within a plant environment and to potentially affect the growth and development of their plant hosts. Understanding the interactions between endophytic bacteria and their host plants should ultimately result in the design of strategies for improved poplar biomass production on marginal soils as a feedstock for biofuels.

  6. The role of wood-inhabiting bacteria in pine wilt disease

    PubMed Central

    Zhao, Bo Guang; Tao, Jian; Ju, Yun Wei; Wang, Peng Kai; Ye, Jian Ling

    2011-01-01

    The pathogenicity of the pine wood nematode (PWN), Bursaphelenchus xylophilus together with the bacteria isolated from black pine (Pinus thunbergii) bark inoculated to axenic black pine seedlings, significantly exceeded that of the axenic PWNs alone, demonstrating that the bacteria play an important role in pine wilt disease. Inoculation of seedlings with bacteria-free culture filtrates of the seven isolates from the dead seedlings from the above experiment showed that all isolate filtrates killed the seedlings within 8 days. Identification of the bacteria using 16S rDNA sequencing showed that the isolates belonged to strains By253Ydz-fq, S209, 210-50 and 210-50 in Bacillus and the DN1.1 strain of Stenotrophomonas maltophilia, respectively. Completing Koch’s postulates using the seven bacterial isolates to inoculate pine seedlings showed that all the seedlings that received aseptic PWNs mixed with the seven bacterial isolates died within 18 days post inoculation, while those inoculated with ‘wild’ PWNs died 16 days post inoculation. No disease symptoms developed on seedlings that received sterile water or aseptic PWNs. The horizontal transfer of the pathogenic bacteria may explain differences in bacterial species carried by PWN in different geographic areas. PMID:23430766

  7. Risk factors for hospital-acquired pneumonia caused by carbapenem-resistant Gram-negative bacteria in critically ill patients: a multicenter study in Korea.

    PubMed

    Kim, Tark; Chong, Yong Pil; Park, Seong Yeon; Jeon, Min-Hyok; Choo, Eun Joo; Chung, Jin-Won; Lee, Hyun Kyung; Moon, Chisook; Kim, Dong-Min; Peck, Kyong Ran; Kim, Yang Soo

    2014-04-01

    We performed a case-control study to identify risk factors of carbapenem-resistant Gram-negative bacteria (CRGNB) as an increasing cause of hospital-acquired pneumonia (HAP). The study included critically ill adult patients with HAP whose microbial etiology was identified at eight tertiary centers in Korea between June 2008 and December 2009. Eighty two patients with 86 isolates of CRGNB (62 Acinetobacter baumannii, 14 Pseudomonas aeruginosa, and 10 Stenotrophomonas maltophilia) were included in the case group, and 122 patients with carbapenem-susceptible Gram-negative bacteria were included in the control group. Diabetes mellitus (adjusted odds ratio [aOR] 2.82, 95% confidence interval [95% CI] 1.25-6.38), radiologic score ≥5 (aOR 4.56, 95% CI 2.36-8.81), prior fluoroquinolone (aOR 2.39. 95% CI = 1.07-5.35), or carbapenem usage (aOR 2.82, 95% CI 1.75-17.83) were found to be independent risk factors. Fluoroquinolone and carbapenem should be cautiously used to avoid HAP caused by CRGNB.

  8. Stent hypersensitivity and infection in sinus cavities

    PubMed Central

    Soufras, George D.; Hahalis, George

    2013-01-01

    Persistent mucosal inflammation, granulation tissue formation, hypersensitivity, and multifactorial infection are newly described complications of retained drug-eluting stents from endoscopic sinus surgery for refractory rhinosinusitis. In an important report published in Allergy and Rhinology, a 45-year-old male patient suffering from recalcitrant chronic rhinosinusitis underwent functional endoscopic sinus surgery and was found, for the first time, to have steroid-eluting catheters that were inadvertently left in the ethmoid and frontal sinuses. The retained catheters had caused persistent mucosal inflammation and formation of granulation tissue denoting hypersensitivity reaction. These consequences had induced perpetuation of symptoms of chronic rhinosinusitis. Meticulous removal of the retained stents with the nitinol wings from inflamed tissues of the frontal, ethmoidal, and sphenoethmoidal recesses in which they were completely imbedded was successfully performed without polypoid regrowth. Cultures of specimens taken from both left and right stents showed heavy growth of Stenotrophomonas maltophilia and moderate growth of Klebsiella oxytoca, coagulase negative Staphylococcus, and beta-hemolytic Streptococcus anginosus. Fungal infection was not detected. The current knowledge and experience regarding stent hypersensitivity and infection in relation with the use of stents in sinus cavities is reviewed. PMID:24498522

  9. The antibacterial properties of Malaysian tualang honey against wound and enteric microorganisms in comparison to manuka honey

    PubMed Central

    Tan, Hern Tze; Rahman, Rosliza Abdul; Gan, Siew Hua; Halim, Ahmad Sukari; Hassan, Siti Asma'; Sulaiman, Siti Amrah; BS, Kirnpal-Kaur

    2009-01-01

    Background Antibiotic resistance of bacteria is on the rise, thus the discovery of alternative therapeutic agents is urgently needed. Honey possesses therapeutic potential, including wound healing properties and antimicrobial activity. Although the antimicrobial activity of honey has been effectively established against an extensive spectrum of microorganisms, it differs depending on the type of honey. To date, no extensive studies of the antibacterial properties of tualang (Koompassia excelsa) honey on wound and enteric microorganisms have been conducted. The objectives of this study were to conduct such studies and to compare the antibacterial activity of tualang honey with that of manuka honey. Methods Using a broth dilution method, the antibacterial activity of tualang honey against 13 wound and enteric microorganisms was determined; manuka honey was used as the control. Different concentrations of honey [6.25-25% (w/v)] were tested against each type of microorganism. Briefly, two-fold dilutions of honey solutions were tested to determine the minimum inhibitory concentration (MIC) against each type of microorganism, followed by more assays within a narrower dilution range to obtain more precise MIC values. MICs were determined by both visual inspection and spectrophotometric assay at 620 nm. Minimum bactericidal concentration (MBC) also was determined by culturing on blood agar plates. Results By visual inspection, the MICs of tualang honey ranged from 8.75% to 25% compared to manuka honey (8.75-20%). Spectrophotometric readings of at least 95% inhibition yielded MIC values ranging between 10% and 25% for both types of honey. The lowest MBC for tualang honey was 20%, whereas that for manuka honey was 11.25% for the microorganisms tested. The lowest MIC value (8.75%) for both types of honey was against Stenotrophomonas maltophilia. Tualang honey had a lower MIC (11.25%) against Acinetobacter baumannii compared to manuka honey (12.5%). Conclusion Tualang honey

  10. Fourier Transform Infrared Spectroscopy for Rapid Identification of Nonfermenting Gram-Negative Bacteria Isolated from Sputum Samples from Cystic Fibrosis Patients▿

    PubMed Central

    Bosch, Alejandra; Miñán, Alejandro; Vescina, Cecilia; Degrossi, José; Gatti, Blanca; Montanaro, Patricia; Messina, Matías; Franco, Mirta; Vay, Carlos; Schmitt, Juergen; Naumann, Dieter; Yantorno, Osvaldo

    2008-01-01

    The accurate and rapid identification of bacteria isolated from the respiratory tract of patients with cystic fibrosis (CF) is critical in epidemiological studies, during intrahospital outbreaks, for patient treatment, and for determination of therapeutic options. While the most common organisms isolated from sputum samples are Pseudomonas aeruginosa, Staphylococcus aureus, and Haemophilus influenzae, in recent decades an increasing fraction of CF patients has been colonized by other nonfermenting (NF) gram-negative rods, such as Burkholderia cepacia complex (BCC) bacteria, Stenotrophomonas maltophilia, Ralstonia pickettii, Acinetobacter spp., and Achromobacter spp. In the present study, we developed a novel strategy for the rapid identification of NF rods based on Fourier transform infrared spectroscopy (FTIR) in combination with artificial neural networks (ANNs). A total of 15 reference strains and 169 clinical isolates of NF gram-negative bacteria recovered from sputum samples from 150 CF patients were used in this study. The clinical isolates were identified according to the guidelines for clinical microbiology practices for respiratory tract specimens from CF patients; and particularly, BCC bacteria were further identified by recA-based PCR followed by restriction fragment length polymorphism analysis with HaeIII, and their identities were confirmed by recA species-specific PCR. In addition, some strains belonging to genera different from BCC were identified by 16S rRNA gene sequencing. A standardized experimental protocol was established, and an FTIR spectral database containing more than 2,000 infrared spectra was created. The ANN identification system consisted of two hierarchical levels. The top-level network allowed the identification of P. aeruginosa, S. maltophilia, Achromobacter xylosoxidans, Acinetobacter spp., R. pickettii, and BCC bacteria with an identification success rate of 98.1%. The second-level network was developed to differentiate the four

  11. [Post-marketing surveillance of antibacterial activities of cefozopran against various clinical isolates--II. Gram-negative bacteria].

    PubMed

    Igari, Jun; Oguri, Toyoko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

    2002-02-01

    As a post-marketing surveillance, the in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates were yearly evaluated and compared with those of other cephems, oxacephems, penicillins, monobactams, and carbapenems. Changes in CZOP susceptibility for the bacteria were also evaluated with the bacterial resistance ratio calculated with the breakpoint MIC. Twenty-five species (3,362 strains) of Gram-negative bacteria were isolated from the clinical materials annually collected from 1996 to 2000, and consisted of Moraxella (Branhamella) catarrhalis (n = 136), Haemophilus influenzae (n = 289), Escherichia coli (n = 276), Klebsiella pneumoniae (n = 192), Klebsiella oxytoca (n = 157), Enterobacter cloacae (n = 189), Enterobacter aerogenes (n = 93), Serratia marcescens (n = 172), Serratia liquefaciens (n = 24), Citrobacter freundii (n = 177), Citrobacter koseri (n = 70), Proteus mirabilis (n = 113), Proteus vulgaris (n = 89), Morganella morganii (n = 116), Providencia spp. (n = 41), Pseudomonas aeruginosa (n = 290), Pseudomonas fluorescens (n = 56), Pseudomonas putida (n = 63), Acinetobacter baumannii (n = 146), Acinetobacter lwoffii (n = 34), Burkholderia cepacia (n = 101), Stenotrophomonas maltophilia (n = 169), Bacteroides fragilis group (n = 196), and Prevotella/Porphyromonas (n = 173). An antibacterial activity of CZOP against E. coli, K. pneumoniae, K. oxytoca, and S. marcescens was potent and consistent with or more preferable than the study results obtained until the new drug application approval. MIC90 of CZOP against M.(B.) catarrhalis, C. koseri, and P. aeruginosa was not considerably changed and consistent with the study results obtained until the new drug application approval. MIC90 of CZOP against E. cloacae, E. aerogenes, and P. mirabilis increased year by year. The increase in MIC90 of CZOP against E. aerogenes and P. mirabilis, however, was not considered to be an obvious decline in susceptibility. In

  12. A novel salt-tolerant chitobiosidase discovered by genetic screening of a metagenomic library derived from chitin-amended agricultural soil.

    PubMed

    Cretoiu, Mariana Silvia; Berini, Francesca; Kielak, Anna Maria; Marinelli, Flavia; van Elsas, Jan Dirk

    2015-10-01

    Here, we report on the construction of a metagenomic library from a chitin-amended disease-suppressive agricultural soil and its screening for genes that encode novel chitinolytic enzymes. The library, constructed in fosmids in an Escherichia coli host, comprised 145,000 clones containing inserts of sizes of 21 to 40 kb, yielding a total of approximately 5.8 GB of cloned soil DNA. Using genetic screenings by repeated PCR cycles aimed to detect gene sequences of the bacterial chitinase A-class (hereby named chi A genes), we identified and characterized five fosmids carrying candidate genes for chitinolytic enzymes. The analysis thus allowed access to the genomic (fosmid-borne) context of these genes. Using the chiA-targeted PCR, which is based on degenerate primers, the five fosmids all produced amplicons, of which the sequences were related to predicted chitinolytic enzyme-encoding genes of four different host organisms, including Stenotrophomonas maltophilia. Sequencing and de novo annotation of the fosmid inserts confirmed that each one of these carried one or more open reading frames that were predicted to encode enzymes active on chitin, including one for a chitin deacetylase. Moreover, the genetic contexts in which the putative chitinolytic enzyme-encoding genes were located were unique per fosmid. Specifically, inserts from organisms related to Burkholderia sp., Acidobacterium sp., Aeromonas veronii, and the chloroflexi Nitrolancetus hollandicus and/or Ktedonobacter racemifer were obtained. Remarkably, the S. maltophilia chiA-like gene was found to occur in two different genetic contexts (related to N. hollandicus/K. racemifer), indicating the historical occurrence of genetic reshufflings in this part of the soil microbiota. One fosmid containing the insert composed of DNA from the N. hollandicus-like organism (denoted 53D1) was selected for further work. Using subcloning procedures, its putative gene for a chitinolytic enzyme was successfully brought to

  13. Assessment of the Microbial Constituents of the Home Environment of Individuals with Cystic Fibrosis (CF) and Their Association with Lower Airways Infections

    PubMed Central

    Heirali, Alya; McKeon, Suzanne; Purighalla, Swathi; Storey, Douglas G.; Rossi, Laura; Costilhes, Geoffrey; Drews, Steven J.; Rabin, Harvey R.; Surette, Michael G.; Parkins, Michael D.

    2016-01-01

    Introduction Cystic fibrosis (CF) airways are colonized by a polymicrobial community of organisms, termed the CF microbiota. We sought to define the microbial constituents of the home environment of individuals with CF and determine if it may serve as a latent reservoir for infection. Methods Six patients with newly identified CF pathogens were included. An investigator collected repeat sputum and multiple environmental samples from their homes. Bacteria were cultured under both aerobic and anaerobic conditions. Morphologically distinct colonies were selected, purified and identified to the genus and species level through 16S rRNA gene sequencing. When concordant organisms were identified in sputum and environment, pulsed-field gel electrophoresis (PFGE) was performed to determine relatedness. Culture-independent bacterial profiling of each sample was carried out by Illumina sequencing of the V3 region of the 16s RNA gene. Results New respiratory pathogens prompting investigation included: Mycobacterium abscessus(2), Stenotrophomonas maltophilia(3), Pseudomonas aeruginosa(3), Pseudomonas fluorescens(1), Nocardia spp.(1), and Achromobacter xylosoxidans(1). A median 25 organisms/patient were cultured from sputum. A median 125 organisms/home were cultured from environmental sites. Several organisms commonly found in the CF lung microbiome were identified within the home environments of these patients. Concordant species included members of the following genera: Brevibacterium(1), Microbacterium(1), Staphylococcus(3), Stenotrophomonas(2), Streptococcus(2), Sphingomonas(1), and Pseudomonas(4). PFGE confirmed related strains (one episode each of Sphinogomonas and P. aeruginosa) from the environment and airways were identified in two patients. Culture-independent assessment confirmed that many organisms were not identified using culture-dependent techniques. Conclusions Members of the CF microbiota can be found as constituents of the home environment in individuals with

  14. Bacterial reduction of selenium in coal mine tailings pond sediment

    SciTech Connect

    Siddique, T.; Arocena, J.M.; Thring, R.W.; Zhang, Y.Q.

    2007-05-15

    Sediment from a storage facility for coal tailings solids was assessed for its capacity to reduce selenium (Se) by native bacterial community. One Se{sup 6+}-reducing bacterium Enterobacter hormaechei (Tar11) and four Se{sup 4+}-reducing bacteria, Klebsiella pneumoniae (Tar1), Pseudomonasfluorescens (Tar3), Stenotrophomonas maltophilia (Tar6), and Enterobacter amnigenus (Tar8) were isolated from the sediment. Enterobacter horinaechei removed 96% of the added Se{sup 6+} (0.92 mg L{sup -1} from the effluents when Se6+ was determined after 5 d of incubation. Analysis of the red precipitates showed that Se{sup 6+} reduction resulted in the formation of spherical particles ({lt}1.0 {mu} m) of Se 0 as observed under scanning electron microscope (SEM) and confirmed by EDAX. Selenium speciation was performed to examine the fate of the added Se{sup 6+} in the sediment with or without addition of Enterobacter hormaechei cells. More than 99% of the added Se{sup 6+} (about 2.5 mg L{sup -1}) was transformed in the nonsterilized sediment (without Enterobacter hormaechei cells) as well as in the sterilized (heat-killed) sediment (with Enterobacter hormaechei cells). The results of this study suggest that the lagoon sediments at the mine site harbor Se{sup 6+}- and Se{sup 4+} -reducing bacteria and may be important sinks for soluble Se (Se{sup 6+} and Se{sup 4+}). Enterobacter hormaechei isolated from metal-contaminated sediment may have potential application in removing Se from industrial effluents.

  15. Acquisition and Evolution of Plant Pathogenesis–Associated Gene Clusters and Candidate Determinants of Tissue-Specificity in Xanthomonas

    PubMed Central

    Van Sluys, Marie-Anne; White, Frank F.; Ryan, Robert P.; Dow, J. Maxwell; Rabinowicz, Pablo; Salzberg, Steven L.; Leach, Jan E.; Sonti, Ramesh; Brendel, Volker; Bogdanove, Adam J.

    2008-01-01

    Background Xanthomonas is a large genus of plant-associated and plant-pathogenic bacteria. Collectively, members cause diseases on over 392 plant species. Individually, they exhibit marked host- and tissue-specificity. The determinants of this specificity are unknown. Methodology/Principal Findings To assess potential contributions to host- and tissue-specificity, pathogenesis-associated gene clusters were compared across genomes of eight Xanthomonas strains representing vascular or non-vascular pathogens of rice, brassicas, pepper and tomato, and citrus. The gum cluster for extracellular polysaccharide is conserved except for gumN and sequences downstream. The xcs and xps clusters for type II secretion are conserved, except in the rice pathogens, in which xcs is missing. In the otherwise conserved hrp cluster, sequences flanking the core genes for type III secretion vary with respect to insertion sequence element and putative effector gene content. Variation at the rpf (regulation of pathogenicity factors) cluster is more pronounced, though genes with established functional relevance are conserved. A cluster for synthesis of lipopolysaccharide varies highly, suggesting multiple horizontal gene transfers and reassortments, but this variation does not correlate with host- or tissue-specificity. Phylogenetic trees based on amino acid alignments of gum, xps, xcs, hrp, and rpf cluster products generally reflect strain phylogeny. However, amino acid residues at four positions correlate with tissue specificity, revealing hpaA and xpsD as candidate determinants. Examination of genome sequences of xanthomonads Xylella fastidiosa and Stenotrophomonas maltophilia revealed that the hrp, gum, and xcs clusters are recent acquisitions in the Xanthomonas lineage. Conclusions/Significance Our results provide insight into the ancestral Xanthomonas genome and indicate that differentiation with respect to host- and tissue-specificity involved not major modifications or wholesale

  16. In vitro volatile organic compound profiling using GC×GC-TOFMS to differentiate bacteria associated with lung infections: a proof-of-concept study.

    PubMed

    Nizio, K D; Perrault, K A; Troobnikoff, A N; Ueland, M; Shoma, S; Iredell, J R; Middleton, P G; Forbes, S L

    2016-04-27

    Chronic pulmonary infections are the principal cause of morbidity and mortality in individuals with cystic fibrosis (CF). Due to the polymicrobial nature of these infections, the identification of the particular bacterial species responsible is an essential step in diagnosis and treatment. Current diagnostic procedures are time-consuming, and can also be expensive, invasive and unpleasant in the absence of spontaneously expectorated sputum. The development of a rapid, non-invasive methodology capable of diagnosing and monitoring early bacterial infection is desired. Future visions of real-time, in situ diagnosis via exhaled breath testing rely on the differentiation of bacteria based on their volatile metabolites. The objective of this proof-of-concept study was to investigate whether a range of CF-associated bacterial species (i.e. Pseudomonas aeruginosa, Burkholderia cenocepacia, Haemophilus influenzae, Stenotrophomonas maltophilia, Streptococcus pneumoniae and Streptococcus milleri) could be differentiated based on their in vitro volatile metabolomic profiles. Headspace samples were collected using solid phase microextraction (SPME), analyzed using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) and evaluated using principal component analysis (PCA) in order to assess the multivariate structure of the data. Although it was not possible to effectively differentiate all six bacteria using this method, the results revealed that the presence of a particular pattern of VOCs (rather than a single VOC biomarker) is necessary for bacterial species identification. The particular pattern of VOCs was found to be dependent upon the bacterial growth phase (e.g. logarithmic versus stationary) and sample storage conditions (e.g. short-term versus long-term storage at  -18 °C). Future studies of CF-associated bacteria and exhaled breath condensate will benefit from the approaches presented in this study and further

  17. Antibacterial Activity of Salvadora persica L. (Miswak) Extracts against Multidrug Resistant Bacterial Clinical Isolates.

    PubMed

    Al-Ayed, Mohamed Saeed Zayed; Asaad, Ahmed Morad; Qureshi, Mohamed Ansar; Attia, Hany Goda; AlMarrani, Abduljabbar Hadi

    2016-01-01

    Much effort has focused on examining the inhibitory effect of Salvadora persica (miswak) on oral microorganisms, but information concerning its antibacterial activity against other human pathogens, particularly multidrug resistant (MDR) isolates, is scarce. Therefore, this study aimed to assess the in vitro antibacterial activities of Salvadora persica L. extracts against 10 MDR bacterial clinical isolates other than oral pathogens. The antibacterial activity of aqueous and methanol miswak extracts was assessed using the agar dilution and minimum inhibitory concentration (MIC) methods. Overall, the 400 mg/mL of miswak extract was the most effective on all strains. The methanol extract exhibited a stronger antibacterial activity against Gram-negative (3.3-13.6 mm) than Gram-positive (1.8-8.3 mm) bacteria. The lowest MIC value was seen for E. coli (0.39, 1.56 µg/mL), followed by Streptococcus pyogenes (1.56 µg/mL). The highest MIC value (6.25, 12.5 µg/mL) was recorded for methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii, and Stenotrophomonas maltophilia. This study demonstrates, for the first time, the moderate to strong antibacterial activity of miswak extracts against all tested MDR-pathogens. Methanol extract appears to be a potent antimicrobial agent that could be considered as complementary and alternative medicine against resistant pathogens. Further studies on a large number of MDR organisms are necessary to investigate and standardize the inhibitory effect of miswak extracts against these emerging pathogens.

  18. Antibacterial Activity of Salvadora persica L. (Miswak) Extracts against Multidrug Resistant Bacterial Clinical Isolates

    PubMed Central

    Al-Ayed, Mohamed Saeed Zayed; Asaad, Ahmed Morad; Qureshi, Mohamed Ansar; Attia, Hany Goda; AlMarrani, Abduljabbar Hadi

    2016-01-01

    Much effort has focused on examining the inhibitory effect of Salvadora persica (miswak) on oral microorganisms, but information concerning its antibacterial activity against other human pathogens, particularly multidrug resistant (MDR) isolates, is scarce. Therefore, this study aimed to assess the in vitro antibacterial activities of Salvadora persica L. extracts against 10 MDR bacterial clinical isolates other than oral pathogens. The antibacterial activity of aqueous and methanol miswak extracts was assessed using the agar dilution and minimum inhibitory concentration (MIC) methods. Overall, the 400 mg/mL of miswak extract was the most effective on all strains. The methanol extract exhibited a stronger antibacterial activity against Gram-negative (3.3–13.6 mm) than Gram-positive (1.8–8.3 mm) bacteria. The lowest MIC value was seen for E. coli (0.39, 1.56 µg/mL), followed by Streptococcus pyogenes (1.56 µg/mL). The highest MIC value (6.25, 12.5 µg/mL) was recorded for methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii, and Stenotrophomonas maltophilia. This study demonstrates, for the first time, the moderate to strong antibacterial activity of miswak extracts against all tested MDR-pathogens. Methanol extract appears to be a potent antimicrobial agent that could be considered as complementary and alternative medicine against resistant pathogens. Further studies on a large number of MDR organisms are necessary to investigate and standardize the inhibitory effect of miswak extracts against these emerging pathogens. PMID:26904146

  19. Antibacterial Activity of Blue Light against Nosocomial Wound Pathogens Growing Planktonically and as Mature Biofilms

    PubMed Central

    Thwaite, Joanne E.; Burt, Rebecca; Laws, Thomas R.; Raguse, Marina; Moeller, Ralf; Webber, Mark A.; Oppenheim, Beryl A.

    2016-01-01

    ABSTRACT The blue wavelengths within the visible light spectrum are intrinisically antimicrobial and can photodynamically inactivate the cells of a wide spectrum of bacteria (Gram positive and negative) and fungi. Furthermore, blue light is equally effective against both drug-sensitive and -resistant members of target species and is less detrimental to mammalian cells than is UV radiation. Blue light is currently used for treating acnes vulgaris and Helicobacter pylori infections; the utility for decontamination and treatment of wound infections is in its infancy. Furthermore, limited studies have been performed on bacterial biofilms, the key growth mode of bacteria involved in clinical infections. Here we report the findings of a multicenter in vitro study performed to assess the antimicrobial activity of 400-nm blue light against bacteria in both planktonic and biofilm growth modes. Blue light was tested against a panel of 34 bacterial isolates (clinical and type strains) comprising Acinetobacter baumannii, Enterobacter cloacae, Stenotrophomonas maltophilia, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Enterococcus faecium, Klebsiella pneumoniae, and Elizabethkingia meningoseptica. All planktonic-phase bacteria were susceptible to blue light treatment, with the majority (71%) demonstrating a ≥5-log10 decrease in viability after 15 to 30 min of exposure (54 J/cm2 to 108 J/cm2). Bacterial biofilms were also highly susceptible to blue light, with significant reduction in seeding observed for all isolates at all levels of exposure. These results warrant further investigation of blue light as a novel decontamination strategy for the nosocomial environment, as well as additional wider decontamination applications. IMPORTANCE Blue light shows great promise as a novel decontamination strategy for the nosocomial environment, as well as additional wider decontamination applications (e.g., wound closure during surgery). This warrants further

  20. Biosynthesis and structural characterization of silver nanoparticles from bacterial isolates

    SciTech Connect

    Zaki, Sahar; El Kady, M.F.; Abd-El-Haleem, Desouky

    2011-10-15

    Graphical abstract: In this study five bacterial isolates belong to different genera were found to be able to biosynthesize silver nanoparticles. Biosynthesis and spectral characterization are reported here. Highlights: {yields} About 300 bacterial isolates were screened for their ability to produce nanosilvers {yields} Five of them were potential candidates for synthesis of silver nanoparticles {yields} Production of silver nanoparticles was examined using UV-Vis, XRD, SEM and EDS. {yields} The presence of nanoparticles with all five bacterial isolates was confirmed. -- Abstract: This study aimed to develop a green process for biosynthesis of silver nanomaterials by some Egyptian bacterial isolates. This target was achieved by screening an in-house culture collection consists of 300 bacterial isolates for silver nanoparticle formation. Through screening process, it was observed that strains belonging to Escherichia coli (S30, S78), Bacillus megaterium (S52), Acinetobacter sp. (S7) and Stenotrophomonas maltophilia (S54) were potential candidates for synthesis of silver nanoparticles. The extracellular production of silver nanoparticles by positive isolates was investigated by UV-Vis spectroscopy, X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). The results demonstrated that UV-visible spectrum of the aqueous medium containing silver ion showed a peak at 420 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy micrograph showed formation of silver nanoparticles in the range of 15-50 nm. XRD-spectrum of the silver nanoparticles exhibited 2{theta} values corresponding to the silver nanocrystal that produce in hexagonal and cubic crystal configurations with different plane of orientation. In addition, the signals of the silver atoms were observed by EDS-spectrum analysis that confirms the presence of silver nanoparticles (Ag

  1. New insight into microbial diversity and functions in traditional Vietnamese alcoholic fermentation.

    PubMed

    Thanh, Vu Nguyen; Thuy, Nguyen Thanh; Chi, Nguyen Thuy; Hien, Dinh Duc; Ha, Bui Thi Viet; Luong, Dao Thi; Ngoc, Pham Duc; Ty, Pham Van

    2016-09-02

    The roles of microorganisms in traditional alcoholic fermentation are often assumed based on abundance in the starter and activity in pure culture. There is a serious lack of hard evidence on the behavior and activity of individual microbial species during the actual fermentation process. In this study, microbial succession and metabolite changes during 7days of traditional Vietnamese alcoholic fermentation were monitored. Special attention was devoted to starch degradation. In total, 22 microbial species, including 6 species of filamentous fungi (Rhizopus microsporus, Rhizopus arrhizus, Mucor indicus, Mucor circinelloides, Cunninghamella elegans, Aspergillus niger), 1 yeast-like fungus (Saccharomycopsis fibuligera), 7 yeasts (Saccharomyces cerevisiae, Clavispora lusitaniae, Wickerhamomyces anomalus, Lindnera fabianii, Pichia kudriavzevii, Candida rugosa, Candida tropicalis), and 8 bacteria (Stenotrophomonas maltophilia, Lactobacillus brevis, Lactobacillus helveticus, Acinetobacter baumannii, Staphylococcus hominis, Bacillus megaterium, Enterobacter asburiae, Pediococcus pentosaceus) were identified. Despite the presence of a complex microbiota in the starter, the fermentation process is consistent and involves a limited number of functional species. Rapid change in microbial composition of fermentation mash was observed and it was correlated with ethanol content. Microbial biomass reached maximum during first 2days of solid state fermentation. Acidification of the medium took place in day 1, starch degradation in days 2, 3, 4, and alcohol accumulation from day 3. Although Sm. fibuligera dominated by cell count amongst potential starch degraders, zymography indicated that it did not produce amylase in the fermentation mash. In mixed culture with Rhizopus, amylase production by Sm. fibuligera is regulated by the moisture content of the substrate. Rhizopus was identified as the main starch degrader and S. cerevisiae as the main ethanol producer. Bacterial load was

  2. Air-dust-borne associations of phototrophic and hydrocarbon-utilizing microorganisms: promising consortia in volatile hydrocarbon bioremediation.

    PubMed

    Al-Bader, Dhia; Eliyas, Mohamed; Rayan, Rihab; Radwan, Samir

    2012-11-01

    Aquatic and terrestrial associations of phototrophic and heterotrophic microorganisms active in hydrocarbon bioremediation have been described earlier. The question arises: do similar consortia also occur in the atmosphere? Dust samples at the height of 15 m were collected from Kuwait City air, and analyzed microbiologically for phototrophic and heterotrophic hydrocarbon-utilizing microorganisms, which were subsequently characterized according to their 16S rRNA gene sequences. The hydrocarbon utilization potential of the heterotrophs alone, and in association with the phototrophic partners, was measured quantitatively. The chlorophyte Gloeotila sp. and the two cyanobacteria Nostoc commune and Leptolyngbya thermalis were found associated with dust, and (for comparison) the cynobacteria Leptolyngbya sp. and Acaryochloris sp. were isolated from coastal water. All phototrophic cultures harbored oil vapor-utilizing bacteria in the magnitude of 10(5) g(-1). Each phototrophic culture had its unique oil-utilizing bacteria; however, the bacterial composition in Leptolyngbya cultures from air and water was similar. The hydrocarbon-utilizing bacteria were affiliated with Acinetobacter sp., Aeromonas caviae, Alcanivorax jadensis, Bacillus asahii, Bacillus pumilus, Marinobacter aquaeolei, Paenibacillus sp., and Stenotrophomonas maltophilia. The nonaxenic cultures, when used as inocula in batch cultures, attenuated crude oil in light and dark, and in the presence of antibiotics and absence of nitrogenous compounds. Aqueous and diethyl ether extracts from the phototrophic cultures enhanced the growth of the pertinent oil-utilizing bacteria in batch cultures, with oil vapor as a sole carbon source. It was concluded that the airborne microbial associations may be effective in bioremediating atmospheric hydrocarbon pollutants in situ. Like the aquatic and terrestrial habitats, the atmosphere contains dust-borne associations of phototrophic and heterotrophic hydrocarbon

  3. Species distribution and antimicrobial susceptibility of gram-negative aerobic bacteria in hospitalized cancer patients

    PubMed Central

    Ashour, Hossam M; El-Sharif, Amany

    2009-01-01

    Background Nosocomial infections pose significant threats to hospitalized patients, especially the immunocompromised ones, such as cancer patients. Methods This study examined the microbial spectrum of gram-negative bacteria in various infection sites in patients with leukemia and solid tumors. The antimicrobial resistance patterns of the isolated bacteria were studied. Results The most frequently isolated gram-negative bacteria were Klebsiella pneumonia (31.2%) followed by Escherichia coli (22.2%). We report the isolation and identification of a number of less-frequent gram negative bacteria (Chromobacterium violacum, Burkholderia cepacia, Kluyvera ascorbata, Stenotrophomonas maltophilia, Yersinia pseudotuberculosis, and Salmonella arizona). Most of the gram-negative isolates from Respiratory Tract Infections (RTI), Gastro-intestinal Tract Infections (GITI), Urinary Tract Infections (UTI), and Bloodstream Infections (BSI) were obtained from leukemic patients. All gram-negative isolates from Skin Infections (SI) were obtained from solid-tumor patients. In both leukemic and solid-tumor patients, gram-negative bacteria causing UTI were mainly Escherichia coli and Klebsiella pneumoniae, while gram-negative bacteria causing RTI were mainly Klebsiella pneumoniae. Escherichia coli was the main gram-negative pathogen causing BSI in solid-tumor patients and GITI in leukemic patients. Isolates of Escherichia coli, Klebsiella, Enterobacter, Pseudomonas, and Acinetobacter species were resistant to most antibiotics tested. There was significant imipenem -resistance in Acinetobacter (40.9%), Pseudomonas (40%), and Enterobacter (22.2%) species, and noticeable imipinem-resistance in Klebsiella (13.9%) and Escherichia coli (8%). Conclusion This is the first study to report the evolution of imipenem-resistant gram-negative strains in Egypt. Mortality rates were higher in cancer patients with nosocomial Pseudomonas infections than any other bacterial infections. Policies restricting

  4. Isolation of UV-B resistant bacteria from two high altitude Andean lakes (4,400 m) with saline and non saline conditions.

    PubMed

    Flores, María R; Ordoñez, Omar F; Maldonado, Marcos J; Farías, María E

    2009-12-01

    Laguna (L.) Negra and L. Verde are high altitude Andean lakes located at the 4,400 m altitude in the Andean desert (Puna) in the Argentine northwest. Both lakes are exposed to extreme weather conditions but differ in salinity contents (salinity 6.7% for L. Negra and 0.27% for L. Verde). The aim of this work was to isolate ultraviolet B fraction (UV-B) resistant bacteria under UV-stress in order to determine, a possible connection, between resistance to UV-B and tolerance to salinity. DNA damage was determined by measuring CPDs accumulation. Connection among pigmentation production and UV resistance was also studied. Water samples were exposed to artificial UV-B radiation for 24 h. Water aliquots were plated along the exposition on different media, with different salinity and carbon source content (Lake medium (LM) done with the lake water plus agar and LB). CFU were counted and DNA damage accumulation was determined. Isolated bacteria were identified by 16S rDNA sequence. Their salinity tolerance, were measured at 1, 5 and 10% NaCl and their pigment production in both media was determined. In general it was found that UV resistance and pigment production were the optimum in Lake Medium done with lake water which maintained similar salinity. The most resistant bacteria in L. Negra were different strains of Exiguobacterium sp. and, in L. Verde, Staphylococcus sp. and Stenotrophomonas maltophilia. These bacteria showed the production and increase of UV-Vis absorbing compounds under UV stress and in LM. Bacterial communities from both lakes were well adapted to high UV-B exposure under the experimental conditions, and in many cases UV-B even stimulated growth. The idea that resistance to UV-B could be related to adaptation to high salinity is still an open question that has to be answered with future experiments.

  5. Isolation and Identification of Plant Growth Promoting Rhizobacteria from Cucumber Rhizosphere and Their Effect on Plant Growth Promotion and Disease Suppression

    PubMed Central

    Islam, Shaikhul; Akanda, Abdul M.; Prova, Ananya; Islam, Md. T.; Hossain, Md. M.

    2016-01-01

    Plant growth promoting rhizobacteria (PGPR) are the rhizosphere bacteria that may be utilized to augment plant growth and suppress plant diseases. The objectives of this study were to identify and characterize PGPR indigenous to cucumber rhizosphere in Bangladesh, and to evaluate their ability to suppress Phytophthora crown rot in cucumber. A total of 66 isolates were isolated, out of which 10 (PPB1, PPB2, PPB3, PPB4, PPB5, PPB8, PPB9, PPB10, PPB11, and PPB12) were selected based on their in vitro plant growth promoting attributes and antagonism of phytopathogens. Phylogenetic analysis of 16S rRNA sequences identified these isolates as new strains of Pseudomonas stutzeri, Bacillus subtilis, Stenotrophomonas maltophilia, and Bacillus amyloliquefaciens. The selected isolates produced high levels (26.78–51.28 μg mL-1) of indole-3-acetic acid, while significant acetylene reduction activities (1.79–4.9 μmole C2H4 mg-1 protein h-1) were observed in eight isolates. Cucumber plants grown from seeds that were treated with these PGPR strains displayed significantly higher levels of germination, seedling vigour, growth, and N content in root and shoot tissue compared to non-treated control plants. All selected isolates were able to successfully colonize the cucumber roots. Moreover, treating cucumber seeds with these isolates significantly suppressed Phytophthora crown rot caused by Phytophthora capsici, and characteristic morphological alterations in P. capsici hyphae that grew toward PGPR colonies were observed. Since these PGPR inoculants exhibited multiple traits beneficial to the host plants, they may be applied in the development of new, safe, and effective seed treatments as an alternative to chemical fungicides. PMID:26869996

  6. Insight into the Role of Substrate-binding Residues in Conferring Substrate Specificity for the Multifunctional Polysaccharide Lyase Smlt1473

    PubMed Central

    MacDonald, Logan C.; Berger, Bryan W.

    2014-01-01

    Anionic polysaccharides are of growing interest in the biotechnology industry due to their potential pharmaceutical applications in drug delivery and wound treatment. Chemical composition and polymer length strongly influence the physical and biological properties of the polysaccharide and thus its potential industrial and medical applications. One promising approach to determining monomer composition and controlling the degree of polymerization involves the use of polysaccharide lyases, which catalyze the depolymerization of anionic polysaccharides via a β-elimination mechanism. Utilization of these enzymes for the production of custom-made oligosaccharides requires a high degree of control over substrate specificity. Previously, we characterized a polysaccharide lyase (Smlt1473) from Stenotrophomonas maltophilia k279a, which exhibited significant activity against hyaluronan (HA), poly-β-d-glucuronic acid (poly-GlcUA), and poly-β-d-mannuronic acid (poly-ManA) in a pH-regulated manner. Here, we utilize a sequence structure guided approach based on a homology model of Smlt1473 to identify nine putative substrate-binding residues and examine their effect on substrate specificity via site-directed mutagenesis. Interestingly, single point mutations H221F and R312L resulted in increased activity and specificity toward poly-ManA and poly-GlcUA, respectively. Furthermore, a W171A mutant nearly eliminated HA activity, while increasing poly-ManA and poly-GlcUA activity by at least 35%. The effect of these mutations was analyzed by comparison with the high resolution structure of Sphingomonas sp. A1-III alginate lyase in complex with poly-ManA tetrasaccharide and by taking into account the structural differences between HA, poly-GlcUA, and poly-ManA. Overall, our results demonstrate that even minor changes in active site architecture have a significant effect on the substrate specificity of Smlt1473, whose structural plasticity could be applied to the design of highly

  7. [Utility of prolonged incubation and terminal subcultures of blood cultures from immunocompromised patients].

    PubMed

    Soloaga, R; Procopio, A; Manganello, S; Ivanovic, V; Romay, N; Pirosanto, Y; Fernández, A; Zudiker, R; Echeverría, A; Nagel, C; del Castillo, M; López, E; Gutfraind, Z; Tokumoto, M; Guelfand, L

    2001-01-01

    The value of blind terminal subcultures (7 and 30 days) and prolonged incubation (30 days) of blood cultures from immunosuppressed patients was analyzed in the Fundación Favaloro, the Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia and the Hospital de Niños Ricardo Gutiérrez. A total of 2707 blood cultures and 369 patients were included (transplantation of solid organs 154, oncohematologic disorders 106 and solid tumors 109). Bact-Alert bottles were incubated at 35 degrees C for 30 days in the Bact-Alert System. Bottles with positive signals were routinely removed, and aliquots of the broth were Gram stained and subcultured aerobically in chocolate agar and Sabouraud agar. A total of 136 bacteremic episodes were obtained. The positivization time of blood cultures was 81.6% at 24 h, 93.3% at 48 h, 94.5% at 72 h and 97.7% within 7 days. Only 3 (2.2%) episodes were positive by blind terminal subcultures and 1 (0.75%) by prolonged incubation (14 days). The median time and range of positivization in hours were 13.8 and 2.2-168, respectively. The microorganisms isolated were coagulase negative staphylococci (n = 24), Klebsiella pneumoniae (n = 22), Staphylococcus aureus (n = 21), Escherichia coli (n = 18), Acinetobacter spp (n = 9), Candida spp (n = 8), Pseudomonas aeruginosa (n = 6), Enterobacter cloacae (n = 5), Stenotrophomonas maltophilia (n = 5), Enterococcus faecalis, Salmonella spp and Capnocytophaga sputigena (n = 2), Enterobacter aerogenes, Enterococcus faecium, Citrobacter diversus, Candida albicans, Klebsiella oxytoca, Chryseomonas luteola, Serratia marcescens, Abiotrophia spp, Campylobacter jejuni, Moraxella catarrhalis, Moraxella urethralis, Neisseria sicca, beta hemolytic group G streptococci, Rhodococcus equi, Micrococcus spp, Cryptococcus neoformans and Streptococcus mitis (n = 1). In our experience, blind terminal subcultures and prolonged incubation of blood cultures from immunosuppressed patients are unnecessary and

  8. [Diagnostic and therapeutic management of Gram-negative infections].

    PubMed

    Bassetti, Matteo; Repetto, Ernestina

    2008-04-01

    Among Gram negative bacteria, Pseudomonas aeruginosa, the extended spectrum beta-lactamases (ESBL)-producing strains, Acinetobacter spp, in particular the multiresistant Acinetobacter baumannii, and Stenotrophomonas maltophilia are the most implicated micrororganisms in the ever more increasing problem of bacterial resistance. Possible solutions have to be searched, on one hand, in the use of new drugs but, on the other hand, in the re-evaluation of those already available drugs, possibly considering a new role for old drugs such as colistine and fosfomycin. Concerning ESBL-producing strains, the most recent data provided by EARSS report, in Italy, an incidence rate of 10-25 percent. The insurgence of an infection sustained by an ESBL+ve strain is strictly related to some well known risk factors, like the hospital stay itself, the disease severity, the length of stay in ICU, intubation and mechanical ventilation, catheterization, urinary or artery, and the past exposure to antibiotics. The raise in ESBL producing strains is closely related to the increasing use of cephalosporins. In the setting of a Gram negative infection, the combination therapy guarantees a higher coverage by reducing insurgence of possible resistance mechanisms, possibly resulting synergistic, and allowing a de-escalation therapy, although to this latter other problems, such as tolerability, costs and compliance, can be related. Another basic aspect to take into account of, in order to achieve the maximal efficacy of the antibiotic treatment, is the right dosage. In the idea to look for the best approach for the antibiotic treatment of a severe infection in a hospital setting, when a Gram negative aetiology is implicated, it can be possibly presumed that the right way consists in avoiding inappropriate antibiotic therapies, making therapeutic choices based on guidelines resulted from local epidemiological data, initiating the therapy promptly, avoiding excessive use of antibiotics, possibly

  9. Insight into the role of substrate-binding residues in conferring substrate specificity for the multifunctional polysaccharide lyase Smlt1473.

    PubMed

    MacDonald, Logan C; Berger, Bryan W

    2014-06-27

    Anionic polysaccharides are of growing interest in the biotechnology industry due to their potential pharmaceutical applications in drug delivery and wound treatment. Chemical composition and polymer length strongly influence the physical and biological properties of the polysaccharide and thus its potential industrial and medical applications. One promising approach to determining monomer composition and controlling the degree of polymerization involves the use of polysaccharide lyases, which catalyze the depolymerization of anionic polysaccharides via a β-elimination mechanism. Utilization of these enzymes for the production of custom-made oligosaccharides requires a high degree of control over substrate specificity. Previously, we characterized a polysaccharide lyase (Smlt1473) from Stenotrophomonas maltophilia k279a, which exhibited significant activity against hyaluronan (HA), poly-β-d-glucuronic acid (poly-GlcUA), and poly-β-d-mannuronic acid (poly-ManA) in a pH-regulated manner. Here, we utilize a sequence structure guided approach based on a homology model of Smlt1473 to identify nine putative substrate-binding residues and examine their effect on substrate specificity via site-directed mutagenesis. Interestingly, single point mutations H221F and R312L resulted in increased activity and specificity toward poly-ManA and poly-GlcUA, respectively. Furthermore, a W171A mutant nearly eliminated HA activity, while increasing poly-ManA and poly-GlcUA activity by at least 35%. The effect of these mutations was analyzed by comparison with the high resolution structure of Sphingomonas sp. A1-III alginate lyase in complex with poly-ManA tetrasaccharide and by taking into account the structural differences between HA, poly-GlcUA, and poly-ManA. Overall, our results demonstrate that even minor changes in active site architecture have a significant effect on the substrate specificity of Smlt1473, whose structural plasticity could be applied to the design of highly

  10. Nitrate treatment effects on bacterial community biofilm formed on carbon steel in produced water stirred tank bioreactor.

    PubMed

    Marques, Joana Montezano; de Almeida, Fernando Pereira; Lins, Ulysses; Seldin, Lucy; Korenblum, Elisa

    2012-06-01

    To better understand the impact of nitrate in Brazilian oil reservoirs under souring processes and corrosion, the goal of this study was to analyse the effect of nitrate on bacterial biofilms formed on carbon steel coupons using reactors containing produced water from a Brazilian oil platform. Three independent experiments were carried out (E1, E2 and E3) using the same experimental conditions and different incubation times (5, 45 and 80 days, respectively). In every experiment, two biofilm-reactors were operated: one was treated with continuous nitrate flow (N reactor), and the other was a control reactor without nitrate (C reactor). A Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis approach using the 16S rRNA gene was performed to compare the bacterial groups involved in biofilm formation in the N and C reactors. DGGE profiles showed remarkable changes in community structure only in experiments E2 and E3. Five bands extracted from the gel that represented the predominant bacterial groups were identified as Bacillus aquimaris, B. licheniformis, Marinobacter sp., Stenotrophomonas maltophilia and Thioclava sp. A reduction in the sulfate-reducing bacteria (SRB) most probable number counts was observed only during the longer nitrate treatment (E3). Carbon steel coupons used for biofilm formation had a slightly higher weight loss in N reactors in all experiments. When the coupon surfaces were analysed by scanning electron microscopy, an increase in corrosion was observed in the N reactors compared with the C reactors. In conclusion, nitrate reduced the viable SRB counts. Nevertheless, the nitrate dosing increased the pitting of coupons.

  11. Count, identification and antimicrobial susceptibility of bacteria recovered from dental solid waste in Brazil.

    PubMed

    Vieira, Cristina Dutra; de Carvalho, Maria Auxiliadora Roque; Cussiol, Noil Amorim de Menezes; Alvarez-Leite, Maria Eugênia; dos Santos, Simone Gonçalves; Gomes, Renata Maria da Fonseca; Silva, Marcos Xavier; Nicoli, Jacques Robert; Farias, Luiz de Macêdo

    2011-06-01

    In Brazil, few studies on microbial content of dental solid waste and its antibiotic susceptibility are available. An effort has been made through this study to evaluate the hazardous status of dental solid waste, keeping in mind its possible role in cross-infection chain. Six samples of solid waste were collected at different times and seasons from three dental health services. The microbial content was evaluated in different culture media and atmospheric conditions, and the isolates were submitted to antibiotic susceptibility testing. A total of 766 bacterial strains were isolated and identified during the study period. Gram-positive cocci were the most frequent morphotype isolated (48.0%), followed by Gram-negative rods (46.2%), Gram-positive rods (5.0%), Gram-negative-cocci (0.4%), and Gram-positive coccobacillus (0.1%). Only two anaerobic bacteria were isolated (0.3%). The most frequently isolated species was Staphylococcus epidermidis (29.9%), followed by Stenotrophomonas maltophilia (8.2%), and Enterococcus faecalis (6.7%). High resistance rate to ampicillin was observed among Gram-negative rods (59.4%) and Gram-positive cocci (44.4%). For Gram-negative rods, high resistance was also noted to aztreonam (47.7%), cefotaxime (47.4%), ceftriaxone and cefazolin (43.7%), and ticarcillin-clavulanic acid (38.2%). Against Gram-positive cocci penicillin exhibit a higher resistance rate (45.0%), followed by ampicillin, erythromycin (27.2%), and tetracycline (22.0%). The present study demonstrated that several pathogenic bacteria are present in dental solid waste and can survive after 48 h from the waste generation time and harbor resistance profiles against several clinical recommended antibiotics.

  12. Diagnostic value of PCR analysis of bacteria and fungi from blood in empiric-therapy-resistant febrile neutropenia.

    PubMed

    Nakamura, Akiko; Sugimoto, Yuka; Ohishi, Kohshi; Sugawara, Yumiko; Fujieda, Atsushi; Monma, Fumihiko; Suzuki, Kei; Masuya, Masahiro; Nakase, Kazunori; Matsushima, Yoshiko; Wada, Hideo; Katayama, Naoyuki; Nobori, Tsutomu

    2010-06-01

    This study aimed to assess the clinical utility of PCR for the analysis of bacteria and fungi from blood for the management of febrile neutropenic patients with hematologic malignancies. Using a PCR system able to detect a broad range of bacteria and fungi, we conducted a prospective pilot study of periodic analyses of blood from patients following intensive chemotherapy. When fever occurred, it was treated with empirical antibiotic therapy, basically without knowledge of the PCR results. In 23 febrile episodes during the neutropenic period, bacteria were detected by PCR in 11 cases, while the same species were identified by blood culture in 3 cases. In 10 out of 11 PCR-positive cases, fever could be managed by empirical therapy. In the empirical-therapy-resistant case, the identification of Stenotrophomonas maltophilia by PCR led to improvement of fever. No fungi were detected by PCR in febrile cases, while Aspergillus fumigatus was detected in one afebrile patient, several days before a clinical diagnosis was made. In subsequent sporadic PCR analyses in 15 cases of febrile neutropenia, bacteria were detected by both PCR and blood culture in 7 cases and by PCR alone in 6. Fungi were not detected. While fever was improved by empirical therapy in 12 out of the 13 PCR-positive cases, the identification of Pseudomonas aeruginosa by PCR in one therapy-resistant case contributed to the successful treatment of persistent fever. Our results indicate that PCR analysis of bacteria from blood provides essential information for managing empirical-therapy-resistant febrile neutropenia.

  13. Microbe associated molecular patterns from rhizosphere bacteria trigger germination and Papaver somniferum metabolism under greenhouse conditions.

    PubMed

    Bonilla, A; Sarria, A L F; Algar, E; Muñoz Ledesma, F J; Ramos Solano, B; Fernandes, J B; Gutierrez Mañero, F J

    2014-01-01

    Ten PGPR from different backgrounds were assayed on Papaver somniferum var. Madrigal to evaluate their potential as biotic elicitors to increase alkaloid content under the rationale that some microbe associated molecular patterns (MAMPs) are able to trigger plant metabolism. First, the 10 strains and their culture media at two different concentrations were tested for their ability to trigger seed germination. Then, the best three strains were tested for their ability to increase seedling growth and alkaloid levels under greenhouse conditions. Only three strains and their culture media enhanced germination. Then, germination enhancing capacity of these best three strains, N5.18 Stenotrophomonas maltophilia, Aur9 Chryseobacterium balustinum and N21.4 Pseudomonas fluorescens was evaluated in soil. Finally, the three strains were applied on seedlings at two time points, by soil drench or by foliar spray. Photosynthesis was measured, plant height was recorded, capsules were weighted and alkaloids analyzed by HPLC. Only N5.18 delivered by foliar spray significantly increased plant height coupled to an increase in total alkaloids and a significant increase in opium poppy straw dry weight; these increases were supported by a better photosynthetic efficiency. The relative contents of morphine, thebaine, codeine and oripavine were affected by this treatment causing a significant increase in morphine coupled to a decrease in thebaine, demonstrating the effectivity of MAMPs from N5.18 in this plant species. Considering the increase in capsule biomass and alkaloids together with the acceleration of germination, strain N5.18 appears as a good candidate to elicit plant metabolism and consequently, to increase productivity of Papaver somniferum.

  14. Effects of Agaricus lilaceps fairy rings on soil aggregation and microbial community structure in relation to growth stimulation of western wheatgrass (Pascopyrum smithii) in Eastern Montana rangeland.

    PubMed

    Caesar-Tonthat, The Can; Espeland, Erin; Caesar, Anthony J; Sainju, Upendra M; Lartey, Robert T; Gaskin, John F

    2013-07-01

    Stimulation of plant productivity caused by Agaricus fairy rings has been reported, but little is known about the effects of these fungi on soil aggregation and the microbial community structure, particularly the communities that can bind soil particles. We studied three concentric zones of Agaricus lilaceps fairy rings in Eastern Montana that stimulate western wheatgrass (Pascopyrum smithii): outside the ring (OUT), inside the ring (IN), and stimulated zone adjacent to the fungal fruiting bodies (SZ) to determine (1) soil aggregate proportion and stability, (2) the microbial community composition and the N-acetyl-β-D-glucosaminidase activity associated with bulk soil at 0-15 cm depth, (3) the predominant culturable bacterial communities that can bind to soil adhering to wheatgrass roots, and (4) the stimulation of wheatgrass production. In bulk soil, macroaggregates (4.75-2.00 and 2.00-0.25 mm) and aggregate stability increased in SZ compared to IN and OUT. The high ratio of fungal to bacteria (fatty acid methyl ester) and N-acetyl-β-D-glucosaminidase activity in SZ compared to IN and OUT suggest high fungal biomass. A soil sedimentation assay performed on the predominant isolates from root-adhering soil indicated more soil-binding bacteria in SZ than IN and OUT; Pseudomonas fluorescens and Stenotrophomonas maltophilia isolates predominated in SZ, whereas Bacillus spp. isolates predominated in IN and OUT. This study suggests that growth stimulation of wheatgrass in A. lilaceps fairy rings may be attributed to the activity of the fungus by enhancing soil aggregation of bulk soil at 0-15 cm depth and influencing the amount and functionality of specific predominant microbial communities in the wheatgrass root-adhering soil.

  15. Microbial Diversity in the Early In Vivo-Formed Dental Biofilm

    PubMed Central

    Heller, D.; Helmerhorst, E. J.; Gower, A. C.; Siqueira, W. L.; Paster, B. J.

    2016-01-01

    Although the mature dental biofilm composition is well studied, there is very little information on the earliest phase of in vivo tooth colonization. Progress in dental biofilm collection methodologies and techniques of large-scale microbial identification have made new studies in this field of oral biology feasible. The aim of this study was to characterize the temporal changes and diversity of the cultivable and noncultivable microbes in the early dental biofilm. Samples of early dental biofilm were collected from 11 healthy subjects at 0, 2, 4, and 6 h after removal of plaque and pellicle from tooth surfaces. With the semiquantitative Human Oral Microbiome Identification Microarray (HOMIM) technique, which is based on 16S rRNA sequence hybridizations, plaque samples were analyzed with the currently available 407 HOMIM microbial probes. This led to the identification of at least 92 species, with streptococci being the most abundant bacteria across all time points in all subjects. High-frequency detection was also made with Haemophilus parainfluenzae, Gemella haemolysans, Slackia exigua, and Rothia species. Abundance changes over time were noted for Streptococcus anginosus and Streptococcus intermedius (P = 0.02), Streptococcus mitis bv. 2 (P = 0.0002), Streptococcus oralis (P = 0.0002), Streptococcus cluster I (P = 0.003), G. haemolysans (P = 0.0005), and Stenotrophomonas maltophilia (P = 0.02). Among the currently uncultivable microbiota, eight phylotypes were detected in the early stages of biofilm formation, one belonging to the candidate bacterial division TM7, which has attracted attention due to its potential association with periodontal disease. PMID:26746720

  16. Faropenem: review of a new oral penem.

    PubMed

    Schurek, Kristen N; Wiebe, Ryan; Karlowsky, James A; Rubinstein, Ethan; Hoban, Daryl J; Zhanel, George G

    2007-04-01

    Faropenem medoxomil is a new orally administered penem antibiotic. Its chiral tetrahydrofuran substituent at position C2 is responsible for its improved chemical stability and reduced CNS effects, compared with imipenem. Faropenem demonstrates broad-spectrum in vitro antimicrobial activity against many Gram-positive and -negative aerobes and anaerobes, and is resistant to hydrolysis by nearly all beta-lactamases, including extended-spectrum beta-lactamases and AmpC beta-lactamases. However, faropenem is not active against methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, Pseudomonas aeruginosa or Stenotrophomonas maltophilia. Prospective, multicenter, randomized, double-blind, comparative (not vs placebo) clinical trials of acute bacterial sinusitis (ABS), acute exacerbations of chronic bronchitis (AECB), community-acquired pneumonia (CAP) and uncomplicated skin and skin structure infections (uSSSIs) have demonstrated that faropenem medoxomil has equivalent efficacy and safety compared with cefuroxime, clarithromycin, azithromycin, amoxicillin, cefpodoxime and amoxicillin-clavulanate. The evidence supports faropenem medoxomil as a promising new oral beta-lactam with proven efficacy and safety for the treatment of a variety of community-acquired infections. However, the US FDA recently rejected faropenem for all four indications stating that the clinical trials in ABS and AECB should have been performed versus a placebo. In the CAP studies, the FDA stated that they could not be certain of the validity of the study population actually having the disease and for uSSSI, the FDA stated that only a single trial was not adequate evidence of efficacy for this indication.

  17. Coexistence of free-living amoebae and bacteria in selected South African hospital water distribution systems.

    PubMed

    Muchesa, P; Leifels, M; Jurzik, L; Hoorzook, K B; Barnard, T G; Bartie, C

    2017-01-01

    Pathogenic free-living amoebae (FLA), such as Naegleria fowleri, Balamuthia mandrillaris and Acanthamoeba species isolated from aquatic environments have been implicated in central nervous system, eye and skin human infections. They also allow the survival, growth and transmission of bacteria such as Legionella, Mycobacteria and Vibrio species in water systems. The purpose of this study was to investigate the co-occurrence of potentially pathogenic FLA and their associated bacteria in hospital water networks in Johannesburg, South Africa. A total of 178 water (n = 95) and swab (n = 83) samples were collected from two hospital water distribution systems. FLA were isolated using the amoebal enrichment technique and identified using PCR and 18S rDNA sequencing. Amoebae potentially containing intra-amoebal bacteria were lysed and cultured on blood agar plates. Bacterial isolates were characterized using the VITEK®2 compact System. Free-living amoebae were isolated from 77 (43.3 %) of the samples. Using microscopy, PCR and 18S rRNA sequencing, Acanthamoeba spp. (T3 and T20 genotypes), Vermamoeba vermiformis and Naegleria gruberi specie were identified. The Acanthamoeba T3 and T20 genotypes have been implicated in eye and central nervous system infections. The most commonly detected bacterial species were Serratia marcescens, Stenotrophomonas maltophilia, Delftia acidovorans, Sphingomonas paucimobilis and Comamonas testosteroni. These nosocomial pathogenic bacteria are associated with systematic blood, respiratory tract, the urinary tract, surgical wounds and soft tissues infections. The detection of FLA and their associated opportunistic bacteria in the hospital water systems point out to a potential health risk to immune-compromised individuals.

  18. Yersinia enterocolitica and Photorhabdus asymbiotica β-lactamases BlaA are exported by the twin-arginine translocation pathway.

    PubMed

    Schriefer, Eva-Maria; Hoffmann-Thoms, Stephanie; Schmid, Franz X; Schmid, Annika; Heesemann, Jürgen

    2013-01-01

    In general, β-lactamases of medically important Gram-negative bacteria are Sec-dependently translocated into the periplasm. In contrast, β-lactamases of Mycobacteria spp. (BlaC, BlaS) and the Gram-negative environmental bacteria Stenotrophomonas maltophilia (L2) and Xanthomonas campestris (Bla(XCC-1)) have been reported to be secreted by the twin-arginine translocation (Tat) system. Yersinia enterocolitica carries 2 distinct β-lactamase genes (blaA and blaB) encoding BlaA(Ye) and the AmpC-like β-lactamase BlaB, respectively. By using the software PRED-TAT for prediction and discrimination of Sec from Tat signal peptides, we identified a functional Tat signal sequence for Yersinia BlaA(Ye). The Tat-dependent translocation of BlaA(Ye) could be clearly demonstrated by using a Y. enterocolitica tatC-mutant and cell fractionation. Moreover, we could demonstrate a unique unusual temperature-dependent activity profile of BlaA(Ye) ranging from 15 to 60 °C and a high 'melting temperature' (T(M)=44.3°) in comparison to the related Sec-dependent β-lactamase TEM-1 (20-50°C, T(M)=34.9 °C). Strikingly, the blaA gene of Y. enterocolitica is present in diverse environmental Yersinia spp. and a blaA homolog gene could be identified in the closely related Photorhabdus asymbiotica (BlaA(Pa); 69% identity to BlaA(Ye)). For BlaA(Pa) of P. asymbiotica, we could also demonstrate Tat-dependent secretion. These results suggest that Yersinia BlaA-related β-lactamases may be the prototype of a large Tat-dependent β-lactamase family, which originated from environmental bacteria.

  19. Isolation and Identification of Plant Growth Promoting Rhizobacteria from Cucumber Rhizosphere and Their Effect on Plant Growth Promotion and Disease Suppression.

    PubMed

    Islam, Shaikhul; Akanda, Abdul M; Prova, Ananya; Islam, Md T; Hossain, Md M

    2015-01-01

    Plant growth promoting rhizobacteria (PGPR) are the rhizosphere bacteria that may be utilized to augment plant growth and suppress plant diseases. The objectives of this study were to identify and characterize PGPR indigenous to cucumber rhizosphere in Bangladesh, and to evaluate their ability to suppress Phytophthora crown rot in cucumber. A total of 66 isolates were isolated, out of which 10 (PPB1, PPB2, PPB3, PPB4, PPB5, PPB8, PPB9, PPB10, PPB11, and PPB12) were selected based on their in vitro plant growth promoting attributes and antagonism of phytopathogens. Phylogenetic analysis of 16S rRNA sequences identified these isolates as new strains of Pseudomonas stutzeri, Bacillus subtilis, Stenotrophomonas maltophilia, and Bacillus amyloliquefaciens. The selected isolates produced high levels (26.78-51.28 μg mL(-1)) of indole-3-acetic acid, while significant acetylene reduction activities (1.79-4.9 μmole C2H4 mg(-1) protein h(-1)) were observed in eight isolates. Cucumber plants grown from seeds that were treated with these PGPR strains displayed significantly higher levels of germination, seedling vigour, growth, and N content in root and shoot tissue compared to non-treated control plants. All selected isolates were able to successfully colonize the cucumber roots. Moreover, treating cucumber seeds with these isolates significantly suppressed Phytophthora crown rot caused by Phytophthora capsici, and characteristic morphological alterations in P. capsici hyphae that grew toward PGPR colonies were observed. Since these PGPR inoculants exhibited multiple traits beneficial to the host plants, they may be applied in the development of new, safe, and effective seed treatments as an alternative to chemical fungicides.

  20. Microbiological investigation of Raphanus sativus L. grown hydroponically in nutrient solutions contaminated with spoilage and pathogenic bacteria.

    PubMed

    Settanni, Luca; Miceli, Alessandro; Francesca, Nicola; Cruciata, Margherita; Moschetti, Giancarlo

    2013-01-01

    The survival of eight undesired (spoilage/pathogenic) food related bacteria (Citrobacter freundii PSS60, Enterobacter spp. PSS11, Escherichia coli PSS2, Klebsiella oxytoca PSS82, Serratia grimesii PSS72, Pseudomonas putida PSS21, Stenotrophomonas maltophilia PSS52 and Listeria monocytogenes ATCC 19114(T)) was investigated in mineral nutrient solution (MNS) during the crop cycle of radishes (Raphanus sativus L.) cultivated in hydroponics in a greenhouse. MNSs were microbiologically analyzed weekly by plate count. The evolution of the pure cultures was also evaluated in sterile MNS in test tubes. The inoculated trials contained an initial total mesophilic count (TMC) ranging between 6.69 and 7.78Log CFU/mL, while non-sterile and sterile control trials showed levels of 4.39 and 0.97Log CFU/mL, respectively. In general, all inoculated trials showed similar levels of TMC in MNS during the experimentation, even though the levels of the inoculated bacteria decreased. The presence of the inoculums was ascertained by randomly amplified polymorphic DNA (RAPD) analysis applied on the isolates collected at 7-day intervals. At harvest, MNSs were also analyzed by denaturing gradient gel electrophoresis (DGGE). The last analysis, except P. putida PSS21 in the corresponding trial, did not detect the other bacteria, but confirmed that pseudomonads were present in un-inoculated MNSs. Despite the high counts detected (6.44 and 7.24CFU/g), only C. freundii PSS60, Enterobacter spp. PSS11 and K. oxytoca PSS82 were detected in radishes in a living form, suggesting their internalization.

  1. Prevalent bacterial species and novel phylotypes in advanced noma lesions.

    PubMed

    Paster, B J; Falkler Jr, W A; Enwonwu, C O; Idigbe, E O; Savage, K O; Levanos, V A; Tamer, M A; Ericson, R L; Lau, C N; Dewhirst, F E

    2002-06-01

    The purpose of this study was to determine the bacterial diversity in advanced noma lesions using culture-independent molecular methods. 16S ribosomal DNA bacterial genes from DNA isolated from advanced noma lesions of four Nigerian children were PCR amplified with universally conserved primers and spirochetal selective primers and cloned into Escherichia coli. Partial 16S rRNA sequences of approximately 500 bases from 212 cloned inserts were used initially to determine species identity or closest relatives by comparison with sequences of known species or phylotypes. Nearly complete sequences of approximately 1,500 bases were obtained for most of the potentially novel species. A total of 67 bacterial species or phylotypes were detected, 25 of which have not yet been grown in vitro. Nineteen of the species or phylotypes, including Propionibacterium acnes, Staphylococcus spp., and the opportunistic pathogens Stenotrophomonas maltophilia and Ochrobactrum anthropi were detected in more than one subject. Other known species that were detected included Achromobacter spp., Afipia spp., Brevundimonas diminuta, Capnocytophaga spp., Cardiobacterium sp., Eikenella corrodens, Fusobacterium spp., Gemella haemoylsans, and Neisseria spp. Phylotypes that were unique to noma infections included those in the genera Eubacterium, Flavobacterium, Kocuria, Microbacterium, and Porphyromonas and the related Streptococcus salivarius and genera Sphingomonas and TREPONEMA: Since advanced noma lesions are infections open to the environment, it was not surprising to detect species not commonly associated with the oral cavity, e.g., from soil. Several species previously implicated as putative pathogens of noma, such as spirochetes and Fusobacterium spp., were detected in at least one subject. However, due to the limited number of available noma subjects, it was not possible at this time to associate specific species with the disease.

  2. Detection of DNA from a range of bacterial species in the knee joints of dogs with inflammatory knee arthritis and associated degenerative anterior cruciate ligament rupture.

    PubMed

    Muir, Peter; Oldenhoff, William E; Hudson, Alan P; Manley, Paul A; Schaefer, Susan L; Markel, Mark D; Hao, Zhengling

    2007-01-01

    Mixtures of bacterial nucleic acids can often be detected in synovial joints affected with arthritis. We investigated the potential role of such mixtures of bacterial nucleic acids in the pathogenesis of arthritis in a naturally occurring canine model. Dogs with a common inflammatory knee arthritis in which associated pathological degenerative anterior cruciate ligament (ACL) rupture often develops were studied. Synovial biopsies were obtained from 43 dogs with the naturally occurring ACL rupture arthropathy, 12 dogs with normal knees and intact ACL, and 16 dogs with normal knees and experimentally induced ACL rupture. Using PCR, specimens were tested for Borrelia burgdorferi OspA and p66 gene sequences. Broad-ranging 16S rRNA primers were also used; 'panbacterial' PCR products were cloned and multiple clones were sequenced for bacterial identification. Synovium was also studied histologically. The presence of bacterial DNA within the synovium was significantly associated with the naturally occurring ACL rupture arthropathy (p<0.05); knee joints from 37% of these dogs were PCR-positive. Mixtures of bacterial DNA were common and often included environmental bacteria; predominant organisms included Borrelia burgdorferi and Stenotrophomonas maltophilia. DNA from environmental bacteria was only found in dogs with the naturally occurring ACL rupture arthopathy; joints from 33% of affected dogs contained such bacterial DNA. Synovial inflammation developed in dogs with both naturally occurring and experimentally induced ACL rupture, when compared with intact ACL controls (p<0.01). These results indicate that mixtures of DNA derived from environmental bacteria are commonly found in the knee joint of a naturally occurring canine arthropathy, often in association with a recognized joint pathogen. Our results also suggest that knee instability alone is not responsible for this finding and have led us to hypothesize that mixtures of bacterial DNA are an important causative

  3. Composting and vermicomposting experiences in the treatment and bioconversion of asphaltens from the Prestige oil spill.

    PubMed

    Martín-Gil, Jesús; Navas-Gracia, Luís Manuel; Gómez-Sobrino, Ernesto; Correa-Guimaraes, Adriana; Hernández-Navarro, Salvador; Sánchez-Báscones, Mercedes; del Carmen Ramos-Sánchez, María

    2008-04-01

    This work illustrates the effectiveness of composting and vermicomposting in degrading fuel-in-water emulsions from oil spills (chapapote), and the isolation of potentially useful microorganisms for its biodegradation. Firstly, an alternative to the biodegradation of asphaltens from the Prestige oil spill (still present in some chapapote rafts in the Cantabrian coast) by means of the application of composting techniques to a microbial partnership acclimated to fuel-oil is offered. Our aim is that, after a relatively short period of time, the microorganisms can obtain its source of carbon and energy from asphaltens. The addition of metabolic co-substrates, like cow bed and potato peelings, allows the fragmentation of complex compounds into smaller structures, susceptible to further degradation. Afterwards, a maturation of the compost by means of a treatment with earthworms (Eisenia foetida) is necessary. Thus, through the vermicomposting it will be possible to obtain a valued product, useful in the processes of ground amendment, with little presence of asphaltens and occluded polycyclic aromatic hydrocarbons, rich in humus, and with an important bacterial flora of Bacillus genera, so that it can be typical of co-activators and accelerating products in composting processes. Along with this article, we show some parameters that control the evolution of the compost products (evolved gases, acidity, temperature and humidity); the chemical and microbiological analytical results; and the germination assays of vermicomposting. Results reveal that by using microorganisms living in either earthworm intestines (Stenotrophomonas maltophilia) or vermiculture substrates (Scedosporium apiospermium), it is possible to degrade and to eliminate the polycyclic asphaltens into CO(2) and H(2)O, helped by evaporation, dissolution and/or photo-oxidation processes. The obtained end product has contents of interesting vegetal nutrients and, mainly, it displays very high germination indices.

  4. Evaluation of microorganisms cultured from injured and repressed tissue regeneration sites in endangered giant aquatic Ozark Hellbender salamanders.

    PubMed

    Nickerson, Cheryl A; Ott, C Mark; Castro, Sarah L; Garcia, Veronica M; Molina, Thomas C; Briggler, Jeffrey T; Pitt, Amber L; Tavano, Joseph J; Byram, J Kelly; Barrila, Jennifer; Nickerson, Max A

    2011-01-01

    Investigation into the causes underlying the rapid, global amphibian decline provides critical insight into the effects of changing ecosystems. Hypothesized and confirmed links between amphibian declines, disease, and environmental changes are increasingly represented in published literature. However, there are few long-term amphibian studies that include data on population size, abnormality/injury rates, disease, and habitat variables to adequately assess changes through time. We cultured and identified microorganisms isolated from abnormal/injured and repressed tissue regeneration sites of the endangered Ozark Hellbender, Cryptobranchus alleganiensis bishopi, to discover potential causative agents responsible for their significant decline in health and population. This organism and our study site were chosen because the population and habitat of C. a. bishopi have been intensively studied from 1969-2009, and the abnormality/injury rate and apparent lack of regeneration were established. Although many bacterial and fungal isolates recovered were common environmental organisms, several opportunistic pathogens were identified in association with only the injured tissues of C.a. bishopi. Bacterial isolates included Aeromonas hydrophila, a known amphibian pathogen, Granulicetella adiacens, Gordonai terrae, Stenotrophomonas maltophilia, Aerococcus viridans, Streptococcus pneumoniae and a variety of Pseudomonads, including Pseudomonas aeruginosa, P. stutzeri, and P. alcaligenes. Fungal isolates included species in the genera Penicillium, Acremonium, Cladosporium, Curvularia, Fusarium, Streptomycetes, and the Class Hyphomycetes. Many of the opportunistic pathogens identified are known to form biofilms. Lack of isolation of the same organism from all wounds suggests that the etiological agent responsible for the damage to C. a. bishopi may not be a single organism. To our knowledge, this is the first study to profile the external microbial consortia cultured from a

  5. Update on Acinetobacter species: mechanisms of antimicrobial resistance and contemporary in vitro activity of minocycline and other treatment options.

    PubMed

    Castanheira, Mariana; Mendes, Rodrigo E; Jones, Ronald N

    2014-12-01

    Among Acinetobacter species, A. baumannii and other closely related species are commonly implicated in nosocomial infections. These organisms are usually multidrug resistant (MDR), and therapeutic options to treat A. baumannii infections are very limited. Clinicians have been resorting to older antimicrobial agents to treat infections caused by MDR A. baumannii, and some of these agents have documented toxicity and/or are not optimized for the infection type to be treated. Recent clinical experience supported by antimicrobial susceptibility data suggests that minocycline has greater activity than other tetracyclines and glycylcyclines against various MDR pathogens that have limited therapeutic options available, including Acinetobacter species. An intravenous formulation of minocycline has recently become available for clinical use, and in contrast to most older tetracyclines, minocycline has high activity against Acinetobacter species. In this report, we summarized some of the characteristics of the tetracycline class, and quantified the minocycline activity against contemporary (2007-2011) isolates and its potential therapeutic role against a collection of 5477 A. baumannii and other relevant gram-negative organisms when compared directly with tetracycline, doxycycline, and other broad-spectrum antimicrobial agents. Acinetobacter baumannii strains were highly resistant to all agents tested, with the exception of minocycline (79.1% susceptible) and colistin (98.8% susceptible). Minocycline (minimum inhibitory concentration that inhibits 50% and 90% of the isolates [MIC(50/90)]: 1/8 µg/mL) displayed greater activity than doxycycline (MIC(50/90): 2/>8 µg/mL) and tetracycline hydrochloride (HCL) (only 30.2% susceptible) against A. baumannii isolates, and was significantly more active than other tetracyclines against Burkholderia cepacia, Escherichia coli, Serratia marcescens, and Stenotrophomonas maltophilia isolates. In vitro susceptibility testing using

  6. Microbial Contamination of Glaucoma Eyedrops Used by Patients Compared With Ocular Medications Used in the Hospital

    PubMed Central

    Teuchner, Barbara; Wagner, Julia; Bechrakis, Nikolaos E.; Orth-Höller, Dorothea; Nagl, Markus

    2015-01-01

    Abstract The aim of this study was to compare the percentage of contamination of multiuse eyedrops applied by glaucoma patients at home and by the medical personnel at the outpatient department, the ward, and the operating room of our Department of Ophthalmology. Eyedrops were collected over a period of 11 months. Samples were taken from the dropper tip (smear), drops, and the residual fluid inside the bottle and cultivated on blood agar. Colony forming units were counted and identified by mass spectrometry. The percentage of contamination was significantly higher in eyedrops applied by the patients (29/119; 24.4%, P < 0.01), used in the ward (26/133; 19.5%, P < 0.01), and in the outpatient unit (6/35; 17.1%, P = 0.036) compared with that in the operating room (6/113; 5.3%). The median period of use was 1 week in the operating room compared with 4 weeks in the other groups (P < 0.01). Glaucoma medications were significantly more frequently contaminated than antibiotic and anesthetic eyedrops (P < 0.05). For eyedrops applied by the patients, the tip was more frequently contaminated than the drops and the residual internal fluid. For eyedrops from the ward, the opposite was true. Pathogenic strains (Pseudomonas aeruginosa, Serratia marcescens, Acinetobacter lwoffii, Stenotrophomonas maltophilia, and Staphylococcus aureus) were found only in 6 bottles (1.5%), whereas most of the detected microbes belonged to human or environmental flora. This study underlines the importance of hygienic handling of eyedrops and raises the question of whether single-use glaucoma medication might be preferred to reduce the risk of contamination. PMID:25715262

  7. Microbial consortia that degrade 2,4-DNT by interspecies metabolism: isolation and characterisation.

    PubMed

    Snellinx, Zita; Taghavi, Safieh; Vangronsveld, Jaco; van der Lelie, Daniël

    2003-01-01

    Two consortia, isolated by selective enrichment from a soil sample of a nitroaromatic-contaminated site, degraded 2,4-DNT as their sole nitrogen source without accumulating one or more detectable intermediates. Though originating from the same sample, the optimised consortia had no common members, indicating that selective enrichment resulted in different end points. Consortium 1 and consortium 2 contained four and six bacterial species respectively, but both had two members that were able to collectively degrade 2,4-DNT. Variovorax paradoxus VM685 (consortium 1) and Pseudomonas sp. VM908 (consortium 2) initiate the catabolism of 2,4-DNT by an oxidation step, thereby releasing nitrite and forming 4-methyl-5-nitrocatechol (4M5NC). Both strains contained a gene similar to the dntAa gene encoding 2,4-DNT dioxygenase. They subsequently metabolised 4M5NC to 2-hydroxy-5-methylquinone (2H5MQ) and nitrite, indicative of DntB or 4M5NC monooxygenase activity. A second consortium member, Pseudomonas marginalis VM683 (consortium 1) and P. aeruginosa VM903, Sphingomonas sp. VM904, Stenotrophomonas maltophilia VM905 or P. viridiflava VM907 (consortium 2), was found to be indispensable for efficient growth of the consortia on 2,4-DNT and for efficient metabolisation of the intermediates 4M5NC and 2H5MQ. Knowledge about the interactions in this step of the degradation pathway is rather limited. In addition, both consortia can use 2,4-DNT as sole nitrogen and carbon source. A gene similar to the dntD gene of Burkholderia sp. strain DNT that catalyses ring fission was demonstrated by DNA hybridisation in the second member strains. To our knowledge, this is the first time that consortia are shown to be necessary for 2,4-DNT degradation.

  8. ['In vitro' activity of different antimicrobial agents on Gram-negative nonfermentative bacilli, excluding Pseudomonas aeruginosa and Acinetobacter spp].

    PubMed

    Vay, C A; Almuzara, M N; Rodríguez, C H; Pugliese, M L; Lorenzo Barba, F; Mattera, J C; Famiglietti, A M R

    2005-01-01

    Gram-negative nonfermentative bacilli (NFB) are widely spread in the environment. Besides of difficulties for identification, they often have a marked multiresistance to antimicrobial agents, including those active against Pseudomonas aeruginosa. The objective of this study was to evaluate the 'in vitro' activity of different antimicrobial agents on 177 gram-negative nonfermentative bacilli isolates (excluding Pseudomonas aeruginosa and Acinetobacter spp.) isolated from clinical specimens. Minimum inhibitory concentrations (MIC) were determined according to the Mueller Hinton agar dilution method against the following antibacterial agents: ampicillin, piperacillin, piperacillin-tazobactam, sulbactam, cefoperazone, cefoperazone-sulbactam, ceftazidime, cefepime, aztreonam, imipenem, meropenem, colistin, gentamicin, amikacin, trimethoprim-sulfamethoxazole, chloramphenicol, erythromycin, rifampin, norfloxacin, ciprofloxacin and minocycline. Seven isolates: Sphingobacterium multivorum (2), Sphingobacteriumspiritivorum (1), Empedobacterbrevis (1), Weeksella virosa (1), Bergeyella zoohelcum (1) and Oligella urethralis (1), were tested for amoxicillin-clavulanic acid and ampicillin-sulbactam susceptibility, and susceptibility to cefoperazone or sulbactam was not determined. Multiresistance was generally found in Stenotrophomonas maltophilia, Burkholderia cepacia, Chryseobacterium spp., Myroides spp., Achromobacter xylosoxidans, and Ochrobactrum anthropi isolates. On the other hand, Pseudomonas stutzeri, Shewanella putrefaciens-algae, Sphingomonas paucimobilis, and Pseudomonas oryzihabitans, Bergeyella zoohelcum, Weeksella virosa and Oligella urethralis were widely susceptible to the antibacterial agents tested. As a result of the wide variation in antimicrobial susceptibility shown by different species, a test on susceptibility to different antibacterial agents is essential in order to select an adequate therapy. The marked multiresistance evidenced by some species

  9. AmpG is required for BlaXc beta-lactamase expression in Xanthomonas campestris pv. campestris str. 17.

    PubMed

    Yang, Tsuey-Ching; Chen, Tzu-Fan; Tsai, Jeffrey J P; Hu, Rouh-Mei

    2013-03-01

    The chromosomal ampR(Xc) -bla(Xc) module is essential for the β-lactam resistance of Xanthomonas campestris pv. campestris. Bla(Xc) β-lactamase is expressed at a high basal level in the absence of an inducer and its expression can be further induced by β-lactam. In enterobacteria, ampG encodes an inner membrane facilitator involved in the recycling of murein degradation compounds. An isogenic ampG mutant (XcampG) of X. campestris pv. campestris str. 17 (Xc17) was constructed to investigate the link between murein recycling and bla(Xc) expression. Our data demonstrate that (1) XcampG is susceptible to β-lactam antibiotics; (2) AmpG(Xc) is essential for expression of bla(Xc) ; (3) AmpGs of Xc17, Stenotrophomonas maltophilia KJ (SmKJ) and Escherichia coli DH5α can complement the defect of XcampG; (4) overexpression of AmpG(X) (c) significantly increased bla(Xc) expression; and (5) AmpG(Xc) from Xc17 is able to restore β-lactamase induction of the ampN(Xc) -ampG(Xc) double mutant of SmKJ. In Xc17, ampG(Xc) can be expressed from the promoter residing in the intergenic region of ampN(Xc) -ampG(Xc) and the expression is independent of β-lactam induction. AmpN, which is required for β-lactamases induction in SmKJ, is not required for the β-lactam antibiotic resistance of Xc17.

  10. MALDI-TOF: a useful tool for laboratory identification of uncommon glucose non-fermenting Gram-negative bacteria associated with cystic fibrosis.

    PubMed

    Homem de Mello de Souza, Helena Aguilar Peres; Dalla-Costa, Libera Maria; Vicenzi, Fernando José; Camargo de Souza, Dilair; Riedi, Carlos Antônio; Filho, Nelson Augusto Rosario; Pilonetto, Marcelo

    2014-09-01

    The predisposition of patients with cystic fibrosis (CF) for recurrent pulmonary infections can result in poor prognosis of the disease. Although the clinical significance in CF of micro-organisms, such as Staphylococcus aureus, Haemophilus influenzae and Pseudomonas aeruginosa, is well established, the implication of uncommon glucose non-fermenting Gram-negative bacilli (UGNF-GNB) in respiratory samples from CF patients is still unclear. Because of limitations of traditional methods used in most clinical laboratories, the accurate identification of these microbes is a challenge. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) is an alternative tool for efficient identification of bacteria. This was a retrospective study to evaluate different identification methods in a collection of UGNF-GNB isolated from children with CF during a period of three years. The performance of MALDI-TOF was compared to that of 16S rDNA gene sequencing and to a conventional and automated phenotypic identification. The discriminatory power of MALDI-TOF (75.0 % agreement) was superior to automated techniques (67.1 % agreement) and to conventional phenotypical identification (50.0 % agreement). MALDI-TOF also demonstrated high accuracy in identifying Stenotrophomonas maltophilia, Achromobacter xylosoxidans and Chryseobacterium indologenes, but had limited utility in identifying Pandoraea spp. and some species of Acinetobacter and Chryseobacterium (other than C. indologenes). Although MALDI-TOF identified only 75 % of the isolates in comparison with 16S rDNA gene sequencing, the prompt identification and high discriminatory power exhibited by MALDI-TOF make it a useful tool for the characterization of micro-organisms that are difficult to identify using routine methods.

  11. Metagenomic analysis of medicinal Cannabis samples; pathogenic bacteria, toxigenic fungi, and beneficial microbes grow in culture-based yeast and mold tests

    PubMed Central

    McKernan, Kevin; Spangler, Jessica; Helbert, Yvonne; Lynch, Ryan C.; Devitt-Lee, Adrian; Zhang, Lei; Orphe, Wendell; Warner, Jason; Foss, Theodore; Hudalla, Christopher J.; Silva, Matthew; Smith, Douglas R.

    2016-01-01

    Background: The presence of bacteria and fungi in medicinal or recreational Cannabis poses a potential threat to consumers if those microbes include pathogenic or toxigenic species. This study evaluated two widely used culture-based platforms for total yeast and mold (TYM) testing marketed by 3M Corporation and Biomérieux, in comparison with a quantitative PCR (qPCR) approach marketed by Medicinal Genomics Corporation. Methods: A set of 15 medicinal Cannabis samples were analyzed using 3M and Biomérieux culture-based platforms and by qPCR to quantify microbial DNA. All samples were then subjected to next-generation sequencing and metagenomics analysis to enumerate the bacteria and fungi present before and after growth on culture-based media. Results: Several pathogenic or toxigenic bacterial and fungal species were identified in proportions of >5% of classified reads on the samples, including Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa, Ralstonia pickettii, Salmonella enterica, Stenotrophomonas maltophilia, Aspergillus ostianus, Aspergillus sydowii, Penicillium citrinum and Penicillium steckii. Samples subjected to culture showed substantial shifts in the number and diversity of species present, including the failure of Aspergillus species to grow well on either platform. Substantial growth of Clostridium botulinum and other bacteria were frequently observed on one or both of the culture-based TYM platforms. The presence of plant growth promoting (beneficial) fungal species further influenced the differential growth of species in the microbiome of each sample. Conclusions: These findings have important implications for the Cannabis and food safety testing industries. PMID:27853518

  12. Chlor-alkali plant contamination of Aussa River sediments induced a large Hg-resistant bacterial community

    NASA Astrophysics Data System (ADS)

    Baldi, Franco; Marchetto, Davide; Gallo, Michele; Fani, Renato; Maida, Isabel; Covelli, Stefano; Fajon, Vesna; Zizek, Suzana; Hines, Mark; Horvat, Milena

    2012-11-01

    A closed chlor-alkali plant (CAP) discharged Hg for decades into the Aussa River, which flows into Marano Lagoon, resulting in the large-scale pollution of the lagoon. In order to get information on the role of bacteria as mercury detoxifying agents, analyses of anions in the superficial part (0-1 cm) of sediments were conducted at four stations in the Aussa River. In addition, measurements of biopolymeric carbon (BPC) as a sum of the carbon equivalent of proteins (PRT), lipids (LIP), and carbohydrates (CHO) were performed to correlate with bacterial biomass such as the number of aerobic heterotrophic cultivable bacteria and their percentage of Hg-resistant bacteria. All these parameters were used to assess the bioavailable Hg fraction in sediments and the potential detoxification activity of bacteria. In addition, fifteen isolates were characterized by a combination of molecular techniques, which permitted their assignment into six different genera. Four out of fifteen were Gram negative with two strains of Stenotrophomonas maltophilia, one Enterobacter sp., and one strain of Brevibacterium frigoritolerans. The remaining strains (11) were Gram positive belonging to the genera Bacillus and Staphylococcus. We found merA genes in only a few isolates. Mercury volatilization from added HgCl2 and the presence of plasmids with the merA gene were also used to confirm Hg reductase activity. We found the highest number of aerobic heterotrophic Hg-resistant bacteria (one order magnitude higher) and the highest number of Hg-resistant species (11 species out of 15) at the confluence of the River Aussa and Banduzzi's channel, which transport Hg from the CAP, suggesting that Hg is strongly detoxified [reduced to Hg(0)] at this location.

  13. Increasing Incidence of Multidrug Resistance Among Cystic Fibrosis Respiratory Bacterial Isolates.

    PubMed

    Rutter, W Cliff; Burgess, Donna R; Burgess, David S

    2017-01-01

    Pseudomonas aeruginosa and Staphylococcus aureus are common pathogens in cystic fibrosis (CF) patients with increasing multidrug resistance (MDR). This study characterized antimicrobial susceptibility trends among organisms isolated from the respiratory tract of CF patients. Microbiological culture and sensitivity results for all CF patients were collected from January 2010 through December 2014. Minimum inhibitory concentrations were obtained using Phoenix(®) and Etest(®) methods. Clinical and Laboratory Standards Institute guidelines were used to remove duplicate isolates and develop antimicrobial susceptibility reports. MDR was defined as resistance to one agent in three or more antibiotic classes or oxacillin resistance in S. aureus. Overall, 542 bacterial isolates from 376 cultures were analyzed for trends. P. aeruginosa (41%), S. aureus (40%), and Stenotrophomonas maltophilia (8%) were the most commonly isolated organisms. Multidrug-resistant organism isolation increased from 39% to 49% (r = 0.76, p = 0.13), while representing 47.6% of all isolates. Multidrug-resistant P. aeruginosa incidence increased each year from 26% to 43% (r = 0.89, p = 0.046), while P. aeruginosa isolation decreased from 47% to 38% over the study period (r = -0.93, p = 0.02). MRSA accounted for 62.6% of all S. aureus isolated, while overall multidrug-resistant S. aureus incidence was 73.1% in all cultures. MDR among common pathogens in CF continues to increase. Empiric therapy for CF exacerbations should be targeted to previous antimicrobial susceptibility, and P. aeruginosa and S. aureus should be empirically covered.

  14. Additional observations and notes on the natural history of the prairie rattlesnake (Crotalus viridis) in Colorado.

    PubMed

    Fitzgerald, Kevin T; Shipley, Bryon K; Newquist, Kristin L; Vera, Rebecca; Flood, Aryn A

    2013-11-01

    On account of their unique anatomy, physiology, natural history, ecology, and behavior, rattlesnakes make ideal subjects for a variety of different scientific disciplines. The prairie rattlesnake (Crotalus viridis) in Colorado was selected for investigation of its relationship to colonies of black-tailed prairie dogs (Cynomys ludovicianus) with regard to spatial ecology. A total of 31 snakes were anesthetized and had radiotransmitters surgically implanted. In addition, at the time of their capture, all snakes underwent the following: (1) they had bacterial culture taken from their mouths for potential isolation of pathogenic bacteria; (2) similarly, they had cloacal bacterial cultures taken to assess potentially harmful bacteria passed in the feces; and (3) they had blood samples drawn to investigate the presence of any zoonotic agents in the serum of the snakes. The results of the study and their implications are discussed here. Traditionally, a low incidence of bacterial wound infection has been reported following snakebite. Nevertheless, the oral cavity of snakes has long been known to house a wide variety of bacterial flora. In our study, 10 different bacterial species were isolated from the mouths of the rattlesnakes, 6 of which are capable of being zoonotic pathogens and inducing human disease. More studies are necessary to see why more rattlesnake bites do not become infected despite the presence of such pathogenic bacteria. The results of fecal bacteria isolated revealed 13 bacterial species, 12 of which can cause disease in humans. Of the snakes whose samples were cultured, 26% were positive for the presence of the pathogen Salmonella arizonae, one of the causative agents of reptile-related salmonellosis in humans. It has long been reported that captive reptiles have a much higher incidence than wild, free-ranging species. This study shows the incidence of Salmonella in a wild, free-ranging population of rattlesnakes. In addition, Stenotrophomonas

  15. [Post-marketing surveillance of antibacterial activities of cefozopran against various clinical isolates--II. Gram-negative bacteria].

    PubMed

    Igari, Jun; Oguri, Toyoko; Hiramatsu, Nobuyoshi; Akiyama, Kazumitsu; Koyama, Tsuneo

    2003-10-01

    As a post-marketing surveillance, the in vitro antibacterial activities of cefozopran (CZOP), an agent of cephems, against various clinical isolates were yearly evaluated and compared with those of other cephems, oxacephems, carbapenems, monobactams, and penicillins. Changes in CZOP susceptibility among bacteria were also evaluated with the bacterial resistance ratio calculated from the breakpoint MIC. Twenty-five species (4,154 strains) of Gram-negative bacteria were isolated from the clinical materials annually collected from 1996 to 2001, and consisted of Moraxella (Branhamella) catarrhalis, Haemophilus influenzae, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens, Serratia liquefaciens, Citrobacter freundii, Citrobacter koseri, Proteus mirabilis, Proteus vulgaris, Morganella morganii, Providencia spp., Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida, Acinetobacter baumannii, Acinetobacter Iwoffii, Burkholderia cepacia, Stenotrophomonas maltophilia, Bacteroides fragilis group, and Prevotella/Porphyromonas. CZOP preserved its antibacterial activity against M. (B.) catarrhalis (MIC90: 4 micrograms/mL) and showed comparable activity to carbapenems against H. influenzae (MIC90: 1 microgram/mL). The antibacterial activity of CZOP against E. coli was preferable (MIC90: 0.125 microgram/mL) and comparable to those of cefpirome (CPR), cefepime (CFPM), and imipenem (IPM). The MIC90 of CZOP against K. pneumoniae and K. oxytoca was 1 and 0.25 microgram/mL, respectively. The MIC90 of CZOP against E. cloacae increased during 6 years (32 to 128 micrograms/mL). The antibacterial activity of CZOP against E. aerogenes was preferable (MIC90: 1 microgram/mL). The antibacterial activities of CZOP against S. marcescens and S. liquefaciens were relatively potent (MIC90: 0.5 and 0.25 microgram/mL) and comparable to those of CPR, CFPM, and carumonam. CZOP preserved comparable antibacterial

  16. Nosocomial pneumonia : rationalizing the approach to empirical therapy.

    PubMed

    Andriesse, Gunnar I; Verhoef, Jan

    2006-01-01

    Nosocomial pneumonia or hospital-acquired pneumonia (HAP) causes considerable morbidity and mortality. It is the second most common nosocomial infection and the leading cause of death from hospital-acquired infections. In 1996 the American Thoracic Society (ATS) published guidelines for empirical therapy of HAP. This review focuses on the literature that has appeared since the ATS statement. Early diagnosis of HAP and its etiology is crucial in guiding empirical therapy. Since 1996, it has become clear that differentiating mere colonization from etiologic pathogens infecting the lower respiratory tract is best achieved by employing bronchoalveolar lavage (BAL) or protected specimen brush (PSB) in combination with quantitative culture and detection of intracellular microorganisms. Endotracheal aspirate and non-bronchoscopic BAL/PSB in combination with quantitative culture provide a good alternative in patients suspected of ventilator-associated pneumonia. Since culture results take 2-3 days, initial therapy of HAP is by definition empirical. Epidemiologic studies have identified the most frequently involved pathogens: Enterobacteriaceae, Haemophilus influenzae, Streptococcus pneumoniae and Staphylococcus aureus ('core pathogens'). Empirical therapy covering only the 'core pathogens' will suffice in patients without risk factors for resistant microorganisms. Studies that have appeared since the ATS statement issued in 1996, demonstrate several new risk factors for HAP with multiresistant pathogens. In patients with risk factors, empirical therapy should consist of antibacterials with a broader spectrum. The most important risk factors for resistant microorganisms are late onset of HAP (>/=5 days after admission), recent use of antibacterial therapy, and mechanical ventilation. Multiresistant bacteria of specific interest are methicillin-resistant S. aureus (MRSA), Pseudomonas aeruginosa, Acinetobacter calcoaceticus-baumannii, Stenotrophomonas maltophilia and extended

  17. Biochemical Sequence Analyses of GES-1, a Novel Class A Extended-Spectrum β-Lactamase, and the Class 1 Integron In52 from Klebsiella pneumoniae

    PubMed Central

    Poirel, Laurent; Le Thomas, Isabelle; Naas, Thierry; Karim, Amal; Nordmann, Patrice

    2000-01-01

    Klebsiella pneumoniae ORI-1 was isolated in 1998 in France from a rectal swab of a 1-month-old girl who was previously hospitalized in Cayenne Hospital, Cayenne, French Guiana. This strain harbored a ca. 140-kb nontransferable plasmid, pTK1, that conferred an extended-spectrum cephalosporin resistance profile antagonized by the addition of clavulanic acid, tazobactam, or imipenem. The gene for GES-1 (Guiana extended-spectrum β-lactamase) was cloned, and its protein was expressed in Escherichia coli DH10B, where this pI-5.8 β-lactamase of a ca. 31-kDa molecular mass conferred resistance to oxyimino cephalosporins (mostly to ceftazidime). GES-1 is weakly related to the other plasmid-located Ambler class A extended-spectrum β-lactamases (ESBLs). The highest percentage of amino acid identity was obtained with the carbenicillinase GN79 from Proteus mirabilis; with YENT, a chromosome-borne penicillinase from Yersinia enterocolitica; and with L-2, a chromosome-borne class A cephalosporinase from Stenotrophomonas maltophilia (36% amino acid identity each). However, a dendrogram analysis showed that GES-1 clustered within a class A ESBL subgroup together with ESBLs VEB-1 and PER-1. Sequencing of a 7,098-bp DNA fragment from plasmid pTK1 revealed that the GES-1 gene was located on a novel class 1 integron named In52 that was characterized by (i) a 5′ conserved segment containing an intI1 gene possessing two putative promoters, P1 and P2, for coordinated expression of the downstream antibiotic resistance genes and an attI1 recombination site; (ii) five antibiotic gene cassettes, blaGES-1, aac(6′)Ib′ (gentamicin resistance and amikacin susceptibility), dfrXVb (trimethoprim resistance), a novel chloramphenicol resistance gene (cmlA4), and aadA2 (streptomycin-spectinomycin resistance); and (iii) a 3′ conserved segment consisting of qacEΔ1 and sulI. The blaGES-1 and aadA2 gene cassettes were peculiar, since they lacked a typical 59-base element. This work identified

  18. Application of a constructed wetland system for polluted stream remediation

    NASA Astrophysics Data System (ADS)

    Tu, Y. T.; Chiang, P. C.; Yang, J.; Chen, S. H.; Kao, C. M.

    2014-03-01

    ., Steroidobacter denitrificans, Hydrocarboniphaga effuse were responsible for nitrogen removal, and the dominant carbon degrading bacteria (Stenotrophomonas maltophilia, H. effuse, Alcaligenes sp., Pseudomonas sp., Fusibacter sp., Chlofoflexi, Guggenheimella bovis, Bacillus pumilus) were responsible for carbon reduction. The denaturing gradient gel electrophoresis (DGGE) and nucleotide sequence techniques provide a guide for microbial ecology evaluation, which can be used as an indication of contaminants removal. Results from this study show that constructed wetlands have the potential to be developed into an environmentally acceptable river water quality improvement and wastewater polishment alternative for practical application.

  19. Bacterial constituents of indoor air in a high throughput building in the tropics.

    PubMed

    Li, Tee Chin; Ambu, Stephen; Mohandas, Kavitha; Wah, Mak Joon; Sulaiman, Lokman Hakim; Murgaiyah, Malathi

    2014-09-01

    Airborne bacteria are significant biotic constituents of bioaerosol. Bacteria at high concentrations in the air can compromise indoor air quality (IAQ) and result in many diseases. In tropical environments like Malaysia that extensively utilize air-conditioning systems, this is particularly significant due to continuous recirculation of indoor air and the potential implications for human health. Currently, there is a lack of knowledge regarding the impact of airborne bacteria on IAQ in Malaysia. This study was prompted by a need for reliable baseline data on airborne bacteria in the indoor environment of tropical equatorial Malaysia, that may be used as a reference for further investigations on the potential role played by airborne bacteria as an agent of disease in this region. It was further necessitated due to the threat of bioterrorism with the potentiality of release of exotic pathogenic microorganisms into indoor or outdoor air. Before scientists can detect the latter, a gauge of the common microorganisms in indoor (as well as outdoor) air needs to be ascertained, hence the expediency of this study. Bacterial counts from the broad-based and targeted study were generally in the order of 10(2) colony-forming units (CFU) per m(3) of air. The most prevalent airborne bacteria found in the broad-based study that encompassed all five levels of the building were Gram-positive cocci (67.73%), followed by Gram-positive rods (24.26%) and Gram-negative rods (7.10%). Gram-negative cocci were rarely detected (0.71%). Amongst the genera identified, Kytococcus sp., Micrococcus sp., Staphylococcus sp., Leifsonia sp., Bacillus sp. and Corynebacterium sp. predominated in indoor air. The most dominant bacterial species were Kytococcus sedentarius, Staphylococcus epidermidis and Micrococcus luteus. The opportunistic and nosocomial pathogen, Stenotrophomonas maltophilia was also discovered at a high percentage in the cafeteria. The bacteria isolated in this study have been

  20. In vivo development of daptomycin resistance in vancomycin-susceptible methicillin-resistant Staphylococcus aureus severe infections previously treated with glycopeptides.

    PubMed

    Capone, A; Cafiso, V; Campanile, F; Parisi, G; Mariani, B; Petrosillo, N; Stefani, S

    2016-04-01

    Our aim was to describe the clinical and microbiological features of four cases of severe vancomycin-susceptible methicillin-resistant Staphylococcus aureus (MRSA) infections in which the vancomycin non-susceptibility development and daptomycin resistance occurred under therapy with teicoplanin (three cases) and daptomycin switched to vancomycin (one case). Clinical data were retrospectively reviewed. On nine clinical epidemiologically unrelated daptomycin-susceptible (DAP-S) and daptomycin-resistant (DAP-R) MRSA, we performed: (i) DAP-VAN-TEC-CFX-RIF minimum inhibitory concentrations (MICs); (ii) glycopeptide resistance detection (GRD) by δ-hemolysis; (iii) glycopeptide population analysis; (iv) molecular characterization by PFGE-MLST-SCCmec-agr-typing; (v) rpoB and mprF single nucleotide polymorphisms (SNPs); (vi) dltA-mprF-atl-sceD expression by real-time quantitative polymerase chain reaction (qPCR). Three out of the four patients did not survive despite salvage treatment; two died with active MRSA infection and one died because of Stenotrophomonas maltophilia sepsis. The fourth patient, in which a reversion to a DAP-S phenotype occurred, survived with daptomycin plus trimethoprim/sulfamethoxazole and oxacillin treatment, and endovascular device removal. Daptomycin resistance development was preceded by a stable heterogeneous vancomycin-intermediate S. aureus (hVISA) or VISA phenotype acquisition, while in one case, daptomycin resistance was preceded by an unstable daptomycin heteroresistance (hDAP) behavior reverting to DAP-S during vancomycin plus rifampin therapy followed by high doses of daptomycin. All DAP-R strains showed hVISA or DAP-R traits, including mutations and/or up-regulation of genes involved in cell wall turnover and cell membrane perturbation. In our study, daptomycin resistance arose during glycopeptide therapy. The emergence of DAP-R isolates was preceded by a stable VISA or hVISA phenotype or by instability reverting to a DAP

  1. Ceftobiprole Activity against over 60,000 Clinical Bacterial Pathogens Isolated in Europe, Turkey, and Israel from 2005 to 2010

    PubMed Central

    Flamm, Robert K.; Sader, Helio S.; Jones, Ronald N.

    2014-01-01

    Ceftobiprole medocaril is a newly approved drug in Europe for the treatment of hospital-acquired pneumonia (HAP) (excluding patients with ventilator-associated pneumonia but including ventilated HAP patients) and community-acquired pneumonia in adults. The aim of this study was to evaluate the in vitro antimicrobial activity of ceftobiprole against prevalent Gram-positive and -negative pathogens isolated in Europe, Turkey, and Israel during 2005 through 2010. A total of 60,084 consecutive, nonduplicate isolates from a wide variety of infections were collected from 33 medical centers. Species identification was confirmed, and all isolates were susceptibility tested using reference broth microdilution methods. Ceftobiprole had high activity against methicillin-susceptible Staphylococcus aureus (MSSA) (100.0% susceptible), methicillin-susceptible coagulase-negative staphylococci (CoNS), beta-hemolytic streptococci, and Streptococcus pneumoniae (99.3% susceptible), with MIC90 values of 0.25, 0.12, ≤0.06, and 0.5 μg/ml, respectively. Ceftobiprole was active against methicillin-resistant S. aureus (MRSA) (98.3% susceptible) and methicillin-resistant CoNS, having a MIC90 of 2 μg/ml. Ceftobiprole was active against Enterococcus faecalis (MIC50/90, 0.5/4 μg/ml) but not against most Enterococcus faecium isolates. Ceftobiprole was very potent against the majority of Enterobacteriaceae (87.3% susceptible), with >80% inhibited at ≤0.12 μg/ml. The potency of ceftobiprole against Pseudomonas aeruginosa (MIC50/90, 2/>8 μg/ml; 64.6% at MIC values of ≤4 μg/ml) was similar to that of ceftazidime (MIC50/90, 2/>16 μg/ml; 75.4% susceptible), but limited activity was observed against Acinetobacter spp. and Stenotrophomonas maltophilia. High activity was also observed against all Haemophilus influenzae (MIC90, ≤0.06 μg/ml) and Moraxella catarrhalis (MIC50/90, ≤0.06/0.25 μg/ml) isolates. Ceftobiprole demonstrated a wide spectrum of antimicrobial activity against this

  2. Intravenous Colistin Use for Multidrug-Resistant Gram-Negative Infections in Pediatric Patients

    PubMed Central

    Karaaslan, Ayşe; Çağan, Eren; Kadayifci, Eda Kepenekli; Atıcı, Serkan; Akkoç, Gülşen; Yakut, Nurhayat; Demir, Sevliya Öcal; Soysal, Ahmet; Bakır, Mustafa

    2016-01-01

    Background The emergence of infections due to multidrug-resistant Gram-negative bacilli (MDR-GNB) has led to the resurrection of colistin use. The data on colistin use and drug-related adverse effects in children are scarce. Aims In this study, we aimed to evaluate the clinical efficacy and safety of colistin use in critically ill pediatric patients. Study Design This study has a retrospective study design. Methods Sixty-one critically ill children were identified through the department’s patient files archive during the period from January 2011 to November 2014. Results Twenty-nine females and thirty-two males with a mean±standard deviation (SD) age of 61±9 months (range 0–216, median 12 months) received IV colistin due to MDR-GNB infections. Bacteremia (n=23, 37.7%) was the leading diagnosis, followed by pneumonia (n=19, 31%), clinical sepsis (n=7, 11.4%), wound infection (n=6, 9.8%), urinary tract infection (n=5, 8.1%) and meningitis (n=1, 1.6%). All of the isolates were resistant to carbapenems; however, all were susceptible to colistin. The isolated microorganisms in decreasing order of frequency were: Acinetobacter baumanni (n=27, 44.2%), Pseudomonas aeruginosa (n=17, 27.8%), Klebsiella pneumoniae (n=6, 9.8%), K. pneumoniae and Stenotrophomonas maltophilia (n=1, 1.6%), K. pneumoniae and A. baumanni (n=1, 1.6%), K. oxytoca (n=1, 1.6%) and Enterobacter cloacae (n=1, 1.6%). In seven patients, no microorganisms were detected; however, five of these patients were colonized by carbapenem-resistant K. pneumoniae. The mean duration of colistin therapy was 12 days (range 3–45). Colistin was administered concomitantly with one of the following antibiotics: carbapenem (n=50, %82), ampicillin-sulbactam (n=5, 8%), quinolones (n=5, 8%), rifampicin (n=1, 1.6%). Carbapenem was the most frequently used antibiotic. Nephrotoxicity was observed in only 1 patient, and we did not observe neurotoxicity in this study. All the patients received intravenous colistin

  3. Persistence of microbial communities including Pseudomonas aeruginosa in a hospital environment: a potential health hazard

    PubMed Central

    2014-01-01

    Background The persistence of microbial communities and how they change in indoor environments is of immense interest to public health. Moreover, hospital acquired infections are significant contributors to morbidity and mortality. Evidence suggests that, in hospital environments agent transfer between surfaces causes healthcare associated infections in humans, and that surfaces are an important transmission route and may act as a reservoir for some of the pathogens. This study aimed to evaluate the diversity of microorganisms that persist on noncritical equipment and surfaces in a main hospital in Portugal, and are able to grow in selective media for Pseudomonas, and relate them with the presence of Pseudomonas aeruginosa. Results During 2 years, a total of 290 environmental samples were analyzed, in 3 different wards. The percentage of equipment in each ward that showed low contamination level varied between 22% and 38%, and more than 50% of the equipment sampled was highly contaminated. P. aeruginosa was repeatedly isolated from sinks (10 times), from the taps’ biofilm (16 times), and from the showers and bedside tables (two times). Two ERIC clones were isolated more than once. The contamination level of the different taps analyzed showed correlation with the contamination level of the hand gels support, soaps and sinks. Ten different bacteria genera were frequently isolated in the selective media for Pseudomonas. Organisms usually associated with nosocomial infections as Stenotrophomonas maltophilia, Enterococcus feacalis, Serratia nematodiphila were also repeatedly isolated on the same equipment. Conclusions The environment may act as a reservoir for at least some of the pathogens implicated in nosocomial infections. The bacterial contamination level was related to the presence of humidity on the surfaces, and tap water (biofilm) was a point of dispersion of bacterial species, including potentially pathogenic organisms. The materials of the equipment

  4. Prevalence of Resistant Gram-Negative Bacilli in Bloodstream Infection in Febrile Neutropenia Patients Undergoing Hematopoietic Stem Cell Transplantation

    PubMed Central

    Wang, Ling; Wang, Ying; Fan, Xing; Tang, Wei; Hu, Jiong

    2015-01-01

    Abstract Bloodstream infection (BSI) is an important cause of morbidity and mortality in patients undergoing hematopoietic stem cell transplantation (HSCT). To evaluate the causative bacteria and identify risk factors for BSI associated mortality in febrile neutropenia patients undergoing HSCT, we collected the clinical and microbiological data from patients underwent HSCT between 2008 and 2014 and performed a retrospective analysis. Throughout the study period, among 348 episodes of neutropenic fever in patients underwent HSCT, 89 episodes in 85 patients had microbiological defined BSI with a total of 108 isolates. Gram-negative bacteria (GNB) were the most common isolates (76, 70.3%) followed by gram-positive bacteria (GPB, 29, 26.9%) and fungus (3, 2.8%). As to the drug resistance, 26 multiple drug resistance (MDR) isolates were identified. Resistant isolates (n = 23) were more common documented in GNB, mostly Escherichia coli (9/36, 25%) and Klebsiella pneumonia (6/24, 25%). A total of 12 isolated were resistant to carbapenem including 4 K pneumoniae (4/24, 16.7%), 3 Stenotrophomonas maltophilia, and 1 Pseudomonas aeruginosa and other 4 GNB isolates (Citrobacter freumdii, Pseudomonas stutzeri, Acinetobacter baumanii, and Chryseobacterium indologenes). As to the GPB, only 3 resistant isolates were documented including 2 methicillin-resistant isolates (Staphylococcus hominis and Arcanobacterium hemolysis) and 1 vancomycin-resistant Enterococcus faecium. Among these 85 patients with documented BSI, 11 patients died of BSI as primary or associated cause with a BSI-related mortality of 13.1 ± 3.7% and 90-day overall survival after transplantation at 80.0 ± 4.3%. Patients with high-risk disease undergoing allo-HSCT, prolonged neutropenia (≥15 days) and infection with carbapenem-resistant GNB were associated with BSI associated mortality in univariate and multivariate analyses. Our report revealed a prevalence of GNB in BSI of neutropenic patients

  5. Cross-class metallo-β-lactamase inhibition by bisthiazolidines reveals multiple binding modes

    PubMed Central

    Hinchliffe, Philip; González, Mariano M.; Mojica, Maria F.; González, Javier M.; Castillo, Valerie; Saiz, Cecilia; Kosmopoulou, Magda; Tooke, Catherine L.; Llarrull, Leticia I.; Mahler, Graciela; Bonomo, Robert A.; Vila, Alejandro J.; Spencer, James

    2016-01-01

    Metallo-β-lactamases (MBLs) hydrolyze almost all β-lactam antibiotics and are unaffected by clinically available β-lactamase inhibitors (βLIs). Active-site architecture divides MBLs into three classes (B1, B2, and B3), complicating development of βLIs effective against all enzymes. Bisthiazolidines (BTZs) are carboxylate-containing, bicyclic compounds, considered as penicillin analogs with an additional free thiol. Here, we show both l- and d-BTZ enantiomers are micromolar competitive βLIs of all MBL classes in vitro, with Kis of 6–15 µM or 36–84 µM for subclass B1 MBLs (IMP-1 and BcII, respectively), and 10–12 µM for the B3 enzyme L1. Against the B2 MBL Sfh-I, the l-BTZ enantiomers exhibit 100-fold lower Kis (0.26–0.36 µM) than d-BTZs (26–29 µM). Importantly, cell-based time-kill assays show BTZs restore β-lactam susceptibility of Escherichia coli-producing MBLs (IMP-1, Sfh-1, BcII, and GOB-18) and, significantly, an extensively drug-resistant Stenotrophomonas maltophilia clinical isolate expressing L1. BTZs therefore inhibit the full range of MBLs and potentiate β-lactam activity against producer pathogens. X-ray crystal structures reveal insights into diverse BTZ binding modes, varying with orientation of the carboxylate and thiol moieties. BTZs bind the di-zinc centers of B1 (IMP-1; BcII) and B3 (L1) MBLs via the free thiol, but orient differently depending upon stereochemistry. In contrast, the l-BTZ carboxylate dominates interactions with the monozinc B2 MBL Sfh-I, with the thiol uninvolved. d-BTZ complexes most closely resemble β-lactam binding to B1 MBLs, but feature an unprecedented disruption of the D120–zinc interaction. Cross-class MBL inhibition therefore arises from the unexpected versatility of BTZ binding. PMID:27303030

  6. Nile Red Detection of Bacterial Hydrocarbons and Ketones in a High-Throughput Format

    SciTech Connect

    Pinzon, NM; Aukema, KG; Gralnick, JA; Wackett, LP

    2011-06-28

    A method for use in high-throughput screening of bacteria for the production of long-chain hydrocarbons and ketones by monitoring fluorescent light emission in the presence of Nile red is described. Nile red has previously been used to screen for polyhydroxybutyrate (PHB) and fatty acid esters, but this is the first report of screening for recombinant bacteria making hydrocarbons or ketones. The microtiter plate assay was evaluated using wild-type and recombinant strains of Shewanella oneidensis and Escherichia coli expressing the enzyme OleA, previously shown to initiate hydrocarbon biosynthesis. The strains expressing exogenous Stenotrophomonas maltophilia oleA, with increased levels of ketone production as determined by gas chromatography-mass spectrometry, were distinguished with Nile red fluorescence. Confocal microscopy images of S. oneidensis oleA-expressing strains stained with Nile red were consistent with a membrane localization of the ketones. This differed from Nile red staining of bacterial PHB or algal lipid droplets that showed intracellular inclusion bodies. These results demonstrated the applicability of Nile red in a high-throughput technique for the detection of bacterial hydrocarbons and ketones. IMPORTANCE In recent years, there has been renewed interest in advanced biofuel sources such as bacterial hydrocarbon production. Previous studies used solvent extraction of bacterial cultures followed by gas chromatography-mass spectrometry (GC-MS) to detect and quantify ketones and hydrocarbons (Beller HR, Goh EB, Keasling JD, Appl. Environ. Microbiol. 76: 1212-1223, 2010; Sukovich DJ, Seffernick JL, Richman JE, Gralnick JA, Wackett LP, Appl. Environ. Microbiol. 76: 3850-3862, 2010). While these analyses are powerful and accurate, their labor-intensive nature makes them intractable to high-throughput screening; therefore, methods for rapid identification of bacterial strains that are overproducing hydrocarbons are needed. The use of high

  7. Bulgecin A as a β-lactam enhancer for carbapenem-resistant Pseudomonas aeruginosa and carbapenem-resistant Acinetobacter baumannii clinical isolates containing various resistance mechanisms

    PubMed Central

    Skalweit, Marion J; Li, Mei

    2016-01-01

    Genetic screening of Pseudomonas aeruginosa (PSDA) and Acinetobacter baumannii (ACB) reveals genes that confer increased susceptibility to β-lactams when disrupted, suggesting novel drug targets. One such target is lytic transglycosylase. Bulgecin A (BlgA) is a natural product of Pseudomonas mesoacidophila and a lytic transglycosolase inhibitor that works synergistically with β-lactams targeting PBP3 for Enterobacteriaceae. BlgA also weakly inhibits di-Zn2+ metallo-β-lactamases like L1 of Stenotrophomonas maltophilia. We hypothesized that because of its unique mechanism of action, BlgA could restore susceptibility to carbapenems in carbapenem-resistant PSDA (CR-PSDA) and carbapenem-resistant ACB, as well as ACB resistant to sulbactam. A BlgA-containing extract was prepared using a previously published protocol. CR-PSDA clinical isolates demonstrating a variety of carbapenem resistance mechanisms (VIM-2 carbapenemases, efflux mechanisms, and AmpC producer expression) were characterized with agar dilution minimum inhibitory concentration (MIC) testing and polymerase chain reaction. Growth curves using these strains were prepared using meropenem, BlgA extract, and meropenem plus BlgA extract. A concentrated Blg A extract combined with low concentrations of meropenem, was able to inhibit the growth of clinical strains of CR-PSDA for strains that had meropenem MICs ≥8 mg/L by agar dilution, and a clinical strain of an OXA-24 producing ACB that had a meropenem MIC >32 mg/L and intermediate ampicillin/sulbactam susceptibility. Similar experiments were conducted on a TEM-1 producing ACB strain resistant to sulbactam. BlgA with ampicillin/sulbactam inhibited the growth of this organism. As in Enterobacteriaceae, BlgA appears to restore the efficacy of meropenem in suppressing the growth of CR-PSDA and carbapenem-resistant ACB strains with a variety of common carbapenem resistance mechanisms. BlgA extract also inhibits VIM-2 β-lactamase in vitro. BlgA may prove to be

  8. Multispecies Biofilm Development on Space Station Heat Exhanger Core Material

    NASA Technical Reports Server (NTRS)

    Pyle, B. H.; Roth, S. R.; Vega, L. M.; Pickering, K. D.; Alvarez, Pedro J. J.; Roman, M. C.

    2007-01-01

    Investigations of microbial contamination of the cooling system aboard the International Space Station (ISS) suggested that there may be a relationship between heat exchanger (HX) materials and the degree of microbial colonization and biofilm formation. Experiments were undertaken to test the hypothesis that biofilm formation is influenced by the type and previous exposure of HX surfaces. Acidovorax delafieldii, Comamonas acidovorans, Hydrogenophaga pseudoflava, Pseudomonas stutzeri, Sphingomonas paucimobilis, and Stenotrophomonas maltophilia, originally isolated from ISS cooling system fluid, were cultured on R2A agar and suspended separately in fresh filter-sterilized ISS cooling fluid, pH 8.3. Initial numbers in each suspension ranged from 10(exp 6)-10(exp 7) CFU/ml, and a mixture contained greater than 10(exp 7) CFU/ml. Coupons of ISS HX material, previously used on orbit (HXOO) or unused (HXUU), polycarbonate (PC) and 316L polished stainless steel (SS) were autoclaved, covered with multispecies suspension in sterile tubes and incubated in the dark at ambient (22-25 C). Original HX material contained greater than 90% Ni, 4.5% Si, and 3.2% B, with a borate buffer. For approximately 10 weeks, samples of fluid were plated on R2A agar, and surface colonization assessed by SYBR green or BacLight staining and microscopy. Suspension counts for the PC and SC samples remained steady at around 10(exp 7) CFU/ml. HXUU counts declined about 1 log in 21 d then remained steady, and HXOO counts declined 2 logs in 28 d, fluctuated and stabilized about 10(exp 3) CFU/ml from 47-54 d. Predominantly yellow S. paucimobilis predominated on plates from HXOO samples up to 26 d, then white or translucent colonies of other species appeared. All colony types were seen on plates from other samples throughout the trial. Epifluorescence microscopy indicated microbial growth on all surfaces by 21 d, followed by variable colonization. After 54 d, all but the HXOO samples had well

  9. BSAC standardized disc susceptibility testing method (version 8).

    PubMed

    Andrews, J M

    2009-09-01

    There have been considerable changes to the format of the recommendations since the previous version (version 7). The majority of the footnotes to the tables have been removed and the notations added to the end column; it is hoped that this change will avoid confusion in interpretation. Antibiotics have been separated into groups, e.g. beta-lactams, aminoglycosides, etc. Recommendations for urinary tract infections (UTIs) have been removed for most agents except for those that are administered solely for the treatment of uncomplicated UTIs or where there are limited recommendations for specific organisms, e.g. trimethoprim. For agents that previously had dual recommendations, systemic recommendations remain and the intermediate category can be used for interpretation for UTIs because intermediate susceptibility infers that the infection may respond as the agent is concentrated at the site of infection. This change will also avoid errors in interpretation when an organism is isolated from multiple sites, e.g. blood and urine. The changes that have been made to version 7 are as follows: MIC and zone diameter breakpoints (BPs) for trimethoprim, fosfomycin and nitrofurantoin for UTIs (Table 7); MIC and zone diameter breakpoints (BPs) for doripenem (Tables 7-9); colistin MIC BPs for Pseudomonas spp. (Table 9), co-trimoxazole MIC BPs for Stenotrophomonas maltophilia (Table 10); staphylococci MIC and zone diameter BPs for clarithromycin, clindamycin, erythromycin, quinupristin/dalfopristin, trimethoprim UTI, nitrofurantoin UTI and rifampicin (Table 11); Streptococcus pneumoniae MIC and zone diameter BPs for azithromycin, clarithromycin, erythromycin, co-trimoxazole, linezolid, rifampicin and telithromycin (Table 12); addition of streptomycin recommendations for enterococci (Table 13); enterococcal MIC and zone diameter BPs for quinupristin/dalfopristin, nitrofurantoin UTI and trimethoprim UTI (Table 13); beta-haemolytic streptococci MIC and zone diameter BPs for

  10. BSAC standardized disc susceptibility testing method (version 7).

    PubMed

    Andrews, J M

    2008-08-01

    The changes that have been made to the previous version of the recommendations (version 6) are as follows: medium and incubation condition for testing Acinetobacter spp. (Tables 1 and 6); use of cefoxitin as an indicator antibiotic for detecting methicillin/oxacillin/cefoxitin resistance in coagulase-negative staphylococci (Tables 1, 6 and 11); MIC breakpoint for co-trimoxazole based on the trimethoprim concentration in a 1:19 combination with sulfamethoxazole (Tables 7, 10, 11, 12, 15, 16 and 19); advice on the use of azithromycin for the treatment of infections with Salmonella typhi (footnote to Table 7); amendment to the recommendation for cefuroxime for the treatment of infections with Proteus mirabilis (footnote Table 7); MIC and zone diameter breakpoints for Stenotrophomonas maltophilia only (Table 10); MIC breakpoints for daptomycin (Tables 11 and 15); clarification for staphylococci that the neomycin zone diameter breakpoints are for topical use only and differentiate the isolates outside the 'wild-type' population in Table 11; clarification for beta-haemolytic streptococci that the linezolid zone diameter breakpoints relate to an MIC breakpoint of 2 mg/L as no data for the intermediate category are currently available (Table 15); clarification that strains with reduced susceptibility to fluoroquinolones give no zone of inhibition with a 30 microg nalidixic acid disc (Tables 16 and 21); erythromycin is no longer used for therapy of Neisseria gonorrhoeae, but may be tested for epidemiological purposes (Table 17); clarification that the ciprofloxacin zone diameter breakpoint for Neisseria meningitidis relates to the MIC breakpoint of 0.03 mg/L as no data for the intermediate category are currently available; clarification that the ciprofloxacin zone diameter breakpoints for Campylobacter spp. relate to an MIC breakpoint of 0.5 mg/L as no data for the intermediate category are currently available; clarification that for ciprofloxacin and vancomycin zone

  11. Cryobacterium levicorallinum sp. nov., a psychrophilic bacterium isolated from glacier ice.

    PubMed

    Liu, Qing; Liu, Hongcan; Zhang, Jianli; Zhou, Yuguang; Xin, Yuhua

    2013-08-01

    In this study, two psychrophilic bacterial strains were isolated from the China No. 1 glacier in Xinjiang, north-west China. Cells were Gram-positive rods. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains belonged to the genus Cryobacterium. Phylogenetic analysis showed that they clustered together and are most closely related to Cryobacterium luteum CGMCC 1.11210(T), Cryobacterium flavum CGMCC 1.11215(T), Cryobacterium psychrophilum CGMCC 1.4292(T), Cryobacterium psychrotolerans CGMCC 1.5382(T) and Cryobacterium roopkundense CGMCC 1.10672(T). The major cellular fatty acids of the novel strains were anteiso-C15 : 0, anteiso-C15 : 1 A, iso-C16 : 0 and iso-C15 : 0. Both strains contained diphosphatidylglycerol, phosphatidylglycerol and one unidentified glycolipid in the cell membrane. The results of DNA-DNA hybridization and physiological tests allowed the genotypic and phenotypic differentiation of strains Hh34(T) and Hh28 from related species. However, their high DNA-DNA relatedness showed that they belong to the same novel species. Strain Hh34(T) (= NBRC 107883(T) = CGMCC 1.11211(T)) was selected as the type strain to represent this novel species, for which the name Cryobacterium levicorallinum sp. nov. is proposed.

  12. In vitro antibacterial activity and beta-lactamase stability of CP-70,429 a new penem antibiotic.

    PubMed Central

    Minamimura, M; Taniyama, Y; Inoue, E; Mitsuhashi, S

    1993-01-01

    In in vitro susceptibility tests, the new penem CP-70,429 showed potent antibacterial activity against gram-positive and gram-negative bacteria except Pseudomonas aeruginosa and Xanthomonas maltophilia. CP-70,429 was stable to various types of beta-lactamases except for the enzyme from X. maltophilia and was 16- to 128-fold more active than the other compounds against beta-lactamase-producing strains of Enterobacter cloacae and Citrobacter freundii. PMID:8363389

  13. In vitro antibacterial activity and beta-lactamase stability of CP-70,429 a new penem antibiotic.

    PubMed

    Minamimura, M; Taniyama, Y; Inoue, E; Mitsuhashi, S

    1993-07-01

    In in vitro susceptibility tests, the new penem CP-70,429 showed potent antibacterial activity against gram-positive and gram-negative bacteria except Pseudomonas aeruginosa and Xanthomonas maltophilia. CP-70,429 was stable to various types of beta-lactamases except for the enzyme from X. maltophilia and was 16- to 128-fold more active than the other compounds against beta-lactamase-producing strains of Enterobacter cloacae and Citrobacter freundii.

  14. Nematode-trapping fungi and fungus-associated bacteria interactions: the role of bacterial diketopiperazines and biofilms on Arthrobotrys oligospora surface in hyphal morphogenesis.

    PubMed

    Li, Lei; Yang, Min; Luo, Jun; Qu, Qing; Chen, Ying; Liang, Lianming; Zhang, Keqin

    2016-11-01

    In soil, nematode-trapping fungi and bacteria often share microhabitats and interact with each other, but effects of fungus-associated bacteria on its trap formation are underestimated. We have ascertained the presence of Stenotrophomonas and Rhizobium genera associated with A. oligospora GJ-1. After A. oligospora GJ-1 without associated bacteria (cured Arthrobotrys) was co-cultivated with Stenotrophomonas and its supernatant extract, microscopic study of hyphae from co-cultivation indicated that bacterial biofilm formation on hyphae was related to trap formation in fungi and Stenotrophomonas supernatant extract. Four diketopiperazines (DKPs) were purified from Stenotrophomonas supernatant extract that could not induce traps in the cured Arthrobotrys. When cured Arthrobotrys was cultured with Stenotrophomonas and one of DKPs, polar attachment, bacterial biofilms on hyphae and trap formation in fungi were observed. After cured Arthrobotrys with bacterial biofilms was consecutively transferred several times on nutrient poor medium, trap formation disappeared with the disappearance of bacterial biofilms on hyphae. DKPs could facilitate chemotaxis of Stenotrophomonas towards fungal extract which was suggested to contribute to bacterial biofilms on hyphae. Furthermore, when cured Arthrobotrys was cultured with Stenotrophomonas and DKPs in soil, trap formation in fungi and bacterial biofilms on hyphae were also observed, and the fungal activity against nematode was enhanced.

  15. In vitro evaluation of the probiotic and functional potential of Lactobacillus strains isolated from fermented food and human intestine.

    PubMed

    Ren, Dayong; Li, Chang; Qin, Yanqing; Yin, Ronglan; Du, Shouwen; Ye, Fei; Liu, Cunxia; Liu, Hongfeng; Wang, Maopeng; Li, Yi; Sun, Yang; Li, Xiao; Tian, Mingyao; Jin, Ningyi

    2014-12-01

    This study aims to evaluate the functional and probiotic characteristics of eight indigenous Lactobacillus strains in vitro. The selected lactobacilli include strains of Lactobacillus casei subsp. casei, Lactobacillus salivarius subsp. salicinius, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus delbrueckii subsp. lactis, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus rhamnosus. All strains tolerated both pH 2 for 3 h and 1% bile salt for 24 h. The strains CICC 23174 and CGMCC 1.557 were the most adhesive strains producing the highest quantity of EPS. Although a wide variation in the ability of the eight strains to deplete cholesterol and nitrite, antagonize pathogens, scavenge free radical, and stimulate innate immune response were observed, the strains CICC 23174 and CGMCC 1.557 showed the widest range of these useful traits. Taken together, the strains CICC 23174 and CGMCC 1.557 exhibited the best probiotic properties with the potential for use in the production of probiotic fermented foods.

  16. Phylogenomic analysis shows that ‘Bacillus vanillea’ is a later heterotypic synonym of Bacillus siamensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Bacillus vanillea’ XY18T (=CGMCC 8629 T =NCCB 100507 T) was isolated from cured vanilla beans and involved in the formation of vanilla aroma compounds. A draft genome of this type strain was assembled and yielded a length of 3.72 Mbp and a GC content of 46.3%. Comparative genomic analysis with its ...

  17. Amycolatopsis flava sp. nov., a halophilic actinomycete isolated from Dead Sea.

    PubMed

    Wei, Xiaomin; Jiang, Yingying; Chen, Xiu; Jiang, Yi; Lai, Hangxian

    2015-10-01

    A novel halophilic, filamentous actinomycete, designated strain AFM 10111(T), was isolated from a sediment sample collected from the Dead Sea of Israel and its taxonomic position was established by using a polyphasic taxonomic approach. The isolate grew at 20-35 °C, pH 5-12 and with 1-30 % NaCl. The substrate mycelium is white or yellow, well developed, branched and fragments into squarish, rod-like elements. The isolate contained meso-diaminopimelic acid as cell-wall diamino acid, and arabinose and galactose as whole-cell sugars. The major menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine phosphatidylmethylethanolamine and one unidentified phospholipid. Major fatty acids were iso-C16:0, iso-C16:1 H, C17:1 ω6c. The DNA G + C content was 67.7 mol %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain AFM 10111(T) belongs to the genus Amycolatopsis, and formed a distinct clade with Amycolatopsis marina CGMCC 4.3568(T) and Amycolatopsis palatopharyngis CGMCC 4.1729(T), with the sequence similarity 98.4 and 98.6 %. The level of DNA-DNA relatedness between the strain AFM 10111(T) and A. marina CGMCC 4.3568(T) and A. palatopharyngis CGMCC 4.1729(T) were 46.9 ± 3.08 and 49.4 ± 1.25 %. The combined genotypic and phenotypic data indicate that strain AFM 10111(T) represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis flava sp. nov. is proposed. The type strain is AFM 10111(T) (= DSM 46658(T) = CGMCC 4.7123(T)).

  18. Salisediminibacterium haloalkalitolerans sp. nov., isolated from Lonar soda lake, India, and a proposal for reclassification of Bacillus locisalis as Salisediminibacterium locisalis comb. nov., and the emended description of the genus Salisediminibacterium and of the species Salisediminibacterium halotolerans.

    PubMed

    Sultanpuram, Vishnuvardhan Reddy; Mothe, Thirumala; Mohammed, Farooq

    2015-05-01

    A Gram stain-positive, rod-shaped, non-motile, orange-pigmented and non-endospore-forming novel bacterial strain 10nlg(T) was isolated from Lonar soda lake in India. Based on the 16S rRNA gene sequence analysis, it was belonged to the genus Salisediminibacterium and was most closely related to Salisediminibacterium halotolerans CGMCC 1.7654(T) (99.9 %), Bacillus locisalis CGMCC 1.6286(T) (99.1 %) and other members in the Bacillaceae (<93.6 %). Further, DNA-DNA hybridization results demonstrated that strain 10nlg(T) was distantly (<70 %) related to S. halotolerans halo-2(T) (59.6 %) and B. locisalis CGMCC 1.6286(T) (53.2 %). Strain 10nlg(T) was catalase positive and oxidase negative. The cell wall of the strain 10nlg(T) contains meso-diaminopimelic acid as the diagnostic amino acid. Polar lipids include diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, five phospholipids and two unknown lipids. The predominant isoprenoid quinone was MK-7. Anteiso-C15:0 (32.6 %) was the predominant fatty acid, and significant proportions of iso-C15:0 (9.3 %), anteiso-C17:0 (8.3 %), C16:0 (4.3 %) were also detected in the strain 10nlg(T). The DNA G+C content of the strain 10nlg(T) was 45.6 mol %. The results of phylogenetic, physiological and biochemical tests allowed a distinct differentiation of strain 10nlg(T) not only from S. halotolerans halo-2(T), the only member of the genus Salisediminibacterium, but also from the related Bacillaceae members. Strain 10nlg(T) represents a novel species of the genus Salisediminibacterium for which the name Salisediminibacterium haloalkalitolerans sp. nov. is proposed. The type strain is 10nlg(T) (=KCTC 33414(T) = CGMCC 1.12818(T)). B. locisalis CGMCC 1.6286(T) also represents a member of the genus Salisediminibacterium for which the name Salisediminibacterium locisalis CG1(T) is proposed. The type strain is CG1(T) = (CCM 7370(T) = CECT 7152(T) = CGMCC 1.6286(T) = DSM 18085(T)). In accordance with the new polar lipid data

  19. Comparison of bactericidal effects of commonly used antiseptics against pathogens causing nosocomial infections. Part 2.

    PubMed

    Yasuda, T; Yoshimura, Y; Takada, H; Kawaguchi, S; Ito, M; Yamazaki, F; Iriyama, J; Ishigo, S; Asano, Y

    1997-01-01

    Opportunistic infections caused by gram-negative rods (GNR), conventionally regarded as organisms with low or no pathogenicity, and intractable infections caused by various resistant organisms pose a great problem now. In view of this, we determined the bactericidal effects of 5 commonly used disinfectants using as the test strains Xanthomonas maltophilia and Serratia marcescens, chosen among other GNR since they often cause nosocomial infections. Regarding the bactericidal activities against X. maltophilia and S. marcescens, both sensitive strains and resistant strains were killed within 20 s of exposure to povidone-iodine and sodium hypochlorite. With chlorhexidine, 1 strain each of both species was not killed within 10 min of exposure at a concentration of 0.2%. Both sensitive strains and resistant strains of X. maltophilia were killed within 20 s of exposure to benzalkonium at 0.02%, while a concentration of 0.1% was required for benzalkonium to kill S. marcescens within 20 s. With Tego-51, both sensitive strains and resistant strains of X. maltophilia were killed within 20 s at 0.02%, while 1 strain of S. marcescens was not killed within 20 s at a concentration of 0.1%. In the use of disinfectants, comparative bactericidal effects of various disinfectants against clinical isolates should be taken into consideration.

  20. Bensingtonia rectispora sp. nov. and Bensingtonia bomiensis sp. nov., ballistoconidium-forming yeast species from Tibetan plant leaves.

    PubMed

    Wang, Qi-Ming; Boekhout, Teun; Bai, Feng-Yan

    2012-08-01

    Five yeast strains isolated from plant leaves collected in south-east Tibet formed cream to brownish colonies and produced asymmetrical ballistoconidia and CoQ-9 as the major ubiquinone. Sequence analysis of the 26S rRNA D1/D2 domain and the internal transcribed spacer region indicated that these strains represented two novel species of the genus Bensingtonia. The names Bensingtonia rectispora sp. nov. (type strain XZ 4C5(T) = CGMCC 2.02635(T) = CBS 10710(T)) and Bensingtonia bomiensis sp. nov. (type strain XZ 33D1(T) = CGMCC 2.02670(T) = CBS 10713(T)) are proposed for the two novel species, which are phylogenetically closely related to Bensingtonia naganoensis, Bensingtonia pseudonaganoensis and the type species of the genus, Bensingtonia ciliata.

  1. Ballistosporomyces changbaiensis sp. nov. and Ballistosporomyces bomiensis sp. nov., two novel species isolated from shrub plant leaves.

    PubMed

    Han, Pei-Jie; Li, Ai-Hua; Wang, Qi-Ming; Bai, Feng-Yan

    2016-07-01

    Four strains, CB 266(T), CB 272, XZ 44D1(T) and XZ 49D2, isolated from shrub plant leaves in China were identified as two novel species of the genus Ballistosporomyces by the sequence analysis of the small subunit of ribosomal RNA (SSU rRNA), the D1/D2 domains of the large subunit of rRNA (LSU rRNA) and internal transcribed spacer (ITS) + 5.8S rRNA region, and physiological comparisons. Ballistosporomyces changbaiensis sp. nov. (type strain CB 266(T) = CGMCC 2.02298(T) = CBS 10124(T), Mycobank number MB 815700) and Ballistosporomyces bomiensis sp. nov. (type strain XZ 44D1(T) = CGMCC 2.02661(T) = CBS 12512(T), Mycobank number MB 815701) are proposed to accommodate these two new species.

  2. Evaluation of immunomodulatory activity of two potential probiotic Lactobacillus strains by in vivo tests.

    PubMed

    Ren, Dayong; Li, Chang; Qin, Yanqing; Yin, Ronglan; Du, Shouwen; Liu, Hongfeng; Zhang, Yanfang; Wang, Cuiyan; Rong, Fengjun; Jin, Ningyi

    2015-10-01

    Here we evaluate the immunomodulatory function of two potential probiotic strains, Lactobacillus salivarius CICC 23174 and Lactobacillus plantarum CGMCC 1.557. Mice were fed with each Lactobacillus strain at different doses for several consecutive days. The effects of the two probiotic strains on immune organs, immune cells and immune molecules were investigated on days 10 and 20. Both Lactobacillus strains increased the spleen index, improved the spleen lymphocyte transformation rate, enhanced sIgA production and improved the number of CD11c(+) CD80(+) double-positive cells. L. plantarum CGMCC 1.557 was the more active strain in enhancing the phagocytic activity of macrophages, while, L. salivarius CICC 23174 was the more effective strain at maintaining the Th1/Th2 balance. This study suggests that these two Lactobacillus strains have beneficial effects on regulation of immune responses, which has promising implications for the development of ecological agents and functional foods.

  3. Characterization of Inulin Hydrolyzing Enzyme(s) in Oleaginous Yeast Trichosporon cutaneum in Consolidated Bioprocessing of Microbial Lipid Fermentation.

    PubMed

    Wang, Juan; Zhang, Huizhan; Bao, Jie

    2015-11-01

    Oleaginous yeast Trichosporon cutaneum CGMCC 2.1374 was found to utilize inulin directly for microbial lipid fermentation without a hydrolysis step. The potential inulinase-like enzyme(s) in T. cutaneum CGMCC 2.1374 were characterized and compared with other inulinase enzymes produced by varied yeast strains. The consolidated bioprocessing (CBP) for lipid accumulated using inulin was optimized with 4.79 g/L of lipid produced from 50 g/L inulin with the lipid content of 33.6% in dry cells. The molecular weight of the enzyme was measured which was close to invertase in Saccharomyces cerevisiae. The study provided information for inulin hydrolyzing enzyme(s) in oleaginous yeasts, as well as a preliminary CBP process for lipid production from inulin feedstock.

  4. Complete genome sequence of Lactobacillus paracasei L9, a new probiotic strain with high lactic acid-producing capacity.

    PubMed

    Jiang, Yunyun; Li, Zhuanyu; Ren, Fazheng; Liu, Songling; Zhao, Liang; Sun, Erna; Zhang, Ming; Guo, Huiyuan; Zhang, Hao; Jiang, Lu; Hou, Caiyun

    2015-12-20

    Lactobaillus paracasei L9 (CGMCC No. 9800) is a new strain with probiotic properties originating from healthy human intestine. Previous studies evidenced that the strain regulates immune modulation and contributes to the production of high amounts of lactic acid. The genome of L. paracasei L9 contains a circular 3076,437-bp chromosome, encoding 3044 CDSs, 15 rRNA genes and 59 tRNA genes.

  5. Streptomyces bryophytorum sp. nov., an endophytic actinomycete isolated from moss (Bryophyta).

    PubMed

    Li, Chuang; Jin, Pinjiao; Liu, Chongxi; Ma, Zhaoxu; Zhao, Junwei; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng

    2016-09-01

    A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).

  6. Derxomyces amylogenes sp. nov., Derxomyces bambusicola sp. nov. and Derxomyces corylopsis sp. nov., three ballistoconidium-forming yeast species isolated from subtropical plant leaves.

    PubMed

    Liu, Xin-Zhan; Wang, Qi-Ming; Boekhout, Teun; Bai, Feng-Yan

    2012-04-01

    Among ballistoconidium-forming yeast strains isolated from various plant leaves collected from subtropical forests in eastern and central China, four strains forming cream to yellowish coloured colonies were revealed to represent three novel Derxomyces species by conventional and molecular characterization. Phylogenetic analysis based on combined sequences of the internal transcribed spacer (ITS) and 26S rRNA gene D1/D2 domain showed that strains GT-753 and ZJJ-890T were conspecific and closely related to Derxomyces boninensis, Derxomyces mrakii and Derxomyces qinlingensis. Strain ZJJ-394T was basal to the branch formed by Derxomyces komagatae, Derxomyces pseudoschimicola and Derxomyces schimicola with strong bootstrap support. Strain GT-475T was closely related to Derxomyces linzhiensis. The strains differed significantly from their close relatives in D1/D2 and ITS sequences and in physiological criteria. Three novel species are proposed: Derxomyces amylogenes sp. nov. (type strain ZJJ-890T=CGMCC 2.4407T=CBS 12233T), Derxomyces bambusicola sp. nov. (type strain GT-475T=CGMCC 2.4411T=CBS 12234T) and Derxomyces corylopsis sp. nov. (type strain ZJJ-394T=CGMCC 2.4409T=CBS 12259T).

  7. Nocardioides glacieisoli sp. nov., isolated from a glacier.

    PubMed

    Liu, Qing; Liu, Hong-Can; Zhang, Jian-Li; Zhou, Yu-Guang; Xin, Yu-Hua

    2015-12-01

    A Gram-stain-positive, rod-shaped, non-spore-forming bacterium (strain HLT3-15T) was isolated from the ice tongue surface of the Hailuogou glacier in Szechwan Province, PR China. Phylogenetic analysis, based on 16S rRNA gene sequences, revealed that strain HLT3-15T belonged to the genus Nocardioides. The highest levels of sequence similarities were found with Nocardioides hwasunensis CGMCC 4.6881T and Nocardioides ganghwensis CGMCC 4.6875T (98.5 % and 98.3 %, respectively). However, DNA-DNA relatedness demonstrated that strain HLT3-15T was distinct from its closest phylogenetic neighbours. The major cellular fatty acids of strain HLT3-15T were C17 : 1ω8c and iso-C16 : 0. Strain HLT3-15T contained ll-2,6-diaminopimelic acid as the diamino acid in the cell-wall peptidoglycan and MK-8(H4) as the predominant menaquinone. On the basis of a polyphasic approach, a novel species, Nocardioides glacieisoli sp. nov., is proposed with HLT3-15T ( = CGMCC 1.11097T = NBRC 109781T) as the type strain.

  8. Nitrile-hydrolyzing enzyme from Meyerozyma guilliermondii and its potential in biosynthesis of 3-hydroxypropionic acid.

    PubMed

    Zhang, Qiang; Gong, Jin-Song; Dong, Ting-Ting; Liu, Ting-Ting; Li, Heng; Dou, Wen-Fang; Lu, Zhen-Ming; Shi, Jin-Song; Xu, Zheng-Hong

    2017-03-11

    3-Hydroxypropionic acid (3-HP) is an important platform chemical in organic synthesis. Traditionally, 3-HP was produced by chemical methods and fermentation process. In this work, a novel enzymatic method was developed for green synthesis of 3-HP. A yeast strain harboring nitrile-hydrolyzing enzyme was newly isolated from environmental samples using 3-hydroxypropionitrile (3-HPN) as the sole nitrogen source. It was identified to be Meyerozyma guilliermondii CGMCC12935 by sequencing of the 18S ribosomal DNA and internal transcribed spacer, together with analysis of the morphology characteristics. The catalytic properties of M. guilliermondii CGMCC12935 resting cells were determined, and the optimum activity was achieved at 55 °C and pH 7.5. The enzyme showed broad substrate specificity towards nitriles, especially 3-HPN, aminoacetonitrile and 3-cyanopyridine. The presence of Ag(+), Pb(2+) and excess substrate inhibited the enzyme activity, whereas 5% (v/v) ethyl acetate had a positive effect on the enzyme activity. M. guilliermondii CGMCC12935 resting cells by addition of 3% glucose could thoroughly hydrolyze 500 mM 3-HPN into 3-HP within 100 h and the maximal accumulative production of 3-HP reached 216.33 mM, which was over twofolds than the control group with no additional glucose. And this work would lay the foundation for biological production of 3-HP in industry.

  9. Consequences of cps mutation of Klebsiella pneumoniae on 1,3-propanediol fermentation.

    PubMed

    Guo, Ni-Ni; Zheng, Zong-Ming; Mai, Yu-Lin; Liu, Hong-Juan; Liu, De-Hua

    2010-03-01

    The filtration in 1,3-propanediol (1,3-PD) downstream process is influenced by the large amounts of capsular polysaccharides (CPS) produced by Klebsiella pneumoniae CGMCC 1.6366. The morphological and fermentation properties were investigated with the CPS-deficient mutant K. pneumoniae CGMCC 1.6366 CPS. Similar biomass was obtained with CGMCC 1.6366, and the mutant strain in batch cultures indicating the cell growth was slightly inhibited by CPS defection. The viscosity of fermentation broth by mutant strain decreased by 27.45%. The flux with ceramic membrane filter was enhanced from 168.12 to 303.6 l h(-1) m(-2), exhibiting the great importance for downstream processing of 1,3-PD fermentation. The products spectrum of mutant isolate changed remarkably regarding to the concentration of fermentation products. The synthesis of important 1,3-PD and 2,3-butanediol was enhanced from 9.73 and 4.06 g l(-1) to 10.37 and 4.77 g l(-1) in batch cultures. The noncapsuled K. pneumoniae provided higher 1,3-PD yield of 0.54 mol mol(-1) than that of encapsuled wild parent in batch cultures. The fed-batch fermentation of mutant strain resulted in 1,3-PD concentration, yield, and productivity of 78.13 g l(-1), 0.53 mol mol(-1), and 1.95 g l(-1) h(-1), respectively.

  10. Effects of different osmolarities on bacterial biofilm formation

    PubMed Central

    Kavamura, Vanessa Nessner; de Melo, Itamar Soares

    2014-01-01

    Biofilm formation depends on several factors. The influence of different osmolarities on bacterial biofilm formation was studied. Two strains (Enterobacter sp. and Stenotrophomonas sp.) exhibited the most remarkable alterations. Biofilm formation is an important trait and its use has been associated to the protection of organisms against environmental stresses. PMID:25242950

  11. Carbapenem-based prodrugs. Design, synthesis, and biological evaluation of carbapenems.

    PubMed

    Hakimelahi, Gholam Hossein; Moosavi-Movahedi, Ali Akbar; Saboury, Ali Akbar; Osetrov, Valeriy; Khodarahmi, Ghadam Ali; Shia, Kak-Shan

    2005-04-01

    Syntheses of racemic trans-3-hydroxycarbonyl-6-(phenylacetamido)carbapenem (13), trans-3-phosphono-6-(phenylacetamido)carbapenem (17), and beta-lactam based prodrugs 19 and 22 were accomplished. Carbapenem 13 was found to possess antibacterial activity, comparable with imipenem (+)-3, against Staphylococcus aureus FDA 209P, S. aureus 95, Escherichia coli ATCC 39188, Klebsiella pneumoniae NCTC 418, Pseudomonas aeruginosa 1101-75, P. aeruginosa 18S-H, and Xanthomonas maltophilia GN 12873. Like imipenem ((+)-3), carbapenem 13 was not stable to X. maltophilia oxyiminocephalosporinase type II. Its phosphonate analog 17, however, was neither a significant antibacterial agent nor a good beta-lactamase inhibitor. Chemical combinations of trans carbapenem 13 with cis carbapenem 6 (compound 19) as well as clavulanic acid (20) with cis carbapenem 6 (compound 22) via a tetrachloroethane linker exhibited remarkable activity against beta-lactamase producing microorganisms in vitro.

  12. Flavobacterium xueshanense sp. nov. and Flavobacterium urumqiense sp. nov., two psychrophilic bacteria isolated from glacier ice.

    PubMed

    Dong, Kun; Liu, Hongcan; Zhang, Jianli; Zhou, Yuguang; Xin, Yuhua

    2012-05-01

    Two Gram-stain-negative, rod-shaped bacteria, designated strains Sr22(T) and Sr25(T), were isolated from water of melted ice from the China No.1 glacier, Xinjiang Uygur Autonomous Region, China. Cells formed yellow, circular, convex colonies. 16S rRNA gene sequence analysis indicated that strains Sr22(T) and Sr25(T) belong to the genus Flavobacterium, sharing ≤99.1  and ≤99.6 % similarity, respectively, with the type strains of recognized species of the genus. Strain Sr22(T) shared highest 16S rRNA gene sequence similarity with Flavobacterium tiangeerense CGMCC 1.6847(T) (98.6 %), Flavobacterium fryxellicola LMG 22022(T) (98.1 %) and Flavobacterium omnivorum CGMCC 1.2747(T) (99.1 %). Strain Sr25(T) shared highest similarity with Flavobacterium sinopsychrotolerans CGMCC 1.8704(T) (98.5 %), Flavobacterium degerlachei NBRC 102677(T) (98.4 %) and Flavobacterium xinjiangense CGMCC 1.2749(T) (99.5 %). The predominant fatty acids of strain Sr22(T) were iso-C(15 : 1) G (6.01 %), iso-C(15 : 0) (8.93 %), iso-C(16 : 1) H (12.68 %), iso-C(16 : 0) (10.4 %), C(15 : 1)ω6c (8.97 %), C(17 : 1)ω6c (5.96 %), iso-C(16 : 0) 3-OH (11.14 %) and summed feature 3 (comprising C(16 : 1)ω7c and/or C(16 : 1)ω6c, 12.33 %). The major fatty acids of strain Sr25(T) were iso-C(15 : 0) (10.8 %), anteiso-C(15 : 0) (5.23 %), C(15 : 1)ω6c (11.79 %), C(17 : 1)ω6c (5.43 %), iso-C(16 : 0) 3-OH (7.04 %) and summed feature 3 (20.42 %). The genomic DNA G+C contents of strains Sr22(T) and Sr25(T) were 37.2 and 35.1 mol%. On the basis of differential phenotypic and phylogenetic characteristics, these strains are considered to represent two novel species of the genus Flavobacterium, for which the names Flavobacterium xueshanense sp. nov. (type strain Sr22(T)  = CGMCC 1.9227(T)  = NBRC 106479(T)) and Flavobacterium urumqiense sp. nov. (type strain Sr25(T)  = CGMCC 1.9230(T)  = NBRC 106480

  13. The opposite roles of agdA and glaA on citric acid production in Aspergillus niger.

    PubMed

    Wang, Lu; Cao, Zhanglei; Hou, Li; Yin, Liuhua; Wang, Dawei; Gao, Qiang; Wu, Zhenqiang; Wang, Depei

    2016-07-01

    Citric acid is produced by an industrial-scale process of fermentation using Aspergillus niger as a microbial cell factory. However, citric acid production was hindered by the non-fermentable isomaltose and insufficient saccharification ability in A. niger when liquefied corn starch was used as a raw material. In this study, A. niger TNA 101ΔagdA was constructed by deletion of the α-glucosidase-encoding agdA gene in A. niger CGMCC 10142 genome using Agrobacterium tumefaciens-mediated transformation. The transformants A. niger OG 1, OG 17, and OG 31 then underwent overexpression of glucoamylase in A. niger TNA 101ΔagdA. The results showed that the α-glucosidase activity of TNA 101ΔagdA was decreased by 62.5 % compared with CGMCC 10142, and isomaltose was almost undetectable in the fermentation broth. The glucoamylase activity of the transformants OG 1 and OG 17 increased by 34.5 and 16.89 % compared with that of TNA 101ΔagdA, respectively. In addition, for the recombinants TNA 101ΔagdA, OG 1 and OG 17, there were no apparent defects in the growth development. Consequently, in comparison with CGMCC 10142, TNA 101ΔagdA and OG 1 decreased the residual reducing sugar by 52.95 and 88.24 %, respectively, and correspondingly increased citric acid production at the end of fermentation by 8.68 and 16.87 %. Citric acid production was further improved by decreasing the non-fermentable residual sugar and increasing utilization rate of corn starch material in A. niger. Besides, the successive saccharification and citric acid fermentation processes were successfully integrated into one step.

  14. Flavobacterium phragmitis sp. nov., an endophyte of reed (Phragmites australis).

    PubMed

    Liu, Min; Li, Yan Hong; Liu, Yang; Zhu, Jing Nan; Liu, Qun Fang; Liu, Yin; Gu, Jin Gang; Zhang, Xiao Xia; Li, Chun Li

    2011-11-01

    A Gram-staining-negative bacterium, designated strain BLN2(T), was isolated from within the roots of reeds (Phragmites australis) in Beijing Cuihu Wetland (China) and characterized using a polyphasic taxonomic approach. The cells were yellow-pigmented, rod-shaped, strictly aerobic and devoid of flagella, but showed gliding motility. Strain BLN2(T) produced yellow, translucent, circular and convex colonies, with optimal growth at 30 °C and pH 7.0. The major respiratory quinone was menaquinone 6 (MK-6) and the predominant fatty acids were iso-C(15 : 0), summed feature 3 (comprising C(16 : 1)ω7c and/or C(16 : 1)ω6c), C(16 : 0) 3-OH, C(16 : 0,) iso-C(17 : 0) 3-OH and iso-C(15 : 0) 3-OH. The G+C content of the genomic DNA was 34.8 mol%. The 16S rRNA gene sequence analysis showed that strain BLN2(T) belonged to the genus Flavobacterium and was most closely related to Flavobacterium anhuiense CGMCC 1.6859(T) (97.0 % sequence similarity). The DNA-DNA relatedness between strain BLN2(T) and F. anhuiense CGMCC 1.6859(T) was 25.7 %. Based on the phenotypic data and phylogenetic inference presented, it is concluded that strain BLN2(T) represents a novel species within the genus Flavobacterium, for which the name Flavobacterium phragmitis sp. nov. is proposed. The type strain is BLN2(T) ( = DSM 23314(T) = CGMCC 1.10370(T)).

  15. Salibacterium halotolerans gen. nov., sp. nov., a bacterium isolated from a salt pan, reclassification of Bacillus qingdaonensis as Salibacterium qingdaonense comb. nov. and Bacillus halochares as Salibacterium halochares comb. nov.

    PubMed

    Vishnuvardhan Reddy, Sultanpuram; Thirumala, Mothe; Sasikala, Chintalapati; Venkata Ramana, Chintalapati

    2015-11-01

    Two novel Gram-stain-positive, rod-shaped, non-motile, non-endospore-forming bacterial strains, S7T and IB5, were isolated from Khavda, India. Based on 16S rRNA gene sequence analysis they were identified as belonging to the class Bacilli, order Bacillales, family Bacillaceae, and were most closely related to Bacillus qingdaonensis CGMCC 1.6134T (97.3 %, sequence similarity), Bacillus halochares LMG 24571T (96.9 %), Bacillus salarius KCTC 3912T (95.6 %) and Bacillus aidingensis DSM 18341T (95.3 %). However, these strains shared only 88.2 % 16S rRNA gene sequence similarity with Bacillus subtilis subsp. subtilis DSM 10T, indicating that strains S7T and IB5 might not be members of the genus Bacillus. The DNA-DNA relatedness of these strains with B. qingdaonensis CGMCC 1.6134T was 42.9 ± 0.8. The cell-wall peptidoglycan of strains S7T and IB5 contained meso-diaminopimelic acid, while the polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, a phospholipid and three unknown lipids. The predominant isoprenoid quinone was MK-7. anteiso-C15 : 0 was the predominant fatty acid. The results of the phylogenetic, chemotaxonomic and biochemical tests allowed a clear differentiation of strains S7T and IB5, suggesting that they represent a novel member of the family Bacillaceae, for which the name Salibacterium halotolerans gen. nov., sp. nov. is proposed. The type strain of Salibacterium halotolerans is S7T ( = KCTC 33658T = CGMCC 1.15324T). Based on the results of the present study, it is also suggested that B. qingdaonensis and B. halochares should be transferred to this novel genus, as Salibacterium qingdaonense comb. nov. and Salibacterium halochares comb. nov., respectively.

  16. Complete genome sequence of endophytic nitrogen-fixing Klebsiella variicola strain DX120E

    PubMed Central

    2015-01-01

    Klebsiella variicola strain DX120E (=CGMCC 1.14935) is an endophytic nitrogen-fixing bacterium isolated from sugarcane crops grown in Guangxi, China and promotes sugarcane growth. Here we summarize the features of the strain DX120E and describe its complete genome sequence. The genome contains one circular chromosome and two plasmids, and contains 5,718,434 nucleotides with 57.1% GC content, 5,172 protein-coding genes, 25 rRNA genes, 87 tRNA genes, 7 ncRNA genes, 25 pseudo genes, and 2 CRISPR repeats. PMID:26203334

  17. Effect of surfactants on PAH biodegradation by a bacterial consortium and on the dynamics of the bacterial community during the process.

    PubMed

    González, N; Simarro, R; Molina, M C; Bautista, L F; Delgado, L; Villa, J A

    2011-10-01

    The aim of this work was to evaluate the effect of a non-biodegradable (Tergitol NP-10) and a biodegradable (Tween-80) surfactant on growth, degradation rate and microbial dynamics of a polycyclic aromatic hydrocarbon (PAHs) degrading consortium (C2PL05) from a petroleum polluted soil, applying cultivable and non cultivable techniques. Growth and degradation rate were significantly lower with Tergitol NP-10 than that with Tween-80. Toxicity did not show any significant reduction with Tergitol NP-10 whereas with Tween-80 toxicity was almost depleted (30%) after 40 days. Regarding to the cultured bacteria, Pseudomonas and Stenotrophomonas groups were dominant during PAH degradation with Tergitol NP-10, whereas Enterobacter and Stenotrophomonas were dominant with Tween-80. DGGE analyses (PRIMER and MDS) showed that bacteria composition was more similar between treatments when PAHs were consumed than when PAHs concentration was still high. Community changes between treatments were a consequence of Pseudomonas sp., Sphingomonas sp., Sphingobium sp. and Agromonas sp.

  18. Diversity of bacterial endophytes in roots of Mexican husk tomato plants (Physalis ixocarpa) and their detection in the rhizosphere.

    PubMed

    Marquez-Santacruz, H A; Hernandez-Leon, R; Orozco-Mosqueda, M C; Velazquez-Sepulveda, I; Santoyo, G

    2010-12-07

    Endophytic bacterial diversity was estimated in Mexican husk tomato plant roots by amplified rDNA restriction analysis and sequence homology comparison of the 16S rDNA genes. Sixteen operational taxonomic units from the 16S rDNA root library were identified based on sequence analysis, including the classes Gammaproteobacteria, Betaproteobacteria, Actinobacteria, and Bacilli. The predominant genera were Stenotrophomonas (21.9%), Microbacterium (17.1%), Burkholderia (14.3%), Bacillus (14.3%), and Pseudomonas (10.5%). In a 16S rDNA gene library of the same plant species' rhizosphere, only common soil bacteria, including Stenotrophomonas, Burkholderia, Bacillus, and Pseudomonas, were detected. We suggest that the endophytic bacterial diversity within the roots of Mexican husk tomato plants is a subset of the rhizosphere bacterial population, dominated by a few genera.

  19. Community dynamics of a mixed-bacterial culture growing on petroleum hydrocarbons in batch culture.

    PubMed

    Van Hamme, J D; Odumeru, J A; Ward, O P

    2000-05-01

    The effects of various hydrocarbon substrates, and a chemical surfactant capable of enhancing crude-oil biodegradation, on the community structure of a mixed-bacterial inoculum were examined in batch culture. Of 1000 TSA-culturable isolates, 68.6% were identified at the genus level or better by phospholipid fatty acid analysis over 7-day time course experiments. Cultures were exposed to 20 g/L Bow River crude oil with and without 0.625 g/L Igepal CO-630 (a nonylphenol ethoxylate surfactant), 5 g/L saturates, 5 g/L aromatics, or 125 g/L refinery sludge. A group of six genera dominated the cultures: Acinetobacter, Alcaligenes, Ochrobactrum, Pseudomonas/Flavimonas, Stenotrophomonas, and Yersinia. Species from four of the genera were shown to be capable of hydrocarbon degradation, and counts of hydrocarbon degrading and total heterotrophic bacteria over time were nearly identical. Pseudomonas/Flavimonas and Stenotrophomonas normally dominated during the early portions of cultures, although the lag phase of Stenotrophomonas appears to have been increased by surfactant addition. Acinetobacter calcoaceticus was the most frequently isolated microorganism during exposure to the saturate fraction of crude oil. Regardless of substrate, the culture medium supported a greater variety of organisms during the latter portions of cultures. Understanding the community structure and dynamics of mixed bacterial cultures involved in treatment of heterogeneous waste substrates may assist in process development and optimization studies.

  20. Transcriptional characteristics associated with lichenysin biosynthesis in Bacillus licheniformis from Chinese Maotai-flavor liquor making.

    PubMed

    Wu, Qun; Zhang, Rong; Peng, Suqin; Xu, Yan

    2015-01-28

    This work investigated the biosynthetic mechanism of lichenysin, the newly identified nonvolatile matrix component in Chinese liquors. Transcriptomes were analyzed in three producers, Bacillus licheniformis CGMCC 3961, 3962, and 3963, which were isolated from Maotai-flavor liquor-making process and produced 386.3, 553.5, and 795.2 μg/L lichenysin in a simulative liquor fermentation process. Lichenysin synthetase genes lchAA-AD in these three producers were expressed much more highly than those of the nonproducer B. licheniformis ATCC 14580 (>18.4-fold). In addition, ABC transporters were the most significant responsive metabolic pathway, and the expression levels of peptide transporter genes dppABCDE all increased more than 19.2-fold. When B. licheniformis CGMCC 3963 was cultured in synthetic medium, the expression of dppABCDE and lichenysin both increased with the addition of casein hydrolysate (containing various peptides). This indicated that peptide would act as a substrate for lichenysin synthesis. This work sheds new light on the mechanism for lichenysin biosynthesis.

  1. Proteomic profiling of Bacillus licheniformis reveals a stress response mechanism in the synthesis of extracellular polymeric flocculants.

    PubMed

    Yu, Wencheng; Chen, Zhen; Shen, Liang; Wang, Yuanpeng; Li, Qingbiao; Yan, Shan; Zhong, Chuan-Jian; He, Ning

    2016-04-01

    Some bioflocculants composed of extracellular polymeric substances are produced under peculiar conditions. Bacillus licheniformis CGMCC2876 is a microorganism that secretes both extracellular polysaccharides (EPS) and poly-gamma-glutamic acid (γ-PGA) under stress conditions. In this work, SWATH acquisition LC-MS/MS method was adopted for differential proteomic analysis of B. licheniformis, aiming at determining the bacterial stress mechanism. Compared with LB culture, 190 differentially expressed proteins were identified in B. licheniformis CGMCC2876 cultivated in EPS culture, including 117 up-regulated and 73 down-regulated proteins. In γ-PGA culture, 151 differentially expressed proteins, 89 up-regulated and 62 down-regulated, were found in the cells. Up-regulated proteins involved in amino acid biosynthesis were found to account for 43% and 41% of the proteomes in EPS and γ-PGA cultivated cells, respectively. Additionally, a series of proteins associated with amino acid degradation were found to be repressed under EPS and γ-PGA culture conditions. Transcriptional profiling via the qPCR detection of selected genes verified the proteomic analysis. Analysis of free amino acids in the bacterial cells further suggested the presence of amino acid starvation conditions. EPS or γ-PGA was synthesized to alleviate the effect of amino acid limitation in B. licheniformis. This study identified a stress response mechanism in the synthesis of macromolecules in B. licheniformis, providing potential culture strategies to improve the production of two promising bioflocculants.

  2. Cryobacterium flavum sp. nov. and Cryobacterium luteum sp. nov., isolated from glacier ice.

    PubMed

    Liu, Qing; Liu, Hongcan; Wen, Ying; Zhou, Yuguang; Xin, Yuhua

    2012-06-01

    Gram-positive, rod-shaped bacteria, strains Hh8(T), Hh15(T) and Hh40-2, were isolated from the No. 1 glacier in Xinjiang, north-west China. Colonies of strain Hh8(T) were orange-yellow, convex and round on PYG plates. Strain Hh8(T) grew at 0-19 °C and pH 5.5-10.5. Colonies of strain Hh15(T), which was able to grow at 0-20 °C and pH 5.5-12, were lemon yellow, convex and round on PYG plates. Phylogenetic analysis based on 16S rRNA gene sequences showed that these three strains were related to members of the genus Cryobacterium. The major cellular fatty acids of the novel strains were anteiso-C(15:0), iso-C(16:0), iso-C(15:0) and anteiso-C(15:1) A. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, two novel species, Cryobacterium flavum sp. nov. (type strain Hh8(T) = CGMCC 1.11215(T) = NBRC 107879(T)) and Cryobacterium luteum sp. nov. (type strain Hh15(T) = CGMCC 1.11210(T) = NBRC 107880(T)), are proposed.

  3. Optimization of culture medium compositions for gellan gum production by a halobacterium Sphingomonas paucimobilis.

    PubMed

    Zhang, Jun; Dong, Ya-chen; Fan, Lin-lin; Jiao, Zhi-hua; Chen, Qi-he

    2015-01-22

    The effect of culture medium compositions on gellan gum production produced by fermentation with a halobacterium Sphingomonas paucimobilis QHZJUJW CGMCC2428 was studied. In this work, a fractional factorial design was applied to investigate the main factors that affected gellan gum production by S. paucimobilis QHZJUJW CGMCC2428. Sucrose was the best carbon source for gellan gum and peptone displayed better inducing effect. Central composite design and response surface methodology were adopted to derive a statistical model for optimizing submerged culture medium composition. These experimental results showed that the optimum culture medium for producing gellan gum was composed of 40.00 (w/v) sucrose, 3.00% peptone (w/v), MgSO4 (w/v), 9.20% KH2PO4 (w/v), 7.50% Na2HPO4 (w/v), 4.30% K2SO4 (w/v), pH 6.8-7.0. The maximal gellan gum was 19.89±0.68 g/L, which was agreed closely with the predicated value (20.12 g/L). After incubated for 72 h under the optimized culture medium in 5-L bioreactor, the gellan gum fermentation reached about 19.90±0.68 g/L, which was higher than that in the initial cultivation medium.

  4. Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing.

    PubMed

    Hu, Nan; Yuan, Bo; Sun, Juan; Wang, Shi-An; Li, Fu-Li

    2012-09-01

    Thermotolerant inulin-utilizing yeast strains are desirable for ethanol production from Jerusalem artichoke tubers by consolidated bioprocessing (CBP). To obtain such strains, 21 naturally occurring yeast strains isolated by using an enrichment method and 65 previously isolated Saccharomyces cerevisiae strains were investigated in inulin utilization, extracellular inulinase activity, and ethanol fermentation from inulin and Jerusalem artichoke tuber flour at 40 °C. The strains Kluyveromyces marxianus PT-1 (CGMCC AS2.4515) and S. cerevisiae JZ1C (CGMCC AS2.3878) presented the highest extracellular inulinase activity and ethanol yield in this study. The highest ethanol concentration in Jerusalem artichoke tuber flour fermentation (200 g L(-1)) at 40 °C achieved by K. marxianus PT-1 and S. cerevisiae JZ1C was 73.6 and 65.2 g L(-1), which corresponded to the theoretical ethanol yield of 90.0 and 79.7 %, respectively. In the range of 30 to 40 °C, temperature did not have a significant effect on ethanol production for both strains. This study displayed the distinctive superiority of K. marxianus PT-1 and S. cerevisiae JZ1C in the thermotolerance and utilization of inulin-type oligosaccharides reserved in Jerusalem artichoke tubers. It is proposed that both K. marxianus and S. cerevisiae have considerable potential in ethanol production from Jerusalem artichoke tubers by a high temperature CBP.

  5. Streptomyces chlorus sp. nov. and Streptomyces viridis sp. nov., isolated from soil.

    PubMed

    Kim, Byung-Yong; Rong, Xiaoying; Zucchi, Tiago D; Huang, Ying; Goodfellow, Michael

    2013-05-01

    Two actinomycete strains, BK125(T) and BK199(T), isolated from a hay meadow soil sample were investigated to determine their taxonomic position using a polyphasic approach. The isolates produced greenish-yellow and light green aerial mycelium on oatmeal agar, respectively. They contained anteiso-C15 : 0, iso-C15 : 0 and C16 : 0 as the major fatty acids, and MK-9 (H6) and MK-9 (H8) as the predominant isoprenoid quinones. Phylogenetic analysis of the 16S rRNA gene sequences showed that the isolates formed distinct phyletic lines towards the periphery of the Streptomyces prasinus subclade. Analysis of DNA-DNA relatedness between the two isolates showed that they belonged to different genomic species. The organisms were also distinguished from one another and from type strains of species classified in the S. prasinus subclade using a combination of genotypic and phenotypic properties. On the basis of these data, it is proposed that the isolates be assigned to the genus Streptomyces as Streptomyces chlorus sp. nov. and Streptomyces viridis sp. nov. with isolates BK125(T) ( = KACC 20902(T) = CGMCC 4.5798(T)) and BK199(T) ( = KACC 21003(T) = CGMCC 4.6824(T)) as the respective type strains.

  6. Pseudomonas linyingensis sp. nov.: a novel bacterium isolated from wheat soil subjected to long-term herbicides application.

    PubMed

    He, Wei-Hong; Wang, Ya-Nan; Du, Xun; Zhou, Yang; Jia, Bin; Bian, Jiang; Liu, Shuang-Jiang; Chen, Guo-Can

    2012-11-01

    A strain of genus Pseudomonas, LYBRD3-7(T) was isolated from long-term sulfonylurea herbicides applied wheat-field soil in Linying located in Henan province of China. This strain is a strictly aerobic and Gram-negative short rod-shaped bacterium with single flagellum. Phylogenetic evaluation based on 16S rRNA gene sequence analysis placed this isolate as a member of Pseudomonas, and most closely to Pseudomonas tuomuerensis CGMCC 1.1365(T) (97.1 %) and P. alcaligenes IAM12411(T) (97.1 %). Morphological characters and chemotaxonomic data confirmed the affiliation of strain LYBRD3-7(T) to the genus Pseudomonas. The results of phylogenetic analysis, physiological and biochemical studies, and DNA-DNA hybridization allowed the differentiation of genotype and phenotype between strain LYBRD3-7(T) and the phylogenetic closest species with valid names. The name proposed for the new species is Pseudomonas linyingensis sp. nov. The type strain is LYBRD3-7(T) (=CGMCC 1.10701(T ) =LMG 25967(T)).

  7. Global insights into acetic acid resistance mechanisms and genetic stability of Acetobacter pasteurianus strains by comparative genomics

    NASA Astrophysics Data System (ADS)

    Wang, Bin; Shao, Yanchun; Chen, Tao; Chen, Wanping; Chen, Fusheng

    2015-12-01

    Acetobacter pasteurianus (Ap) CICC 20001 and CGMCC 1.41 are two acetic acid bacteria strains that, because of their strong abilities to produce and tolerate high concentrations of acetic acid, have been widely used to brew vinegar in China. To globally understand the fermentation characteristics, acid-tolerant mechanisms and genetic stabilities, their genomes were sequenced. Genomic comparisons with 9 other sequenced Ap strains revealed that their chromosomes were evolutionarily conserved, whereas the plasmids were unique compared with other Ap strains. Analysis of the acid-tolerant metabolic pathway at the genomic level indicated that the metabolism of some amino acids and the known mechanisms of acetic acid tolerance, might collaboratively contribute to acetic acid resistance in Ap strains. The balance of instability factors and stability factors in the genomes of Ap CICC 20001 and CGMCC 1.41 strains might be the basis for their genetic stability, consistent with their stable industrial performances. These observations provide important insights into the acid resistance mechanism and the genetic stability of Ap strains and lay a foundation for future genetic manipulation and engineering of these two strains.

  8. Global insights into acetic acid resistance mechanisms and genetic stability of Acetobacter pasteurianus strains by comparative genomics.

    PubMed

    Wang, Bin; Shao, Yanchun; Chen, Tao; Chen, Wanping; Chen, Fusheng

    2015-12-22

    Acetobacter pasteurianus (Ap) CICC 20001 and CGMCC 1.41 are two acetic acid bacteria strains that, because of their strong abilities to produce and tolerate high concentrations of acetic acid, have been widely used to brew vinegar in China. To globally understand the fermentation characteristics, acid-tolerant mechanisms and genetic stabilities, their genomes were sequenced. Genomic comparisons with 9 other sequenced Ap strains revealed that their chromosomes were evolutionarily conserved, whereas the plasmids were unique compared with other Ap strains. Analysis of the acid-tolerant metabolic pathway at the genomic level indicated that the metabolism of some amino acids and the known mechanisms of acetic acid tolerance, might collaboratively contribute to acetic acid resistance in Ap strains. The balance of instability factors and stability factors in the genomes of Ap CICC 20001 and CGMCC 1.41 strains might be the basis for their genetic stability, consistent with their stable industrial performances. These observations provide important insights into the acid resistance mechanism and the genetic stability of Ap strains and lay a foundation for future genetic manipulation and engineering of these two strains.

  9. Preparation, characterization, and antifungal activity of hymexazol-linked chitosan derivatives

    NASA Astrophysics Data System (ADS)

    Li, Yan; Qin, Yukun; Liu, Song; Li, Pengcheng; Xing, Rong'e.

    2016-09-01

    In this study, three hymexazol-linked chitosan derivatives (HML-CS) were synthesized and their structures confirmed by Fourier transform infrared and elemental analysis. Linkage ratios were measured by high performance liquid chromatography. The derivatives' antifungal activity against the plant pathogenic fungi Rhizoctonia solani CGMCC 3.28 and Gibberella zeae CGMCC 3.42 were investigated at concentrations of 100, 200, and 400 mg/L. These HML-CS derivatives exhibited stronger antifungal activity than CS alone. HML-CS-1 showed the best antifungal activity against G. zeae, whose antifungal index was 65.9% at 400 mg/L, and also showed the best antifungal activity against R. solani, whose antifungal index was 52.7% at 400 mg/L. This conjugation of CS and HML suggested the presence of synergistic effects between the moieties and indicated that these derivatives possessed great potential as novel fungicides and require further research for the development of applications in crop protection.

  10. Highly efficient production of D-lactic acid from chicory-derived inulin by Lactobacillus bulgaricus.

    PubMed

    Xu, Qianqian; Zang, Ying; Zhou, Jie; Liu, Peng; Li, Xin; Yong, Qiang; Ouyang, Jia

    2016-11-01

    Inulin is a readily available feedstock for cost-effective production of biochemicals. To date, several studies have explored the production of bioethanol, high-fructose syrup and fructooligosaccharide, but there are no studies regarding the production of D-lactic acid using inulin as a carbon source. In the present study, chicory-derived inulin was used for D-lactic acid biosynthesis by Lactobacillus bulgaricus CGMCC 1.6970. Compared with separate hydrolysis and fermentation processes, simultaneous saccharification and fermentation (SSF) has demonstrated the best performance of D-lactic acid production. Because it prevents fructose inhibition and promotes the complete hydrolysis of inulin, the highest D-lactic acid concentration (123.6 ± 0.9 g/L) with a yield of 97.9 % was obtained from 120 g/L inulin by SSF. Moreover, SSF by L. bulgaricus CGMCC 1.6970 offered another distinct advantage with respect to the higher optical purity of D-lactic acid (>99.9 %) and reduced number of residual sugars. The excellent performance of D-lactic acid production from inulin by SSF represents a high-yield method for D-lactic acid production from non-food grains.

  11. Flavobacterium qiangtangensis sp. nov., isolated from Qiangtang basin in Qinghai-Tibetan Plateau, China.

    PubMed

    Huang, Faqi; Zhang, Yali; Zhu, Youhai; Wang, Pingkang; Lu, Jun; Lv, Jie

    2014-09-01

    A flexirubin-type yellow-pigmented, non-gliding, non-flagellated, gram-negative bacterium strain, designated F3(T), was isolated from a drilling core sample of the Qiangtang basin, Qinghai-Tibetan plateau, China. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain F3(T) belongs to the genus Flavobacterium, with the highest 16S rRNA gene sequence similarity to the Flavobacterium noncentrifugens CGMCC 1.10076(T) (94.92 %). Strain F3(T) grew optimally at temperature about 20 °C, at pH about 7.0-8.0, at NaCl concentration 0 % (w/v). The DNA G+C content of the isolate was 35.5 mol%. The major polar lipid was phosphatidylethanolamine, predominant cellular fatty acids of the strain was iso-C15:0 (22.02 %), while the major menaquinone was menaquinone 6. Due to the phenotypic and genetic distinctiveness and several other characteristic studied in this article, we consider F3(T) as a novel species of the genus Flavobacterium, and propose to name it Flavobacterium qiangtangensis sp. nov. The type strain is F3(T) (=CGMCC 1.12706(T) = JCM 19739(T)).

  12. Global insights into acetic acid resistance mechanisms and genetic stability of Acetobacter pasteurianus strains by comparative genomics

    PubMed Central

    Wang, Bin; Shao, Yanchun; Chen, Tao; Chen, Wanping; Chen, Fusheng

    2015-01-01

    Acetobacter pasteurianus (Ap) CICC 20001 and CGMCC 1.41 are two acetic acid bacteria strains that, because of their strong abilities to produce and tolerate high concentrations of acetic acid, have been widely used to brew vinegar in China. To globally understand the fermentation characteristics, acid-tolerant mechanisms and genetic stabilities, their genomes were sequenced. Genomic comparisons with 9 other sequenced Ap strains revealed that their chromosomes were evolutionarily conserved, whereas the plasmids were unique compared with other Ap strains. Analysis of the acid-tolerant metabolic pathway at the genomic level indicated that the metabolism of some amino acids and the known mechanisms of acetic acid tolerance, might collaboratively contribute to acetic acid resistance in Ap strains. The balance of instability factors and stability factors in the genomes of Ap CICC 20001 and CGMCC 1.41 strains might be the basis for their genetic stability, consistent with their stable industrial performances. These observations provide important insights into the acid resistance mechanism and the genetic stability of Ap strains and lay a foundation for future genetic manipulation and engineering of these two strains. PMID:26691589

  13. Microbial killing activity of peracetic acid.

    PubMed

    Thamlikitkul, V; Trakulsomboon, S; Louisirirotchanakul, S; Chaiprasert, A; Foongladda, S; Thipsuvan, K; Arjratanakool, W; Kunyok, R; Wasi, C; Santiprasitkul, S; Danchaivijitr, S

    2001-10-01

    In vitro killing activity of peracetic acid (Perasafe) at a concentration of 0.26 per cent w/v was tested against Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Salmonella paratyphi A, Acinetobacter baumannii, Sternotrophomonas maltophilia, Enterococcus faecium, Enterococcus faecalis, methicillin-resistant Staphylococcus aureus (MRSA), Bacillus subtilis spore, Mycobacterium tuberculosis and human immuno-deficiency virus type I. Exposure to Peracetic acid (0.26% w/v) for 10 minutes resulted in massive killing of all the aforementioned organisms and spore.

  14. In vitro activity of amiloride combined with tobramycin against Pseudomonas isolates from patients with cystic fibrosis.

    PubMed Central

    Cohn, R C; Jacobs, M; Aronoff, S C

    1988-01-01

    The diuretic amiloride has been under recent investigation as adjunctive therapy for pulmonary disease in children with cystic fibrosis (CF). In preliminary studies, the antimicrobial activity of this agent alone or combined with beta-lactam agents against reference strains of Pseudomonas aeruginosa and P. cepacia was poor; however, amiloride was markedly synergistic with tobramycin against P. cepacia. The purpose of this study was to determine the extent of amiloride-tobramycin synergy against CF respiratory isolates of P. aeruginosa, P. cepacia, and P. maltophilia. The MICs of tobramycin and amiloride alone against the Pseudomonas test strains were determined by agar dilution. Synergy was determined by combining each of four subinhibitory concentrations of amiloride (at least fourfold below the MIC) with doubling dilutions of tobramycin and comparing the MIC of tobramycin alone and in combination for each strain. At the highest concentration tested, the drug combination synergistically inhibited 50% of the P. cepacia strains tested; the combination was synergistic against fewer isolates of P. aeruginosa and P. maltophilia. Only P. cepacia was inhibited by tobramycin combined with amiloride at achievable airway concentrations. We conclude that the combination of tobramycin and amiloride may be potentially useful in the treatment of P. cepacia infections in children with CF. PMID:3364958

  15. Death Becomes Them: Bacterial Community Dynamics and Stilbene Antibiotic Production in Cadavers of Galleria mellonella Killed by Heterorhabditis and Photorhabdus spp.

    PubMed Central

    Wollenberg, Amanda C.; Slough, Greg; Hoinville, Megan E.

    2016-01-01

    ABSTRACT Insect larvae killed by entomopathogenic nematodes are thought to contain bacterial communities dominated by a single bacterial genus, that of the nematode's bacterial symbiont. In this study, we used next-generation sequencing to profile bacterial community dynamics in greater wax moth (Galleria mellonella) larvae cadavers killed by Heterorhabditis nematodes and their Photorhabdus symbionts. We found that, although Photorhabdus strains did initially displace an Enterococcus-dominated community present in uninfected G. mellonella insect larvae, the cadaver community was not static. Twelve days postinfection, Photorhabdus shared the cadaver with Stenotrophomonas species. Consistent with this result, Stenotrophomonas strains isolated from infected cadavers were resistant to Photorhabdus-mediated toxicity in solid coculture assays. We isolated and characterized a Photorhabdus-produced antibiotic from G. mellonella cadavers, produced it synthetically, and demonstrated that both the natural and synthetic compounds decreased G. mellonella-associated Enterococcus growth, but not Stenotrophomonas growth, in vitro. Finally, we showed that the Stenotrophomonas strains described here negatively affected Photorhabdus growth in vitro. Our results add an important dimension to a broader understanding of Heterorhabditis-Photorhabdus biology and also demonstrate that interspecific bacterial competition likely characterizes even a theoretically monoxenic environment, such as a Heterorhabditis-Photorhabdus-parasitized insect cadaver. IMPORTANCE Understanding, and eventually manipulating, both human and environmental health depends on a complete accounting of the forces that act on and shape microbial communities. One of these underlying forces is hypothesized to be resource competition. A resource that has received little attention in the general microbiological literature, but likely has ecological and evolutionary importance, is dead/decaying multicellular organisms

  16. Characterization of copper-resistant bacteria and bacterial communities from copper-polluted agricultural soils of central Chile

    PubMed Central

    2012-01-01

    Background Copper mining has led to Cu pollution in agricultural soils. In this report, the effects of Cu pollution on bacterial communities of agricultural soils from Valparaiso region, central Chile, were studied. Denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA genes was used for the characterization of bacterial communities from Cu-polluted and non-polluted soils. Cu-resistant bacterial strains were isolated from Cu-polluted soils and characterized. Results DGGE showed a similar high number of bands and banding pattern of the bacterial communities from Cu-polluted and non-polluted soils. The presence of copA genes encoding the multi-copper oxidase that confers Cu-resistance in bacteria was detected by PCR in metagenomic DNA from the three Cu-polluted soils, but not in the non-polluted soil. The number of Cu-tolerant heterotrophic cultivable bacteria was significantly higher in Cu-polluted soils than in the non-polluted soil. Ninety two Cu-resistant bacterial strains were isolated from three Cu-polluted agricultural soils. Five isolated strains showed high resistance to copper (MIC ranged from 3.1 to 4.7 mM) and also resistance to other heavy metals. 16S rRNA gene sequence analyses indicate that these isolates belong to the genera Sphingomonas, Stenotrophomonas and Arthrobacter. The Sphingomonas sp. strains O12, A32 and A55 and Stenotrophomonas sp. C21 possess plasmids containing the Cu-resistance copA genes. Arthrobacter sp. O4 possesses the copA gene, but plasmids were not detected in this strain. The amino acid sequences of CopA from Sphingomonas isolates (O12, A32 and A55), Stenotrophomonas strain (C21) and Arthrobacter strain (O4) are closely related to CopA from Sphingomonas, Stenotrophomonas and Arthrobacter strains, respectively. Conclusions This study suggests that bacterial communities of agricultural soils from central Chile exposed to long-term Cu-pollution have been adapted by acquiring Cu genetic determinants. Five bacterial isolates

  17. Complete genome sequence of Streptococcus thermophilus MN-BM-A01, a strain with high exopolysaccharides production.

    PubMed

    Bai, Ying; Sun, Erna; Shi, Yudong; Jiang, Yunyun; Chen, Yun; Liu, Songling; Zhao, Liang; Zhang, Ming; Guo, Huiyuan; Zhang, Hao; Mu, Zhishen; Ren, Fazheng

    2016-04-20

    Streptococcus thermophilus MN-BM-A01 (ST MN-BM-A01) (CGMCC No. 11383) was a strain isolated from Yogurt Block in Gansu, China. The yogurt fermented with this strain has good flavor, acidity, and viscosity. Moreover, ST MN-BM-A01 could produce a high level of EPS which can confer the yogurt with improved rheological properties. We reported the complete genome sequence of ST MN-BM-A01 that contains 1,876,516bp encoding 1704 coding sequences (CDSs), 67 tRNA genes and 6 rRNA operons. The genomic sequence indicated that this strain included a 35.3-kb gene cluster involved in EPS biosynthesis.

  18. Complete genome sequence of Lactobacillus salivarius Ren, a probiotic strain with anti-tumor activity.

    PubMed

    Sun, Erna; Ren, Fazheng; Liu, Songling; Ge, Shaoyang; Zhang, Ming; Guo, Huiyuan; Jiang, Lu; Zhang, Hao; Zhao, Liang

    2015-09-20

    Lactobacillus salivarius Ren (LsR) (CGMCC No. 3606) is a probiotic strain that was isolated from the feces of a healthy centenarian living in Bama, Guangxi, China. Previous studies have shown that this strain decreases 4-nitroquinoline 1-oxide (4-NQO)-induced genotoxicity in vitro. It also suppresses 4-NQO-induced oral carcinogenesis and 1,2-dimethylhydrazine (DMH)-induced colorectal carcinogenesis, and therefore may be used as an adjuvant therapeutic agent for cancer. Here, we report the complete genome sequence of LsR that consists of a circular chromosome of 1751,565 bp and two plasmids (pR1, 176,951 bp; pR2, 49,848 bp).

  19. Ogataea ganodermae sp. nov., a methanol-assimilating yeast species isolated from basidiocarps of Ganoderma sp.

    PubMed

    Ji, Zhao-Hui; Bai, Feng-Yan

    2008-06-01

    Three methanol-utilizing yeast strains were isolated from basidiocarps of Ganoderma sp. collected from a tree trunk in Mangshan Mountain, Hunan Province, southern China. These strains formed hat-shaped ascospores in unconjugated and deliquescent asci. Sequence analysis of the large-subunit rRNA gene D1/D2 domain and internal transcribed spacer (ITS) region, electrophoretic karyotype comparison and phenotypic characterization demonstrated that the three strains represent a novel species of the genus Ogataea, which is described as Ogataea ganodermae sp. nov. (type strain SHS 2.1(T) =CGMCC AS 2.3435(T) =CBS 10646(T)). Phylogenetically, the novel species was closely related to Ogataea pini and Ogataea henricii. The latter two taxa with similar D1/D2 sequences were confirmed to represent separate species by ITS sequence and electrophoretic karyotype comparisons.

  20. Production of nano bacterial cellulose from waste water of candied jujube-processing industry using Acetobacter xylinum.

    PubMed

    Li, Zheng; Wang, Lifen; Hua, Jiachuan; Jia, Shiru; Zhang, Jianfei; Liu, Hao

    2015-04-20

    The work is aimed to investigate the suitability of waste water of candied jujube-processing industry for the production of bacterial cellulose (BC) by Gluconacetobacter xylinum CGMCC No.2955 and to study the structure properties of bacterial cellulose membranes. After acid pretreatment, the glucose of hydrolysate was higher than that of waste water of candied jujube. The volumetric yield of bacterial cellulose in hydrolysate was 2.25 g/L, which was 1.5-folds of that in waste water of candied jujube. The structures indicated that the fiber size distribution was 3-14 nm in those media with an average diameter being around 5.9 nm. The crystallinity index of BC from pretreatment medium was lower than that of without pretreatment medium and BCs from various media had similar chemical binding. Ammonium citrate was a key factor for improving production yield and the crystallinity index of BC.

  1. Evaluation of the formation of volatiles and sensory characteristics of persimmon (Diospyros kaki L.f.) fruit wines using different commercial yeast strains of Saccharomyces cerevisiae.

    PubMed

    Zhu, Jian Cai; Niu, Yun Wei; Feng, Tao; Liu, Sheng Jiang; Cheng, He Xing; Xu, Na; Yu, Hai Yan; Xiao, Zuo Bing

    2014-01-01

    This study evaluated the effects of five strains (IFFI 1346, IFFI 1363, CICC 31482, D254 and CGMCC2.346) of the yeast Saccharomyces cerevisiae on the aromatic profiles of fermented persimmon (Diospyros kaki L.f.) musts. A total of 50 and 60 compounds were identified in persimmon wine by stir bar sorptive extraction coupled with gas chromatography-mass spectrometry. According to odour activity values (OAVs), 26 detected compounds showed an OAV above 1. Principal component analysis explained the distribution of these persimmon wines on the basis of volatile compounds with OAV>1. The volatile compounds with high OAV included ethyl hexanoate, ethyl octanoate, methyl decanoate, linalool and geraniol. Quantitative descriptive analysis was employed. The result showed that persimmon wines fermented with strains IFFI 1363 and D254 were strongly correlated with persimmon, aroma harmony, fruity, fusel and taste balanced, fullness, hedonic scale. Therefore, the two yeast strains could be used as starter culture for persimmon wine production.

  2. A high-throughput screening strategy for nitrile-hydrolyzing enzymes based on ferric hydroxamate spectrophotometry.

    PubMed

    He, Yu-Cai; Ma, Cui-Luan; Xu, Jian-He; Zhou, Li

    2011-02-01

    Nitrile-hydrolyzing enzymes (nitrilase or nitrile hydratase/amidase) have been widely used in the pharmaceutical industry for the production of carboxylic acids and their derivatives, and it is important to build a method for screening for nitrile-hydrolyzing enzymes. In this paper, a simple, rapid, and high-throughput screening method based on the ferric hydroxamate spectrophotometry has been proposed. To validate the accuracy of this screening strategy, the nitrilases from Rhodococcus erythropolis CGMCC 1.2362 and Alcaligenes sp. ECU0401 were used for evaluating the method. As a result, the accuracy for assaying aliphatic and aromatic carboxylic acids was as high as the HPLC-based method. Therefore, the method may be potentially used in the selection of microorganisms or engineered proteins with nitrile-hydrolyzing enzymes.

  3. Efficient production of dihydroxyacetone from biodiesel-derived crude glycerol by newly isolated Gluconobacter frateurii.

    PubMed

    Liu, Yu-Peng; Sun, Yang; Tan, Cong; Li, Hua; Zheng, Xiao-Juan; Jin, Kui-Qi; Wang, Gang

    2013-08-01

    The efficient production of dihydroxyacetone (DHA) on biodiesel-derived glycerol based media was developed. A newly isolated strain, Gluconobacter frateurii CGMCC 5397, could convert crude glycerol to DHA with high yield and productivity. In shake-flask fermentation, the DHA concentration of 73.1 gl(-1) was attained at 48 h using an optimum medium containing biodiesel-derived crude glycerol. When fed-batch fermentation was carried out in a 7-l stirred bioreactor with crude glycerol, the DHA concentration, productivity, and yield were 125.8 gl(-1), 2.6 gl(-1)h(-1), and 90.5% at 48 h, respectively. This study suggests that the inexpensive biodiesel-derived crude glycerol could be utilized for efficient production of DHA by G. frateurii.

  4. Streptomyces yunnanensis sp. nov., a mesophile from soils in Yunnan, China.

    PubMed

    Zhang, Qi; Li, Wen-Jun; Cui, Xiao-Long; Li, Ming-Gang; Xu, Li-Hua; Jiang, Cheng-Lin

    2003-01-01

    A strain was isolated from red soil from the suburb of Kunming in Yunnan, China, during the screening of agricultural antibiotics which prevented and cured wheat-stem rust. This isolate, designated YIM 41004T (= CGMCC 4.1004T = DSM 41793), was identified by a polyphasic approach. The test results suggested that this strain was clearly assigned to the genus Streptomyces and found to be marginally close to Williams cluster 32 based on the morphological and physiological data. The almost-complete 16S rRNA gene sequence of the strain was determined and compared with those of representative streptomycetes. The phylogenetic tree confirmed its membership in the genus Streptomyces and demonstrated that this strain represented a separate phyletic line in a clade encompassed by streptomycetes within cluster 32. Based on the polyphasic evidence, it is therefore proposed that strain YIM 41004T should be classified as Streptomyces yunnanensis sp. nov.

  5. Phylogenomic analysis shows that ‘Bacillus vanillea’ is a later heterotypic synonym of Bacillus siamensis.

    PubMed

    Dunlap, Christopher A

    2015-10-01

    ‘Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) was isolated from cured vanilla beans and involved in the formation of vanilla aroma compounds. A draft genome of this strain was assembled and yielded a length of 3.71 Mbp with a DNA G+C content of 46.3 mol%. Comparative genomic analysis with its nearest relatives showed only minor differences between this strain and the genome of the Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T), with a calculated DNA–DNA hybridization (DDH) value of 91.2 % and an average nucleotide identity (ANI) of 98.9 %. This DDH value is well above the recommended 70 % threshold for species delineation, as well as the ANI threshold of 95 %. In addition, the results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the type strains of these two taxa are highly similar with phenotype coherence. A core genome multi-locus sequencing analysis was conducted for the strains and the results show that ‘Bacillus vanillea’ XY18 clusters closely to the type strain of Bacillus siamensis. Therefore, it is proposed that the species ‘Bacillus vanillea’ XY18 ( = CGMCC 8629 = NCCB 100507) should be reclassified as a later heterotypic synonym of Bacillus siamensis KCTC 13613T ( = BCC 22614T = KACC 16244T). An emended description of Bacillus siamensis is provided.

  6. Identification of key genes involved in polysaccharide bioflocculant synthesis in Bacillus licheniformis.

    PubMed

    Chen, Zhen; Liu, Peize; Li, Zhipeng; Yu, Wencheng; Wang, Zhi; Yao, Haosheng; Wang, Yuanpeng; Li, Qingbiao; Deng, Xu; He, Ning

    2017-03-01

    The present study reports the sequenced genome of Bacillus licheniformis CGMCC 2876, which is composed of a 4,284,461 bp chromosome that contains 4,188 protein-coding genes, 72 tRNA genes, and 21 rRNA genes. Additional analysis revealed an eps gene cluster with 16 open reading frames. Conserved Domains Database analysis combined with qPCR experiments indicated that all genes in this cluster were involved in polysaccharide bioflocculant synthesis. Phosphoglucomutase and UDP-glucose pyrophosphorylase were supposed to be key enzymes in polysaccharide secretion in B. licheniformis. A biosynthesis pathway for the production of polysaccharide bioflocculant involving the integration of individual genes was proposed based on functional analysis. Overexpression of epsDEF from the eps gene cluster in B. licheniformis CGMCC 2876 increased the flocculating activity of the recombinant strain by 90% compared to the original strain. Similarly, the crude yield of polysaccharide bioflocculant was enhanced by 27.8%. Overexpression of the UDP-glucose pyrophosphorylase gene not only increased the flocculating activity by 71% but also increased bioflocculant yield by 13.3%. Independent of UDP-N-acetyl-D-mannosamine dehydrogenase gene, flocculating activity, and polysaccharide yield were negatively impacted by overexpression of the UDP-N-acetylglucosamine 2-epimerase gene. Overall, epsDEF and gtaB2 were identified as key genes for polysaccharide bioflocculant synthesis in B. licheniformis. These results will be useful for further engineering of B. licheniformis for industrial bioflocculant production. Biotechnol. Bioeng. 2017;114: 645-655. © 2016 Wiley Periodicals, Inc.

  7. Streptosporangium jiaoheense sp. nov. and Streptosporangium taraxaci sp. nov., actinobacteria isolated from soil and dandelion root (Taraxacum mongolicum Hand.-Mazz.).

    PubMed

    Zhao, Junwei; Guo, Lifeng; Li, Zhilei; Piao, Chenyu; Li, Yao; Li, Jiansong; Liu, Chongxi; Wang, Xiangjing; Xiang, Wensheng

    2016-06-01

    Two novel actinobacteria, designated strains NEAU-Jh1-4T and NEAU-Wp2-0T, were isolated from muddy soil collected from a riverbank in Jiaohe and a dandelion root collected from Harbin, respectively. A polyphasic study was carried out to establish the taxonomic positions of these two strains. The phylogenetic analysis based on the 16S rRNA gene sequences of strains NEAU-Jh1-4T and NEAU-Wp2-0T indicated that strain NEAU-Jh1-4T clustered with Streptosporangium nanhuense NEAU-NH11T (99.32 % 16S rRNA gene sequence similarity), Streptosporangium purpuratum CY-15110T (98.30 %) and Streptosporangium yunnanense CY-11007T (97.95 %) and strain NEAU-Wp2-0T clustered with 'Streptosporangium sonchi  ' NEAU-QS7 (99.39 %), 'Streptosporangium kronopolitis' NEAU-ML10 (99.26 %), 'Streptosporangium shengliense' NEAU-GH7 (98.85 %) and Streptosporangium longisporum DSM 43180T (98.69 %). Moreover, morphological and chemotaxonomic properties of the two isolates also confirmed their affiliation to the genus Streptosporangium. However, the low level of DNA-DNA hybridization and some phenotypic characteristics allowed the isolates to be differentiated from the most closely related species. Therefore, it is proposed that strains NEAU-Jh1-4T and NEAU-Wp2-0T represent two novel species of the genus Streptosporangium, for which the name Streptosporangium jiaoheense sp. nov. and Streptosporangium taraxaci sp. nov. are proposed. The type strains are NEAU-Jh1-4T (=CGMCC 4.7213T=JCM 30348T) and NEAU-Wp2-0T (=CGMCC 4.7217T=JCM 30349T), respectively.

  8. Sphingomonas qilianensis sp. nov., Isolated from Surface Soil in the Permafrost Region of Qilian Mountains, China.

    PubMed

    Piao, Ai-Lian; Feng, Xiao-Min; Nogi, Yuichi; Han, Lu; Li, Yonghong; Lv, Jie

    2016-04-01

    A Gram-stain-negative, strictly aerobic, non-motile and rod-shaped bacterial strain, designated X1(T), was isolated from the permafrost region of Qilian Mountains in northwest of China. Phylogenetic analyses of 16S rRNA gene sequence revealed that strain X1(T) was a member of the genus Sphingomonas and shared the highest 16S rRNA gene sequence similarity with Sphingomonas oligophenolica JCM 12082(T) (96.9%), followed by Sphingomonas glacialis CGMCC 1.8957(T) (96.7%) and Sphingomonas alpina DSM 22537(T) (96.4%). Strain X1(T) was able to grow at 15-30 °C, pH 6.0-10.0 and with 0-0.3% NaCl (w/v). The DNA G+C content of the isolate was 64.8 mol%. Strain X1(T)-contained Q-10 as the dominant ubiquinone and C(18:1)ω7c, C(16:1)ω7c, C(16:0) and C(14:0) 2-OH as the dominant fatty acids. The polar lipid profile of strain XI(T)-contained sphingoglycolipid, phosphatidylglycerol, phosphatidylethanolamine, one unidentified glycolipid and two unidentified phospholipid. Due to the phenotypic and genetic distinctiveness and other characteristic studied in this article, we consider X1(T) as a novel species of the genus Sphingomonas and propose to name it Sphingomonas qilianensis sp. nov. The type strain is X1(T) (=CGMCC 1.15349(T) = KCTC 42862(T)).

  9. Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov., two novel actinomycetes isolated from muddy soil and emended description of the genus Actinomadura.

    PubMed

    Zhao, Junwei; Guo, Lifeng; Sun, Pengyu; Han, Chuanyu; Bai, Lu; Liu, Chongxi; Li, Yunxi; Xiang, Wensheng; Wang, Xiangjing

    2015-12-01

    Two novel actinomycetes, designated strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T), were isolated from muddy soil collected from a riverbank in Jiaohe, Jilin Province, north China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibited 99.9 % 16S rRNA gene sequence similarity with each other and that they are closely related to Actinomadura viridis IFO 15238(T) (99.6, 99.6 %) and Actinomadura vinacea IFO 14688(T) (99.3, 99.3 %). Phylogenetic analysis based on the 16S rRNA gene sequences indicated that the two cultures clustered together and formed a cluster with A. viridis IFO 15238(T), A. vinacea IFO 14688(T) and Actinomadura rugatobispora IFO 14382(T). However, the DNA-DNA hybridization value between strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) was 63.6 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-Jh1-3(T) and NEAU-Jh2-5(T) represent two novel species of the genus Actinomadura, for which the names Actinomadura jiaoheensis sp. nov. and Actinomadura sporangiiformans sp. nov. are proposed. The type strains are NEAU-Jh1-3(T) (=CGMCC 4.7197(T) = JCM 30341(T)) and NEAU-Jh2-5(T) (=CGMCC 4.7211(T) = JCM 30342(T)), respectively.

  10. A new strategy for strain improvement of Aurantiochytrium sp. based on heavy-ions mutagenesis and synergistic effects of cold stress and inhibitors of enoyl-ACP reductase.

    PubMed

    Cheng, Yu-Rong; Sun, Zhi-Jie; Cui, Gu-Zhen; Song, Xiaojin; Cui, Qiu

    2016-11-01

    Developing a strain with high docosahexaenoic acid (DHA) yield and stable fermenting-performance is an imperative way to improve DHA production using Aurantiochytrium sp., a microorganism with two fatty acid synthesis pathways: polyketide synthase (PKS) pathway and Type I fatty acid synthase (FAS) pathway. This study investigated the growth and metabolism response of Aurantiochytrium sp. CGMCC 6208 to two inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan), and proposed a method of screening high DHA yield Aurantiochytrium sp. strains with heavy ion mutagenesis and pre-selection by synergistic usage of cold stress (4°C) and FAS inhibitors (triclosan and isoniazid). Results showed that (1) isoniazid and triclosan have positive effects on improving DHA level of cells; (2) mutants from irradiation dosage of 120Gy yielded more DHA compared with cells from 40Gy, 80Gy treatment and wild type; (3) DHA contents of mutants pre-selected by inhibitors of enoyl-ACP reductase of Type II FAS pathway (isoniazid and triclosan)at 4°C, were significantly higher than that of wild type; (4) compared to the wild type, the DHA productivity and yield of a mutant (T-99) obtained from Aurantiochytrium sp. CGMCC 6208 by the proposed method increased by 50% from 0.18 to 0.27g/Lh and 30% from 21 to 27g/L, respectively. In conclusion, this study developed a feasible method to screen Aurantiochytrium sp. with high DHA yield by a combination of heavy-ion mutagenesis and mutant-preselection by FAS inhibitors and cold stress.

  11. Streptosporangium sonchi sp. nov. and Streptosporangium kronopolitis sp. nov., two novel actinobacteria isolated from a root of common sowthistle (Sonchus oleraceus L.) and a millipede (Kronopolites svenhedind Verhoeff).

    PubMed

    Ma, Zhaoxu; Liu, Hui; Liu, Chongxi; He, Hairong; Zhao, Junwei; Wang, Xin; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng

    2015-06-01

    Two novel actinobacteria, designated strains NEAU-QS7(T) and NEAU-ML10(T), were isolated from a root of Sonchus oleraceus L. and a Kronopolites svenhedind Verhoeff specimen, respectively, collected from Wuchang, Heilongjiang Province, China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The two strains were observed to form abundant aerial hyphae that differentiated into spherical spore vesicles. The phylogenetic analysis based on the 16S rRNA gene sequences of strains NEAU-QS7(T) and NEAU-ML10(T) showed that the two novel isolates exhibited 99.7 % 16S rRNA gene sequence similarity with each other and that they are most closely related to Streptosporangium shengliense NEAU-GH7(T) (99.1, 99.0 %) and Streptosporangium longisporum DSM 43180(T) (99.1, 99.0 %). However, the DNA-DNA hybridization value between strains NEAU-QS7(T) and NEAU-ML10(T) was 46.5 %, and the values between the two strains and their closest phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their closest phylogenetic relatives. Thus, strains NEAU-QS7(T) and NEAU-ML10(T) represent two novel species of the genus Streptosporangium, for which the names Streptosporangium sonchi sp. nov. and Streptosporangium kronopolitis sp. nov. are proposed. The type strains are NEAU-QS7(T) (=CGMCC 4.7142(T) =DSM 46717(T)) and NEAU-ML10(T) (=CGMCC 4.7153(T) =DSM 46720(T)), respectively.

  12. Diversity, metal resistance and uranium sequestration abilities of bacteria from uranium ore deposit in deep earth stratum.

    PubMed

    Islam, Ekramul; Sar, Pinaki

    2016-05-01

    Metal resistance and uranium (U) sequestration abilities of bacteria residing in subsurface U ore was investigated using 122 pure culture strains isolated through enrichment. The cumulative frequencies of isolates resistant to each metal tested were as follows: As(V), 74%; Zn, 58%; Ni, 53%; Cd, 47%; Cr(VI), 41%; Co, 40%; Cu, 20%; and Hg, 4%. 16S rRNA gene analysis revealed that isolated bacteria belonged to 14 genera with abundance of Arthrobacter, Microbacterium, Acinetobacter and Stenotrophomonas. Cobalt did not interfere with the growth of most of the bacterial isolates belonging to different groups while U allowed growth of four different genera of which Stenotrophomonas and Microbacterium showed high U tolerance. Interestingly, tolerance to Ni, Zn, Cu, and Hg was observed only in Microbacterium, Arthrobacter, Paenibacillus¸ and Acinetobacter, respectively. However, Microbacterium was found to be dominant when isolated from other five different metal enrichments including U. Uranium removal study showed that 84% of the test bacteria could remove more than 50mgUg(-1) dry weight from 80 or 160mgL(-1) U within 48h. In general, Microbacterium, Arthrobacter and Acinetobacter could remove a higher amount of U. High resolution transmission electron microscopy (HRTEM) study of U exposed cells revealed that accumulated U sequestered mostly around the cell periphery. The study highlights that indigenous U ore deposit bacteria have the potential to interact with U, and thus could be applied for bioremediation of U contaminated sites or wastes.

  13. Restricted streptomycin use in apple orchards did not adversely alter the soil bacteria communities.

    PubMed

    Walsh, Fiona; Smith, Daniel P; Owens, Sarah M; Duffy, Brion; Frey, Jürg E

    2013-01-01

    Streptomycin has been authorized for restricted use in the prevention of the fire blight disease of pome fruit orchards in the EU and Switzerland. This study addresses the important topic of the influence of the use of streptomycin in agriculture on the total bacteria community within the soil ecosystem. Soil samples were taken from soils under apple trees, prior to streptomycin application and 2 weeks post streptomycin application or water application (untreated control). High throughput 16S rRNA gene amplicon sequencing was used to generate datasets from the soils under apple trees in apple orchards from three different locations in Switzerland. We hypothesized that the use of streptomycin would reduce the bacterial diversity within the soil samples and enhance a reduction in the variety of taxa present. Bacterial species such as Pseudomonas, Burkholderia, and Stenotrophomonas are intrinsically resistant to many antibiotics and as such it is of interest to investigate if the use of streptomycin provided a selective advantage for these bacteria in the soil ecosystem. The application of streptomycin did not influence the abundance and diversities of major bacteria taxa of the soils or the Pseudomonas, Burkholderia, and Stenotrophomonas species. We also discovered that apple orchards under the same management practices, did not harbor the same bacterial communities. The restricted application of streptomycin in the protection of apple orchards from the fire blight pathogen Erwinia amylovora under the guidelines in Switzerland did not alter either the bacterial diversity or abundance within these soil ecosystems.

  14. Biodegradation of geosmin in drinking water by novel bacteria isolated from biologically active carbon.

    PubMed

    Zhou, Beihai; Yuan, Rongfang; Shi, Chunhong; Yu, Liying; Gu, Junnong; Zhang, Chunlei

    2011-01-01

    Three strains of Gram-negative bacteria capable of removing geosmin from drinking water were isolated from biologically active carbon and identified to be Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp. based on physio-biochemistry analysis and 16S rRNA gene sequence analysis. Removal efficiencies of 2 mg/L geosmin in mineral salts medium were 84.0%, 80.2% and 74.4% for Chryseobacterium sp., Sinorhizobium sp. and Stenotrophomonas sp., respectively, while removal efficiencies of 560 ng/L geosmin in filter influent were 84.8%, 82.3% and 82.5%, respectively. The biodegradation of geosmin was determined to be a pseudo first-order reaction, with rate constants at 2 mg/L and 560 ng/L being 0.097 and 0.086 day(-1), 0.089 and 0.084 day(-1), 0.074 and 0.098 day(-1) for the above mentioned degraders, respectively. The biomass of culture in the presence of geosmin was much higher than that in the absence of geosmin.

  15. Antimicrobial Activity of Medicinal Plants Correlates with the Proportion of Antagonistic Endophytes

    PubMed Central

    Egamberdieva, Dilfuza; Wirth, Stephan; Behrendt, Undine; Ahmad, Parvaiz; Berg, Gabriele

    2017-01-01

    Medicinal plants are known to harbor potential endophytic microbes, due to their bioactive compounds. In a first study of ongoing research, endophytic bacteria were isolated from two medicinal plants, Hypericum perforatum and Ziziphora capitata with contrasting antimicrobial activities from the Chatkal Biosphere Reserve of Uzbekistan, and their plant-specific traits involved in biocontrol and plant growth promotion were evaluated. Plant extracts of H. perforatum exhibited a remarkable activity against bacterial and fungal pathogens, whereas extracts of Z. capitata did not exhibit any potential antimicrobial activity. Matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) was used to identify plant associated culturable endophytic bacteria. The isolated culturable endophytes associated with H. perforatum belong to eight genera (Arthrobacter, Achromobacter, Bacillus, Enterobacter, Erwinia, Pseudomonas, Pantoea, Serratia, and Stenotrophomonas). The endophytic isolates from Z. capitata also contain those genera except Arthrobacter, Serratia, and Stenotrophomonas. H. perforatum with antibacterial activity supported more bacteria with antagonistic activity, as compared to Z. capitata. The antagonistic isolates were able to control tomato root rot caused by Fusarium oxysporum and stimulated plant growth under greenhouse conditions and could thus be a cost-effective source for agro-based biological control agents. PMID:28232827

  16. Physiological and phylogenetic characterization of a stable benzene-degrading, chlorate-reducing microbial community.

    PubMed

    Weelink, Sander A B; Tan, Nico C G; Ten Broeke, Harm; van Doesburg, Wim; Langenhoff, Alette A M; Gerritse, Jan; Stams, Alfons J M

    2007-05-01

    A stable anoxic enrichment culture was obtained that degraded benzene with chlorate as an electron acceptor. The benzene degradation rate was 1.65 mM benzene per day, which is similar to reported aerobic benzene degradation rates but 20-1650 times higher than reported for anaerobic benzene degradation. Denaturing gradient gel electrophoresis of part of the 16S rRNA gene, cloning and sequencing showed that the culture had a stable composition after the seventh transfer. Five bacterial clones were further analyzed. Two clones corresponded to bacteria closely related to Alicycliphilus denitrificans K601. The three other clones corresponded to bacteria closely related to Zoogloea resiniphila PIV-3A2w, Mesorhizobium sp. WG and Stenotrophomonas acidaminiphila. DGGE analysis of cultures grown with different electron donors and acceptors indicated that the bacterium related to Alicycliphilus denitrificans K601 is able to degrade benzene coupled to chlorate reduction. The role of the other bacteria could not be conclusively determined. The bacterium related to Mesorhizobium sp. WG can be enriched with benzene and oxygen, but not with acetate and chlorate, while the bacterium related to Stenotrophomonas acidaminophila grows with acetate and chlorate, but not with benzene and oxygen. As oxygen is produced during chlorate reduction, an aerobic pathway of benzene degradation is most likely.

  17. [The identification of nonfermentative gram-negative bacteria. Experiences with 676 apyocyaninogenic strains (author's transl)].

    PubMed

    Berger, U; Piotrowski, H D

    1981-02-01

    During a period of 16 months 1757 strains of nonfermentative gram-negative rods have been isolated from clinical material. Of the, 1205 (69%) were P. aeruginosa, 124 (10%) of which failed to produce pyocyanin. The apyocyaninogenic strains as well as the remaining 552 isolates were differentiated by steps according to a diagnostic scheme developed by us. For identification of species two or three steps were needed. By this procedure, 530 of the 552 strains could be assigned to nineteen species within the genera Pseudomonas, Achromobacter, Alcaligenes, Flavobacterium, Agrobacterium and Acinetobacter. 17 strains could not be identified below the genus level, one strain belonged to CDC-group VE-2 and four strains were not identifiable. 72% of the 552 strains belonged to only four species: Pseudomonas putida, P. maltophilia, Acinetobacter lwoffii and A. anitratus.

  18. A bacterial protein has homology with human chorionic gonadotropin (hCG).

    PubMed

    Grover, S; Woodward, S R; Odell, W D

    1993-06-30

    Studies from our laboratory have demonstrated the presence of a 48.5 kD cell wall protein in the bacterium, Xanthomonas maltophilia, which immunologically resembles the beta subunit of human chorionic gonadotropin. Primers were designed from the amino acid sequences of enzymatically cleaved peptide fragments of this protein. These primers were used to obtain PCR amplified products, which were subsequently cloned in a PCR11TA cloning vector, and a 492 base pair nucleotide sequence was obtained with a 164 amino acid open reading frame. When this nucleotide sequence was aligned with exon 2 of genes 5 and 6 of the beta hCG gene, a 53% homology was observed. The translated protein sequence had a 35% homology with hCG and a 25% homology with human luteinizing hormone.

  19. Meteorite organics in planetary environments: hydrothermal release, surface activity, and microbial utilization

    NASA Technical Reports Server (NTRS)

    Mautner, M. N.; Leonard, R. L.; Deamer, D. W.

    1995-01-01

    Up to 50% of the organics in the Murchison meteorite, possibly including some of the polymer, is released in high temperature and pressure aqueous environments, to 350 degrees C and 250 bar, that simulate submarine volcanic, hydrothermal or impact-induced conditions. Meteorite organics of prebiotic significance, such as nonanoic acid, glycine, and pyrene survive the hydrothermal conditions. The released material is surface active with surface pressures up to 19.8 x 10(-3) N m-1, and exhibits an extended surface tension isotherm which suggests a mixture of amphiphilic components. One component, nonanoic acid, is shown to form vesicles. The materials extracted under mild conditions, at 120 degrees C, are nutrients for the humic acid bacterium Pseudomonas maltophilia and efficient nutrients for the oligotroph Flavobacterium oryzihabitans, demonstrating the capability of microorganisms to metabolize extraterrestrial organics.

  20. Patterns of oral bacterial infection in captive snakes.

    PubMed

    Draper, C S; Walker, R D; Lawler, H E

    1981-12-01

    The bacterial isolates from culture specimens of snakes with infectious stomatitis were compared with those from culture specimens of the oral cavity of healthy captive snakes. Cloacal swab specimens were also taken from healthy snakes to compare their intestinal and oral bacterial populations. The healthy snakes had a predominantly gram-positive oral flora, with Corynebacterium spp and coagulase-negative Staphylococcus spp being the organisms isolated most frequently. The specimens from snakes with infectious stomatitis yielded predominantly gram-negative bacteria. The organisms most frequently isolated from these specimens were Pseudomonas aeruginosa, Providencia rettgeri, and P maltophilia. The cloacal swabbing of healthy snakes also resulted in the isolation of predominantly gram-negative organisms, suggesting that these bacteria are not exogenous pathogens but opportunistic invaders.

  1. Motion of the Zinc Ions in Catalysis by a di-Zinc Metallo-beta-Lactamase

    SciTech Connect

    R Breece; Z Hu; M Crowder; D Tierney

    2011-12-31

    We report rapid-freeze-quench X-ray absorption spectroscopy of a dizinc metallo-beta-lactamase (MbetaL) reaction intermediate. The Zn(II) ions in the dinuclear active site of the S. maltophilia Class B3 MbetaL move away from each other, by approximately 0.3 A after 10 ms of reaction with nitrocefin, from 3.4 to 3.7 A. Together with our previous characterization of the resting enzyme and its nitrocefin product complex, where the Zn(II) ion separation relaxes to 3.6 A, these data indicate a scissoring motion of the active site that accompanies the ring-opening step. The average Zn(II) coordination number of 4.5 in the resting enzyme appears to be maintained throughout the reaction with nitrocefin. This is the first direct structural information available on early stage dizinc metallo-beta-lactamase catalysis.

  2. Meteorite organics in planetary environments: hydrothermal release, surface activity, and microbial utilization

    NASA Astrophysics Data System (ADS)

    Mautner, Michael N.; Leonard, Robert L.; Deamer, David W.

    1995-02-01

    Up to 50% of the organics in the Murchison meteorite, possibly including some of the polymer, is released in high temperature and pressure aqueous environments, to 350°C and 250 bar, that simulate submarine volcanic, hydrothermal or impact-induced conditions. Meteorite organics of prebiotic significance, such as nonanoic acid, glycine, and pyrene survive the hydrothermal conditions. The released material is surface active with surface pressures up to 19.8 × 10 -3 N m -1, and exhibits an extended surface tension isotherm which suggests a mixture of amphiphilic components. One component, nonanoic acid, is shown to form vesicles. The materials extracted under mild conditions, at 120°C, are nutrients for the humic acid bacterium Pseudomonas maltophilia and efficient nutrients for the oligotroph Flavobacterium oryzihabitans, demonstrating the capability of micro-organisms to metabolize extraterrestrial organics.

  3. Shewanella psychrophila sp. nov. and Shewanella piezotolerans sp. nov., isolated from west Pacific deep-sea sediment.

    PubMed

    Xiao, Xiang; Wang, Peng; Zeng, Xiang; Bartlett, Douglas Hoyt; Wang, Fengping

    2007-01-01

    Two Shewanella-like bacterial strains, WP2(T) and WP3(T), which were isolated from west Pacific deep-sea sediment, were studied to determine their taxonomic position. Cells of the two bacteria were facultatively anaerobic, Gram-negative rods and motile by means of a single polar flagellum. Strain WP2(T) was psychrophilic, growing optimally at about 10-15 degrees C, whereas strain WP3(T) was psychrotolerant, growing optimally at 15-20 degrees C. The two strains grew in the pressure range 0.1-50 MPa, with optimal growth at 20 MPa. Strain WP3(T) was able to use nitrate, fumarate, trimethylamine N-oxide (TMAO), DMSO and insoluble Fe(III) as terminal electron acceptors during anaerobic growth, whereas strain WP2(T) was able to use only nitrate, TMAO and DMSO. The 16S rRNA gene sequences of strains WP2(T) and WP3(T) were 97 % identical, and showed highest similarity (97 %) to those of Shewanella fidelis KMM 3589 and Shewanella benthica ATCC 43992(T), respectively. The gyrB gene sequences of strains WP2(T)and WP3 (T) were also determined, and showed highest similarity to those of Shewanella violacea JCM 10179(T) (90 %) and Shewanella sairae SM2-1(T) (87 %), respectively. Contrary to the 16S rRNA gene sequence results, the phylogeny based on gyrB gene sequence analysis placed strain WP2(T), S. violacea and S. benthica in one group, while strain WP3(T) grouped with S. fidelis and S. sairae. DNA-DNA hybridization experiments supported the placement of strain WP2(T) with S. violacea and S. benthica. Phylogenetic evidence, together with DNA-DNA relatedness and phenotypic characteristics, indicated that the two new strains represented two novel deep-sea Shewanella species. The names Shewanella psychrophila sp. nov. (type strain WP2(T)=JCM 13876(T)=CGMCC 1.6159(T)) and Shewanella piezotolerans (type strain WP3(T)=JCM 13877(T)=CGMCC 1.6160(T)) are proposed.

  4. Bacillus lindianensis sp. nov., a novel alkaliphilic and moderately halotolerant bacterium isolated from saline and alkaline soils.

    PubMed

    Dou, Guiming; Liu, Hongcan; He, Wei; Ma, Yuchao

    2016-01-01

    Two alkaliphilic and halotolerant Gram-stain positive, rod-shaped and endospore-forming bacteria, designated strains 12-3(T) and 12-4, were isolated from saline and alkaline soils collected in Lindian county, Heilongjiang province, China. Both strains were observed to grow well at a wide range of temperature and pH values, 10-45 °C and pH 8-12, with optimal growth at 37 °C and pH 9.0, respectively. Growth of the two strains was found to occur at total salt concentrations of 0-12 % (w/v), with an optimum at 4 % (w/v). The G+C contents of the genomic DNA of strains 12-3(T) and 12-4 were determined to be 42.7 and 42.4 mol%, respectively, and the major cellular fatty acids were identified as anteiso-C15:0 and anteiso-C17:0. In isolate 12-3(T), meso-diaminopimelic acid was found to be the diagnostic diamino acid of the cell wall peptidoglycan; diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were identified as the major cellular polar lipids; and menaquinone-7 was identified as the predominant isoprenoid quinone. Strains 12-3(T) and 12-4 share very close 16S rRNA gene sequence similarity (99.74 %) and their DNA-DNA relatedness was 95.3 ± 0.63 %, meaning that the two strains can be considered to belong to the same species. 16S rRNA gene sequence-based phylogenetic analysis revealed strains 12-3(T) and 12-4 exhibit high similarities to Bacillus pseudofirmus DSM 8715(T) (98.7 %), Bacillus marmarensis DSM 21297(T) (97.2 %) and Bacillus nanhaiisediminis CGMCC 1.10116(T) (97.1 and 97.0 %, respectively). DNA-DNA hybridization values between isolate 12-3(T) and the type strains of closely related Bacillus species were below 30 %. On the basis of the polyphasic evidence presented, strains 12-3(T) and 12-4 are considered to represent a novel species of the genus Bacillus, for which the name Bacillus lindianensis sp. nov. is proposed. The type strain is 12-3(T) (DSM 26864(T) = CGMCC 1.12717(T)).

  5. Larsenimonas suaedae sp. nov., a moderately halophilic, endophytic bacterium isolated from the euhalophyte Suaeda salsa.

    PubMed

    Xia, Zhi-Jie; Wu, Hong-Zhen; Cui, Chun-Xiao; Chen, Qi; Zhao, Guo-Yan; Wang, Hai-Xia; Dai, Mei-Xue

    2016-08-01

    A moderately halophilic, Gram-stain-negative, non-endospore-forming endophytic bacterium designated strain ST307T was isolated from the euhalophyte Suaeda salsa in Dongying, China. Strain ST307T was aerobic, rod-shaped, motile and orange-yellow-pigmented. The organism grew at NaCl concentrations of 0.6-20 % (w/v) (optimum 5-6 %, w/v), at temperatures of 5-45 °C (optimum 35 °C) and at pH 5-9 (optimum pH 7-8). It accumulated poly-β-hydroxybutyric acid and produced exopolysaccharides. The major fatty acids were C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C16 : 1ω7c/C16 : 1ω6c. The predominant lipoquinone was ubiquinone Q-9. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, a glycoaminolipid and a phosphoglycoaminolipid. The DNA G+C content was 60.5 mol%. Phylogenetic analyses of 16S rRNA gene sequences and concatenated atpA, rpoD and secA gene sequences revealed that the strain represents a member of the genus Larsenimonas. The closest related type strain was Larsenimonas salina M1-18T. Mean DNA-DNA relatedness values between strain ST307T and the related species L. salina M1-18T, Chromohalobacter beijerinckii DSM 7218T, C. canadensis DSM 6769T, C. israelensis DSM 6768T, C. marismortui CGMCC 1.2321T, C. nigrandesensis DSM 14323T, C. salexigens DSM 3043T and C. sarecensis DSM 15547T were 15±2-45±1 %. On the basis of phenotypic, chemotaxonomic and molecular features, strain ST307T clearly represents a novel species of the genus Larsenimonas. The name Larsenimonassuaedae sp. nov. is proposed, with ST307T (=CGMCC 1.8902T=DSM 22428T) as the type strain.

  6. Catellatospora vulcania sp. nov. and Catellatospora paridis sp. nov., two novel actinobacteria isolated from volcanic sediment and the rhizosphere of Paris polyphylla.

    PubMed

    Jia, Feiyu; Guo, Siyu; Shen, Yue; Gao, Meiyue; Liu, Chongxi; Zhou, Shuyu; Li, Jiansong; Guan, Xuejiao; Wang, Xiangjing; Xiang, Wensheng

    2016-01-01

    Two novel actinobacteria, designated strains NEAU-JM1(T) and NEAU-CL2(T), were isolated from volcanic sediment and the rhizosphere soil of Paris polyphylla, respectively, collected from Jiling and Heilongjiang Province, China. A polyphasic study was carried out to establish the taxonomic positions of these strains. The 16S rRNA gene sequence analysis showed that the two novel isolates exhibit 99.5 % 16S rRNA gene sequence similarity with each other. The phylogenetic analysis based on 16S rRNA gene sequence of strain NEAU-JM1(T) showed it to be closely related to Catellatospora methionotrophica JCM 7543(T) (99.4 % sequence similarity), Catellatospora coxensis DSM 44901(T) (99.4 %), Catellatospora citrea DSM 44097(T) (99.3 %) and Catellatospora chokoriensis JCM 12950(T) (99.2 %), and that of strain NEAU-CL2(T) to C. citrea DSM 44097(T) (99.4 %), C. methionotrophica JCM 7543(T) (99.3 %), C. chokoriensis JCM 12950(T) (99.3 %) and C. coxensis DSM 44901(T) (99.2 %). However, the DNA-DNA hybridization value between strains NEAU-JM1(T) and NEAU-CL2(T) was 62.2 %, and the values between the two strains and their close phylogenetic relatives were also below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, the two strains can be distinguished from each other and their close phylogenetic relatives. Thus, strains NEAU-JM1(T) and NEAU-CL2(T) can be concluded to represent two novel species of the genus Catellatospora, for which the names Catellatospora vulcania sp. nov. and Catellatospora paridis sp. nov. are proposed. The type strains are NEAU-JM1(T) (=CGMCC 4.7174(T) = JCM 30054(T)) and NEAU-CL2(T) (=CGMCC 4.7236(T) = DSM 100519(T)), respectively.

  7. Psychroflexus salis sp. nov. and Psychroflexus planctonicus sp. nov., isolated from a salt lake.

    PubMed

    Zhong, Zhi-Ping; Liu, Ying; Wang, Fang; Zhou, Yu-Guang; Liu, Hong-Can; Liu, Zhi-Pei

    2016-01-01

    Two Gram-stain-negative, catalase- and oxidase-positive, strictly aerobic, non-motile, moderately halophilic bacteria (strains X15M-6T and X15M-8T) were isolated from Lake Xiaochaidan, a salt lake in Qaidam basin, Qinghai Province, China. Cells of X15M-6T were rod-like or coccoid, 0.5-0.9 μm wide and 0.9-1.5 μm long; cells of X15M-8T were rods, 0.3-0.6 μm wide and 1.2-2.2 μm long. Growth was observed in the presence of 0.5-14.0 % (w/v) NaCl (optimum, 3.0 %) and at pH 6.5-10.0 (optimum, pH 7.0-7.5) for both. X15M-6T and X15M-8T grew at 10-35 °C (optimum, 20-25 °C) and 4-35 °C (optimum, 25 °C), respectively. Both contained iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0 3-OH as the major fatty acids, phosphatidylethanolamine and an unknown lipid as the major polar lipids, and menaquinone MK-6 as the major respiratory quinone. The DNA G+C contents were 32.8 and 35.0 mol% for X15M-6T and X15M-8T, respectively. Phylogenetic trees based on 16S rRNA gene sequences showed that both strains belonged to the genus Psychroflexus and formed a separate lineage. In addition, strains X15M-6T and X15M-8T shared 96.8 % 16S rRNA gene sequence similarity and showed highest similarities to members of the genus Psychroflexus (92.7-93.5 and 91.8-93.1 %, respectively). Based on the above data, it is concluded that strains X15M-6T and X15M-8T represent two novel species of the genus Psychroflexus, for which the names Psychroflexus salis sp. nov. (type strain X15M-6T = CGMCC 1.12925T = JCM 30615T) and Psychroflexus planctonicus sp. nov. (type strain X15M-8T = CGMCC 1.12931T = JCM 30616T) are proposed.

  8. Pseudomonas zhaodongensis sp. nov., isolated from saline and alkaline soils.

    PubMed

    Zhang, Lei; Pan, Yuanyuan; Wang, Kaibiao; Zhang, Xiaoxia; Zhang, Cheng; Zhang, Shuang; Fu, Xiaowei; Jiang, Juquan

    2015-03-01

    Strain NEAU-ST5-21(T) was isolated from saline and alkaline soils in Zhaodong City, Heilongjiang Province, China. It was aerobic, Gram-stain-negative, rod-shaped and motile with a polar flagellum. It produced yellow-orange colonies with a smooth surface, and grew in the presence of 0-5 % (w/v) NaCl (optimum 0 %, w/v), at temperatures of 20-40 °C (optimum 28 °C) and at pH 7-11 (optimum pH 7). Phylogenetic analyses based on the separate 16S rRNA gene sequences and concatenated 16S rRNA, gyrB and rpoD gene sequences indicated that strain NEAU-ST5-21(T) belongs to the genus Pseudomonas in the class Gammaproteobacteria. The most closely related species is Pseudomonas xanthomarina, whose type strain (KMM 1447(T)) showed gene sequence similarities of 99.0 % for 16S rRNA, 81.8 % for gyrB and 85.0 % for rpoD with strain NEAU-ST5-21(T). DNA-DNA hybridization values between strain NEAU-ST5-21(T) and P. xanthomarina DSM 18231(T), Pseudomonas kunmingensis CGMCC 1.12273(T), Pseudomonas stutzeri DSM 5190(T), Pseudomonas oleovorans subsp. lubricantis DSM 21016(T), Pseudomomas chengduensis CGMCC 2318(T), Pseudomonas alcaliphila DSM 17744(T) and Pseudomonas toyotomiensis DSM 26169(T) were 52±0 % to 25±2 %. The DNA G+C content of strain NEAU-ST5-21(T) was 65 mol%. The major fatty acids (>10 %) were C18 : 1ω7c and/or C18 : 1ω6c, C16 : 1ω7c and/or C16 : 1ω6c and C16 : 0, the predominant respiratory quinone was ubiquinone 9, and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, one unknown phospholipid, phosphatidylglycerol, one unknown aminolipid, one unknown lipid and a glycolipid. The proposed name is Pseudomonas zhaodongensis sp. nov., NEAU-ST5-21(T) ( = ACCC 06362(T) = DSM 27559(T)) being the type strain.

  9. Flavobacterium aquaticum sp. nov., isolated from a water sample of a rice field.

    PubMed

    Subhash, Y; Sasikala, Ch; Ramana, Ch V

    2013-09-01

    Strain JC164(T) was isolated from a water sample from a rice field at Jamdih, Mau, Uttar Pradesh, India. Colonies of strain JC164(T) were brown-yellow and cells were Gram-stain-negative. Catalase, oxidase and amylase were present. iso-C(15:0), iso-C(16:0), iso-C15 1 G, iso-C(15:0) 3-OH and iso-C(14:0) were the predominant fatty acids with minor amounts of iso-C(16:0) 3-OH, anteiso-C(15:0), C(16:0), iso-C(16:1) H, iso-C(14:0) 3-OH and iso-C(13:0). Strain JC164(T) contained phosphatidylethanolamine and a few unidentified lipids (L1, L3 and L6) as major polar lipids. Bacteriohopane derivative 1 (BHD1) and diplopterol (DPL) were the major hopanoids. β-Carotene was one among the several spirilloxanthin series carotenoids present in strain JC164(T). Genomic DNA G+C content was 39.6 mol%. 16S rRNA gene sequence comparisons indicated that strain JC164(T) represents a member of the genus Flavobacterium (family Flavobacteriaceae, class Flavobacteriia). The most closely related taxa to strain JC164(T) were Flavobacterium sasangense YC6274(T) (98.5%), Flavobacterium cucumis R2A45-3(T) (98.1%), Flavobacterium cheniae NJ-26(T) (97.2%) and the novel strain possessed <95.1% sequence similarity with other members of the genus Flavobacterium. However, strain JC164(T) showed 12.5 ± 2, 13.6 ± 1 and 17.4 ± 2% genomic DNA association (based on DNA-DNA hybridization) with Flavobacterium sasangense KCTC 22246(T), Flavobacterium cucumis DSM 18830(T) and Flavobacterium cheniae CGMCC 1.6844(T), respectively. The distinct genomic difference and morphological, physiological and chemotaxonomic differences from the previously described taxa support the classification of strain JC164(T) as a representative of a novel species of the genus Flavobacterium, for which the name Flavobacterium aquaticum sp. nov. is proposed. The type strain is JC164(T) ( = KCTC 32196(T) = CGMCC 1.12398=LMG 27251(T)).

  10. Kinetics of carbendazim degradation in a horizontal tubular biofilm reactor.

    PubMed

    Alvarado-Gutiérrez, María Luisa; Ruiz-Ordaz, Nora; Galíndez-Mayer, Juvencio; Santoyo-Tepole, Fortunata; Curiel-Quesada, Everardo; García-Mena, Jaime; Ahuatzi-Chacón, Deifilia

    2016-12-22

    The fungicide carbendazim is an ecotoxic agent affecting aquatic biota. Due to its suspected hormone-disrupting effects, it is considered a "priority hazard substance" by the Water Framework Directive of the European Commission, and its degradation is of major concern. In this work, a horizontal tubular biofilm reactor (HTBR) operating in plug-flow regime was used to study the kinetics of carbendazim removal by an acclimated microbial consortium. The reactor was operated in steady state continuous culture at eight different carbendazim loading rates. The concentrations of the fungicide were determined at several distances of the HTBR. At the loading rates tested, the highest instantaneous removal rates were observed in the first section of the tubular biofilm reactor. No evidence of inhibition of the catabolic activity of the microbial community was found. Strains of the genera Flectobacillus, Klebsiella, Stenotrophomonas, and Flavobacterium were identified in the biofilm; the last three degrade carbendazim in axenic culture.

  11. Treatment of seafood processing wastewater using upflow microbial fuel cell for power generation and identification of bacterial community in anodic biofilm.

    PubMed

    Jayashree, C; Tamilarasan, K; Rajkumar, M; Arulazhagan, P; Yogalakshmi, K N; Srikanth, M; Banu, J Rajesh

    2016-09-15

    Tubular upflow microbial fuel cell (MFC) utilizing sea food processing wastewater was evaluated for wastewater treatment efficiency and power generation. At an organic loading rate (OLR) of 0.6 g d(-1), the MFC accomplished total and soluble chemical oxygen demand (COD) removal of 83 and 95%, respectively. A maximum power density of 105 mW m(-2) (2.21 W m(-3)) was achieved at an OLR of 2.57 g d(-1). The predominant bacterial communities of anode biofilm were identified as RB1A (LC035455), RB1B (LC035456), RB1C (LC035457) and RB1E (LC035458). All the four strains belonged to genera Stenotrophomonas. The results of the study reaffirms that the seafood processing wastewater can be treated in an upflow MFC for simultaneous power generation and wastewater treatment.

  12. Complex bacterial infections pre- and posttransplant.

    PubMed

    Lease, Erika D; Zaas, David W

    2010-04-01

    Infections complications following lung transplantation are associated with significant morbidity and mortality. Management of infections is most challenging in patients with cystic fibrosis (CF), but all lung transplant recipients are at heightened risk for opportunistic infections. Particularly in CF, pretransplant infections with PSEUDOMONAS AERUGINOSA, other highly resistant bacteria (e.g., STENOTROPHOMONAS, BURKHOLDERIA), and mycobacteria play a major role in recipient selection and post-lung transplant outcomes. Understanding the clinical impact and management strategies for each of these different pathogens is critical to maximizing the benefit of lung transplantation. In the review, we discuss each of these infections both as pretransplant risk factors as well as posttransplant pathogens and the individual issues that arise with each infection.

  13. Highly sensitive determination of ectoine and other compatible solutes by anion-exchange chromatography and pulsed amperometric detection.

    PubMed

    Riis, Volker; Maskow, Thomas; Babel, Wolfgang

    2003-09-01

    In saline media prokaryotes compensate for the osmotic pressure of the surrounding medium by producing osmolytes. Although these osmolytes or osmoprotectors have quite diverse structures, most of them can be determined by anion-exchange chromatography combined with integrated pulsed amperometric detection. This technique offers the advantages of very high sensitivity and new opportunities to determine ectoine and 5-hydroxyectoine-two important osmolytes -after hydrolytic cleavage of the pyrimidine ring. It can even be used to screen bacterial colonies on agar for compatible solutes. Furthermore, it allows amino acids and osmolytes of this type to be determined without derivatization. To test the method we applied it to two halotolerant bacterial strains: Stenotrophomonas rhizophila DSM 14405(T) and Halomonas elongata DSM 2581(T). The first strain produced trehalose and glucosylglycerol, and the second ectoine, as the main osmotic counterweight. The relationship between the content of these osmolytes in the bacterial biomass and the external salinity is described.

  14. Synthesis and structural characterization of Pd(II) complexes derived from perimidine ligand and their in vitro antimicrobial studies

    NASA Astrophysics Data System (ADS)

    Azam, Mohammad; Warad, Ismail; Al-Resayes, Saud I.; Alzaqri, Nabil; Khan, Mohammad Rizwan; Pallepogu, Raghavaiah; Dwivedi, Sourabh; Musarrat, Javed; Shakir, Mohammad

    2013-09-01

    A novel series of Pd(II) complexes derived from 2-thiophenecarboxaldehyde and 1,8-diaminonaphthalene has been synthesized and characterized by various physico-chemical and spectroscopic techniques viz., elemental analyses, IR, UV-vis, 1H and 13C NMR spectroscopy, and ESI-mass spectrometry. The structure of ligand, 2-(2-thienyl)-2,3-dihydro-1H-perimidine has been ascertained on the basis of single crystal X-ray diffraction. All Pd(II) complexes together with the corresponding ligand have been evaluated for their ability to suppress the in vitro growth of microbes, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Citrobacter sp., Bacillus subtilis and Stenotrophomonas acidaminiphila and results show that Pd(II) complexes have more significant antimicrobial activity than their corresponding ligand. Fluorescence spectroscopic measurements clearly support that both of the Pd(II) complexes show significant DNA binding with calf thymus DNA.

  15. Biodegradation of antibiotic ciprofloxacin: pathways, influential factors, and bacterial community structure.

    PubMed

    Liao, Xiaobin; Li, Bingxin; Zou, Rusen; Dai, Yu; Xie, Shuguang; Yuan, Baoling

    2016-04-01

    Antibiotic ciprofloxacin is ubiquitous in the environment. However, little is known about ciprofloxacin dissipation by microbial community. The present study investigated the biodegradation potential of ciprofloxacin by mixed culture and the influential factors and depicted the structure of ciprofloxacin-degrading microbial community. Both the original microbiota from drinking water biofilter and the microbiota previously acclimated to high levels of ciprofloxacin could utilize ciprofloxacin as sole carbon and nitrogen sources, while the acclimated microbiota had a much stronger removal capacity. Temperature rise and the presence of carbon or nitrogen sources favored ciprofloxacin biodegradation. Many novel biotransformation products were identified, and four different metabolic pathways for ciprofloxacin were proposed. Bacterial community structure illustrated a profound shift with ciprofloxacin biodegradation. The ciprofloxacin-degrading bacterial community was mainly composed of classes Gammaproteobacteria, Bacteroidia, and Betaproteobacteria. Microorganisms from genera Pseudoxanthomonas, Stenotrophomonas, Phenylobacterium, and Leucobacter might have links with the dissipation of ciprofloxacin. This work can provide some new insights towards ciprofloxacin biodegradation.

  16. Biodegradation and decolourization of anaerobically treated distillery spent wash by a novel bacterial consortium.

    PubMed

    Mohana, Sarayu; Desai, Chirayu; Madamwar, Datta

    2007-01-01

    The aim of this study was to isolate microorganisms capable of decolourizing and degrading anaerobically treated distillery spent wash. A bacterial consortium DMC comprising of three bacterial cultures was selected on the basis of rapid effluent decolourization and degradation, which exhibited 67 +/- 2% decolourization within 24 h and 51 +/- 2% chemical oxygen demand reduction within 72 h when incubated at 37 degrees C under static condition in effluent supplemented with 0.5% glucose, 0.1% KH(2)PO(4), 0.05% KCl and 0.05% MgSO(4) x 7H(2)O. Addition of organic or inorganic nitrogen sources did not support decolourization. The cultures were identified as Pseudomonas aeruginosa PAO1, Stenotrophomonas maltophila and Proteus mirabilis by the 16S rDNA analysis.

  17. Responses of earthworms and microbial communities in their guts to Triclosan.

    PubMed

    Ma, Lili; Xie, Yuwei; Han, Zhihua; Giesy, John P; Zhang, Xiaowei

    2017-02-01

    Responses of the earthworm (Eisenia fetida) and compositions of associated microbial communities were determined after exposure to various concentrations of Triclosan (TCS) for 7 d. Concentrations of TCS were greater in intestines than in epidermis of earthworms, which suggested that earthworms accumulate TCS mainly by ingestion rather than by epidermic penetration. Exposure to TCS caused a concentration-dependent increase in activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) and in malondialdehyde (MDA) in E. fetida. Analyses of both the bacterial and eukaryotic community by next generation sequencing (NGS), demonstrated that TCS caused a concentration-dependent decrease in sensitive genera. While relative abundances of Pseudomonas, Stenotrophomonas, and Achromobacter were increased. Nine susceptible microbial groups were more sensitive to exposure to TCS, than were activities of enzymes in earthworms. Thus, rapid genomic measures of gut flora can be used as indicators to assess adverse effects of chemicals on earthworms.

  18. Carbon utilization profiles of bacteria colonizing the headbox water of two paper machines in a Canadian mill.

    PubMed

    Kashama, Johnny; Prince, Véronique; Simao-Beaunoir, Anne-Marie; Beaulieu, Carole

    2009-03-01

    Forty-one bacterial strains isolated from the headbox water of two machines in a Canadian paper mill were associated with the genera Asticcacaulis, Acidovorax, Bacillus, Exiguobacterium, Hydrogenophaga, Pseudomonas, Pseudoxanthomonas, Staphylococcus, Stenotrophomonas based on the sequence of their 16S rRNA genes. The metabolic profile of these strains were determined using Biolog EcoPlate, and the bacteria were divided into four metabolic groups. Metabolic profiles of the bacterial communities colonizing the headbox water of two paper machines was also determined weekly over a 1 year period. The only compound that was not reduced by the bacterial community was 2-hydroxybenzoic acid. Utilization frequency of the other carbon sources in the Biolog EcoPlate ranged from 3 to 100%. The metabolic profiles of the bacterial community did not vary considerably between the two paper machines. However, the metabolic profile varied among the sampling dates.

  19. Isolation and genetic identification of PAH degrading bacteria from a microbial consortium.

    PubMed

    Molina, M Carmen; González, Natalia; Bautista, L Fernando; Sanz, Raquel; Simarro, Raquel; Sánchez, Irene; Sanz, José L

    2009-11-01

    Polycyclic aromatic hydrocarbons (PAH; naphthalene, anthracene and phenanthrene) degrading microbial consortium C2PL05 was obtained from a sandy soil chronically exposed to petroleum products, collected from a petrochemical complex in Puertollano (Ciudad Real, Spain). The consortium C2PL05 was highly efficient degrading completely naphthalene, phenanthrene and anthracene in around 18 days of cultivation. The toxicity (Microtox method) generated by the PAH and by the intermediate metabolites was reduced to levels close to non-toxic in almost 40 days of cultivation. The identified bacteria from the contaminated soil belonged to gamma-proteobacteria and could be include in Enterobacter and Pseudomonas genus. DGGE analysis revealed uncultured Stenotrophomonas ribotypes as a possible PAH degrader in the microbial consortium. The present work shows the potential use of these microorganisms and the total consortium for the bioremediation of PAH polluted areas since the biodegradation of these chemicals takes place along with a significant decrease in toxicity.

  20. Biogeochemical characterization of MC252 oil:sand aggregates on a coastal headland beach.

    PubMed

    Urbano, Marilany; Elango, Vijaikrishnah; Pardue, John H

    2013-12-15

    MC252 oil:sand aggregates, termed surface residue balls (SRBs), were sampled for physical, chemical and microbial characteristics from different tidal zones on a coastal headland beach in Louisiana, USA. Supratidal SRBs were smaller, had low moisture content, and salinities that were <2 ppt. Intertidal SRBs were hypersaline and had higher N and sulfate concentrations, consistent with regular tidal inundation. Crude oil components were highest in the intertidal "oil mat" SRBs with C1- and C2-phenanthrenes, C2- and C3-dibenzothiophenes comprising the majority of the PAH concentrations. In the other SRB categories, PAHs and alkanes were depleted and profiles were skewed toward higher molecular weight compounds. Oxygen microelectrode measurements demonstrated that saturated O2 is present immediately after wetting, but O2 consumption in the interior of the aggregate occurs after a few days. Microbial populations varied with position on the beach but sequences similar to known PAH-degrading taxa (Mycobacterium sp. and Stenotrophomonas sp.) were observed.

  1. Molecular characterization of nitrogen-fixing bacteria isolated from brazilian agricultural plants at São Paulo state

    PubMed Central

    Reinhardt, Érica. L.; Ramos, Patrícia L.; Manfio, Gilson P.; Barbosa, Heloiza R.; Pavan, Crodowaldo; Moreira-Filho, Carlos A.

    2008-01-01

    Fourteen strains of nitrogen-fixing bacteria were isolated from different agricultural plant species, including cassava, maize and sugarcane, using nitrogen-deprived selective isolation conditions. Ability to fix nitrogen was verified by the acetylene reduction assay. All potentially nitrogen-fixing strains tested showed positive hybridization signals with a nifH probe derived from Azospirillum brasilense. The strains were characterized by RAPD, ARDRA and 16S rDNA sequence analysis. RAPD analyses revealed 8 unique genotypes, the remaining 6 strains clustered into 3 RAPD groups, suggesting a clonal origin. ARDRA and 16S rDNA sequence analyses allowed the assignment of 13 strains to known groups of nitrogen-fixing bacteria, including organisms from the genera Azospirillum, Herbaspirillum, Pseudomonas and Enterobacteriaceae. Two strains were classified as Stenotrophomonas ssp. Molecular identification results from 16S rDNA analyses were also corroborated by morphological and biochemical data. PMID:24031239

  2. Wastewater Irrigation Increases the Abundance of Potentially Harmful Gammaproteobacteria in Soils in Mezquital Valley, Mexico

    PubMed Central

    Broszat, Melanie; Nacke, Heiko; Blasi, Ronja; Siebe, Christina; Huebner, Johannes; Daniel, Rolf

    2014-01-01

    Wastewater contains large amounts of pharmaceuticals, pathogens, and antimicrobial resistance determinants. Only a little is known about the dissemination of resistance determinants and changes in soil microbial communities affected by wastewater irrigation. Community DNAs from Mezquital Valley soils under irrigation with untreated wastewater for 0 to 100 years were analyzed by quantitative real-time PCR for the presence of sul genes, encoding resistance to sulfonamides. Amplicon sequencing of bacterial 16S rRNA genes from community DNAs from soils irrigated for 0, 8, 10, 85, and 100 years was performed and revealed a 14% increase of the relative abundance of Proteobacteria in rainy season soils and a 26.7% increase in dry season soils for soils irrigated for 100 years with wastewater. In particular, Gammaproteobacteria, including potential pathogens, such as Pseudomonas, Stenotrophomonas, and Acinetobacter spp., were found in wastewater-irrigated fields. 16S rRNA gene sequencing of 96 isolates from soils irrigated with wastewater for 100 years (48 from dry and 48 from rainy season soils) revealed that 46% were affiliated with the Gammaproteobacteria (mainly potentially pathogenic Stenotrophomonas strains) and 50% with the Bacilli, whereas all 96 isolates from rain-fed soils (48 from dry and 48 from rainy season soils) were affiliated with the Bacilli. Up to six types of antibiotic resistance were found in isolates from wastewater-irrigated soils; sulfamethoxazole resistance was the most abundant (33.3% of the isolates), followed by oxacillin resistance (21.9% of the isolates). In summary, we detected an increase of potentially harmful bacteria and a larger incidence of resistance determinants in wastewater-irrigated soils, which might result in health risks for farm workers and consumers of wastewater-irrigated crops. PMID:24951788

  3. Wastewater irrigation increases the abundance of potentially harmful gammaproteobacteria in soils in Mezquital Valley, Mexico.

    PubMed

    Broszat, Melanie; Nacke, Heiko; Blasi, Ronja; Siebe, Christina; Huebner, Johannes; Daniel, Rolf; Grohmann, Elisabeth

    2014-09-01

    Wastewater contains large amounts of pharmaceuticals, pathogens, and antimicrobial resistance determinants. Only a little is known about the dissemination of resistance determinants and changes in soil microbial communities affected by wastewater irrigation. Community DNAs from Mezquital Valley soils under irrigation with untreated wastewater for 0 to 100 years were analyzed by quantitative real-time PCR for the presence of sul genes, encoding resistance to sulfonamides. Amplicon sequencing of bacterial 16S rRNA genes from community DNAs from soils irrigated for 0, 8, 10, 85, and 100 years was performed and revealed a 14% increase of the relative abundance of Proteobacteria in rainy season soils and a 26.7% increase in dry season soils for soils irrigated for 100 years with wastewater. In particular, Gammaproteobacteria, including potential pathogens, such as Pseudomonas, Stenotrophomonas, and Acinetobacter spp., were found in wastewater-irrigated fields. 16S rRNA gene sequencing of 96 isolates from soils irrigated with wastewater for 100 years (48 from dry and 48 from rainy season soils) revealed that 46% were affiliated with the Gammaproteobacteria (mainly potentially pathogenic Stenotrophomonas strains) and 50% with the Bacilli, whereas all 96 isolates from rain-fed soils (48 from dry and 48 from rainy season soils) were affiliated with the Bacilli. Up to six types of antibiotic resistance were found in isolates from wastewater-irrigated soils; sulfamethoxazole resistance was the most abundant (33.3% of the isolates), followed by oxacillin resistance (21.9% of the isolates). In summary, we detected an increase of potentially harmful bacteria and a larger incidence of resistance determinants in wastewater-irrigated soils, which might result in health risks for farm workers and consumers of wastewater-irrigated crops.

  4. The Effects of Mary Rose Conservation Treatment on Iron Oxidation Processes and Microbial Communities Contributing to Acid Production in Marine Archaeological Timbers

    PubMed Central

    Preston, Joanne; Smith, Andrew D.; Schofield, Eleanor J.; Chadwick, Alan V.; Jones, Mark A.; Watts, Joy E. M.

    2014-01-01

    The Tudor warship the Mary Rose has reached an important transition point in her conservation. The 19 year long process of spraying with polyethylene glycol (PEG) has been completed (April 29th 2013) and the hull is air drying under tightly controlled conditions. Acidophilic bacteria capable of oxidising iron and sulfur have been previously identified and enriched from unpreserved timbers of the Mary Rose, demonstrating that biological pathways of iron and sulfur oxidization existed potentially in this wood, before preservation with PEG. This study was designed to establish if the recycled PEG spray system was a reservoir of microorganisms capable of iron and sulfur oxidization during preservation of the Mary Rose. Microbial enrichments derived from PEG impregnated biofilm collected from underneath the Mary Rose hull, were examined to better understand the processes of cycling of iron. X-ray absorption spectroscopy was utilised to demonstrate the biological contribution to production of sulfuric acid in the wood. Using molecular microbiological techniques to examine these enrichment cultures, PEG was found to mediate a shift in the microbial community from a co-culture of Stenotrophomonas and Brevunidimonas sp, to a co-culture of Stenotrophomonas and the iron oxidising Alicyclobacillus sp. Evidence is presented that PEG is not an inert substance in relation to the redox cycling of iron. This is the first demonstration that solutions of PEG used in the conservation of the Mary Rose are promoting the oxidation of ferrous iron in acidic solutions, in which spontaneous abiotic oxidation does not occur in water. Critically, these results suggest PEG mediated redox cycling of iron between valence states in solutions of 75% PEG 200 and 50% PEG 2000 (v/v) at pH 3.0, with serious implications for the future use of PEG as a conservation material of iron rich wooden archaeological artefacts. PMID:24586230

  5. Isolation and characterization of plant growth-promoting rhizobacteria from wheat rhizosphere and their effect on plant growth promotion

    PubMed Central

    Majeed, Afshan; Hameed, Sohail; Imran, Asma; Rahim, Nasir

    2015-01-01

    The present study was conducted to characterize the native plant growth promoting (PGP) bacteria from wheat rhizosphere and root-endosphere in the Himalayan region of Rawalakot, Azad Jammu and Kashmir (AJK), Pakistan. Nine bacterial isolates were purified, screened in vitro for PGP characteristics and evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum L.). Among nine bacterial isolates, seven were able to produce indole-3- acetic acid in tryptophan-supplemented medium; seven were nitrogen fixer, and four were able to solubilize inorganic phosphate in vitro. Four different morphotypes were genotypically identified based on IGS-RFLP fingerprinting and representative of each morphotype was identified by 16S rRNA gene sequencing analysis except Gram-positive putative Bacillus sp. Based on 16S rRNA gene sequence analysis, bacterial isolates AJK-3 and AJK-9 showing multiple PGP-traits were identified as Stenotrophomonas spp. while AJK-7 showed equal homologies to Acetobacter pasteurianus and Stenotrophomonas specie. Plant inoculation studies indicated that these Plant growth-promoting rhizobacteria (PGPR) strains provided a significant increase in shoot and root length, and shoot and root biomass. A significant increase in shoot N contents (up to 76%) and root N contents (up to 32%) was observed over the un-inoculated control. The study indicates the potential of these PGPR for inoculums production or biofertilizers for enhancing growth and nutrient content of wheat and other crops under field conditions. The study is the first report of wheat associated bacterial diversity in the Himalayan region of Rawalakot, AJK. PMID:25852661

  6. Isolation and characterization of a hydrocarbonoclastic bacterial enrichment from total petroleum hydrocarbon contaminated sediments: potential candidates for bioaugmentation in bio-based processes.

    PubMed

    Di Gregorio, Simona; Siracusa, Giovanna; Becarelli, Simone; Mariotti, Lorenzo; Gentini, Alessandro; Lorenzi, Roberto

    2016-06-01

    Seven hydrocarbonoclastic new bacterial isolates were isolated from dredged sediments of a river estuary in Italy. The sediments were contaminated by shipyard activities since decades, mainly ascribable to the exploitation of diesel oil as the fuel for recreational and commercial navigation of watercrafts. The bacterial isolates were able to utilize diesel oil as sole carbon source. Their metabolic capacities were evaluated by GC-MS analysis, with reference to the depletion of both the normal and branched alkanes, the nC18 fatty acid methyl ester and the unresolved complex mixture of organic compounds. They were taxonomically identified as different species of Stenotrophomonas and Pseudomonas spp. by the combination of amplified ribosomal DNA restriction analysis (ARDRA) and repetitive sequence-based PCR (REP-PCR) analysis. The metabolic activities of interest were analyzed both in relation to the single bacterial strains and to the combination of the latter as a multibacterial species system. After 6 days of incubation in mineral medium with diesel oil as sole carbon source, the Stenotrophomonas sp. M1 strain depleted 43-46 % of Cn-alkane from C28 up to C30, 70 % of the nC18 fatty acid methyl ester and the 46 % of the unresolved complex mixture of organic compounds. On the other hand, the Pseudomonas sp. NM1 strain depleted the 76 % of the nC18 fatty acid methyl ester, the 50 % of the unresolved complex mixture of organic compounds. The bacterial multispecies system was able to completely deplete Cn-alkane from C28 up to C30 and to deplete the 95 % of the unresolved complex mixture of organic compounds. The isolates, either as single strains and as a bacterial multispecies system, were proposed as candidates for bioaugmentation in bio-based processes for the decontamination of dredged sediments.

  7. Bacterial strains isolated from river water having the ability to split alcohol ethoxylates by central fission.

    PubMed

    Budnik, Irena; Zembrzuska, Joanna; Lukaszewski, Zenon

    2016-07-01

    Alcohol ethoxylates (AE) are a major component of the surfactant stream discharged into surface water. The "central fission" of AE with the formation of poly(ethylene glycols) (PEG) is considered to be the dominant biodegradation pathway. However, information as to which bacterial strains are able to perform this reaction is very limited. The aim of this work was to establish whether such an ability is unique or common, and which bacterial strains are able to split AE used as a sole source of organic carbon. Four bacterial strains were isolated from river water and were identified on the basis of phylogenetic trees as Enterobacter strain Z2, Enterobacter strain Z3, Citrobacter freundii strain Z4, and Stenotrophomonas strain Z5. Sterilized river water and "artificial sewage" were used for augmentation of the isolated bacteria. The test was performed in bottles filled with a mineral salt medium spiked with surfactant C12E10 (10 mg L(-1)) and an inoculating suspension of the investigated bacterial strain. Sequential extraction of the tested samples by ethyl acetate and chloroform was used for separation of PEG from the water matrix. LC-MS was used for PEG determination on the basis of single-ion chromatograms. All four selected and investigated bacterial strains exhibit the ability to split fatty alcohol ethoxylates with the production of PEG, which is evidence that this property is a common one rather than specific to certain bacterial strains. However, this ability increases in the sequence: Stenotrophomonas strain Z5 < Enterobacter strain Z2 < Enterobacter strain Z3 = Citrobacter freundii strain Z4. Graphical Abstract Biodegradation by central fission of alcohol ethoxylates by bacterial strains isolated from river water.

  8. Characterization of Co(III) EDTA-Reducing Bacteria in Metal- and Radionuclide-Contaminated Groundwater

    SciTech Connect

    Gao, Weimin; Gentry, Terry J; Mehlhorn, Tonia L; Carroll, Sue L; Jardine, Philip M; Zhou, Jizhong

    2010-01-01

    The Waste Area Grouping 5 (WAG5) site at Oak Ridge National Laboratory has a potential to be a field site for evaluating the effectiveness of various bioremediation approaches and strategies. The site has been well studied in terms of its geological and geochemical properties over the past decade. However, despite the importance of microorganisms in bioremediation processes, the microbiological populations at the WAG5 site and their potential in bioremediation have not been similarly evaluated. In this study, we initiated research to characterize the microbial populations in WAG5 groundwater. Approximately 100 isolates from WAG5 groundwater were isolated and selected based on colony morphology. Fifty-five unique isolates were identified by BOX-PCR and subjected to further characterization. 16S rRNA sequences indicated that these isolates belong to seventeen bacterial genera including Alcaligenes (1 isolate), Aquamonas (1), Aquaspirillum (1), Bacillus (10), Brevundimonas (5), Caulobacter (7), Dechloromonas (2), Janibacter (1), Janthinobacterium (2), Lactobacillus (1), Paenibacillus (4), Pseudomonas (9), Rhodoferax (1), Sphingomonas (1), Stenotrophomonas (6), Variovorax (2), and Zoogloea (1). Metal respiration assays identified several isolates, which phylogenically belong or are close to Caulobacter, Stenotrophomonas, Bacillus, Paenibacillus and Pseudomonas, capable of reducing Co(III)EDTA- to Co(II)EDTA{sup 2-} using the defined M1 medium under anaerobic conditions. In addition, using WAG5 groundwater directly as the inoculants, we found that organisms associated with WAG5 groundwater can reduce both Fe(III) and Co(III) under anaerobic conditions. Further assays were then performed to determine the optimal conditions for Co(III) reduction. These assays indicated that addition of various electron donors including ethanol, lactate, methanol, pyruvate, and acetate resulted in metal reduction. These experiments will provide useful background information for future

  9. Interactions of Freshwater Cyanobacteria with Bacterial Antagonists

    PubMed Central

    Beier, Sara; Grabherr, Manfred

    2017-01-01

    ABSTRACT Cyanobacterial and algal mass development, or blooms, have severe effects on freshwater and marine systems around the world. Many of these phototrophs produce a variety of potent toxins, contribute to oxygen depletion, and affect water quality in several ways. Coexisting antagonists, such as cyanolytic bacteria, hold the potential to suppress, or even terminate, such blooms, yet the nature of this interaction is not well studied. We isolated 31 cyanolytic bacteria affiliated with the genera Pseudomonas, Stenotrophomonas, Acinetobacter, and Delftia from three eutrophic freshwater lakes in Sweden and selected four phylogenetically diverse bacterial strains with strong-to-moderate lytic activity. To characterize their functional responses to the presence of cyanobacteria, we performed RNA sequencing (RNA-Seq) experiments on coculture incubations, with an initial predator-prey ratio of 1:1. Genes involved in central cellular pathways, stress-related heat or cold shock proteins, and antitoxin genes were highly expressed in both heterotrophs and cyanobacteria. Heterotrophs in coculture expressed genes involved in cell motility, signal transduction, and putative lytic activity. l,d-Transpeptidase was the only significantly upregulated lytic gene in Stenotrophomonas rhizophila EK20. Heterotrophs also shifted their central metabolism from the tricarboxylic acid cycle to the glyoxylate shunt. Concurrently, cyanobacteria clearly show contrasting antagonistic interactions with the four tested heterotrophic strains, which is also reflected in the physical attachment to their cells. In conclusion, antagonistic interactions with cyanobacteria were initiated within 24 h, and expression profiles suggest varied responses for the different cyanobacteria and studied cyanolytes. IMPORTANCE Here, we present how gene expression profiles can be used to reveal interactions between bloom-forming freshwater cyanobacteria and antagonistic heterotrophic bacteria. Species

  10. Soluble Expression of (+)-γ-Lactamase in Bacillus subtilis for the Enantioselective Preparation of Abacavir Precursor.

    PubMed

    Xue, Tian-Yun; Xu, Guo-Chao; Han, Rui-Zhi; Ni, Ye

    2015-07-01

    Chiral Vince lactam (γ-lactam) is an important precursor of many carbocyclic nucleoside analogues and pharmaceuticals. Here, a (+)-γ-lactamase encoding gene delm from Delftia sp. CGMCC 5755 was identified through genome hunting. To achieve its soluble and functional expression, Escherichia coli and Bacillus subtilis expression systems were introduced. Compared with E. coli system, recombinant (+)-γ-lactamase showed improved protein solubility and catalytic activity in B. subtilis 168. Reaction conditions for enantioselective resolution of γ-lactam were optimized to be at 30 °C, pH 9.0, and 300 rpm when employing the recombinant B. subtilis 168/pMA5-delm whole cells. Kinetic analysis showed that the apparent V max and K m were 0.595 mmol/(min · gDCW) and 378 mmol/L, respectively. No obvious substrate inhibition was observed. In a 500-mL reaction system, enantioselective resolution of 100 g/L γ-lactam was achieved with 10 g/L dry cells, resulting in 55.2 % conversion and 99 % ee of (-)-γ-lactam. All above suggested that recombinant B. subtilis 168/pMA5-delm could potentially be applied in the preparation of optically pure (-)-γ-lactam.

  11. Enhanced deacidification activity in Schizosaccharomyces pombe by genome shuffling.

    PubMed

    Ding, Su; Zhang, Ying; Zhang, Jing; Zeng, Wei; Yang, Ying; Guan, Jingxi; Pan, Lixia; Li, Wei

    2015-02-01

    A problem frequently occurring in making some kinds of wines, particularly Vitis quinquangularis Rehd wine, is the presence of malic acid at high concentrations, which is detrimental to the quality of wines. Thus, there is a need of the ways for effectively reducing the malic acid levels in wine. This study aimed to generate shuffled fusants of Schizosaccharomyces pombe with enhanced deacidification activity for reducing the excessive malic acid content in wine. Sz. pombe CGMCC 2.1628 was used as the original strain. The starting mutant population was generated by UV treatment. The mutants with higher deacidification activity were selected and subjected to recursive protoplast fusion. The resulting fusants were screened by using the indicator of malic acid concentration of fermentation supernatants on 96-well microtitre plates, measured with bromocresol green. After three rounds of genome shuffling, the best-performing fusant, named GS3-1, was obtained. Its deacidification activity (consumed 4.78 g/l malic acid within 10 days) was increased by 225.2% as compared to that of original strain. In the Vitis quinquangularis Rehd wine fermentation test, GS3-1 consumed 4.0 g/l malic acid during the whole cycle of fermentation, providing up to 185.7% improvement in malic acid consumption compared with that of the original strain. This study shows that GS3-1 has great potential for improving the quality of Vitis quinquangularis Rehd wine.

  12. [Ethanol production from sweet sorghum stalks by advanced solid state fermentation (ASSF) technology].

    PubMed

    Han, Bing; Wang, Li; Li, Shizhong; Wang, Erqiang; Zhang, Lei; Li, Tiancheng

    2010-07-01

    A robust strain of the species Saccharomyces cerevisiae CGMCC1949 was screened and identified, and advanced solid state fermentation (ASSF) technology for fuel ethanol production from sweet sorghum stalks was thus developed. The fermentation time was shortened to less than 30 h, and ethanol yield was 92% of its theoretical maximum. And in the meantime, the cost-effective storage was established for sweet sorghum stalks, with less than 5% sugar loss after 200 days of storage, making the plant operation could extend up to 200 days without feedstock shortage. With the fermentation kinetics and heat-mass transfer models, modeling of the ASSF process was investigated, and the rotating drum bioreactor was designed. Furthermore, the ASSF technology was successfully applied in the pilot plant in which the rotating drum bioreactor was scaled up to 127 m3, and ethanol yield of 91% was achieved. At the end, techno-economic analysis (TEA) conducted by ASPEN indicated that ethanol production from sweet sorghum stalks by the ASSF is economically competitive.

  13. Massilia eurypsychrophila sp. nov. a facultatively psychrophilic bacteria isolated from ice core.

    PubMed

    Shen, Liang; Liu, Yongqin; Gu, Zhengquan; Xu, Baiqing; Wang, Ninglian; Jiao, Nianzhi; Liu, Hongcan; Zhou, Yuguang

    2015-07-01

    Strain B528-3(T), a Gram-stain-negative, rod-shaped, aerobic, facultatively psychrophilic bacterium with polar flagella, was isolated from an ice core drilled from Muztagh Glacier, Xinjiang, China. The novel isolate was classified into the genus Massilia. The 16S rRNA gene sequence of the novel isolate shares a pairwise similarity of less than 97% with those of all the type strains of the genus Massilia. The major fatty acids of strain B528-3(T) were summed feature 3 (C16:1ω7c and/or iso-C15:0 2-OH) (57.31%), C16:0 (11.46%) and C18:1ω7c (14.72%). The predominant isoprenoid quinone was Q-8. The DNA G + C content was 62.2 mol% (Tm). The major polar lipids of this bacterium were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. From the genotypic and phenotypic data, it is evident that strain B528-3(T) represents a novel species of the genus Massilia, for which the name Massilia eurypsychrophila sp. nov. is proposed. The type strain is B528-3(T) ( = JCM 30074(T) = CGMCC 1.12828(T)).

  14. Expression of POX2 gene and disruption of POX3 genes in the industrial Yarrowia lipolytica on the γ-decalactone production.

    PubMed

    Guo, Yanqiong; Song, Huanlu; Wang, Zhaoyue; Ding, Yongzhi

    2012-04-20

    The yeast Yarrowia lipolytica growing on methyl ricinoleate can produce γ-decalactone, the worthy aroma compound, which can exhibit fruity and creamy sensorial notes, and recognized internationally as a safe food additive. Unfortunately, the yield is poor because of lactone degradation by enzyme Aox3 (POX3 gene encoded), which was responsible for continuation of oxidation after C(10) level and lactone reconsumption. In this paper, we chose the industrial Y. lipolytica (CGMCC accession number 2.1405), which is the diploid strain as the starting strain and constructed the recombinant strain Tp-12 by targeting the POX3 locus of the wild type, one copy of POX3 was deleted by CRF1+POX2 insertion. The other recombinant strain Tpp-11, which was a null mutant possessing multiple copies of POX2 and disrupted POX3 genes on two chromosomes, was constructed by inserting XPR2+hpt into the other copy of POX3 of Tp-12. The growth ability of the recombinants was changed after genetic modification in the fermentation medium. The production of γ-decalactone was increased, resulting from blocking β-oxidation at the C(10) Aox level and POX2 overexpression. The recombinant strain Tpp-11 was stable. Because there was no reconsumption of γ-decalactone, the mutant strain could be grown in continuous fermentation of methyl ricinoleate to produce γ-decalactone.

  15. Marinomonas arctica sp. nov., a psychrotolerant bacterium isolated from the Arctic.

    PubMed

    Zhang, De-Chao; Li, Hui-Rong; Xin, Yu-Hua; Liu, Hong-Can; Chen, Bo; Chi, Zhen-Ming; Zhou, Pei-Jin; Yu, Yong

    2008-07-01

    A novel psychrotolerant, Gram-negative, motile bacterium, designated strain 328(T), was isolated from sea-ice samples collected off the Canadian Basin of the Arctic Ocean (7 degrees 23' 14'' N 14 degrees 06' 55'' W). Strain 328(T) was able to grow at 0-37 degrees C, with optimum growth at 25-27 degrees C. It possessed phosphatidylethanolamine and phosphatidylglycerol as major phospholipids and C(10 : 0) 3-OH (31.78 %), C(18 : 1)omega7c (27.50 %) and iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c (19.22 %) as predominant cellular fatty acids. The DNA G+C content of strain 328(T) was 45.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 328(T) was a member of the genus Marinomonas (92.7-96.0 % 16S rRNA gene sequence similarity). On the basis of phenotypic, chemotaxonomic and phylogenetic distinctiveness, strain 328(T) was considered to represent a novel species of the genus Marinomonas. The name Marinomonas arctica sp. nov. is proposed, with strain 328(T) (=CGMCC 1.6498(T)=JCM 14976(T)) as the type strain.

  16. Marinobacter psychrophilus sp. nov., a psychrophilic bacterium isolated from the Arctic.

    PubMed

    Zhang, De-Chao; Li, Hui-Rong; Xin, Yu-Hua; Chi, Zhen-Ming; Zhou, Pei-Jin; Yu, Yong

    2008-06-01

    Strain 20041(T) was isolated from sea-ice of the Canadian Basin (7 degrees 23' 14'' N 14 degrees 06' 55'' W). Phylogenetic analysis based on 16S rRNA gene homology showed that strain 20041(T) was related to members of the genus Marinobacter and had highest 16S rRNA gene sequence similarity with the type strain of Marinobacter maritimus. Cells were Gram-negative, rod-shaped, psychrophilic and motile. The temperature range for growth was 0-22 degrees C, with optimum growth occurring at 16-18 degrees C and at approximately pH 6.0-9.0. Strain 20041(T) had ubiquinone-9 as the major respiratory quinone and iso-C(15 : 0) 2OH and/or C(16 : 1)omega7c, C(16 : 0), C(18 : 1)omega9c and C(12 : 0) 3OH as major fatty acids. The genomic DNA G+C content was 55.4 mol%. On the basis of the phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain 20041(T) is considered to represent a novel species, for which the name Marinobacter psychrophilus sp. nov. is proposed. The type strain is 20041(T) (=CGMCC 1.6499(T)=JCM 14643(T)).

  17. Complete genome sequence of the heavy metal resistant bacterium Altererythrobacter atlanticus 26DY36(T), isolated from deep-sea sediment of the North Atlantic Mid-ocean ridge.

    PubMed

    Wu, Yue-Hong; Cheng, Hong; Zhou, Peng; Huo, Ying-Yi; Wang, Chun-Sheng; Xu, Xue-Wei

    2015-12-01

    Altererythrobacter atlanticus 26DY36(T) (CGMCC 1.12411(T)=JCM 18865(T)) was isolated from the North Atlantic Mid-Ocean Ridge. The strain is resistant to heavy metals, such as Mn(2+) (200 mM), Co(2+) (2.0mM), Cu(2+) (1mM), Zn(2+) (1mM), Hg(2+) (0.1mM) and Cd(2+) (0.5mM). Here we describe the genome sequence and annotation, as well as the features of the organism. A. atlanticus 26DY36(T) harbors a chromosome (3,386,291 bp) and a circular plasmid (88,815 bp). The genome contains 3322 protein-coding genes (2483 with predicted functions), 47 tRNA genes and 6 rRNA genes. A. atlanticus 26DY36(T) encodes dozens of genes related to heavy metal resistance and has potential applications in the bioremediation of heavy metal-contaminated environments.

  18. Dickeyafangzhongdai sp. nov., a plant-pathogenic bacterium isolated from pear trees (Pyrus pyrifolia).

    PubMed

    Tian, Yanli; Zhao, Yuqiang; Yuan, Xiaoli; Yi, Jianping; Fan, Jiaqin; Xu, Zhigang; Hu, Baishi; De Boer, Solke H; Li, Xiang

    2016-09-01

    Gram-stain-negative, pectinolytic bacteria were repeatedly isolated from pear trees displaying symptoms of bleeding canker in China. Three strains, JS5T, LN1 and QZH3, had identical 16S rRNA gene sequences that shared 99 % similarity to the type strain of Dickeya dadantii. Phylogenetic analysis of strains JS5T, LN1 and QZH3 with isolates representing all species of the genus Dickeya and related Pectobacterium species supported their affiliation to Dickeya. Multi-locus sequence typing employing concatenated sequences encoding recA, fusA, gapA, purA, rplB, dnaX and the intergenic spacer illustrated a phylogeny which placed strains JS5T, LN1 and QZH3 as a distinct clade, separate from all other species of the genus Dickeya. Average nucleotide identity values obtained in comparison with all species of the genus Dickeya supported the distinctiveness of strain JS5T within the genus Dickeya. Additionally, all three strains were phenotypically distinguished from other species of the genus Dickeya by failing to hydrolyse casein, and by producing acids from (-)-d-arabinose, (+)melibiose, (+)raffinose, mannitol and myo-inositol, but not from 5-keto-d-gluconate or β-gentiobiose. The name Dickeya fangzhongdai sp. nov. is proposed to accommodate these strains; the type strain is JS5T (=CGMCC 1.15464T=DSM 101947T).

  19. Lysinibacillus endophyticus sp. nov., an indole-3-acetic acid producing endophytic bacterium isolated from corn root (Zea mays cv. Xinken-5).

    PubMed

    Yu, Jiang; Guan, Xuejiao; Liu, Chongxi; Xiang, Wensheng; Yu, Zhenhua; Liu, Xiaobing; Wang, Guanghua

    2016-10-01

    A Gram-positive, aerobic, motile, rod-shaped bacterium, designated strain C9(T), was isolated from surface sterilised corn roots (Zea mays cv. Xinken-5) and found to be able to produce indole-3-acetic acid. A polyphasic taxonomic study was carried out to determine the status of strain C9(T). The major cellular fatty acids were found to contain iso-C15:0, anteiso-C15:0 and anteiso-C17:0, and the only menaquinone was identified as MK-7. The polar lipid profile was found to contain diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids and an unidentified lipid. The cell wall peptidoglycan was found to be of the A4α L-Lys-D-Asp type and the whole cell sugar was found to be glucose. Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain C9(T) belongs to the genus Lysinibacillus and is closely related to Lysinibacillus chungkukjangi NBRC 108948(T) (98.1 % similarity) and Lysinibacillus sinduriensis DSM 27595(T) (98.0 %). However, the low levels of DNA-DNA relatedness and some differential phenotypic characteristics allowed the strain to be distinguished from its close relatives. Therefore, it is concluded that strain C9(T) represents a novel species of the genus Lysinibacillus, for which the name Lysinibacillus endophyticus sp. nov. is proposed. The type strain is C9(T) (=DSM 100506(T) = CGMCC 1.15291(T)).

  20. Sporosarcina antarctica sp. nov., a psychrophilic bacterium isolated from the Antarctic.

    PubMed

    Yu, Yong; Xin, Yu-Hua; Liu, Hong-Can; Chen, Bo; Sheng, Jun; Chi, Zhen-Ming; Zhou, Pei-Jin; Zhang, De-Chao

    2008-09-01

    A Gram-positive, psychrophilic, rod-shaped bacterium, designated strain N-05(T), was isolated from soil samples collected off King George Island, west Antarctica (6 degrees 13' 31'' S 5 degrees 57' 08'' W). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain N-05(T) was related to members of the genus Sporosarcina and had highest 16S rRNA gene sequence similarity with the type strain of Sporosarcina macmurdoensis (98.0%). The temperature range for growth of strain N-05(T) was 0-23 degrees C, with optimum growth occurring at 17-18 degrees C and approximately pH 6.0-8.0. Strain N-05(T) had MK-7 as the major menaquinone and anteiso-C(15:0) and C(16:1)omega7c alcohol as major fatty acids. The genomic DNA G+C content was 39.2 mol%. On the basis of phenotypic characteristics, phylogenetic analysis and DNA-DNA relatedness data, strain N-05(T) is considered to represent a novel species of the genus Sporosarcina, for which the name Sporosarcina antarctica is proposed. The type strain is N-05(T) (=CGMCC 1.6503(T)=JCM 14646(T)).

  1. Friedmanniella flava sp. nov., a soil actinomycete.

    PubMed

    Zhang, Xuefang; Zhang, Jianli; Zhang, Yabo; Xin, Yuhua; He, Hongju

    2013-05-01

    A novel actinomycete, strain W6(T), was isolated from a soil sample of Yunnan Province, China. The bacterium was aerobic, non-motile, non-spore-forming and Gram-stain-positive. Genetic, phenotypic and chemical properties of the isolate were studied. 16S rRNA gene sequence data suggested that the novel isolate belonged to the genus Friedmanniella and shared 98.6% sequence similarity with Friedmanniella antarctica DSM 11053(T) and Friedmanniella okinawensis DSM 21744(T), the most closely related species. The cell-wall peptidoglycan contained ll-diaminopimelic acid, and mycolic acids were absent. The main menaquinone was MK-9(H4) and the predominant fatty acids were anteiso-C15:0 and iso-C15:0. The phospholipid profile contained phosphatidylglycerol, phosphatidylinositol, phosphatidylcholine and diphosphatidylglycerol. The DNA G+C content of strain W6(T) was 72 mol%. Strain W6(T) showed 30.0% and 28.5% DNA-DNA relatedness, respectively, to F. antarctica DSM 11053(T) and F. okinawensis DSM 21744(T). The combined genotypic and phenotypic data showed that strain W6(T) should be assigned to the genus Friedmanniella as a representative of a novel species, for which the name Friedmanniella flava sp. nov. is proposed. The type strain is W6(T) ( = CGMCC 4.6856(T) =JCM 17701(T)).

  2. Arcticibacter pallidicorallinus sp. nov. isolated from glacier ice.

    PubMed

    Liu, Qing; Kim, Song-gun; Liu, Hong-can; Xin, Yu-hua; Zhou, Yu-guang

    2014-07-01

    A Gram-stain-negative, rod-shaped bacterium (strain Hh36(T)) was isolated from the No. 1 glacier in Xinjiang, north-west China. Colonies of strain Hh36(T) were pink, convex and round on PYG medium plates. Strain Hh36(T) was able to grow at 4-30 °C and pH 6.0-8.0. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Hh36(T) was related to members of the genus Arcticibacter. The major cellular fatty acids of the novel strain were iso-C15 : 0, summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and iso-C17 : 0 3-OH. The G+C content of the genomic DNA was 44.0 mol%. On the basis of phenotypic characteristics and phylogenetic analysis, strain Hh36(T) is considered to represent a novel species of the genus Arcticibacter, for which the name Arcticibacter pallidicorallinus sp. nov. is proposed. The type strain is Hh36(T) ( = CGMCC 1.9313(T)  = KCTC 32542(T)).

  3. Sphingomonas psychrolutea sp. nov., a psychrotolerant bacterium isolated from glacier ice.