Molecular characterization of Burkholderia cepacia isolates from cystic fibrosis (CF) patients in an Italian CF center

Petrucca, Andrea; Cipriani, Paola; Valenti, Piera; Santapaola, Daniela; Cimmino, Carmen; Scoarughi, Gian Luca; Santino, Iolanda; Stefani, Stefania; Sessa, Rosa; Nicoletti, Mauro

doi:10.1016/s0923-2508(03)00145-1

Cited by 17 publications

(16 citation statements)

References 32 publications

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“…The other genomovars accounted for 1% or fewer of the isolates received (B. vietnamiensis and B. ambifaria) or were absent (B. dolosa and B. anthina). These data confirm and extend results from previous studies of B. cepacia complex bacteria infecting Italian CF patients (1,44,49). About 13% of the B. cenocepacia isolates belonged to HaeIII-recA RFLP type U, which was reported as a major problem in the Genoa CF center (44).…”

Section: Discussionsupporting

confidence: 89%

“…Some CF patients may be transiently colonized, and others may be chronically or asymptomatically colonized, suggesting differences in host response or pathogenic potential among B. cepacia complex bacteria. Prevention of pulmonary infection is vital for CF patients; hence, the importance of stringent infection control policies, including patient segregation, has been strongly emphasized (24,36,49,54,63). However, it is also recognized that segregation places considerable social and personal burdens on CF patients and their families.…”

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confidence: 99%

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Transmission of Burkholderia cepacia Complex: Evidence for New Epidemic Clones Infecting Cystic Fibrosis Patients in Italy

et al. 2005

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To analyze national prevalence, genomovar distribution, and epidemiology of the Burkholderia cepacia complex in Italy, 225 putative B. cepacia complex isolates were obtained from 225 cystic fibrosis (CF) patients attending 18 CF centers. The genomovar status of these isolates was determined by a polyphasic approach, which included whole-cell protein electrophoresis and recA restriction fragment length polymorphism (RFLP) analysis. Two approaches were used to genotype B. cepacia complex isolates: BOX-PCR fingerprinting and pulsedfield gel electrophoresis (PFGE) of genomic macrorestriction fragments. A total of 208 (92%) of 225 isolates belonged to the B. cepacia complex, with Burkholderia cenocepacia as the most prevalent species (61.1%). Clones delineated by PFGE were predominantly linked to a single center; in contrast, BOX-PCR clones were composed of isolates collected either from the same center or from different CF centers and comprised multiple PFGE clusters. Three BOX-PCR clones appeared of special interest. One clone was composed of 17 B. cenocepacia isolates belonging to recA RFLP type H. These isolates were collected from six centers and represented three PFGE clusters. The presence of insertion sequence IS1363 in all isolates and the comparison with PHDC reference isolates identified this clone as PHDC, an epidemic clone prominent in North American CF patients. The second clone included 22 isolates from eight centers and belonged to recA RFLP type AT. The genomovar status of strains with the latter RFLP type is not known. Most of these isolates belonged to four different PFGE clusters. Finally, a third clone comprised nine B. pyrrocinia isolates belonging to recA RFLP type Se13. They represented three PFGE clusters and were collected in three CF centers.In the late 1970s and 1980s, reports on the recovery of Burkholderia cepacia from cystic fibrosis (CF) specimens began to appear (29), and the emergence of this pathogen was subsequently reviewed (25,35,36). Polyphasic taxonomic studies identified bacteria tentatively classified as B. cepacia as a complex of at least nine closely related species (genomovars). This B. cepacia complex consists of B. cepacia,

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Section: Discussionsupporting

confidence: 89%

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confidence: 99%

Transmission of Burkholderia cepacia Complex: Evidence for New Epidemic Clones Infecting Cystic Fibrosis Patients in Italy

et al. 2005

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“…Bacteria, media and culture conditions P. aeruginosa ATCC 15692 (PAOl) and B. cenocepacia PVl clinical isolate (8) were maintained in Brain Heart Infusion broth (BHI) (Difco Laboratories, MD, USA) with glycerol (25%) at -80°C and streaked on plates ofTryptone Soy Agar (TSA) (Oxoid LTD, England) and Burkholderia cepacia medium (BCM) (Oxoid LTD, England), respectively, before the experiments. In all experiments bacteria were grown on Chemically Defined Medium (CDM), containing 48 mM glucose, 7.4 mM KCI, 6 mM NaCI, 48 mM(NH4)2S04' 0.5 mM MgS0 4x7HP, 60 mM MOPS, 3.8 mM K 2HP04x3HP and 0.1% Casamino acid (9).…”

Section: Methodsmentioning

confidence: 99%

Bovine Lactoferrin Inhibits the Efficiency of Invasion of Respiratory A549 Cells of Different Iron-Regulated Morphological Forms of Pseudomonas Aeruginosa and Burkholderia Cenocepacia

Berlutti¹,

Superti

Nicoletti

et al. 2008

Int J Immunopathol Pharmacol

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Pseudomonas aeruginosa and Burkholderia cenocepacia are two important opportunistic respiratory pathogens of cystic fibrosis (CF) patients. Infections caused by these microorganisms are particularly difficult to eradicate because they are usually highly resistant to several currently available broad-spectrum antibiotics. Lactoferrin (Lf), a glycoprotein found in physiological fluids of mammals and present at high concentrations in infected and inflamed tissues, plays an important role in the natural defence mechanism against pathogens and in immune regulation. In the present study, we evaluate the ability of bovine lactoferrin (bLf) to influence P. aeruginosa PAO1 and B. cenocepacia PV1 adhesiveness and invasiveness, using the A549 human bronchial cell line. Three different iron-induced morphological forms of bacteria (free-living, aggregates and biofilm) were assayed. The addition of bLf to cells just before infection had little influence on adhesion efficiency for all three of the morphological forms of B. cenocepacia PV1, while a slight increase in adhesion efficiency by P. aeruginosa PAO1 was noticed. Conversely, invasion of all three morphological forms of both P. aeruginosa and B. cenocepacia was strongly inhibited by the presence of bLf, independently of its degree of iron-binding activity. This is the first report demonstrating an anti-invasive property of bLf for strains of P. aeruginosa and B. cenocepacia.

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“…Bacteria, media and culture conditions Pseudomonas aeruginosa ATCC 15692 (PAOl) and Burkholderia cenocepacia PVl clinical isolate (17) were maintained in Brain Heart Infusion broth (BHI) (Difco Laboratories, MD, USA) with glycerol (25%) at -80°C and checked for purity on Tryptone Soy Agar (TSA) (Oxoid LTD, England) and Burkholderia cepacia medium (BCM) (Oxoid LTD, England), respectively. In all experiments Chemically Defined Medium (CDM) containing glucose 48 mM, KCl 7.4 mM, NaCI 6 mM, (NH4)2S04 48 mM, MgS0 4 x 7H 20 0.5 mM, MOPS 60 mM, K 2HP04 X 3H 20 3.8 mM, Casamino acid 0.1% was used (18).…”

Section: Methodsmentioning

confidence: 99%

Iron Availability Influences Aggregation, Biofilm, Adhesion and Invasion of Pseudomonas Aeruginosa and Burkholderia Cenocepacia

Berlutti

Morea²,

Battistoni

et al. 2005

Int J Immunopathol Pharmacol

111

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Pseudomonas aeruginosa and Burkholderia cenocepacia are predominant opportunistic pathogens in cystic fibrosis (CF) patients. In healthy humans the lower respiratory tract as well as all mucosa, contains a very low free iron concentration (10-18 M), while in CF patients' sputum iron concentration is very high, showing a median value of 63x10-6 M. Accumulation of catalytic reactive iron heavily contributes to subsequent clinical complications in the lung disorders by the production of reactive oxygen species and increases bacterial growth and virulence. The data reported in this study indicate that low iron concentration (Fe 3 + 1 JA.M) induced free-living forms and motility both in P. aeruginosa and B. cenocepacia, while high iron concentrations (Fe 3 + 10 and 100 tJ.M) stimulated aggregation and biofilm formation already in the fluid phases, so demonstrating that aggregation and biofllm formation are positively iron-modulated in these bacteria. Moreover, the different morphological forms (free-living, aggregates and biofllm) showed different capabilities of adhering and invading the bronchial cell line A549. P. aeruginosa PA01 aggregates, and mostly biofllm, exerted the highest adhesion efficiency, while B. cenocepacia PV1 aggregates or biofilm the lowest. A significant reduction in invasion efficiency by P. aeruginosa biofilm and a significant increase in cell internalization by B. cenocepacia biofllm has been reported. Therefore, the iron availability is an important signal to which P. aeruginosa and B. cenocepacia counteract by leaving the motile free-living forms and entering into a new lifestyle, i.e, biofllm. These data could contribute to explain that the ironoverload of the sputum of CF patients, inducing nonmotile forms, aggregates and biofllm, may facilitate penetration of host epithelial barriers contributing to the establishment of infection, colonization, persistence and systemic spread of these opportunistic pathogens.Pseudomonas aeruginosa is a ubiquitous Gramnegative motile bacterium found in different environments such as soil, freshwater and marine habitat. Moreover, P. aeruginosa is the predominant opportunistic pathogen in cystic fibrosis (CF) patients, as the lungs of >90% of all CF patients are colonized by this bacterium (1). Chronic colonization of P. aeruginosa in CF patient airways is a major source of morbidity and mortality (2) leading to epithelial surface damage and airway plugging which results in a decrease of pulmonary function (3).Although not as prevalent as P. aeruginosa, Burkholderia cepacia is another important opportunistic respiratory pathogen in CF patients (2),

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Molecular characterization of Burkholderia cepacia isolates from cystic fibrosis (CF) patients in an Italian CF center

Cited by 17 publications

References 32 publications

Transmission of Burkholderia cepacia Complex: Evidence for New Epidemic Clones Infecting Cystic Fibrosis Patients in Italy

Transmission of Burkholderia cepacia Complex: Evidence for New Epidemic Clones Infecting Cystic Fibrosis Patients in Italy

Bovine Lactoferrin Inhibits the Efficiency of Invasion of Respiratory A549 Cells of Different Iron-Regulated Morphological Forms of Pseudomonas Aeruginosa and Burkholderia Cenocepacia

Iron Availability Influences Aggregation, Biofilm, Adhesion and Invasion of Pseudomonas Aeruginosa and Burkholderia Cenocepacia

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