Longitudinal bacterial prevalence in cystic fibrosis airways: Fact and artifact.

BACKGROUND: Opportunistic bacterial infection is a hallmark of cystic fibrosis (CF) lung disease and early mortality. Poorly characterized prevalence changes have accompanied two decades of health improvements, with CFTR modulators likely to further affect infection epidemiology. METHODS: Bacterial prevalence change trends across birth cohorts were assessed with linear regression using 2001-2019 US CF Foundation Patient Registry data. Informative missingness was assessed, as was age-to-age infection status. RESULTS: Bacterial prevalence constantly changed from 2001 to 2019, with changes differing across birth cohorts. Informative censoring affected prevalence change for some organisms. Age-to-age infection status changes were greater than net changes in bacterial prevalence and varied by age. CONCLUSIONS: CF infection epidemiology changed over two decades and will continue to do so. Understanding how modulators affect infection epidemiology will require creative designs for longitudinal prevalence change studies emphasizing prevalence changes independent of effects on lung biology.

Rhizobium radiobacter bacteremia in a neonate.

Rhizobium radiobacter bacteremia is an infrequent cause of human infection. We report a rare manifestation of R. radiobacter infection in which bacteremia occurred in a newborn infant without other risk factors.

Induction of immune response to the 17 kDa OMPA Burkholderia cenocepacia polypeptide and protection against pulmonary infection in mice after nasal vaccination with an OMP nanoemulsion-based vaccine.

Burkholderia cepacia complex (Bcc) are opportunistic bacteria associated with life-threatening illness in persons with cystic fibrosis. Once Bcc colonization is established, these antimicrobial-resistant and biofilm-forming bacteria are difficult to eradicate and are associated with increased rates of morbidity and mortality. At present, no vaccines are available to prevent the Bcc infection. There is currently a paucity of published information regarding the development of vaccines designed to prevent Burkholderia colonization. This work expands on the recent studies published by Bertot et al. [Infect Immun 75(6):2740-2752, 2007], where successful protective immune responses were generated in mice using a B. multivorans OMP-based vaccine. Here, we evaluate an experimental mucosal vaccine against Bcc using a novel mucosal adjuvant (nanoemulsion) and a novel B. cenocepacia-based OMP antigen. The OMP antigen derived from B. cenocepacia was mixed with either nanoemulsion or with PBS and delivered intranasally to CD-1 mice. Serum analysis showed robust IgG and mucosal secretory IgA immune responses in vaccinated versus control mice. The antibodies had cross-neutralizing activity against both B. cenocepacia and B. multivorans species. We found that immunized mice were protected against pulmonary colonization with B. cenocepacia. We have also identified that a 17 kDa OmpA-like protein highly conserved between Burkholderia and Ralstonia species as a new immunodominant epitope in mucosal immunization.

Survival after lung transplantation of cystic fibrosis patients infected with Burkholderia cepacia complex.

Within the Burkholderia cepacia complex (Bcc), B. cenocepacia portends increased mortality compared with other species. We investigated the impact of Bcc infection on mortality and re-infection following lung transplant (LT). Species designation for isolates from Bcc-infected patients was determined using 16S rDNA and recA gene analyses. Of 75 cystic fibrosis patients undergoing LT from September 1992 to August 2002, 59 had no Bcc and 16 had Bcc (including 7 B. cenocepacia) isolated in the year before LT. Of the latter, 87.5% had Bcc recovered after transplantation, and all retained their pretransplant strains. Survival was 97%, 92%, 76% and 63% for noninfected patients; 89%, 89%, 67% and 56% for patients infected with Bcc species other than B. cenocepacia; and 71%, 29%, 29% and 29% for patients with B. cenocepacia (p = 0.014) at 1 month, 1 year, 3 years and 5 years, respectively. Patients infected with B. cenocepacia before transplant were six times more likely to die within 1 year of transplant than those infected with other Bcc species (p = 0.04) and eight times than noninfected patients (p < 0.00005). Following LT, infection with Bcc species other than B. cenocepacia does not significantly impact 5-year survival whereas infection with B. cenocepacia pretransplant is associated with decreased survival.

Identification of Bordetella spp. in respiratory specimens from individuals with cystic fibrosis.

Bordetella spp. are not normally included when considering the opportunistic bacterial species that are typically involved in respiratory tract infections in individuals with cystic fibrosis (CF). By using a combination of bacterial genotyping and 16S rDNA sequencing, Bordetella spp. were identified in cultures obtained from 43 individuals with CF. Most (n = 23) patients were infected with Bordetella bronchiseptica/parapertussis; five were infected with Bordetella hinzii, four with Bordetella petrii, three with Bordetella avium, and eight with unidentified Bordetella spp. Consideration should be given to the presence of these organisms in the evaluation of CF sputum cultures.

Cystic Fibrosis Mice Develop Spontaneous Chronic Bordetella Airway Infections.

Chronic pulmonary disease and infection is the primary cause of morbidity and mortality in people with cystic fibrosis (CF). Though Pseudomonas aeruginosa, is most commonly found in the airways of individuals with CF, there is increasing appreciation for the diversity of the CF microbiome, including other taxa such as Bordetella. Here we describe the identification and impact of Bordetella pseudohinzii infection in CF mice, which previously have not been thought to develop spontaneous airway infections. We determined that CF mice are more susceptible to the B. pseudohinzii infections, and less able to resolve the infection than non-CF mice. Moreover, in both CF and non-CF mice, B. pseudohinzii infections lead to markedly reduced respiratory rates and a CF-specific immune response. These results establish the CF mouse model as an important tool for the study of CF-relevant infection and highlight the potential contribution of Bordetella to CF clinical pathology.

Draft Genome Sequence of the Pandoraea apista LMG 16407 Type Strain.

Pandoraea species, in particular Pandoraea apista, are opportunistic, multidrug-resistant pathogens in persons with cystic fibrosis (CF). To aid in understanding the role of P. apista in CF lung disease, we used Illumina MiSeq and nanopore MinION technology to sequence the whole genome of the P. apista LMG 16407(T).

Microbiological and immunologic considerations with aerosolized drug delivery.

The development of drug resistance is a major theoretical concern with the long-term delivery of aerosolized antibiotics via inhalation. A randomized, placebo-controlled, double-blind study, which compared inhaled tobramycin plus standard cystic fibrosis (CF) care to placebo plus standard CF care, examined the following microbiological parameters: percentage of patients with at least one Pseudomonas aeruginosa (PA) strain with a minimal inhibitory concentration (MIC) > 16 microg/mL (ie, the breakpoint for tobramycin resistance delivered by the parenteral route); changes in the levels of the lowest concentration required to inhibit the growth of 50% of strains tested (MIC(50)) and 90% of strains tested (MIC(90)); the percentage of patients with an increase, decrease, or change in the MIC of the most resistant and most prevalent PA strains; and the percentage of patients in whom the PA strain with the highest MIC also was the most prevalent. During the first 6 months, which included three on-drug and off-drug cycles of 4 weeks' duration each, the percentage of tobramycin-treated patients with at least one PA isolate and with an MIC > 16 microg/mL was 13% at baseline, 26% at 20 weeks, and 23% at 24 weeks vs 10%, 17%, and 8%, respectively, for placebo-treated patients. No significant change was observed in MIC(50) at 20 and 24 weeks. The increase in MIC(90) was not statistically significant. At 24 weeks, there was no increase in the percentage of patients in either group in whom the PA strain with the highest MIC became most the prevalent strain. After the third on-drug cycle, 33% of the tobramycin group showed an increase in the MIC of the strain with the highest MIC. This decreased to 26% after 1 month off drug therapy. A preliminary analysis of the 12-month and 18-month data showed a decrease in the proportion of resistant PA isolates after each off-drug cycle. This return to susceptibility following an off-drug cycle was not observed at 24 months. The mechanism of resistance in this setting is believed to be increased impermeability to drug. At all time points, pulmonary function improved even in patients with MICs of > or = 128 microg/mL. At 6 months, no increase was seen in the rates of superinfection with tobramycin-resistant, Gram-negative pathogens. Increases in Stenotrophomonas maltophilia were detected in patients after 18 and 24 months of tobramycin therapy and were similar to those rates in patients receiving placebo. These rates may be independent of inhalation therapy.

Misidentification of Burkholderia cepacia in US cystic fibrosis treatment centers: an analysis of 1,051 recent sputum isolates.

BACKGROUND: Burkholderia cepacia remains a significant pathogen in persons with cystic fibrosis (CF). The medical and psychosocial consequences of pulmonary colonization with this bacterium are enormous. However, B cepacia may be frequently misidentified from CF sputum culture. STUDY OBJECTIVES: To determine the rate of misidentification of B cepacia recently recovered from CF sputum culture of persons receiving care in US treatment centers. DESIGN: Bacterial isolates cultured from CF sputum and putatively identified as B cepacia or other related nonlactose-fermenting Gram-negative species were referred from participating treatment centers. Isolates underwent polyphasic analyses employing phenotypic (selective media and biochemical testing) and genotypic (polymerase chain reaction) assays to determine species identification. Taxonomic evaluations were performed by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins and amplified-fragment length polymorphism analysis. MEASUREMENTS AND RESULTS: A total of 1,051 isolates recovered from 608 patients were received from 115 treatment centers in 91 US cities. Among the isolates identified as B cepacia by referring laboratories, 11% could not be confirmed as B cepacia by polyphasic analyses. In addition, 36% of isolates not specifically identified by the referring laboratory or identified as a species other than B cepacia were, in fact, found to be members of the B cepacia complex. CONCLUSIONS: Rates of misidentification of B cepacia remain unacceptably high among US treatment centers. These data suggest the need for increased awareness of this problem among CF centers and their affiliated laboratories, better adherence to recommended protocols for evaluation of CF sputum, and greater use of reference laboratories equipped to provide advanced analyses.

Inapparent transmission of Pseudomonas (Burkholderia) cepacia among patients with cystic fibrosis.

Pseudomonas cepacia is a significant pathogen in children and young adults with cystic fibrosis, and prevention of its acquisition has become an important goal in patient management. Although it is now clear that this bacterium can be transmitted from person to person, the frequency of this mode of acquisition and the measures required to prevent it are controversial. In this report we describe the use of a novel genotyping method to extend our previous investigation of person to person transmission of P. cepacia among patients with cystic fibrosis attending an educational program. Three (20%) of 15 individuals acquired P. cepacia after contact with a chronically colonized patient. Analysis revealed that the isolates recovered from the three newly colonized patients were the same as that from the index patient. We also demonstrated that pulmonary colonization with P. cepacia may not be detected by currently recommended culture methods for as long as 2 years after acquisition. These data indicate a need to develop more sensitive means of detecting P. cepacia colonization in order better to understand host-pathogen interaction and to optimize preventive strategies.

Recovery of Pseudomonas cepacia and other Pseudomonas species from the environment.

Nine hundred sixteen cultures were obtained from homes of patients with cystic fibrosis, control homes, salad bars, and food markets, and analyzed for the presence of Pseudomonas cepacia and related bacteria. P cepacia was recovered from 5 (18%) of 27 homes, and from 20 (4%) of 509 cultures collected outside of homes. Relative to other pseudomonads, P cepacia is found infrequently in the environment. It is not clear how frequently these sources contribute to acquisition of this bacteria by persons with cystic fibrosis.

An episodic outbreak of genetically related Burkholderia cepacia among non-cystic fibrosis patients at a university hospital.

OBJECTIVE: To investigate an outbreak of Burkholderia cepacia. DESIGN: Observational study and chart review. PATIENTS: Adult non-cystic fibrosis (CF) patients. SETTING: Intensive care units (ICUs) at a university-affiliated teaching hospital. METHODS: As part of the epidemiological investigation, we conducted a chart review and collected environmental samples. A review of work schedules of healthcare workers also was performed. We used B. cepacia selective agar for preliminary screening for all isolates, which subsequently were confirmed as members of the B. cepacia complex by polyphasic analysis employing conventional biochemical reactions and genus- and species-specific polymerase chain reaction assays. Pulsed-field gel electrophoresis, randomly amplified polymorphic DNA typing, and automated ribotyping were used to genotype the isolates. As part of the intervention, contact isolation precautions were initiated for all patients identified as having had a culture positive for B. cepacia. RESULTS: Between September 1997 and September 1999, B. cepacia was isolated from 31 adult patients without CF in ICUs at a university-affiliated teaching hospital. Based on geographic clustering and genotypic analysis, three distinct clusters were observed involving 20 patients. Isolates from 17 of these patients were available for testing and were found to be of the same strain (outbreak strain). Further taxonomic analysis indicated that the outbreak strain was B. cepacia complex genomovar III. Twelve (71%) of the 17 patients were judged to be infected, and 5 (29%) were colonized with this strain. Six of 200 environmental cultures from multiple sources in the hospital's ICUs yielded B. cepacia. Two of these isolates, both recovered from rooms of colonized patients, were the same genotype as the outbreak strain recovered from patients. CONCLUSION: Despite an extensive investigation, the source of the B. cepacia clone involved in this outbreak remains unknown. The spatial and temporal pattern of cases suggests that cross-transmission of a genetically related strain contributed to clustering among patients. The initiation of contact isolation may have limited the extent of this transmission. Additional studies are needed to elucidate better the epidemiology of nosocomial B. cepacia infection among non-CF adult patients.

Antibacterial agents in pediatrics.

The utility of various antibacterial agents for therapy of infectious diseases in children is determined by the unique pharmacokinetics and potential toxicity in children. The important age-related principle of pharmacokinetics is reviewed in the first section of this article; the second section focuses on specific therapeutic agents and their use in children. Particular emphasis is given to the use of new antibiotics in children, including the new oral cephalosporins and macrolides.

Nontypeable Haemophilus influenzae susceptibility: effect of inoculum size and beta-lactamase production.

The activities of cefixime, cefpodoxime, cefprozil, cefuroxime, loracarbef, and amoxicillin/clavulanate against 72 clinical isolates of nontypeable Haemophilus influenzae were determined by using an agar dilution method. The effects of beta-lactamase production and bacterial inoculum size were investigated. All antimicrobials exhibited a significant inoculum effect, demonstrating the importance of accurately determining inoculum size in the performance of antimicrobial susceptibility testing of H. influenzae.

Survival of Burkholderia cepacia on environmental surfaces.

Burkholderia (Pseudomonas) cepacia is an important pathogen amongst persons with cystic fibrosis (CF), and evidence suggests that transmission of strains within CF clinics contributes to pulmonary colonization of some patients. In order to optimize preventive strategies, the survival of B. cepacia on various environmental surfaces, including cotton cloth, stainless steel, latex and polyvinylchloride (PVC) tubing, was investigated. For surface inoculation, bacteria were suspended in phosphate buffered saline, sputum from CF patients, or sputum from persons without CF. The results demonstrate that amongst the strains examined, organisms survived significantly (P < 0.001) longer when suspended in sputum from CF patients than in either non-CF sputum or buffered saline. Significant (P < 0.001) differences in survival on the various surfaces were found; survival was greatest on PVC. Significant (P < 0.001) strain-to-strain differences in survival were also demonstrated; patient isolates representing predominant CF centre ribotypes survived longest. These data demonstrate that (1) B. cepacia can survive for long periods in respiratory droplets on environmental surfaces typically found in CF clinics, (2) undefined factors in sputum from patients with CF may contribute to survival of B. cepacia, and (3) strain-to-strain variation in survival time may affect strain transmissibility.

Epidemiology and natural history of urinary tract infections in children.

Recent retrospective surveys have supported previous investigations in demonstrating the incidence of UTI during infancy; 0.3% to 1.2% of infants develop symptomatic UTI during the first year of life. Boys are more commonly infected during the first 3 months of life. After the first year, symptomatic UTI is much more frequent among girls. Similarly, asymptomatic bacteriuria is more frequently detected in boys than in girls during the first 12 months of life. Thereafter, the incidence decreases markedly in boys but increases in girls. Recent investigations indicate that lack of circumcision is a risk factor for UTI among male infants. Recurrent UTI is common and frequently asymptomatic. The most important microbiologic factor that is associated with E. coli causing acute pyelonephritis is adherence mediated by P fimbriae. Other factors, such as capsule, lipopolysaccharide, aerobactin production, and serum resistance, also determine the invasiveness of E. coli. Vesicoureteral reflux appears to be an important host factor predisposing to UTI. Microbiologic and host factors that are determinants of renal scarring are under investigation.

Burkholderia cepacia. Management issues and new insights.

Although Burkholderia cepacia colonizes a relatively small proportion of individuals with cystic fibrosis (CF), it is associated with significant morbidity and mortality, and has had a profound impact on infection control practices. This article reviews the current understanding of the epidemiology of B. cepacia infection, describes important recent developments in the microbiology and taxonomy of this species, and presents issues that remain obstacles to defining the optimal management of B. cepacia infection in CF.