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ePS06.8 Cigarette smoke-induced changes in phenotype and virulence in Pseudomonas aeruginosa
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ePS6. Triggers of decline?
ePS06.5 Burkholderia cepacia complex acquisition: A threat in all CF patients? J. Willekens1 , S. Wanyama2 , M. Thomas2 , E. De Wachter3 , I. De Schutter3 , A. Malfroot3 . 1 Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel (VUB), Department of Pediatrics, Cystic Fibrosis Clinic, Brussels, Belgium; 2 Scientific Institute of Public Health WIV-ISP, Public Health and Surveillance, Brussels, Belgium; 3 Department of Pediatrics, Cystic Fibrosis Clinic, Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel (VUB), Brussels, Belgium Introduction: Acquisition of Burkholderia cepacia complex (Bcc) bacteria is considered to be associated with worsening of CF lung disease. Moreover, because of its resistance to antibiotics, Bcc is considered a threat in CF. Method: Data from the Belgian CF Registry (year 2000–2010) were obtained. Inclusion: Bcc-infected patients with entries on lung function, BMI and days of IV treatment in at least 3 y after Bcc acquisition. For each case, we included 2 controls, matched for age at index year (year of first Bcc infection), pancreatic status, sex. FEV1 , days of IV antibiotics and hospitalization were used as surrogates for disease burden. Cumulative data up to 2 y before index year were compared to values after infection using Rank sum test. Rate of decline in lung function was adjusted for baseline lung function, age, sex. Results: Prevalence of Bcc in CF is low in Belgium (<3%). We included 183 patients: 61 cases, 122 controls. 59% were F508del homozygous. Among the Bcc, 54% were unspecified, 31% were B. multivorans. FEV1 at index year had lower tendency in the cases, however without significance. The number of days of IV treatment had higher tendency in the cases (p = ns). FEV1 decline after Bcc acquisition was comparable in cases and controls (−1.1%, SD = 0.5, vs −0.99%, SD = 0.4, p = 0.24). BMI slopes were also comparable between groups. Conclusion: Our study suggests that Bcc infected patients are in no worse clinical condition and demonstrate no faster clinical deterioration compared to Bccuninfected patients. However, registry data are collected retrospectively and bear a risk of inaccuracy. Therefore, these results should be confirmed in a prospective, longitudinal study.
Oral Presentations
ePS06.7 Factors associated with failure to eradicate first or newly acquired Pseudomonas aeruginosa in patients with CF M. Cohen-Cymberknoh1 , N. Gilead2 , S. Gartner3 , N. Simanovski4 , E. Canino3 , J.C. Carre˜no5 , H. Blau6 , H. Mussaffi6 , M. Gur7 , J. Rivlin7 , L. Bentur8 , M. Aviram9 , E. Picard10 , N. Hiller4 , S. Armoni1 , O. Breuer1 , D. Shoseyov1 , E. Kerem1 . 1 Hadassah-Hebrew University Medical Center, CF Center and Pediatric Pulmonary Unit, Jerusalem, Israel; 2 Hadassah-Hebrew University, Jerusalem, Israel; 3 Hospital Universitari Vall d’Hebron, CF Center and Pediatric Pulmonary Unit, Barcelona, Spain; 4 Hadassah-Hebrew University Medical Center, Department of Radiology, Jerusalem, Israel; 5 Hospital Universitari Vall d’Hebron, Pediatric Radiology Department, Barcelona, Spain; 6 Schneider Children’s Medical Center, Graub CF Center and Pediatric Pulmonary Unit, Petach-Tikva, Israel; 7 Carmel Medical Center, CF Center and Pediatric Pulmonary Unit, Haifa, Israel; 8 Rambam Medical Center, CF Center and Pediatric Pulmonary Unit, Haifa, Israel; 9 Soroka Medical Center, CF Center and Pediatric Pulmonary Unit, Beer Sheva, Israel; 10 Shaare=Zedek Medical Center, Pediatric Pulmonary Unit, Jerusalem, Israel Respiratory infections with P. aeruginosa (PA) are a leading cause of morbiditymortality in Cystic fibrosis (CF) patients. Several protocols for eradication are in use, however, the optimal antibiotic dosage, mode of delivery and duration of therapy has not been standardized. Still, ~20% of the infections cannot be eradicated leading to chronic infection. Aims: To analyze the rate of eradication failure of 1st /new PA infection and to identify clinical factors associated with eradication failure. Methods: 183 patients’ files from 6 CF Centers in Israel and 1 in Spain were included. Demographic and clinical data for 7 years (2007–2013) were retrospectively analyzed. Results: For 131 patients (72%) eradication was successful, and were divided into 2 groups: Absolute eradication − successful eradication and completed anti-PA treatment (n = 92) and Presumed eradication − successful eradication with ongoing anti-PA inhalations (n = 39). For 52 patients eradication failed: 34/52 chronically infected and 18/52 intermittent infection. Older age at CF diagnosis, older age at 1st /new PA infection, lower FEV1 %, less sputum cultures taken 1 year prior to 1st /new isolation, previous documented PA colonization and multi/pan-resistant PA strains were all found to be associated with failure to eradication. BMI, gender, and CF-related diabetes did not influence successful eradication. No significant difference was seen in terms of HRCT Brody scores. Additionally, the different eradication protocols did not change the outcome. Conclusions: Early diagnosis, increased follow-up and appropriate therapy are all required in order to enhance the rate of efficient eradication of PA in CF patients.
ePS06.6 Full reversibility of lung function decline following clearance of Mycobacterium abscessus complex infection
ePS06.8 Cigarette smoke-induced changes in phenotype and virulence in Pseudomonas aeruginosa
T. Qvist1 , D. Taylor-Robinson2 , E. Waldmann2 , H.V. Olesen3 , C.R. Hansen4 , I.H. Mathiesen1 , N. Høiby5 , T.L. Katzenstein1 , R. Smyth6 , P. Diggle2 , T. Pressler1 . 1 Copenhagen CF Center, Department of Infectious Diseases, Copenhagen, Denmark; 2 University of Liverpool, Department of Public Health and Policy, Liverpool, United Kingdom; 3 CF Center Skejby, Department of Pediatrics, Aarhus, Denmark; 4 Copenhagen CF Center, Department of Pediatrics, Copenhagen, Denmark; 5 Copenhagen CF Center, Department of Clinical Microbiology, Copenhagen, Denmark; 6 Institute of Child Health, University College London, London, United Kingdom
K.-A. McGown1 , S. McGrath1 , M.M. Tunney1 , J.S. Elborn1 , D.F. Gilpin1 . 1 Queen’s University Belfast, CF and Airways Microbiology Research Group, Belfast, United Kingdom
Objectives: To assess the effect of Mycobacterium abscessus complex (MABSC) infection on lung function (%FEV1) in Danish CF patients. Methods: A longitudinal analysis of %FEV1 trajectory was performed on all Danish patients born after 1974 using a linear mixed effects model adjusting for chronic infections and other clinically important co-variates. All patients were seen routinely every month. Onset of MABSC was defined as date of first positive culture and clearing MABSC as consistent culture negativity in 4 cultures. Results: The dataset contained 53,771 %FEV1 measures on 432 patients of which 44 had at least one positive MABSC culture, 20 of whom cleared infection, either following treatment or spontaneously. Median follow-up was 21 years. Onset of MABSC was associated with an excess %FEV1 decline of −2.22 percentage points per year (95% CI −3.21 to −1.23). This translated to end stage lung disease developing approximately 20 years earlier, than would be expected in a continuously uninfected patient. Clearance of MABSC was associated with returning to the preinfection decline in %FEV1. There was no isolated effect of fulfilling the American Thoracic Society’s criteria for MABSC disease. Conclusion: The strength of this analysis was the high visit frequency (every 4 weeks) and long period of follow-up in a complete national population. We demonstrate that infection with MABSC impacts lung function significantly and that clearing MABSC results in %FEV1 returning to its pre-infection slope, pointing to the importance of eradication therapy.
Objectives: The health risks of either direct or passive smoking in Cystic Fibrosis (CF) have been well established (Kopp et al., 2015). Chronic infection with Pseudomonas aeruginosa (PA) is associated with considerable morbidity and mortality in CF, but the direct effect of cigarette smoke (CS) on PA has not been established. This study aimed to determine the impact of CS on PA phenotype and virulence. Methods: Cigarette smoke extract (CSE) was prepared as described previously (Comer et al, 2012). Respiratory PA isolates (n = 4) and type strain (n = 1; ATCC 27853) were grown aerobically at 37o C +/− CSE and total viable count cfu/ml (TVC) determined at a range of CSE concentrations. Pyocyanin production was quantified following chloroform extraction. Biofilm formation was examined using a microtitre tray assay with quantification by crystal violet staining (Stepanovic et al., 2000). Virulence of PA was determined in the Galleria mellonella infection model. Results: Growth of PA isolates was not completely inhibited by any concentration of CSE used. Both pyocyanin production and biofilm formation were significantly greater when isolates were exposed to CSE (biofilm: p < 0.05). Increased melanisation and decreased survival was observed in the G. mellonella model when PA isolates were exposed to CSE, indicating increased virulence. Conclusion: Exposure of PA to CSE in vitro increases PA virulence. Further clinical studies are required to more fully correlate these changes in phenotype with clinical outcomes.
ePS6. Triggers of decline?
ePS06.5 Burkholderia cepacia complex acquisition: A threat in all CF patients? J. Willekens1 , S. Wanyama2 , M. Thomas2 , E. De Wachter3 , I. De Schutter3 , A. Malfroot3 . 1 Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel (VUB), Department of Pediatrics, Cystic Fibrosis Clinic, Brussels, Belgium; 2 Scientific Institute of Public Health WIV-ISP, Public Health and Surveillance, Brussels, Belgium; 3 Department of Pediatrics, Cystic Fibrosis Clinic, Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel (VUB), Brussels, Belgium Introduction: Acquisition of Burkholderia cepacia complex (Bcc) bacteria is considered to be associated with worsening of CF lung disease. Moreover, because of its resistance to antibiotics, Bcc is considered a threat in CF. Method: Data from the Belgian CF Registry (year 2000–2010) were obtained. Inclusion: Bcc-infected patients with entries on lung function, BMI and days of IV treatment in at least 3 y after Bcc acquisition. For each case, we included 2 controls, matched for age at index year (year of first Bcc infection), pancreatic status, sex. FEV1 , days of IV antibiotics and hospitalization were used as surrogates for disease burden. Cumulative data up to 2 y before index year were compared to values after infection using Rank sum test. Rate of decline in lung function was adjusted for baseline lung function, age, sex. Results: Prevalence of Bcc in CF is low in Belgium (<3%). We included 183 patients: 61 cases, 122 controls. 59% were F508del homozygous. Among the Bcc, 54% were unspecified, 31% were B. multivorans. FEV1 at index year had lower tendency in the cases, however without significance. The number of days of IV treatment had higher tendency in the cases (p = ns). FEV1 decline after Bcc acquisition was comparable in cases and controls (−1.1%, SD = 0.5, vs −0.99%, SD = 0.4, p = 0.24). BMI slopes were also comparable between groups. Conclusion: Our study suggests that Bcc infected patients are in no worse clinical condition and demonstrate no faster clinical deterioration compared to Bccuninfected patients. However, registry data are collected retrospectively and bear a risk of inaccuracy. Therefore, these results should be confirmed in a prospective, longitudinal study.
Oral Presentations
ePS06.7 Factors associated with failure to eradicate first or newly acquired Pseudomonas aeruginosa in patients with CF M. Cohen-Cymberknoh1 , N. Gilead2 , S. Gartner3 , N. Simanovski4 , E. Canino3 , J.C. Carre˜no5 , H. Blau6 , H. Mussaffi6 , M. Gur7 , J. Rivlin7 , L. Bentur8 , M. Aviram9 , E. Picard10 , N. Hiller4 , S. Armoni1 , O. Breuer1 , D. Shoseyov1 , E. Kerem1 . 1 Hadassah-Hebrew University Medical Center, CF Center and Pediatric Pulmonary Unit, Jerusalem, Israel; 2 Hadassah-Hebrew University, Jerusalem, Israel; 3 Hospital Universitari Vall d’Hebron, CF Center and Pediatric Pulmonary Unit, Barcelona, Spain; 4 Hadassah-Hebrew University Medical Center, Department of Radiology, Jerusalem, Israel; 5 Hospital Universitari Vall d’Hebron, Pediatric Radiology Department, Barcelona, Spain; 6 Schneider Children’s Medical Center, Graub CF Center and Pediatric Pulmonary Unit, Petach-Tikva, Israel; 7 Carmel Medical Center, CF Center and Pediatric Pulmonary Unit, Haifa, Israel; 8 Rambam Medical Center, CF Center and Pediatric Pulmonary Unit, Haifa, Israel; 9 Soroka Medical Center, CF Center and Pediatric Pulmonary Unit, Beer Sheva, Israel; 10 Shaare=Zedek Medical Center, Pediatric Pulmonary Unit, Jerusalem, Israel Respiratory infections with P. aeruginosa (PA) are a leading cause of morbiditymortality in Cystic fibrosis (CF) patients. Several protocols for eradication are in use, however, the optimal antibiotic dosage, mode of delivery and duration of therapy has not been standardized. Still, ~20% of the infections cannot be eradicated leading to chronic infection. Aims: To analyze the rate of eradication failure of 1st /new PA infection and to identify clinical factors associated with eradication failure. Methods: 183 patients’ files from 6 CF Centers in Israel and 1 in Spain were included. Demographic and clinical data for 7 years (2007–2013) were retrospectively analyzed. Results: For 131 patients (72%) eradication was successful, and were divided into 2 groups: Absolute eradication − successful eradication and completed anti-PA treatment (n = 92) and Presumed eradication − successful eradication with ongoing anti-PA inhalations (n = 39). For 52 patients eradication failed: 34/52 chronically infected and 18/52 intermittent infection. Older age at CF diagnosis, older age at 1st /new PA infection, lower FEV1 %, less sputum cultures taken 1 year prior to 1st /new isolation, previous documented PA colonization and multi/pan-resistant PA strains were all found to be associated with failure to eradication. BMI, gender, and CF-related diabetes did not influence successful eradication. No significant difference was seen in terms of HRCT Brody scores. Additionally, the different eradication protocols did not change the outcome. Conclusions: Early diagnosis, increased follow-up and appropriate therapy are all required in order to enhance the rate of efficient eradication of PA in CF patients.
ePS06.6 Full reversibility of lung function decline following clearance of Mycobacterium abscessus complex infection
ePS06.8 Cigarette smoke-induced changes in phenotype and virulence in Pseudomonas aeruginosa
T. Qvist1 , D. Taylor-Robinson2 , E. Waldmann2 , H.V. Olesen3 , C.R. Hansen4 , I.H. Mathiesen1 , N. Høiby5 , T.L. Katzenstein1 , R. Smyth6 , P. Diggle2 , T. Pressler1 . 1 Copenhagen CF Center, Department of Infectious Diseases, Copenhagen, Denmark; 2 University of Liverpool, Department of Public Health and Policy, Liverpool, United Kingdom; 3 CF Center Skejby, Department of Pediatrics, Aarhus, Denmark; 4 Copenhagen CF Center, Department of Pediatrics, Copenhagen, Denmark; 5 Copenhagen CF Center, Department of Clinical Microbiology, Copenhagen, Denmark; 6 Institute of Child Health, University College London, London, United Kingdom
K.-A. McGown1 , S. McGrath1 , M.M. Tunney1 , J.S. Elborn1 , D.F. Gilpin1 . 1 Queen’s University Belfast, CF and Airways Microbiology Research Group, Belfast, United Kingdom
Objectives: To assess the effect of Mycobacterium abscessus complex (MABSC) infection on lung function (%FEV1) in Danish CF patients. Methods: A longitudinal analysis of %FEV1 trajectory was performed on all Danish patients born after 1974 using a linear mixed effects model adjusting for chronic infections and other clinically important co-variates. All patients were seen routinely every month. Onset of MABSC was defined as date of first positive culture and clearing MABSC as consistent culture negativity in 4 cultures. Results: The dataset contained 53,771 %FEV1 measures on 432 patients of which 44 had at least one positive MABSC culture, 20 of whom cleared infection, either following treatment or spontaneously. Median follow-up was 21 years. Onset of MABSC was associated with an excess %FEV1 decline of −2.22 percentage points per year (95% CI −3.21 to −1.23). This translated to end stage lung disease developing approximately 20 years earlier, than would be expected in a continuously uninfected patient. Clearance of MABSC was associated with returning to the preinfection decline in %FEV1. There was no isolated effect of fulfilling the American Thoracic Society’s criteria for MABSC disease. Conclusion: The strength of this analysis was the high visit frequency (every 4 weeks) and long period of follow-up in a complete national population. We demonstrate that infection with MABSC impacts lung function significantly and that clearing MABSC results in %FEV1 returning to its pre-infection slope, pointing to the importance of eradication therapy.
Objectives: The health risks of either direct or passive smoking in Cystic Fibrosis (CF) have been well established (Kopp et al., 2015). Chronic infection with Pseudomonas aeruginosa (PA) is associated with considerable morbidity and mortality in CF, but the direct effect of cigarette smoke (CS) on PA has not been established. This study aimed to determine the impact of CS on PA phenotype and virulence. Methods: Cigarette smoke extract (CSE) was prepared as described previously (Comer et al, 2012). Respiratory PA isolates (n = 4) and type strain (n = 1; ATCC 27853) were grown aerobically at 37o C +/− CSE and total viable count cfu/ml (TVC) determined at a range of CSE concentrations. Pyocyanin production was quantified following chloroform extraction. Biofilm formation was examined using a microtitre tray assay with quantification by crystal violet staining (Stepanovic et al., 2000). Virulence of PA was determined in the Galleria mellonella infection model. Results: Growth of PA isolates was not completely inhibited by any concentration of CSE used. Both pyocyanin production and biofilm formation were significantly greater when isolates were exposed to CSE (biofilm: p < 0.05). Increased melanisation and decreased survival was observed in the G. mellonella model when PA isolates were exposed to CSE, indicating increased virulence. Conclusion: Exposure of PA to CSE in vitro increases PA virulence. Further clinical studies are required to more fully correlate these changes in phenotype with clinical outcomes.