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71 Pseudomonas aeruginosa in paranasal sinuses and in lower airways in cystic fibrosis patients
Posters
4. Microbiology
69 The Cystic Fibrosis Sputum Induction Trial (CF-SpIT ). Induced sputum in young healthy non-productive children with cystic fibrosis J. Tame1 , K. Ronchetti1 , C. Paisey2 , I. Doull1 , E. Mahenthiralingham2 , R. Howe3 , J.T. Forton1,4 . 1 Children’s Hospital for Wales, Department of Paediatric Respiratory Medicine, Cardiff, United Kingdom; 2 Cardiff University, Department of Biosciences, Cardiff, United Kingdom; 3 University Hospital for Wales, Department of Microbiology, Cardiff, United Kingdom; 4 Institute of Molecular and Experimental Medicine, Cardiff University School of Medicine, Cardiff, United Kingdom Objectives: Young children with CF are often non-productive of sputum even during a chest exacerbation. Effective bacterial surveillance is challenging, but critical as respiratory infection and lung disease begins early in life. CF-SpIT (CF-Sputum Induction Trial: UKCRN 14615) seeks to identify whether using induced sputum (IS) improves bacterial surveillance, in young non-productive children with CF. Methods: 86 children from CF-SpIT are included in this analysis. In all cases, a routine cough swab (CS) was followed by IS. Paired samples were compared for microbiology yield. In 20 children, where bronchoscopy was clinically indicated, CS, IS and extensive 6 lobe bronchoalveolar lavage (BAL) were compared for microbiology yield. Results: IS was tolerated in 98% of patients, and successful in 81% (n = 69). Tolerability and success did not differ in children below or above 6 years of age, despite more of the younger cohort being naive to hypertonic saline (30%), not spontaneously productive (92%) and unwell at the time (68%). Of the 69 patients, 17% had positive cough swabs, 47% had positive IS samples (c2 = 10. P = 0.002). IS identified a pathogen not previously isolated in the previous year in 29% of patients. In 20 patients where CS, IS and BAL were taken, 20 pathogens were grown in 16 patients. CS, IS and BAL identified 5, 13 and 14 pathogens respectively. Pathogens were identified on IS but not on BAL in 4 patients. 16S sequencing of culturenegative BAL was positive for the pathogen identified on IS in 3 of 4 cases. Conclusion: Induced Sputum is a well tolerated, non-invasive, effective way to sample the lower airway in young non-productive healthy children with CF.
D. Dolce1 , S. Bresci2 , E. Masi3 , S. Campana1 , C. Braggion1 , G. Taccetti1 . 1 CF Center, Department of Paediatric Medicine, Anna Meyer Children’s University Hospital, Florence, Italy; 2 Infectious Disease Unit, Careggi University Hospital, Florence, Italy; 3 Rehabilitation Center, Anna Meyer Children’s University Hospital, Florence, Italy Objectives: The paranasal sinuses could be a site of P. aeruginosa (PA) infection and/or reservoir for episodes of pulmonary re-infection in CF patients. The aim of this research was to clarify the role of the paranasal sinuses in the PA pulmonary infection. Methods: Paired nasal lavage and lower airways samples were taken from 65 CF patients (median age 22.7 range 1.8–53.2 years) during 2014. Patients were classified in four group according to Leeds definition: 3 (4%) patients never infected, 18 (28%) PA-free, 24 (37%) intermittently infected, 9 (14%) chronically infected plus 11 (17%) lung transplanted patients. PA isolates from upper airways and lower airways were genotyped using BOX-PCR analysis. Results: We found that 21 (32.3%) out of 65 patients had a positive PA culture from the nasal lavage. The mucoid PA phenotype was isolated in 9 (42.8%) out 21 positive samples. In the 3 never-infected patients all samples were PA-negative. Only 1 (5.5%) out of 18 patients had positive nasal lavage in the PA-free group. In the intermittent PA infection group 4 (16.6%) out of 24 patients had a positive upper respiratory tract. In the chronically infected patients all 9 (100%) nasal lavage samples were positive for PA. Seven (63.6%) out of 11 nasal lavage were positive for PA in patients who had been transplanted. The PA strains from the upper and lower respiratory tract cultures had the same genotype in 19 (90.5%) out of 21 patients except in 1 patient with chronic PA and in 1 transplanted patient. Conclusion: Identical PA genotype and phenotype isolated from the upper and lower respiratory samples suggest that the paranasal sinuses may be a reservoir for pulmonary infection.
70 Dry powder inhalation devices are a safe alternative to nebulizers regarding contamination with CF specific pathogenic germs
72 Potential for airborne bacterial contamination in an outpatient respiratory clinic
P. Opitz1 , C. Schwarz2 , R. Fischer1 , M. Hogardt3 . 1 Pneumologische Praxis M¨unchen-Pasing, M¨unchen, Germany; 2 Charit´e − Universit¨atsmedizin Berlin, Christiane-Herzog-Ambulanz/Sektion Mukoviszidose, Berlin, Germany; 3 Universit¨ atsklinikum Frankfurt, Institut f¨ur Med. Mikrobiologie u. Krankenhaushygiene, Frankfurt, Germany
J. McElderry1 , K. O’Neill1 , M.M. Tunney1 , D.G. Downey2 , J.C. Rendall2 , J.E. Moore2 , J.S. Elborn1 , D.F. Gilpin1 . 1 Queen’s University, Belfast, United Kingdom; 2 Belfast Health and Social Care Trust, Belfast, United Kingdom
Objective: To assess the contamination risk with CF-relevant pathogens of the Tobi Podhaler dry powder inhaler (DPI) device compared to established nebulizers and to obtain “real-world” data from nebulizers of patients with chronic colonization of the lung with Gram-negative bacteria in the patient’s usual inhalation setting, to learn more about patient’s cleaning and inhalation habits. Methods: Microbiological samples from the mouthpiece and medication reservoir were collected using an eSwab® from a device that was prepared in the patient’s personal way before next inhalation. Results: Microbiological analysis showed that the samples of 8/26 dry powder devices were sterile while the other 16 samples were contaminated with nonpathogenic bacteria of the human respiratory flora. Only 1/20 nebulizer devices were sterile. In 4 devices Staphylococcus aureus was detected. The other nebulizers were contaminated with a variety of bacterial species belonging to the human respiratory tract flora and bacteria typically found in humid sources (e.g. Pseudomonas fluorescens, Acinetobacter ursingii, Mycobacterium fortuitum), indicating the potential transmission risk of CF-relevant species such as P. aeruginosa. Conclusion: Tobi Podhaler is used 7 days twice daily. For cleaning it is supposed to be wiped with a dry and clean cloth. The cleaning procedure of a nebulizer is more time consuming and challenging. Under “real-world” conditions we found that the typical cleaning procedure of a nebulizer is failing to sterilize the device completely. Using a DPI with almost zero cleaning effort seems to be a satisfactory alternative regarding the hygienic requirements of an inhalation device.
S75
71 Pseudomonas aeruginosa in paranasal sinuses and in lower airways in cystic fibrosis patients
Objectives: Recent studies have shown that bacteria of clinical significance in respiratory infection, such as Pseudomonas aeruginosa, have potential for airborne dissemination. This study examined airborne bacteria at an outpatient respiratory clinic and to determine levels of antibiotic resistance among airborne isolates. Methods: Air was sampled (Casella slit air sampler, 30litres/min) on two separate days at various locations and time points in a Belfast Respiratory Outpatient Clinic, starting before clinic, until clinic finished. Bronchiectasis (BE) patients attended morning clinic, and Cystic Fibrosis (CF) patients in the afternoon. Air samples were deposited onto M¨uller-Hinton agar, incubated for 24−48 hrs/37o C/O2 , and individual colonies counted, Gram stained and identified by 16s rRNA sequencing. The antibiotic susceptibility of selected isolates (n = 11) was determined by E-test® . Results: The majority of bacteria detected were Gram +ve (184/242; 76%). The most common were Micrococcus, Kocuria and Staphylococcus spp. Moraxella spp. and one Stenotrophomonas maltophilia isolate were also cultured. No apparent differences in bacterial load were observed between either the BE or CF clinics, or at sequential time points. Of the isolates tested, 8/11 (73%) were resistant to clindamycin. Discussion: Although Moraxella, staphylococci and S. maltophilia were observed, the majority of bacteria isolated were those normally designated as skin commensals. Work is currently ongoing, using molecular methods to more fully describe the airborne microbiome and resistome, and to compare patient vs airborne isolates to gauge the potential for transmission within the clinic.
4. Microbiology
69 The Cystic Fibrosis Sputum Induction Trial (CF-SpIT ). Induced sputum in young healthy non-productive children with cystic fibrosis J. Tame1 , K. Ronchetti1 , C. Paisey2 , I. Doull1 , E. Mahenthiralingham2 , R. Howe3 , J.T. Forton1,4 . 1 Children’s Hospital for Wales, Department of Paediatric Respiratory Medicine, Cardiff, United Kingdom; 2 Cardiff University, Department of Biosciences, Cardiff, United Kingdom; 3 University Hospital for Wales, Department of Microbiology, Cardiff, United Kingdom; 4 Institute of Molecular and Experimental Medicine, Cardiff University School of Medicine, Cardiff, United Kingdom Objectives: Young children with CF are often non-productive of sputum even during a chest exacerbation. Effective bacterial surveillance is challenging, but critical as respiratory infection and lung disease begins early in life. CF-SpIT (CF-Sputum Induction Trial: UKCRN 14615) seeks to identify whether using induced sputum (IS) improves bacterial surveillance, in young non-productive children with CF. Methods: 86 children from CF-SpIT are included in this analysis. In all cases, a routine cough swab (CS) was followed by IS. Paired samples were compared for microbiology yield. In 20 children, where bronchoscopy was clinically indicated, CS, IS and extensive 6 lobe bronchoalveolar lavage (BAL) were compared for microbiology yield. Results: IS was tolerated in 98% of patients, and successful in 81% (n = 69). Tolerability and success did not differ in children below or above 6 years of age, despite more of the younger cohort being naive to hypertonic saline (30%), not spontaneously productive (92%) and unwell at the time (68%). Of the 69 patients, 17% had positive cough swabs, 47% had positive IS samples (c2 = 10. P = 0.002). IS identified a pathogen not previously isolated in the previous year in 29% of patients. In 20 patients where CS, IS and BAL were taken, 20 pathogens were grown in 16 patients. CS, IS and BAL identified 5, 13 and 14 pathogens respectively. Pathogens were identified on IS but not on BAL in 4 patients. 16S sequencing of culturenegative BAL was positive for the pathogen identified on IS in 3 of 4 cases. Conclusion: Induced Sputum is a well tolerated, non-invasive, effective way to sample the lower airway in young non-productive healthy children with CF.
D. Dolce1 , S. Bresci2 , E. Masi3 , S. Campana1 , C. Braggion1 , G. Taccetti1 . 1 CF Center, Department of Paediatric Medicine, Anna Meyer Children’s University Hospital, Florence, Italy; 2 Infectious Disease Unit, Careggi University Hospital, Florence, Italy; 3 Rehabilitation Center, Anna Meyer Children’s University Hospital, Florence, Italy Objectives: The paranasal sinuses could be a site of P. aeruginosa (PA) infection and/or reservoir for episodes of pulmonary re-infection in CF patients. The aim of this research was to clarify the role of the paranasal sinuses in the PA pulmonary infection. Methods: Paired nasal lavage and lower airways samples were taken from 65 CF patients (median age 22.7 range 1.8–53.2 years) during 2014. Patients were classified in four group according to Leeds definition: 3 (4%) patients never infected, 18 (28%) PA-free, 24 (37%) intermittently infected, 9 (14%) chronically infected plus 11 (17%) lung transplanted patients. PA isolates from upper airways and lower airways were genotyped using BOX-PCR analysis. Results: We found that 21 (32.3%) out of 65 patients had a positive PA culture from the nasal lavage. The mucoid PA phenotype was isolated in 9 (42.8%) out 21 positive samples. In the 3 never-infected patients all samples were PA-negative. Only 1 (5.5%) out of 18 patients had positive nasal lavage in the PA-free group. In the intermittent PA infection group 4 (16.6%) out of 24 patients had a positive upper respiratory tract. In the chronically infected patients all 9 (100%) nasal lavage samples were positive for PA. Seven (63.6%) out of 11 nasal lavage were positive for PA in patients who had been transplanted. The PA strains from the upper and lower respiratory tract cultures had the same genotype in 19 (90.5%) out of 21 patients except in 1 patient with chronic PA and in 1 transplanted patient. Conclusion: Identical PA genotype and phenotype isolated from the upper and lower respiratory samples suggest that the paranasal sinuses may be a reservoir for pulmonary infection.
70 Dry powder inhalation devices are a safe alternative to nebulizers regarding contamination with CF specific pathogenic germs
72 Potential for airborne bacterial contamination in an outpatient respiratory clinic
P. Opitz1 , C. Schwarz2 , R. Fischer1 , M. Hogardt3 . 1 Pneumologische Praxis M¨unchen-Pasing, M¨unchen, Germany; 2 Charit´e − Universit¨atsmedizin Berlin, Christiane-Herzog-Ambulanz/Sektion Mukoviszidose, Berlin, Germany; 3 Universit¨ atsklinikum Frankfurt, Institut f¨ur Med. Mikrobiologie u. Krankenhaushygiene, Frankfurt, Germany
J. McElderry1 , K. O’Neill1 , M.M. Tunney1 , D.G. Downey2 , J.C. Rendall2 , J.E. Moore2 , J.S. Elborn1 , D.F. Gilpin1 . 1 Queen’s University, Belfast, United Kingdom; 2 Belfast Health and Social Care Trust, Belfast, United Kingdom
Objective: To assess the contamination risk with CF-relevant pathogens of the Tobi Podhaler dry powder inhaler (DPI) device compared to established nebulizers and to obtain “real-world” data from nebulizers of patients with chronic colonization of the lung with Gram-negative bacteria in the patient’s usual inhalation setting, to learn more about patient’s cleaning and inhalation habits. Methods: Microbiological samples from the mouthpiece and medication reservoir were collected using an eSwab® from a device that was prepared in the patient’s personal way before next inhalation. Results: Microbiological analysis showed that the samples of 8/26 dry powder devices were sterile while the other 16 samples were contaminated with nonpathogenic bacteria of the human respiratory flora. Only 1/20 nebulizer devices were sterile. In 4 devices Staphylococcus aureus was detected. The other nebulizers were contaminated with a variety of bacterial species belonging to the human respiratory tract flora and bacteria typically found in humid sources (e.g. Pseudomonas fluorescens, Acinetobacter ursingii, Mycobacterium fortuitum), indicating the potential transmission risk of CF-relevant species such as P. aeruginosa. Conclusion: Tobi Podhaler is used 7 days twice daily. For cleaning it is supposed to be wiped with a dry and clean cloth. The cleaning procedure of a nebulizer is more time consuming and challenging. Under “real-world” conditions we found that the typical cleaning procedure of a nebulizer is failing to sterilize the device completely. Using a DPI with almost zero cleaning effort seems to be a satisfactory alternative regarding the hygienic requirements of an inhalation device.
S75
71 Pseudomonas aeruginosa in paranasal sinuses and in lower airways in cystic fibrosis patients
Objectives: Recent studies have shown that bacteria of clinical significance in respiratory infection, such as Pseudomonas aeruginosa, have potential for airborne dissemination. This study examined airborne bacteria at an outpatient respiratory clinic and to determine levels of antibiotic resistance among airborne isolates. Methods: Air was sampled (Casella slit air sampler, 30litres/min) on two separate days at various locations and time points in a Belfast Respiratory Outpatient Clinic, starting before clinic, until clinic finished. Bronchiectasis (BE) patients attended morning clinic, and Cystic Fibrosis (CF) patients in the afternoon. Air samples were deposited onto M¨uller-Hinton agar, incubated for 24−48 hrs/37o C/O2 , and individual colonies counted, Gram stained and identified by 16s rRNA sequencing. The antibiotic susceptibility of selected isolates (n = 11) was determined by E-test® . Results: The majority of bacteria detected were Gram +ve (184/242; 76%). The most common were Micrococcus, Kocuria and Staphylococcus spp. Moraxella spp. and one Stenotrophomonas maltophilia isolate were also cultured. No apparent differences in bacterial load were observed between either the BE or CF clinics, or at sequential time points. Of the isolates tested, 8/11 (73%) were resistant to clindamycin. Discussion: Although Moraxella, staphylococci and S. maltophilia were observed, the majority of bacteria isolated were those normally designated as skin commensals. Work is currently ongoing, using molecular methods to more fully describe the airborne microbiome and resistome, and to compare patient vs airborne isolates to gauge the potential for transmission within the clinic.