- Email: [email protected]
100 (More) viable cough aerosols from individuals with cystic fibrosis
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Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
97 Nocardia and cystic fibrosis: the impact of Gram staining O. Tuncer1 , S. Olmez1 , B. Sancak1 , G.D. Tugcu2 , N. Emiralioglu2 , B. Otlu3 , 2 1 1 ¨ celik B. Er4 , E. Yalcın ¸ 2 , D. Dogru2 , U. Oz ¸ , N. Kiper2 , B. Sener ¸ . Hacettepe University Medical Faculty, Clinical Microbiology, Ankara, Turkey; 2 Hacettepe University Ihsan Dogramaci Children’s Hospital, Pediatric Pulmonology, Ankara, Turkey; 3 In¨ on¨ u University Medical Faculty, Clinical Microbiology, Malatya, Turkey; 4 Hacettepe University Medical Faculty, Pulmonology, Ankara, Turkey Objectives: We describe Nocardia isolation from multiple respiratory specimens of two CF patients with chronic Pseudomonas colonization. We aimed to investigate the genetic relatedness of the sequential Nocardia isolates in each patient. Methods: When branching Gram-positive bacilli were observed in Gram stained smears, buffered charcoal yeast extract agar was also included to the culture panel and incubation period was extended to minimum 7 days. The plates were incubated at 37o C in 5% CO2 /air environment. Smears were prepared from suspected colonies and stained by Gram and modified Ziehl-Neelsen to confirm the presence of Gram positive, variable staining, branching bacilli that were positive by modified ZiehlNeelsen. 16S rRNA gene sequence analysis was used to identify the Nocardia isolates to species level. The genetic relatedness between the isolates were analysed by pulsed field gel electrophoresis and AP-PCR. Results: In patient A N. transvalensis/N. wallacei and in patient B N. asteroides were identified. After the first isolation of N. transvalensis/ N. wallacei in sputum culture of patient A, she received TMP/SMX for 4 weeks, no new Nocardia-positive culture was obtained. Patient B received piperacillin–tazobactam for two weeks, TMP/SMX treatment still continues now. Last cultures are negative for Nocardia. Conclusion: The presence of Nocardia in sputum cultures does not always imply disease but rather simple colonization. In order to understand the impact of Nocardia in CF lungs, more reports are needed describing the clinical and microbiological features of CF patients who have Nocardia spp.-positive sputum culture. 98 The cystic fibrosis lower airways microbial metagenome P. Moran Losada1 , P. Chouvarine1 , M. Dorda1 , S. Hedtfeld1 , S. Mielke1 , 1 1 A. Schulz1 , L. Wiehlmann1 , B. Tummler ¨ . Medizinische Hochschule Hannover, Clinic for Paediatric Pneumology, Allergology and Neonatology, Hannover, Germany Objectives: Chronic airway infections determine most morbidity in people with cystic fibrosis (CF). Here we present unbiased quantitative data about the frequency and abundance of DNA viruses, archaea, bacteria, molds and fungi in CF lower airways. Methods: The microbial contents of induced sputa collected on several occasions from 10 PS and 15 Phe508del homozygous CF children, adolescents and adults was resolved by high-throughput whole genome sequencing. Results: Deep sputum metagenome sequencing identified on average about ten DNA viruses or fungi and several hundred bacterial taxa. The metagenome of a CF patient was typically found to be made up of an individual signature of multiple lowly abundant species superimposed by few disease-associated pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus as major components. The host-associated signatures ranged from inconspicuous poly-microbial communities in healthy subjects to low-complexity microbiomes dominated by the typical CF pathogens in patients with advanced lung disease. The DNA virus community in CF lungs mainly consisted of phages and occasionally of human pathogens such as adeno- and herpesviruses. The S. aureus and P. aeruginosa populations were composed of one major and numerous minor clone types. The rare clones constitute a low copy genetic resource which could rapidly expand as a response to habitat alterations such as antimicrobial chemotherapy or invasion of novel microbes.
99 Investigating the airway microbiome in cystic fibrosis patients with normal and severe pulmonary function decline: an opportunity for a personalized microbiome-based therapy G. Bacci1 , P. Paganin2 , N. Segata3 , F. Armanini3 , G. Taccetti4 , D. Dolce4 , A. De Alessandri5 , P. Morelli5 , V. Tuccio6 , E.V. Fiscarelli6 , V. Lucidi6 , A. Mengoni1 , A. Bevivino2 . 1 University of Florence, Department of Biology, Florence, Italy; 2 ENEA Casaccia Research Center, Sustainable Territorial and Production Systems Department, Rome, Italy; 3 University of Trento, Centre for Integrative Biology, Trento, Italy; 4 Cystic Fibrosis Center, Anna Meyer Children’s University Hospital, Department of Pediatrics Medicine, Florence, Italy; 5 Cystic Fibrosis Center, G. Gaslini Institute, Department of Pediatrics, Genoa, Italy; 6 Children’s Hospital and Research Institute Bambino Ges` u, CF Microbiology and CF Center, Rome, Italy Objectives: To gain new insights into CF microbiome composition, metabolism and function, trying to unravel the underlying causes of the severe lung disease and identify potential determinants of serious decline in lung function. Methods: Twelve CF subjects attending three Italian CF centres were enrolled. Patients were stratified according to lung function decline rates and disease severity. Shotgun sequencing was performed following standard pipelines in Illumina Hiseq 2000 platform. To infer functional and taxonomic patterns among patients, extrinsic and intrinsic metagenomic data analysis was used. Results: We identified a core set of bacterial metabolic pathways and functions. In particular, the Kegg pathway One carbon pool by folate related to cysteine/methionine metabolism was found more abundant in patients with normal/mild disease. Moreover, antibiotic resistance genes, mainly belonging to efflux and transporter systems, were found to be more present in patients with severe lung disease. A longitudinal analysis on twenty CF patients followed over one-year is ongoing. Conclusion: The different pulmonary function in CF patients co-occurs with potentially different microbiome capabilities, mainly related to an increasing number of genes for multiple antibiotic resistance (efflux systems) and a potential differential presence of metabolic pathways. The ongoing longitudinal analysis will permit us to identify the microbial markers possible related to severe lung function decline in CF patients, and allow a risk assessment and a better focused and personalized therapy of CF patients. Acknowledgement: Supported by grants from the Italian CF Foundation (FFC #10/2014 and FFC #14/2015). 100 (More) viable cough aerosols from individuals with cystic fibrosis M.E. Wood1,2,3 , G.R. Johnson4 , R.E. Stockwell1 , L.J. Sherrard1,5 , N. Jabbour4 , K.A. Ramsay1,2 , L.D. Knibbs2 , T.J. Kidd2,5 , C.E. Wainwright2,6 , L. Morawska4 , S.C. Bell1,3 . 1 QIMR Berghofer Medical Research Institute, Brisbane, Australia; 2 The University of Queensland, Brisbane, Australia; 3 The Prince Charles Hospital, Brisbane, Australia; 4 Queensland University of Technology, Brisbane, Australia; 5 Queen’s University Belfast, Belfast, United Kingdom; 6 Lady Cilento Children’s Hospital, Brisbane, Australia Objectives: Cough-generated airborne P. aeruginosa droplet nuclei can remain viable for up 45 minutes. This prompted changes to infection control guidelines for patients with cystic fibrosis (CF). This study investigated if other CF respiratory pathogens are detectable in cough aerosols and assessed their survival over distance and time. Methods: 27 subjects with CF, mean age 28.6 years (SD 9.7); mean FEV1 63.1% (26.2) predicted were enrolled into 2 groups: 1. non-P. aeruginosa Gram-negative bacteria (GNB), 2. Gram-positive bacteria (GP), Staphylococcus aureus. Subjects performed a series of voluntary coughing into 2 validated aerosol-sampling devices to measure viability at 2 and 4 m and after 5, 15 and 45 min following generation. An Anderson Impactor collected and sized the aerosols, and total CFU (stages 1–6) was measured. Quantitative sputum and cough aerosol cultures were performed. Results: 11/16 (69%) subjects with mean sputum GNB concentration of 1.9×108 CFU/mL (8.0×103 –2.2×109 ) produced viable aerosol containing the target pathogen. The mean (range) CFU identified in cough aerosol at 4 m and at 45 min was 21.6 (0–164) and 15.6 (0–123),
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
respectively. 10/17 (59%) subjects with mean GP sputum concentration of 2.0×107 CFU/mL (1.0×103 –1.0×108 ) produced viable cough aerosol containing S. aureus. A mean (range) of 7.1 CFU (0–55) and 1.0 CFU (0–7) were detected at 4 m and at 45 min, respectively. Conclusion: CF respiratory pathogens, other than P. aeruginosa, are viable at 4 metres and for up to 45 minutes after coughing, which highlights the need for universal and stringent infection control. Acknowledgement: Funding: CFFT (USA), TPCH Foundation 101 Survival of Mycobacterium abscessus in artificially generated aerosols L.A. Fletcher1 , Y. Chen1 , P. Whitaker2 , D.G. Peckham2 , M. Denton3 , I. Clifton2 . 1 University of Leeds, Department of Civil Engineering, Leeds, United Kingdom; 2 St James’s University Hospital, Regional Adult Cystic Fibrosis Unit, Leeds, United Kingdom; 3 Leeds General Infirmary, Department of Medical Microbiology, Leeds, United Kingdom Objectives: There is emerging evidence supporting the possibility of airborne routes for transmission for Pseudomonas aeruginosa and Burkholderia cepacia complex between people with CF. More recently M. abscessus has emerged as a potentially important pathogen in people with CF with evidence of accelerated lung function decline [2]. The aim of this study was to determine whether M. abscessus could survive within artificially demonstrated aerosols using a previously described laminar airflow model [1]. Methods: Five strains of M. abcessus isolated from patients with CF and a reference strain were studied. Aerosols were generated using a Collison 3-jet nebuliser, delivered into an airtight pipe of varying lengths and sampled using an Andersen 6-stage impactor as previously described [1]. Results: All the strains studied were able to produce viable aerosols of M. abscessus which survived 81 s and travelled a distance of 4 m. All the aerosols contained particles which were predominantly less than 2 mm in diameter. Conclusion: This study demonstrates that M. abscessus can survive within artificially generated aerosols in particles within the respirable range. Cross-infection of M. abscessus between people with CF could potentially occur via an airborne route. The survival of M. abscessus within the laminar airflow model was very similar to that of P. aeruginosa [1]. These results have important implications for the care of people with CF and reinforces the need for strict infection control practices to minimise the risk of cross-infection which pathogens such as M. abscessus. Reference(s) [1] Clifton et al. BMC Microbiol 2008. [2] Esther et al. J Cyst Fibros 2010.
102 Transmission of non-tuberculous mycobacteria in patients with cystic fibrosis C. Schwarz1 , J. Roehmel1 , L. Hatzler1 , D. Staab1 , A. Lewin2 . 1 Charit´eUniversit¨ atsmedizin Berlin, Division of Cystic Fibrosis, Berlin, Germany; 2 Robert Koch Institute, Berlin, Germany Objectives: CF patients suffer from bronchopulmonary disease exacerbations due to multiple recurrent and chronic infections of the lung. A contribution of P. aeruginosa in this process is well established but also NTM, such as Mycobacterium avium and Mycobacterium abscessus are becoming increasingly important. The exact role of NTM is not well characterized and ranges from short and long colonization to severe infection. The risk of tansmission is still under debate. Methods: We prospectively analyzed the sputum or bronchoalveolar lavage for NTM in our 330 patients. To assess the species and subspecies of mycobacteria PCR (16S rRNA gen and species specific PCR) was used. In addition, genome sequencing, susceptibility testing (including erm (41)gen), collection of clinical data of the patients with NTM, the treatment courses and side effects of the treatment were evaluated. Results: We analyzed 330 patients with CF. In 19 patients we found NTM. In 5 patients two species of NTM were detected. The leading mycobacterium was M. abscessus followed by M. avium. In two pairs of
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patients we could find the same clusters of NTM isolates. 14 antibiotics were used for susceptibility testing in patients with fast growing species (e.g. M. abscessus) and 13 in patients with slow growing species (e.g. M. avium). Conclusion: We could find in two pairs of patients the same cluster in there NTM isolates. This might underline the potential risk for transmission of NTM in CF. 103 Low prevalence of mycobacteria among Brazilian CF patients: possible explanations T.B. Aiello1 , R.M. Mauch1 , M.P. Arrym1 , T.R. Zaccariotto1 , A.A.D.C. Toro1 , M.C. Pereira1 , I.A. Paschoal1 , J.D. Ribeiro1 , A.F. Ribeiro1 , C.E. Levy1 . 1 Unicamp, Campinas, Brazil Objectives: To investigate the presence of opportunistic mycobacteria in CF patients attended at a reference center in Brazil. Methods: Data of 109 patients were collected in the period from March 2014 to December 2015. At least one culture for mycobacteria was collected per patient per year. Results and Discussion: Seven patients had at least one sputum culture positive to nycobacteria (colonization prevalence 6.4%) and were included in the study. Of these, three patients had at least 2 positive cultures for mycobacteria. The median age of the included patients was 22 years and 4 patients were male. Four patients presented positive sputum culture for M. abscessus, 2 patients for M. avium and 1 patient for M. intracellulare. Two patients are undergoing treatment for atypical mycobacteriosis, because of worsening clinical parameters – FEV1 , CT chest and little response to antibiotical therapy – being one patient with M. avium and other with M. abscessus. Conclusion: Only two patients developed mycobacterial disease and, since the treatment is still ongoing, their clinical outcomes are not known yet. Overall, our findings show that the prevalence of mycobacteria in our center is quite low, when compared with other literature reports. A possible explanation for this scenario is the administration of the BCG vaccine, which is routine for neonates in Brazil, and protects against tuberculosis and, thus, may protect against other mycobacteria. Also, culture-based methods may fail to detect the mycobacterial presence. Thus, other methods, e.g. molecular-based and serological methods, can help to investigate the prevalence of these pathogens. 104 Monitoring and treatment of non tuberculous mycobacteria (NTM): a difficult task V. Galici1 , S. Bresci2 , B. Borchi2 , A. Cavallo2 , G. Taccetti1 , M.T. Simonetti2 , C. Braggion1 . 1 AOU A. Meyer, Florence, Italy; 2 AOU Careggi, Florence, Italy Objectives: To evaluate NTM type and prevalence and therapy efficacy in cystic fibrosis (CF) patients. Methods: We have included all patients able to providing sputum attending to CF Florence Center during 2014. Results: Since 1/1 until 31/12/2014 15 pediatric patients and 100 adult patients cultured at least 1 sputum sample for NTM (44 and 83% of patients providing sputum). The overall prevalence of NTM was 8/115 (6.9%; 6 F, 2 M); mean age was 29.4±11.9 yrs (1 child, 7 adults). Two patients had 1 colture positive for M. xenopi, 2 for MABSC and other 4 patients for MAC. Since 2013 until 2015, 7 patients started therapy for NTM according ATS guidelines: 3 patients with at least 2 positive cultures started triple therapy for MABSC: 1 patient stopped drugs because of side effects after 9 months. Conversion to MAC occurred in 1 patient at the beginning of therapy; eradication failed in 1 patient. 4 patients started therapy for MAC: eradication occurred in 2 patients, eradication failed in 1 patient; 1 patient stopped drugs because of side effects after 2 months. Conclusion: The most frequently isolated NTM were MAC and MABS. Our NTM prevalence is probably underestimated: monitoring for NTM should be improved also using sputum induction procedure. In our experience, eradication failed in 5/7 (71%) patients, in 2 of these 5 patients because of side effects. In our opinion when NTM positive cultures continue to be detected after an eradication failure, an
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
97 Nocardia and cystic fibrosis: the impact of Gram staining O. Tuncer1 , S. Olmez1 , B. Sancak1 , G.D. Tugcu2 , N. Emiralioglu2 , B. Otlu3 , 2 1 1 ¨ celik B. Er4 , E. Yalcın ¸ 2 , D. Dogru2 , U. Oz ¸ , N. Kiper2 , B. Sener ¸ . Hacettepe University Medical Faculty, Clinical Microbiology, Ankara, Turkey; 2 Hacettepe University Ihsan Dogramaci Children’s Hospital, Pediatric Pulmonology, Ankara, Turkey; 3 In¨ on¨ u University Medical Faculty, Clinical Microbiology, Malatya, Turkey; 4 Hacettepe University Medical Faculty, Pulmonology, Ankara, Turkey Objectives: We describe Nocardia isolation from multiple respiratory specimens of two CF patients with chronic Pseudomonas colonization. We aimed to investigate the genetic relatedness of the sequential Nocardia isolates in each patient. Methods: When branching Gram-positive bacilli were observed in Gram stained smears, buffered charcoal yeast extract agar was also included to the culture panel and incubation period was extended to minimum 7 days. The plates were incubated at 37o C in 5% CO2 /air environment. Smears were prepared from suspected colonies and stained by Gram and modified Ziehl-Neelsen to confirm the presence of Gram positive, variable staining, branching bacilli that were positive by modified ZiehlNeelsen. 16S rRNA gene sequence analysis was used to identify the Nocardia isolates to species level. The genetic relatedness between the isolates were analysed by pulsed field gel electrophoresis and AP-PCR. Results: In patient A N. transvalensis/N. wallacei and in patient B N. asteroides were identified. After the first isolation of N. transvalensis/ N. wallacei in sputum culture of patient A, she received TMP/SMX for 4 weeks, no new Nocardia-positive culture was obtained. Patient B received piperacillin–tazobactam for two weeks, TMP/SMX treatment still continues now. Last cultures are negative for Nocardia. Conclusion: The presence of Nocardia in sputum cultures does not always imply disease but rather simple colonization. In order to understand the impact of Nocardia in CF lungs, more reports are needed describing the clinical and microbiological features of CF patients who have Nocardia spp.-positive sputum culture. 98 The cystic fibrosis lower airways microbial metagenome P. Moran Losada1 , P. Chouvarine1 , M. Dorda1 , S. Hedtfeld1 , S. Mielke1 , 1 1 A. Schulz1 , L. Wiehlmann1 , B. Tummler ¨ . Medizinische Hochschule Hannover, Clinic for Paediatric Pneumology, Allergology and Neonatology, Hannover, Germany Objectives: Chronic airway infections determine most morbidity in people with cystic fibrosis (CF). Here we present unbiased quantitative data about the frequency and abundance of DNA viruses, archaea, bacteria, molds and fungi in CF lower airways. Methods: The microbial contents of induced sputa collected on several occasions from 10 PS and 15 Phe508del homozygous CF children, adolescents and adults was resolved by high-throughput whole genome sequencing. Results: Deep sputum metagenome sequencing identified on average about ten DNA viruses or fungi and several hundred bacterial taxa. The metagenome of a CF patient was typically found to be made up of an individual signature of multiple lowly abundant species superimposed by few disease-associated pathogens such as Pseudomonas aeruginosa and Staphylococcus aureus as major components. The host-associated signatures ranged from inconspicuous poly-microbial communities in healthy subjects to low-complexity microbiomes dominated by the typical CF pathogens in patients with advanced lung disease. The DNA virus community in CF lungs mainly consisted of phages and occasionally of human pathogens such as adeno- and herpesviruses. The S. aureus and P. aeruginosa populations were composed of one major and numerous minor clone types. The rare clones constitute a low copy genetic resource which could rapidly expand as a response to habitat alterations such as antimicrobial chemotherapy or invasion of novel microbes.
99 Investigating the airway microbiome in cystic fibrosis patients with normal and severe pulmonary function decline: an opportunity for a personalized microbiome-based therapy G. Bacci1 , P. Paganin2 , N. Segata3 , F. Armanini3 , G. Taccetti4 , D. Dolce4 , A. De Alessandri5 , P. Morelli5 , V. Tuccio6 , E.V. Fiscarelli6 , V. Lucidi6 , A. Mengoni1 , A. Bevivino2 . 1 University of Florence, Department of Biology, Florence, Italy; 2 ENEA Casaccia Research Center, Sustainable Territorial and Production Systems Department, Rome, Italy; 3 University of Trento, Centre for Integrative Biology, Trento, Italy; 4 Cystic Fibrosis Center, Anna Meyer Children’s University Hospital, Department of Pediatrics Medicine, Florence, Italy; 5 Cystic Fibrosis Center, G. Gaslini Institute, Department of Pediatrics, Genoa, Italy; 6 Children’s Hospital and Research Institute Bambino Ges` u, CF Microbiology and CF Center, Rome, Italy Objectives: To gain new insights into CF microbiome composition, metabolism and function, trying to unravel the underlying causes of the severe lung disease and identify potential determinants of serious decline in lung function. Methods: Twelve CF subjects attending three Italian CF centres were enrolled. Patients were stratified according to lung function decline rates and disease severity. Shotgun sequencing was performed following standard pipelines in Illumina Hiseq 2000 platform. To infer functional and taxonomic patterns among patients, extrinsic and intrinsic metagenomic data analysis was used. Results: We identified a core set of bacterial metabolic pathways and functions. In particular, the Kegg pathway One carbon pool by folate related to cysteine/methionine metabolism was found more abundant in patients with normal/mild disease. Moreover, antibiotic resistance genes, mainly belonging to efflux and transporter systems, were found to be more present in patients with severe lung disease. A longitudinal analysis on twenty CF patients followed over one-year is ongoing. Conclusion: The different pulmonary function in CF patients co-occurs with potentially different microbiome capabilities, mainly related to an increasing number of genes for multiple antibiotic resistance (efflux systems) and a potential differential presence of metabolic pathways. The ongoing longitudinal analysis will permit us to identify the microbial markers possible related to severe lung function decline in CF patients, and allow a risk assessment and a better focused and personalized therapy of CF patients. Acknowledgement: Supported by grants from the Italian CF Foundation (FFC #10/2014 and FFC #14/2015). 100 (More) viable cough aerosols from individuals with cystic fibrosis M.E. Wood1,2,3 , G.R. Johnson4 , R.E. Stockwell1 , L.J. Sherrard1,5 , N. Jabbour4 , K.A. Ramsay1,2 , L.D. Knibbs2 , T.J. Kidd2,5 , C.E. Wainwright2,6 , L. Morawska4 , S.C. Bell1,3 . 1 QIMR Berghofer Medical Research Institute, Brisbane, Australia; 2 The University of Queensland, Brisbane, Australia; 3 The Prince Charles Hospital, Brisbane, Australia; 4 Queensland University of Technology, Brisbane, Australia; 5 Queen’s University Belfast, Belfast, United Kingdom; 6 Lady Cilento Children’s Hospital, Brisbane, Australia Objectives: Cough-generated airborne P. aeruginosa droplet nuclei can remain viable for up 45 minutes. This prompted changes to infection control guidelines for patients with cystic fibrosis (CF). This study investigated if other CF respiratory pathogens are detectable in cough aerosols and assessed their survival over distance and time. Methods: 27 subjects with CF, mean age 28.6 years (SD 9.7); mean FEV1 63.1% (26.2) predicted were enrolled into 2 groups: 1. non-P. aeruginosa Gram-negative bacteria (GNB), 2. Gram-positive bacteria (GP), Staphylococcus aureus. Subjects performed a series of voluntary coughing into 2 validated aerosol-sampling devices to measure viability at 2 and 4 m and after 5, 15 and 45 min following generation. An Anderson Impactor collected and sized the aerosols, and total CFU (stages 1–6) was measured. Quantitative sputum and cough aerosol cultures were performed. Results: 11/16 (69%) subjects with mean sputum GNB concentration of 1.9×108 CFU/mL (8.0×103 –2.2×109 ) produced viable aerosol containing the target pathogen. The mean (range) CFU identified in cough aerosol at 4 m and at 45 min was 21.6 (0–164) and 15.6 (0–123),
Posters / Journal of Cystic Fibrosis 15 (2016) S51–S120
respectively. 10/17 (59%) subjects with mean GP sputum concentration of 2.0×107 CFU/mL (1.0×103 –1.0×108 ) produced viable cough aerosol containing S. aureus. A mean (range) of 7.1 CFU (0–55) and 1.0 CFU (0–7) were detected at 4 m and at 45 min, respectively. Conclusion: CF respiratory pathogens, other than P. aeruginosa, are viable at 4 metres and for up to 45 minutes after coughing, which highlights the need for universal and stringent infection control. Acknowledgement: Funding: CFFT (USA), TPCH Foundation 101 Survival of Mycobacterium abscessus in artificially generated aerosols L.A. Fletcher1 , Y. Chen1 , P. Whitaker2 , D.G. Peckham2 , M. Denton3 , I. Clifton2 . 1 University of Leeds, Department of Civil Engineering, Leeds, United Kingdom; 2 St James’s University Hospital, Regional Adult Cystic Fibrosis Unit, Leeds, United Kingdom; 3 Leeds General Infirmary, Department of Medical Microbiology, Leeds, United Kingdom Objectives: There is emerging evidence supporting the possibility of airborne routes for transmission for Pseudomonas aeruginosa and Burkholderia cepacia complex between people with CF. More recently M. abscessus has emerged as a potentially important pathogen in people with CF with evidence of accelerated lung function decline [2]. The aim of this study was to determine whether M. abscessus could survive within artificially demonstrated aerosols using a previously described laminar airflow model [1]. Methods: Five strains of M. abcessus isolated from patients with CF and a reference strain were studied. Aerosols were generated using a Collison 3-jet nebuliser, delivered into an airtight pipe of varying lengths and sampled using an Andersen 6-stage impactor as previously described [1]. Results: All the strains studied were able to produce viable aerosols of M. abscessus which survived 81 s and travelled a distance of 4 m. All the aerosols contained particles which were predominantly less than 2 mm in diameter. Conclusion: This study demonstrates that M. abscessus can survive within artificially generated aerosols in particles within the respirable range. Cross-infection of M. abscessus between people with CF could potentially occur via an airborne route. The survival of M. abscessus within the laminar airflow model was very similar to that of P. aeruginosa [1]. These results have important implications for the care of people with CF and reinforces the need for strict infection control practices to minimise the risk of cross-infection which pathogens such as M. abscessus. Reference(s) [1] Clifton et al. BMC Microbiol 2008. [2] Esther et al. J Cyst Fibros 2010.
102 Transmission of non-tuberculous mycobacteria in patients with cystic fibrosis C. Schwarz1 , J. Roehmel1 , L. Hatzler1 , D. Staab1 , A. Lewin2 . 1 Charit´eUniversit¨ atsmedizin Berlin, Division of Cystic Fibrosis, Berlin, Germany; 2 Robert Koch Institute, Berlin, Germany Objectives: CF patients suffer from bronchopulmonary disease exacerbations due to multiple recurrent and chronic infections of the lung. A contribution of P. aeruginosa in this process is well established but also NTM, such as Mycobacterium avium and Mycobacterium abscessus are becoming increasingly important. The exact role of NTM is not well characterized and ranges from short and long colonization to severe infection. The risk of tansmission is still under debate. Methods: We prospectively analyzed the sputum or bronchoalveolar lavage for NTM in our 330 patients. To assess the species and subspecies of mycobacteria PCR (16S rRNA gen and species specific PCR) was used. In addition, genome sequencing, susceptibility testing (including erm (41)gen), collection of clinical data of the patients with NTM, the treatment courses and side effects of the treatment were evaluated. Results: We analyzed 330 patients with CF. In 19 patients we found NTM. In 5 patients two species of NTM were detected. The leading mycobacterium was M. abscessus followed by M. avium. In two pairs of
S77
patients we could find the same clusters of NTM isolates. 14 antibiotics were used for susceptibility testing in patients with fast growing species (e.g. M. abscessus) and 13 in patients with slow growing species (e.g. M. avium). Conclusion: We could find in two pairs of patients the same cluster in there NTM isolates. This might underline the potential risk for transmission of NTM in CF. 103 Low prevalence of mycobacteria among Brazilian CF patients: possible explanations T.B. Aiello1 , R.M. Mauch1 , M.P. Arrym1 , T.R. Zaccariotto1 , A.A.D.C. Toro1 , M.C. Pereira1 , I.A. Paschoal1 , J.D. Ribeiro1 , A.F. Ribeiro1 , C.E. Levy1 . 1 Unicamp, Campinas, Brazil Objectives: To investigate the presence of opportunistic mycobacteria in CF patients attended at a reference center in Brazil. Methods: Data of 109 patients were collected in the period from March 2014 to December 2015. At least one culture for mycobacteria was collected per patient per year. Results and Discussion: Seven patients had at least one sputum culture positive to nycobacteria (colonization prevalence 6.4%) and were included in the study. Of these, three patients had at least 2 positive cultures for mycobacteria. The median age of the included patients was 22 years and 4 patients were male. Four patients presented positive sputum culture for M. abscessus, 2 patients for M. avium and 1 patient for M. intracellulare. Two patients are undergoing treatment for atypical mycobacteriosis, because of worsening clinical parameters – FEV1 , CT chest and little response to antibiotical therapy – being one patient with M. avium and other with M. abscessus. Conclusion: Only two patients developed mycobacterial disease and, since the treatment is still ongoing, their clinical outcomes are not known yet. Overall, our findings show that the prevalence of mycobacteria in our center is quite low, when compared with other literature reports. A possible explanation for this scenario is the administration of the BCG vaccine, which is routine for neonates in Brazil, and protects against tuberculosis and, thus, may protect against other mycobacteria. Also, culture-based methods may fail to detect the mycobacterial presence. Thus, other methods, e.g. molecular-based and serological methods, can help to investigate the prevalence of these pathogens. 104 Monitoring and treatment of non tuberculous mycobacteria (NTM): a difficult task V. Galici1 , S. Bresci2 , B. Borchi2 , A. Cavallo2 , G. Taccetti1 , M.T. Simonetti2 , C. Braggion1 . 1 AOU A. Meyer, Florence, Italy; 2 AOU Careggi, Florence, Italy Objectives: To evaluate NTM type and prevalence and therapy efficacy in cystic fibrosis (CF) patients. Methods: We have included all patients able to providing sputum attending to CF Florence Center during 2014. Results: Since 1/1 until 31/12/2014 15 pediatric patients and 100 adult patients cultured at least 1 sputum sample for NTM (44 and 83% of patients providing sputum). The overall prevalence of NTM was 8/115 (6.9%; 6 F, 2 M); mean age was 29.4±11.9 yrs (1 child, 7 adults). Two patients had 1 colture positive for M. xenopi, 2 for MABSC and other 4 patients for MAC. Since 2013 until 2015, 7 patients started therapy for NTM according ATS guidelines: 3 patients with at least 2 positive cultures started triple therapy for MABSC: 1 patient stopped drugs because of side effects after 9 months. Conversion to MAC occurred in 1 patient at the beginning of therapy; eradication failed in 1 patient. 4 patients started therapy for MAC: eradication occurred in 2 patients, eradication failed in 1 patient; 1 patient stopped drugs because of side effects after 2 months. Conclusion: The most frequently isolated NTM were MAC and MABS. Our NTM prevalence is probably underestimated: monitoring for NTM should be improved also using sputum induction procedure. In our experience, eradication failed in 5/7 (71%) patients, in 2 of these 5 patients because of side effects. In our opinion when NTM positive cultures continue to be detected after an eradication failure, an