Microbial investigations in throat swab and tracheal aspirate specimens are beneficial to predict the corresponding endotracheal tube biofilm flora among intubated neonates with ventilator‑associated pneumonia

Pan,Yun; Du,Lizhong; Ai,Qing; Song,Sijie; Tang,Xiaoli; Zhu,Danping; Yu,Jialin

doi:10.3892/etm.2017.4631

Microbial investigations in throat swab and tracheal aspirate specimens are beneficial to predict the corresponding endotracheal tube biofilm flora among intubated neonates with ventilator‑associated pneumonia

Authors:
- Yun Pan
- Lizhong Du
- Qing Ai
- Sijie Song
- Xiaoli Tang
- Danping Zhu
- Jialin Yu
View Affiliations

Affiliations: Department of Neonatology, Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China, Department of Pediatrics, Children's Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003, P.R. China, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing 400014, P.R. China
Published online on: June 20, 2017 https://doi.org/10.3892/etm.2017.4631
Pages: 1450-1458

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Ventilator‑associated pneumonia (VAP) is a common nosocomial infection in neonatal intensive care units with high morbidity and mortality. Bacterial biofilm in the endotracheal tube (ET) provides a notable and persistent source of pathogens that may cause VAP, and thus is important for VAP detection. However, during intubation microbial investigations in ET, samples are unavailable due to the infeasibility of collecting ET samples during intubation of neonates. It is therefore of great importance to find alternative sources of samples that can help identify the ET biofilm flora. In the present study, the microbial signatures of throat swabs and tracheal aspirates were compared with ET biofilm samples from VAP neonates using 16S ribosomal RNA gene polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE), cloning and sequencing. Sequences were assigned to phylogenetic species using BLAST. Microbial diversity and richness among the three types of specimens were compared based on their DGGE fingerprints, and taxonomic characteristics based on the BLAST results. The microbial richness and diversity of ET biofilms were similar to tracheal aspirate yet significantly different from throat swab samples (P<0.05). Compared with ET biofilms, the overall constituent ratio of microflora was significantly different in throat swab and tracheal aspirate samples (P<0.05). However tracheal aspirate samples were useful for predicting Staphylococcus sp. in ET biofilms with a sensitivity of 85.7% and a specificity of 83.3%. The sensitivity for the combination of tracheal aspirate and throat swab samples to detect Staphylococcus sp. in ET biofilms was 100%. The detection of Pseudomonas sp. in throat swabs assisted its identification in ET biofilms (sensitivity 33.3% and specificity 100%). The results of the present study suggest that microbial investigations in throat swab and tracheal aspirate samples are beneficial for identifying the ET biofilm flora. There may therefore be clinical applications of using substituent samples to identify pathogens in ET biofilms for VAP surveillance among intubated neonates.

View Figures

View References

1	National Nosocomial Infections Surveillance System, . National Nosocomial Infections Surveillance (NNIS) System Report, data summary from January 1992 through June 2004, issued October 2004. Am J Infect Control. 32:470–485. 2004. View Article : Google Scholar : PubMed/NCBI
2	Aly H, Badawy M, El-Kholy A, Nabil R and Mohamed A: Randomized, controlled trial on tracheal colonization of ventilated infants: Can gravity prevent ventilator-associated pneumonia? Pediatrics. 122:770–774. 2008. View Article : Google Scholar : PubMed/NCBI
3	Tan B, Zhang F, Zhang X, Huang YL, Gao YS, Liu X, Li YL and Qiu JF: Risk factors for ventilator-associated pneumonia in the neonatal intensive care unit: A meta-analysis of observational studies. Eur J Pediatr. 173:427–434. 2014. View Article : Google Scholar : PubMed/NCBI
4	Tan B, Xian-Yang X, Zhang X, Peng-Zhou X, Wang P, Xue J, Ling-Huang Y, Li-Li Y and Fu-Qiu J: Epidemiology of pathogens and drug resistance of ventilator-associated pneumonia in Chinese neonatal intensive care units: A meta-analysis. Am J Infect Control. 42:902–910. 2014. View Article : Google Scholar : PubMed/NCBI
5	Inglis T, Millar M, Jones J and Robinson D: Tracheal tube biofilm as a source of bacterial colonization of the lung. J Clin Microbiol. 27:2014–2018. 1989.PubMed/NCBI
6	Adair C, Gorman S, Feron B, Byers L, Jones D, Goldsmith C, Moore J, Kerr J, Curran M, Hogg G, et al: Implications of endotracheal tube biofilm for ventilator-associated pneumonia. Intensive Care Med. 25:1072–1076. 1999. View Article : Google Scholar : PubMed/NCBI
7	Sottile FD, Marrie TJ, Prough DS, Hobgood CD, Gower DJ, Webb LX, Costerton JW and Gristina AG: Nosocomial pulmonary infection: Possible etiologic significance of bacterial adhesion to endotracheal tubes. Crit Care Med. 14:265–270. 1986. View Article : Google Scholar : PubMed/NCBI
8	Gil-Perotin S, Ramirez P, Marti V, Sahuquillo JM, Gonzalez E, Calleja I, Menendez R and Bonastre J: Implications of endotracheal tube biofilm in ventilator-associated pneumonia response: A state of concept. Crit Care. 16:R932012. View Article : Google Scholar : PubMed/NCBI
9	Cairns S, Thomas JG, Hooper SJ, Wise MP, Frost PJ, Wilson MJ, Lewis MA and Williams DW: Molecular analysis of microbial communities in endotracheal tube biofilms. PLoS One. 6:e147592011. View Article : Google Scholar : PubMed/NCBI
10	Vandecandelaere I, Matthijs N, Van Nieuwerburgh F, Deforce D, Vosters P, De Bus L, Nelis HJ, Depuydt P and Coenye T: Assessment of microbial diversity in biofilms recovered from endotracheal tubes using culture dependent and independent approaches. PLoS One. 7:e384012012. View Article : Google Scholar : PubMed/NCBI
11	Perkins SD, Woeltje KF and Angenent LT: Endotracheal tube biofilm inoculation of oral flora and subsequent colonization of opportunistic pathogens. Int J Med Microbiol. 300:503–511. 2010. View Article : Google Scholar : PubMed/NCBI
12	Charlson ES, Bittinger K, Haas AR, Fitzgerald AS, Frank I, Yadav A, Bushman FD and Collman RG: Topographical continuity of bacterial populations in the healthy human respiratory tract. Am J Respir Crit Care Med. 184:957–963. 2011. View Article : Google Scholar : PubMed/NCBI
13	Pneumatikos IA, Dragoumanis CK and Bouros DE: Ventilator-associated pneumonia or endotracheal tube-associated pneumonia? An approach to the pathogenesis and preventive strategies emphasizing the importance of endotracheal tube. Anesthesiology. 110:673–680. 2009. View Article : Google Scholar : PubMed/NCBI
14	Vandecandelaere I, Matthijs N, Nelis HJ, Depuydt P and Coenye T: The presence of antibiotic-resistant nosocomial pathogens in endotracheal tube biofilms and corresponding surveillance cultures. Pathog Dis. 69:142–148. 2013. View Article : Google Scholar : PubMed/NCBI
15	Costerton JW, Post JC, Ehrlich GD, Hu FZ, Kreft R, Nistico L, Kathju S, Stoodley P, Hall-Stoodley L, Maale G, et al: New methods for the detection of orthopedic and other biofilm infections. FEMS Immunol Med Microbiol. 61:133–140. 2011. View Article : Google Scholar : PubMed/NCBI
16	Wang Y, Hoenig JD, Malin KJ, Qamar S, Petrof EO, Sun J, Antonopoulos DA, Chang EB and Claud EC: 16S rRNA gene-based analysis of fecal microbiota from preterm infants with and without necrotizing enterocolitis. ISME J. 3:944–954. 2009. View Article : Google Scholar : PubMed/NCBI
17	Lu H, Qian G, Ren Z, Zhang C, Zhang H, Xu W, Ye P, Yang Y and Li L: Alterations of Bacteroides sp., Neisseria sp., Actinomyces sp. and Streptococcus sp. populations in the oropharyngeal microbiome are associated with liver cirrhosis and pneumonia. BMC Infect Dis. 15:2392015. View Article : Google Scholar : PubMed/NCBI
18	Liu D, Yu J, Li L, Ai Q, Feng J, Song C and Li H: Bacterial community structure associated with elective cesarean section versus vaginal delivery in Chinese newborns. J Pediatr Gastroenterol Nutr. 60:240–246. 2015. View Article : Google Scholar : PubMed/NCBI
19	Centers for Disease Control and Prevention: Criteria for defining nosocomial pneumonia. http://www cdc gov/ncidod/hip/NNIS/members/pneumonia/Final/PneumoCriteriaV1pdfFebruary 5–2009
20	Cernada M, Brugada M, Golombek S and Vento M: Ventilator-associated pneumonia in neonatal patients: An update. Neonatology. 105:98–107. 2014. View Article : Google Scholar : PubMed/NCBI
21	Lu W, Yu J, Ai Q, Liu D, Song C and Li L: Increased constituent ratios of Klebsiella sp., Acinetobacter sp. and Streptococcus sp. and a decrease in microflora diversity may be indicators of ventilator-associated pneumonia: A prospective study in the respiratory tracts of neonates. PLoS One. 9:e875042014. View Article : Google Scholar : PubMed/NCBI
22	Jones RN: Microbial etiologies of hospital-acquired bacterial pneumonia and ventilator-associated bacterial pneumonia. Clin Infect Dis. 51 Suppl 1:S81–S87. 2010. View Article : Google Scholar : PubMed/NCBI
23	Feldman C, Kassel M, Cantrell J, Kaka S, Morar R, Mahomed AG and Philips J: The presence and sequence of endotracheal tube colonization in patients undergoing mechanical ventilation. Eur Respir J. 13:546–551. 1999. View Article : Google Scholar : PubMed/NCBI
24	Wolcott RD and Ehrlich GD: Biofilms and chronic infections. Jama. 299:2682–2684. 2008. View Article : Google Scholar : PubMed/NCBI
25	Zhu XL, Zhao L, Yang JC, Chen X and Wu XH: Etiology and high risk factors of neonatal ventilator-associated pneumonia. Zhongguo Dang Dai Er Ke Za Zhi. 9:549–552. 2007.(In Chinese). PubMed/NCBI
26	Safdar N, Crnich CJ and Maki DG: The pathogenesis of ventilator-associated pneumonia: Its relevance to developing effective strategies for prevention. Respir Care. 50:725–741. 2005.PubMed/NCBI
27	Brennan MT, Bahrani-Mougeot F, Fox PC, Kennedy TP, Hopkins S, Boucher RC and Lockhart PB: The role of oral microbial colonization in ventilator-associated pneumonia. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 98:665–672. 2004. View Article : Google Scholar : PubMed/NCBI
28	Johanson WG Jr, Pierce AK, Sanford JP and Thomas GD: Nosocomial respiratory infections with gram-negative bacilli: The significance of colonization of the respiratory tract. Ann Intern Med. 77:701–706. 1972. View Article : Google Scholar : PubMed/NCBI
29	Rotimi V and Duerden B: The development of the bacterial flora in normal neonates. J Med Microbiol. 14:51–62. 1981. View Article : Google Scholar : PubMed/NCBI
30	Song S, Du L, Yu J, Ai Q, Pan Y, Fu Y and Wang Z: Does Streptococcus mitis, a neonatal oropharyngeal bacterium, influence the pathogenicity of Pseudomonas aeruginosa? Microbes Infect. 17:710–716. 2015. View Article : Google Scholar : PubMed/NCBI