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Infection and Immunity, June 2005, p. 3210-3218, Vol. 73, No. 6
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.6.3210-3218.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Role of Sialic Acid and Complex Carbohydrate Biosynthesis in Biofilm Formation by Nontypeable Haemophilus influenzae in the Chinchilla Middle Ear

Joseph Jurcisek,1 Laura Greiner,2 Hiroshi Watanabe,2 Anthony Zaleski,2 Michael A. Apicella,2 and Lauren O. Bakaletz1*

Columbus Children's Research Institute and the College of Medicine & Public Health, The Ohio State University, Columbus, Ohio,1 The Department of Microbiology, The University of Iowa, Iowa City, Iowa2

Received 2 February 2005/ Accepted 10 February 2005

Nontypeable Haemophilus influenzae (NTHI) is an important pathogen in respiratory tract infections, including otitis media (OM). NTHI forms biofilms in vitro as well as in the chinchilla middle ear, suggesting that biofilm formation in vivo might play an important role in the pathogenesis and chronicity of OM. We've previously shown that SiaA, SiaB, and WecA are involved in biofilm production by NTHI in vitro. To investigate whether these gene products were also involved in biofilm production in vivo, NTHI strain 2019 and five isogenic mutants with deletions in genes involved in carbohydrate biosynthesis were inoculated into the middle ears of chinchillas. The wild-type strain formed a large, well-organized, and viable biofilm; however, the wecA, lsgB, siaA, pgm, and siaB mutants were either unable to form biofilms or formed biofilms of markedly reduced mass, organization, and viability. Despite their compromised ability to form a biofilm in vivo, wecA, lsgB, and siaA mutants survived in the chinchilla, inducing culture-positive middle ear effusions, whereas pgm and siaB mutants were extremely sensitive to the bactericidal activity of chinchilla serum and thus did not survive. Lectin analysis indicated that sialic acid was an important component of the NTHI 2019 biofilm produced in vivo. Our data suggested that genes involved in carbohydrate biosynthesis and assembly play an important role in the ability of NTHI to form a biofilm in vivo. Collectively, we found that when modeled in a mammalian host, whereas biofilm formation was not essential for survivability of NTHI in vivo, lipooligosaccharide sialylation was indispensable.

* Corresponding author. Mailing address: Center for Microbial Pathogenesis, Columbus Children's Research Institute, Department of Pediatrics, The Ohio State University, College of Medicine and Public Health, 700 Children's Drive, Rm. W591, Columbus, OH 43205-2696. Phone: (614) 722-2915. Fax: (614) 722-2818. E-mail: BakaletL{at}pediatrics.ohio-state.edu.

Editor: D. L. Burns

Infection and Immunity, June 2005, p. 3210-3218, Vol. 73, No. 6
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.6.3210-3218.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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