Center for Biofilm Engineering News Update:  June, 2002 Volume 5, Issue 6   __________________________________________   Research Highlights   Paper Spotlight   Mucosal Biofilm Formation on Middle-Ear Mucosa in the Chinchilla Model  of Otitis Media, by Richard Veeh   For children in the United States, otitis media (OM) is the most prevalent cause of visits to physicians and of prescriptions for antibiotic treatment. Although it is known that OM results from a complex interaction of infectious agent behavior, environmental conditions, and host-related factors, the specific underlying mechanisms of pathophysiology are poorly understood. For many cases of acute OM, middle ear effusions are culture-positive for known pathogenic bacterial species, predominantly Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis. However, for cases of chronic OM with effusion (OME), many culture-negative test results are obtained. Because these chronic cases have also exemplified resistance to antibiotic treatments and positive testing for inflammatory mediators, it has been assumed that OME is not the result of a direct bacterial process but rather is a sterile, inflammatory process directed against residual bacterial metabolites. Several lines of evidence, however, do not support this latter assumption: 1) many "sterile" pediatric effusions contain bacterial DNA. 2) live infectious bacteria persist in culture-negative effusions for several weeks after antibiotic treatment. 3) the presence of bacterial mRNA in culture-negative effusions demonstrates that bacteria are alive and metabolically active. Because evidence is accumulating that bacterial biofilms may enhance antibiotic resistance and confound standard culturing methods, this study was undertaken to determine by direct imaging whether bacterial biofilm formation occurs on middle-ear mucosa. In this study, a total of 48 young adult chinchillas were used for this purpose in two series of experiments. Two animals infected with H. influenzae were euthanized at 10 different times between 3 hours and 22 days after inoculation, with several animals serving as uninoculated controls. Prepared middle-ear specimens were analyzed for biofilm morphology using scanning electron microscopy (SEM) and confocal scanning laser microscopy (CSLM) coupled with different staining procedures. Use of these two imaging techniques revealed the presence of both microcolonies and more extensive biofilm formation on chinchilla middle ear mucosa as early as 24 hours after infection. Our findings provide substantial evidence that mucosal biofilms exist in the chinchilla model of otitis media and support the notion that biofilm formation may be an important factor in the pathogenesis of chronic OME.  Mucosal biofilm formation on middle-ear mucosa in the chinchilla model of otitis media Ehrlich GD, Veeh R, Wang X, Costerton JW, Hayes JD, Hu FZ, Daigle BJ, Ehrlich MD, Post JC JAMA April 3, 2002; 287(13):1710-1715   _______   Recent Media Coverage   Freelance writer, Cathryn Delude, followed the JAMA article on otitis media and published "Culprit in ear infections is a 'biofilm' that protects bacteria," in the Boston Globe, May 28th. She has also written "Microbes Coming into Focus," the Spring 2002 issue of Your World magazine. One article specifically addresses biofilms and is entitled, "Biofilms and Quorum Sensing: The Gang's All Here." Your World is published by the Biotechnology Institute and can be found online at http://www.BiotechInstitute.org. Links to both articles can be found on the CBE Media Coverage web page. http://www.erc.montana.edu/CBEssentials-SW/recent-media-cov/default.html   _______   Technical Advisory Conference   The July 2002 Technical Advisory Conference will include sessions on Medical Biofilms, Cell Signaling, Biofilm Control, Bioremediation, Biofilm Methods, a regulatory update, and others. The Conference will also feature basic and advanced workshops teaching hands-on laboratory techniques to attendees. For further information, please contact Paul Sturman at paul_stu@erc.montana.edu. _______   Biofilm Image Analysis Workshop July 25-26, 2002 Center for Biofilm Engineering Montana State University Bozeman, MT, U.S.A More registrants are needed for the Image Analysis workshop. Please see Workshop details and registration forms on this web-posted pdf file. http://www.erc.montana.edu/CBEssentials-SW/MicrosensorWorkshop/ImageAnalysisWorkshop.pdf _______   Academic Member of the Surfaces in Biomaterials Foundation   The CBE recently joined as an academic member with the Surfaces in Biomaterials Foundation. As a member, the CBE will have the opportunity to interface with industries involved in surface modifications in the medical industry, and also with industries dealing with the prevention of biofilm formation and with biocompatibility issues. Other academic members of the Foundation are the University of Arizona, University of Minnesota, and the University of Washington.     _________________________________________   Education   Summer REU Students and Projects The CBE is looking forward to working with eight, talented undergraduates who were selected for the summer Research Experience for Undergraduates (REU) program. The students began the ten-week program on June 3rd with a week of lab tours and workshops to orient them to the Center. Listed below are the student names, their projects, and project mentors. Assessment of lipophilic stains for multiple stain visualization of bacterial biofilms REU Student: Jessica Janzen Mentor: Luanne Hall-Stoodley, CBE  Determine the effect previous use has on biofilm accumulation in hot tub filters REU Student: Kristin Susens Mentor: Darla Goeres, CBE Develop a protocol for reproducible biofilms that simulates biofilms found in oral cavities REU Student: Ryan Winters Mentors: Mark Pasmore, CBE and Melvin H. Czechowski, Ph.D., Church & Dwight, Inc. Identification and characterization of microbial biofilm populations existing in arsenical-sulfide hot springs of Yellowstone National Park REU Student: William Franck Mentor: Timothy McDermott, Land Resources and Environmental Science Investigating the potential of vaccine development in Staphylococcal biofims  REU Student: Kimberly White Mentor: Jeff Leid, CBE  Evaluation of a method to automate BDOC measurements in drinking water REU Student: Kori Watts Mentor: Phil Butterfield, CBE Rheology of dental plaque biofilms REU Student: Heather Adams Mentors: Paul Stoodley, CBE and Douglas Dudgeon, Philips Oral Healthcare Dual plasmid reporter systems for the elucidation of transcriptional regulator function in gram-positive bacterial species REU Student: Peter Newell Mentor: Mark Shirtliff, CBE _________________________________________   Awards and Honors   College of Engineering Awards   The College of Engineering held the annual staff appreciation banquet in May, where historically, outstanding teaching awards were given. However, this year awards were also given to outstanding researchers and outstanding professional staff. It is our pleasure to announce that three outstanding people from the CBE were among those chosen for these awards. Drs. Zbigniew Lewandowski and Marty Hamilton received Outstanding Researcher Awards, and Peg Dirckx received an Outstanding Professional Staff Award.   _______   Excellence in Surface Science Award   Dr. Bill Costerton received the 2002 Excellence in Surface Science Award from the Surfaces in Biomaterials Foundation. The Foundation recognized Bill's achievements and his early interest in microbial ecology, which led him to study bacteria attached to surfaces or biofilms. He has studied biofilms in many environments ranging from mountain streams, to industrial systems, and to medical devices implanted in humans. He has also held several prestigious positions in his career, and since 1993 has been the Director of the Center for Biofilm Engineering at Montana State University.   _________________________________________   Upcoming CBE Workshops and Meetings     Industrial Associate Conference and TAC Meeting July 23-25, 2002 Montana State University   _______   Biofilm Image Analysis Workshop July 25-26, 2002 Center for Biofilm Engineering Montana State University Bozeman, MT, U.S.A Daily workshop activities will include lectures and laboratory sessions. The lectures will cover biofilm structure and image analysis of biofilms. The lab sessions will cover how to acquire biofilm images and how to use the biofilm image analysis software developed by the Biofilm Structure and Function Research Group. For workshop details and registration forms, see  http://www.erc.montana.edu/CBEssentials-SW/research/Structure_function/default.htm   _______   Microsensors: Manufacture and Applications Workshop August 19-23, 2002 Center for Biofilm Engineering Montana State University Bozeman, MT, U.S.A Microsensors (pH, dissolved oxygen, ion selective microelectrodes) are becoming indispensable tools for studying biofilms and small biological samples. The goal of this workshop is to provide the participants with the knowledge necessary to design, manufacture, and apply such microsensors. Lab sessions and lectures will illustrate the electrochemical principles of the measurements, explain how microsensors are manufactured, and demonstrate how they are applied to study biofilms.  The workshop is sponsored by the Biofilm Structure and Function Research Group at the Center for Biofilm Engineering, Montana State University. The activities include four days of lectures and lab sessions, and a one-day field trip to Yellowstone National Park.  For details on this workshop see the following web address. http://www.erc.montana.edu/CBEssentials-SW/research/Structure_function/default.htm See the Events Calendar for more conference and workshop information at http://www.erc.montana.edu/CBEssentials-SW/whats_new/index.htm   __________________________________________   CBE People in Action   Bill Costerton was an invited speaker at the Canadian Biofilm Meeting, Biofilms - 2002, Working in Partnership, Lake Louise, Alberta, Canada, May 1-2, 2002. He presented "Biofilms in Infections." Anne Camper, American Academy for Microbiology Colloquium, Session Chair, Member of Organizing Committee, Tucson, AZ, May 3-5, 2002. Bill Costerton, presented "The Organization and Function of Microbial Communities," at the American Academy for Microbiology Colloquium, Session Chair, Tucson, AZ, May 3-5, 2002.  Phil Stewart as invited speaker presented “Persistent Infections and Biofilms,” Infectious Disease Association of California Spring Symposium, Carlsbad, California, May 5, 2002. Phil Stewart as invited speaker presented “Microbial Biofilms and Their Control,” to the Diversa Corporation, San Diego, California, May 6, 2002. Thomas Borch presented a poster entitled "Spatial Distribution and Biochemistry of Bacterial Attachment to Basalt Using Synchrotron Radiation-Based Fourier Transform Infrared Spectromicroscopy (SR-FTIR)," at the Synchrotron Environmental Science - II, Argonne National Laboratory, Argonne, Illinois, May 6-8, 2002. Poster authors were Mary Kauffman and Thomas Borch of Montana State University and Michael C. Martin, Lawrence Berkeley National Laboratory, Berkeley, CA.. Anne Camper was invited to present "REU at the CBE: A Rewarding Experience" at the American Control Conference (ACC), Anchorage, AK, May 8, 2002. Bill Costerton and Anne Camper gave a workshop presentation entitled "Engineering Solutions for Dental Biofilms," at the Organization for Safety & Asepsis Procedures (OSAP) 2002 Annual Symposium, Nashville, TN, May 16-17, 2002. Bill Costerton was invited to give a seminar for the Miller Distinguished Lecturer series of The Microbiology Graduate Student Organization (MGSO), Iowa State University, May 16, 2002. He presented, "Biofilms as Complex Multicellular Communities."  Bill Costerton was an invited speaker at the Organization for Safety & Asepsis Procedures (OSAP) 2002 Annual Symposium in Nashville, TN, May 17, 2002. Bill presented "Engineering Solutions for Dental Biofilms." Phil Stewart as invited speaker presented “Microbial Biofilms,” at PPG Industries, Allison Park, Pennsylvania, May 21, 2002. Numerous people from the CBE attended and presented at the 102nd General Meeting of the American Society for Microbiology, Salt Lake City, UT, May 18-24, 2002. Paul Stoodley was invited to present a workshop, "Biofilms IV" and to present a poster entitled "Biofilm Mechanics and Dynamic Surface Transport and Detachment Behavior." Poster authors were C. Rupp, R. Cargo, and P. Stoodley. Mark Shirtliff presented a poster entitled "Interaction of Leukocytes and Staphylococcus aureus Biofilms." Poster authors were M. Shirtliff, P. Stoodley, A.K. Camper, and J. Leid. Suzanne Wilson presented at poster entitled "Detachment Distribution of Single Cells and Cell Clusters from Pseudomonas aeruginosa and Staphylococcus aureus Biofilms." Poster authors were S. Wilson, M. Franklin, J.W. Costerton, and P. Stoodley. Laura Purevdorj presented a poster entitled "The Role of Hydrodynamics in Shaping Quorum Sensing Expression in Pseudomonas aeruginosa Biofilms." Poster authors were L. Purevdorj, M. Kirisits, J.W. Costerton, M. Parsek, and P. Stoodley. __________________________________________   Theses   See theses abstracts at http://www.erc.montana.edu/Res-Lib99-SW/pubs/Theses/default.htm     __________________________________________   Web Watch   Current Opinion in Biotechnology   The June issue of Current Opinion in Biotechnology acknowledged a number of web sites relevant to this issue's topic of "Environmental Biotechnology". Included in the listing were both the web sites of the CBE and BiofilmsOnline.com.   BiofilmsOnline.com   ASM's Education Department invited the Biofilm Institute, Inc. to provide materials for the Science Teacher Day held in conjunction with the ASM General Meeting, May 22, 2002, in Salt Lake City, UT. The Institute recently received funding from the Waksman Foundation for Microbiology to provide educational resources for teachers, consumers and the media. See the June issue of BiofilmsOnline.com at http://www.BiofilmsOnline.com.    _______   Recent Web Updates   Check the Recent Web Updates link located on the CBE home page, to view the latest information posted on the CBE web.   See <http://www.erc.montana.edu/Recent%20Web%20Updates/default.htm>    __________________________________________   Newsletter Listserve   The CBE News Update is a listserve newsletter. If you need to subscribe or unsubscribe from the listserve, follow the directions at the following CBE website.   http://www.erc.montana.edu/Res-Lib99-SW/newsarchives/subscribe.htm   An alternative to subscribing to the listserve is to view the CBE News Update on our web page at http://www.erc.montana.edu/Ind-Col99-SW/Current_Newsletter/default.htm. Newsletter archives can be found at http://www.erc.montana.edu/Res-Lib99-SW/newsarchives/index.htm   __________________________________________   Diane Williams  editor of the CBE News Update