Center for Biofilm Engineering News Update:  April, 2001 Volume 4, Issue 4   __________________________________________   Summer 2001 TAC Meeting   The CBE Technical Advisory Committee (TAC) Meeting has been scheduled for July 24-26, 2001 at the Montana State University-Bozeman campus, in Bozeman, Montana. If you are interested in attending, please contact Paul Sturman, phone: (406) 994-2102 or email: paul_stu@erc.montana.edu.   Workshops will take place on Monday, July 23rd and research presentations will follow on Tuesday through Thursday, July 24th - 26th.     __________________________________________   Research Highlights   Paper Spotlight   Toilet Bowl Biofilm Method   Aims. The purpose of this study was to develop a laboratory biofilm growth reactor system that simulates the toilet bowl environment and can be used for biocide efficacy testing.   Methods & Results. A microbial biofilm reactor system incorporating intermittent flow and nutrient provision was designed. The reactor system was open to the air and was inoculated with organisms collected from toilet bowl biofilms. Once an hour, reactors were supplied with a nutrient solution for a period of five minutes, then drained and refilled with tap water or tap water amended with chlorine. Quantitative measures of the rate and extent of biofilm accumulation were defined. Biofilm accumulated in untreated reactors to cell densities of 108 cfu/cm2 after approximately one week. Biofilm accumulated in laboratory reactors approximately seven times faster than it did in actual toilet bowls. Biofilm accumulated even in the continuous presence of several milligrams per liter of free chlorine. When identical independent experiments were run, the repeatability standard deviation was low for each of the four selected efficacy measures, indicating that the reactor system provided repeatable results. In fact, the repeatability standard deviation of log reduction values was as small as for standard suspension and hard surface disinfection tests. For three different concentrations of chlorine, parallel efficacy tests were run in real toilets and in the laboratory reactors. The lab and field tests produced the same concentration-response relationship with three of the four efficacy measures. For chlorine at least, the reactor system provides efficacy test results that are relevant to the real world application.   Conclusions. This reactor system has been shown to simulate the accumulation of bacterial biofilm that occurs in toilet bowls. The results demonstrate that this system can provide relevant, repeatable, and quick assays of the efficacy of chlorine for inhibiting biofilm growth. Significance and Impact of the study. The laboratory biofilm reactor system described herein can be used to evaluate potential antimicrobial and anti-fouling treatments for control of biofilm formation in toilet bowls.   "A Repeatable Laboratory Method for Testing the Efficacy of Biocides Against Toilet Bowl Biofilms," has been accepted for publication in the Journal of Applied Microbiology. The authors are: BETSEY PITTS1, ALAN WILLSE4, GORDON A MCFETERS3, MARTIN A HAMILTON4, NICHOLAS ZELVER1, and PHILIP S STEWART2* (Center for Biofilm Engineering,1 and Departments of Chemical Engineering,2 Microbiology3, and Mathematical Sciences,4 Montana State University, Bozeman, Montana 59717).   Special Note: Because the results of this project were so encouraging, the CBE subsequently conducted evaluations of the Rotating Disk Reactor system in a different context; namely, (i) as a continuous-flow stirred tank reactor, rather than fill and drain, and (ii) where the biocide is applied after the biofilm has grown to steady state, rather than during growth. A paper presenting those evaluations will appear next month in the Biofilms II edition of Methods in Enzymology.   _______   Collaborative Otitis Media Research   Dr. Bill Costerton and Dr. Richard Veeh have been working collaboratively on otitis media research with Dr. Garth Ehrlich of the Allegheny General Hospital in Pittsburgh, PA. Dr. Ehrlich presented a poster entitled "Direct Electronmicrographic Imaging of Biofilms on the Middle-ear Mucosa," by Dr. Garth Ehrlich, Jay Hayes, Rick Veeh, Fen Hu, Xue Wang, and J. Costerton, at the 24th Annual Association for Research in Otolaryngology MidWinter Meeting, St. Petersburg Beach, Florida, February 6, 2001.   CBE is a subcontractor of the NIH sponsored project, "The Role of Biofilm in the Pathogenesis of Otorrhea". The project is expected to continue for several years. Others involved in the project are CBE researchers Phil Stewart, Betsey Pitts, Karin Sauer, and Dave Davies from Binghamton University, New York.   __________________________________________   Industry Opportunity   Drain and Fill Whirlpool Bathtubs   A new product niche is developing in the area of drain and fill whirlpool bathtubs. Dr. Bill Costerton and Ms. Darla Goeres were involved in a research project which investigated the development of biofilm in a drain and fill whirlpool bathtub. The project was a result of a class action lawsuit filed against Jacuzzi Inc. and Kohler Co. The results showed that biofilm did accumulate in the jet recirculation piping during normal use. In response to these findings and the lawsuit, an opportunity exists for a company to develop a product and cleaning protocol that would more effectively remove biofilm from the piping system of the drain and fill whirlpool bathtub.     _______   Water Quality   Industry Outlook - 2/1/01 12:43:06 PM Water groups urge World Health Organization (WHO) to study heterotrophic plate counts (HPC) in Lisle, IL. The Water Quality Association (WQA) and Aqua Europa, a national federation of water associations in Europe, are urging WHO to hold a forum exploring the issues surrounding heterotrophic bacteria, a range of microorganisms that can be found in drinking water.   European regulatory officials in recent months have been using the heterotrophic bacteria issue as a regulatory threat to the POU water treatment industry, according to WQA Executive Director Peter Censky.   Officials are attempting to require testing and increased monitoring of POU equipment, even though there is no proof that heterotrophic bacteria are health hazards, Censky said.   WQA and Aqua Europa are seeking the forum in hopes that the WHO would reaffirm that heterotrophic bacteria are harmless, according to Censky. "We believe this is a false issue in that HPC poses no threat to human health," Censky said. "Yet they [European officials] are trying to force the industry to add expensive control mechanisms, which could reduce the market and send a message that the equipment is dangerous."   Censky expects the WHO conference could take as long as 18 months to set up. HPCs will be one of the topics on the agenda at WQA's annual convention in Orlando, FL, next month.   __________________________________________   Education   Dr. Luanne Hall-Stoodley was invited to participate on a panel review of the NSF's Integrative Graduate Education and Research Traineeship (IGERT) in Arlington, VA, March 19-20, 2001. Three panels reviewed 68 proposals.   Initiated in 1997, the IGERT program was developed to meet the challenges of educating Ph.D. scientists and engineers with the multidisciplinary backgrounds and the technical, professional, and personal skills needed for the career demands of the future. The program is intended to catalyze a cultural change in graduate education, for students, faculty, and universities, by establishing new, innovative models for graduate education in a fertile environment for collaborative research that transcends traditional disciplinary boundaries. It is also intended to facilitate greater diversity in student participation and preparation and to contribute to the development of a diverse, globally-aware, science and engineering workforce.   IGERT is an NSF-wide endeavor involving the Directorates for Biological Sciences; Computer and Information Science and Engineering; Education and Human Resources; Engineering; Geosciences; Mathematical and Physical Sciences; Social, Behavioral, and Economic Sciences; and the Office of Polar Programs.   _______   Student Awards - Congratulations!   First Place in Paper Competition   Robin Gerlach won first place in the Student Paper Competition for the In Situ and On-Site Bioremediation Symposium which will be held in San Diego, CA, June 4-7, 2001. His paper was titled, "Metal-Reducing Bacteria Facilitate the Geochemical Elimination of Cr(VI) from Contaminated Water." Second and third places were from Oregon State University and the University of Virginia, respectively. Robin competed with over 50 papers submitted to the national competition. He will receive complimentary full registration to the Bioremediation Symposium, including all functions and a copy of the Proceedings. In addition, he will receive $1,500 cash towards travel expenses. The cash award is provided through corporate donations from Probe-Lease, URS Corporation, CH2M Hill, and Battelle Memorial Institute. The conference typically attracts over 2,000 attendees.   Undergraduate Research Fellowship Award   Kate McInnerney, a CBE Undergraduate Scholars Program student, has been selected to participate in the 2001 Undergraduate Research Fellowship (URF) Program sponsored by the American Society for Microbiology (ASM). The award is for a minimum of 8 weeks of summer research beginning in June 2001. She will receive a $2,500 stipend, $600 supply budget and up to $900 for student travel to present her research abstract and to participate in other activities at the 2002 ASM General Meeting in Salt Lake City, UT. The title of her project is "Characterization of Alginate Binding Domains in Alginate Modifying Enzymes."   Undergraduate Scholarship Program Awards   This spring, four CBE students were awarded undergraduate scholarships to work on the following biofilm-related research projects.   Biofilm Growth on Catheters           Student: Janice McKenzie (Chemical Engineering) PI: Ryan Jordan   Characterization of Alginate Binding Domains in Alginate Modifying Enzymes           Student: Kate McInnerney (Biotechnology) PI: Mike Franklin   Characterization of Bioactive Peptides of Pseudomonas aeruginosa           Student: Julie Garcia (Microbiology) PI: Mike Franklin   Indirect Microbial Alteration of Reducible Metal Ions           Student: Kristian Paul (Environmental, Soil & Water Science) PI: Robin Gerlach   __________________________________________   Visiting Student   Aurelie Bonin is working on the W. M. Keck Foundation project on microbial biofilm development. She is developing and testing green fluorescent protein (GFP) reporter gene constructs in Pseudomonas aeruginosa that will be used to map patterns of growth in biofilms. Aurelie is a biochemical engineering student from the Institut National des Sciences Appliques in Lyon, France.   __________________________________________   Visiting Faculty   Dr. Mary Ann deGroote from the National Jewish Medical and Research Center for Respiratory Diseases visited the CBE the week of March 26th.  Dr. deGroote gave a seminar entitled: "Clinical, Microbiological and Epidemiological Correlates of Nontuberculous Mycobacterial Infections: The National Jewish Experience," March 29, 2001. She discussed parallels between CF patients with Pseudomonas infections and patients with nontuberculous mycobacterial infections (mainly M. avium infections). The prestigious National Jewish Medical and Research Center, Denver, Colorado is recognized as the leading respiratory hospital in the US.   _________________________________________   CBE People in Action   Dr. Phil Stewart presented "Antibiotic Resistance of Bacteria in Biofilms," Biofilm Research Group, University of Calgary, Calgary, Alberta, Canada, February 26, 2001.   Dr. Zbigniew Lewandowski co-chaired the Microbially Influenced Corrosion session at the Annual National Association of Corrosion Engineers Meeting, Houston, TX, March 11 - 16, 2001.   Mike Geiser, M.S. candidate chemical engineering, Biomineralization Research Group, presented a paper, "Pit Initiation on 316L Stainless Steel in the Presence of Bacteria Leptothrix Discophora," at the Annual National Association of Corrosion Engineers Meeting, Houston, TX, March 11 - 16, 2001.   Dr. Tim Magnuson presented a poster entitled "Use of Rt-Pcr and In Situ Rt-Pcr Techniques to Detect Functional Gene Expression in Dissimilatory Metal and Sulfate Reducing Bacteria," DOE-NABIR PI meeting, Warrenton, VA, March 12-14, 2001.   Dr. Gill Geesey presented a poster entitled, "Acceptable Endpoints for Metals and Radionuclides: Quantifying the Stability of Uranium and Lead Immobilized under Sulfate Reducing Conditions, " Authors: Brent Peyton, Jim Amonette Gill Geesey and Zbigniew Lewandowski, at the DOE-NABIR PI meeting, Warrenton, VA, March 12-14, 2001.   Dr. Al Cunningham was a course director for a short course "Subsurface Modeling: Multiphase Flow, Transport and Bioremediation," conducted at the Institute for Hydraulic Engineering, University of Stuttgart, Germany, March 12-16, 2001. Other course directors were Dr. Rainer Helmig, University of Stuttgart, and Dr. Richard Ewing, Texas A&M University.   __________________________________________   Theses   See theses abstracts at http://www.erc.montana.edu/Res-Lib99-SW/pubs/Theses/default.htm     __________________________________________   Web Watch   The BiofilmsOnline.com site has been upgraded to a searchable database. The website format remains the same with a new feature of drop down boxes to display archived information. Kudos go to Dale Niemeyer, senior computer science student, for the transparent transition to the database version that provides a powerful search tool of world-wide biofilm information. Dale was also instrumental in developing an electronic database of all projects conducted at the CBE's Biofilm Systems Training Laboratory.   See the April issue at http://www.BiofilmsOnline.com.   Check our Recent Web Updates page created to make it easy for you to locate new information on the CBE web. See http://www.erc.montana.edu/Recent%20Web%20Updates/default.htm     __________________________________________   Upcoming CBE Events & Other Upcoming Workshops and Meetings   http://www.erc.montana.edu/CBEssentials-SW/whats_new/index.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