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1 DEVELOPMENT AND APPLICATION OF A PLATING MEDIA FOR DETECTION OF HELICOBACTER PYLORI IN WATER Alan J. Degnan*, Jon H. Standridge Environmental Health Division Wisconsin State Laboratory of Hygiene 2601 Agriculture Dr. P.O. Box 7996 Madison, WI 53707-7996 ABSTRACT In the U.S. alone, about 5,000,000 people are diagnosed annually with ulcers, 1,000,000 are hospitalized, 40,000 undergo surgery, and 6,500 die from ulcer-related complications (Poms, 2001: Levin, 1998). Once thought to be a result of stress and/or diet, ulcers are now almost exclusively attributed to infection with the bacteria Helicobacter pylori. Laboratory diagnosis of H. pylori has become a standard procedure in the management of dyspeptic patients. Although transmission of the organism through the fecal/oral route is the assumed infection route, the possible mechanisms of human infection such as food, person to person contact, water or fomites are not clearly understood. There are a few reports in the literature suggesting transmission of H. pylori to humans via groundwater (Hegarty, 1999; Hulten, 1995 & 1998). Methods of detection used in those studies were relatively complex and costly (polymerase chain reaction; immunomagnetic separation) and unfortunately, didn't determine if the detected organisms were in fact viable or infectious. The work reported here focused on the development of a microbiological plating media that selects viable H. pylori organisms from samples containing mixed microbial populations, which could then be used for routine screening of ground and/or surface water for the presence of H. pylori. Efforts have resulted in a media formulation that allows the growth of Helicobacter while subsequently excluding common waterborne microbial background contaminants such as gram positive cocci and bacilli, enterobacteriaceae, gram negative bacilli, fungi, and pseudomonads. The laboratory-tested plating media was used to survey a cross section of Wisconsin groundwaters to further evaluate the efficacy of the media for recovering H. pylori from water samples and to begin a data base of H. pylori occurrence. INTRODUCTION A scientific breakthrough occurred in 1982 when J. R. Warren and B. Marshall isolated a bacterium and showed that it caused gastritis and stomach ulcers that affect millions of humans worldwide (Marshall, 1984). Today that claim has been proven to the extent that the National Institute of Health recommends treatment with antibiotics for all patients with peptic ulcers, which are almost exclusively attributed to infection with the bacteria Helicobacter pylori (Graham, 1991). The scope of gastric illnesses around the world is vast. In the U.S. alone, estimates as high as 50% of adult Americans carry the pathogen, most asymptomatically, and in less-developed countries human carriers represent up to 90% of the populations (Munangi, 1997). The source of human infection is not yet known and until recently, the natural reservoir for H. pylori was thought to be the human gastrointestinal tract (Axon, 1996). However, isolation of Helicobacter from non-human sources such as livestock (Vaira, 1992), domestic cats