U.S. Geological Survey, Branch Of Acquisition and Federal Assistance, Box 25046, Denver Federal Center, Denver, CO 80225-0046

B -- PREPARE AND PURIFY BACTERIAL STRAINS FROM WATER SAMPLES, DNA RIBOTYPE ANALYSIS, ANALYZE DNA PATTERNS DUE 011800 POC Diana Romero 303-236-5900 x344 E-MAIL: USGS, CR, Acquisition & Federal Assistance, dlromero@usgs.gov. The U.S. Geological Survey, Water Resources Division, Rolla Missouri is undertaking several watershed scale projects that involve the culturing of bacteria from water samples collected from surface and ground water sources. Selected water samples are to be subjected to state of the art DNA ribotyping techniques to identify the animal source of E. coli bacteria in water samples. A secondary objective is to establish the presence of specific bacteria species (especially the verotoxic E. coli strain O157:H7 and Salmonella species) in the water samples. For the successful completion of these projects, the laboratory also must be operated by an individual or individuals who are knowledgeable or expert in the field of animal intestinal microflora. Although DNA techniques have been used for several years, the techniques have only recently been applied to the study of bacterial contamination in surface and ground water. Identification of bacterial sources in water samples using ribotyping worked on the principle that different warm blooded animal species (i.e. horses, cattle, humans, deer) have E. coli stains that are unique to each species. The ribotype "fingerprint" of E. coli isolated from water samples are compared to fingerprints from know animal hosts to identify the origin of the bacteria. Successful application of the technique requires a thorough understanding of stream hydrology, the distribution of animal reservoirs within the stream watershed. bacterial metabolism, and animal physiology. The initial step of laboratory processing is the culturing of E. coli on selected growth media. Because one of the target organisms (E. coli O157:H7) may be present in water at very low concentrations, it is critical to the project that laboratory culturing begin as quickly as possible to avoid "die-off" of viable cells before they can be cultured. This time constraint requires that the laboratory be with reasonable driving distance (not more that about 4 hours) of the project study areas. All projects are in southern Missouri, and the laboratory at Columbia, Missouri, can be reached within the required time. Before a bacteria sample can be subjected to DNA ribotyping, it is necessary to identify and purify the specific bacteria strain of interest by applying a series extracted from impure colonies will produce ambiguous ribotyping results. This requirement of accepting only pure isolates means that the water samples must be pre-processed by the collector and another laboratory before submittal to the riboprinter for analysis. Ribotypes (based on a specific part of the bacteria chromosomal DNA) are not entirely comparable to serotypes (based on traditional culture type methods and other DNA techniques). Because of this difference, and the secondary USGS objective to identify the presence of E. coli O157:H7 in water samples, purified E. coli isolates from the water samples must be subjected to a second set of selective culture techniques and DNA analysis using eletrophoresis to positively identify E. coli O157:H7. E. coli O157:H7 is know pathogen that can cause serious illness in humans; therefore, positive identification of this organism by standard serologic and electrophoresis techniques is required to ensure integrity of the results. Although manual techniques are somewhat slower, the poorer resolution of commercial riboprinters make them undesirable for the purposes of these surface and ground water investigations. Higher resolution manually produced DNA fingerprint patterns can be interpreted more accurately. A second requirement is that the DNA analysis be performed using a minimum of 16 isolate to ensure that sufficient data is generated to associate a particular E. coli bacterial strain with particular animal group. Analysis 2 isolate per purified sample, are not adequate to answer question addressed in the USGS investigation. Because the application of DNA ribotyping is a new science, few laboratories are capable of conducting the procedure. Based on information available at this time, and in accordance with 41 U.S.C.253 (c)(1), implemented by FAR 6.302-1, the Government considers the services specified as a sole source from: University of Missouri-Columbia, Department of Veterinary Pathobiology, Columbia, Missouri and proposes to negotiate a contract with that firm on that basis. However, should additional sources be identified, they will be considered. Firms who feel that they can furnish the required item or its functional equivalent (based upon the criteria specified herein) are invited to submit in writing an affirmative response to this announcement. An affirmative response would include literature, brochures, and such other materials which correspond to the required items stated herein. This information will be evaluated and used to determine if competitive opportunities exist. Since no solicitation document exists, request for such documents without accompanying information will be considered non-responsive to this request without further consideration. This notice may represent the Government's only official notice of this procurement. See numbered Note 22. Posted 12/02/99 (W-SN405099). (0336)

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