FATE IN WATER OF A RECOMBINANT ESCHERICHIA-COLI K-12 STRAIN USED IN THE COMMERCIAL PRODUCTION OF BOVINE SOMATOTROPIN

The fate in water of Escherichia coli K-12 strain LBB269, both plasmid-free and carrying the recombinant plasmid pBGH1, was studied. E. coli K-12 strain LBB269 (pBGH1) is a nalidixic acid resistant derivative of W3110G (pBGH1), the microorganism used by Monsanto Company for the commercial production of bovine somatotropin. Water samples were obtained from the Missouri River and from the Monsanto Life Sciences Research Center aqueous waste basin. Strains LBB269 and LBB269 (pBGH1) were grown in fermentation vessel under bovine somatotropin (BST) production conditions, and inoculated into the water samples. The inoculated water samples were incubated at 26-degrees-C, and the number of viable E. coli cells was determined as a function of time. In sterile water from both sources, the two strains remained at a constant level for at least 28 days; LBB269 (pBGH1) remained at a constant level in sterile water for at least 300 days. In non-sterile water from both sources, the two strains declined from an initial concentration of about 3.0 x 10(6) cells per ml to less than 10 cells per ml in 147 h. The study conditions did not adversely affect the populations of indigenous microorganisms. The selective loss of strains LBB269 and LBB269 (pBGH1) demonstrates that these E. coli strains do not survive in environmental sources of water. In addition, it was observed that the presence of pBGH1 had essentially no effect on the disappearance of strain LBB269 from either source of water.