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Aquatic nuisance species (ANS) in ship's ballast water are killed by permeating to equilibrium a gaseous mixture consisting essentially of, preferably, ≧84% nitrogen, ≧11% carbon dioxide and ≦4% oxygen through ship's ballast water until the ballast water itself becomes (i) hypercapnic to ≧20 ppm carbon dioxide, and, by association, (ii) acidic to pH ≦7, while preferably further, and also, being rendered (iii) hypoxic to ≦1 ppm oxygen. The permeating is preferably realized by bubbling the gaseous mixture preferably obtained from an inert gas generator through the ballast water over the course of 2+ days while the ballast water is continually maintained a pressure less than atmosphere, preferably −2 p.s.i. or less. The (i) hypercapnic, (ii) acidic and (iii) hypoxic conditions—each of which can be independently realized—synergistically cooperate to kill a broad range of ANS in the ballast water without deleterious effect on the environment when, and if, the ballast water in which the balance of dissolved gases has been changed is discharged.

The present invention relates to compositions comprising conjugation-competent non-dividing cells for delivery of genes to bacterial cells by conjugation. The present invention also relates to methods of making, using, and storing such conjugation competent non-dividing cells.