Preadapted inocula for limiting the risk of errors in biodegradability tests.

Reducing the time for biodegradability tests to 28 days poses a problem when the inoculum contains few biodegraders, as a biodegradable xenobiotic must give a positive result within this time. The influence of initial concentration (X0, number of cells liter-1) on the lag time (hours) of para-nitrophenol biodegradability tests was examined using different concentrations of adapted Pseudomonas putida with para-nitrophenol as the sole carbon and energy source. Lag time decreased as bacterial density increased according to the expression y = 63.5 - 5.7(log10X0). The addition of river water to the P. putida concentrations shortened the lag time-bacterial density relationship and lag time filled the expression y = 52.4 - 5.1(log10X0). The addition of river water also increased the rate of para-nitrophenol biodegradation from 1.29 mgC (liters x hr)-1 to 2.11 mgC (liters x hr)-1. An examination of the effect of the initial adapted P. putida density, expressed as total cell, cultivable bacteria, or biodegraders, was also made on the para-nitrophenol biodegradability test outcome. The model-related cell density and the probability of test response give very similar k constants (kT = 0.56 x 10(-3) liter total cells-1; kv = 0.11 x 10(-3) liter CFU-1, kMPN = 0.16 x 10(-3) liter MPN-1). Comparisons with nonadapted natural mixed culture (activated sludge, river water) indicate that the biodegradability test responses were the same as with adapted cells when the nonadapted cell concentrations were at least 5 x 10(10) total cells liter-1. As this high cell concentration led to carbon contamination, adapting mixed inocula before the test to increase the number of biodegraders appears to be the best solution. Before biodegradability tests, cell density can be adjusted using techniques which are not specific to biodegraders, and only 10(5) total adapted cells liter-1 are needed for a 99.9% chance of a positive response in para-nitrophenol biodegradability tests.