Most Probable Numbers of chemolitho-autotrophic nitrite-oxidizing bacteria in well drained grassland soils: Stimulation by high nitrite concentrations

The results of Most Probable Number (MPN) enumerations of chemolitho-autotrophic nitrite oxidizers are very much dependent on the nitrite concentration applied in the incubation medium. In order to explain this dependency, the influence of pH, nitrite and resultant nitrous acid concentration of the incubation medium on the MPN-enumeration was investigated. It appeared that none of these factors were exclusively responsible for the result of the enumeration. In samples from a well drained grassland soil, highest number have been obtained with a combination of a low pH and a low nitrite concentration in the counting medium. The relation between the MPN-counting results and the nitrous acid concentration showed an optimum with the same soil samples. It was hypothesized that the relatively high numbers of nitrite-oxidizing cells determined in soil samples at a high nitrite concentration and pH 7.3 was due to the presence of dormant cells. However, this hypothesis could not be confirmed with enumerations of aged cell suspensions of different Nitrobacter species. In contrast to the field observations, these resting cells were always enumerated more efficiently at a low nitrite concentration. The importance of the use of more than one incubation medium for the enumeration of nitrite-oxidizing bacteria is emphasized.