Selective depletion of bacteria alters but does not eliminate odors of individuality in Rattus norvegicus

To determine if odors of individuality are influenced by the removal of Gram-negative or Gram-positive gut bacteria, Long-Evans rats were trained in an operant olfactometer to discriminate between the odors of two individual conspecifics and their operant responses to three different odors in randomly presented probe trials were analyzed. Significantly more responses were made to the probe odors from two known individuals than to the probe odors from known individuals with their Gram-negative bacteria eliminated (Experiment 1) or their Gram-positive bacteria eliminated (Experiment 2). Responses to the probe odors from known rats with bacterial selectively depleted did not differ significantly from responses to probe odors from unknown rats. These results support the hypothesis that the urinary odor of an individual rat is altered by the removal of specific gut bacteria. In Experiments 3 and 4, subjects made fewer errors in learning to discriminate between the odors of the familiar rats whose bacteria had been selectively depleted than between the odors of unknown rats. This "savings effect" indicates that some components of the individual urinary odors were retained after the removal of specific gut bacteria. Thus, the eliminated bacteria were not totally responsible for the odors of individuality. The outcome of Experiments 3 and 4 also indicates that conclusions regarding the recognition of odors by rats should not be made on the outcome of probe trial experiments alone.