Adapting the buccal micronucleus cytome assay for use in wild birds: Age and sex affect background frequency in pigeons

Abstract

Micronucleus (MN) formation has been used extensively as a biomarker of damage from genotoxic exposures. The Buccal MN Cytome (BMCyt) assay provides a noninvasive means of quantifying MN frequency in humans, but it has not been developed for use in wildlife. We adapted the BMCyt assay for use in wild birds, with a focus on feral pigeons (Columba livia) as a potential indicator species. Five of six urban bird species sampled using oral cavity swabs produced sufficient buccal cells for the BMCyt assay. The body size of species sampled ranged almost 100-fold (~60 to 5,000 g), but was a not major factor influencing the number of buccal cells collected. Pigeon cells were stained and scored following published BMCyt assay protocols for humans, but with a modified fixation approach using heat and methanol. Pigeons had the same common nuclear abnormalities reported in human studies, and a similar background MN formation frequency of 0.88 MN/1,000 cells. Adult pigeons had on average a threefold higher rate of MN formation than juveniles, and males had a 1.4- to 2.2-fold higher frequency than females. Domestic and feral pigeons did not differ in overall MN frequency. Our results indicate that the BMCyt assay can be used on wild birds, and could provide a means of assessing environmental genotoxicity in pigeons, a useful indicator species. However, bird age and sex are important factors affecting background MN frequency, and thereby the design of environmental studies.