A comparative study was made of the number of spermatozoa trapped on t
he outer perivitelline layer and the number of spermatozoa penetrating
the inner perivitelline layer of the eggs of 27 species of bird. The
total number of spermatozoa (trapped spermatozoa plus holes made by sp
ermatozoa) varied between 29 and 164 000 per egg among species and was
significantly and positively correlated with size of the ovum. In mos
t species, holes formed a 'halo' around the germinal disc area and the
density of holes was much greater in this region than elsewhere, espe
cially in passerine birds. In some species, a high proportion of holes
occurred at some distance from the germinal disc. This seems to be an
artefact due to the fact that some spermatozoa trapped in the outer p
erivitelline layer undergo proteolytic activity between fertilization
and oviposition and create additional holes in the inner perivitelline
layer both at and away from the germinal disc. Across all species and
within most individual species, the number of trapped spermatozoa was
positively correlated with the number of holes in the inner perivitel
line layer. Decreases in the total number of spermatozoa on successive
eggs of a clutch provided an index of the rate at which spermatozoa w
ere used from the sperm storage tubules.