Not much is known about the changes in bone density due to fish reproductive migration. We used micro-CT and inferential statistics to determine whether the relative bone density in the skulls of adult Three-lips fish, that seasonally upstream migrated to a Lake Biwa tributary, changed across their known reproductive season. The relative bone density significantly decreased as standard length and condition factor (K) increased in both sexes. This negative relationship is likely due to age and hormonal effects in the fish. Results from the bone density analysis also revealed that male Three-lips fish had potentially lower relative bone density (although not significantly different) than females during peak reproductive migration, i.e., July to August. On average, male Three-lips fish are larger in length and weight than females, and in many species, females prefer larger males to smaller males, viewing their size as an indicator of genetic fitness and their ability to provide protection. Resources in the skulls of Three-lips males may be distributed in such a way that increases reproductive success, i.e., size at the expense of quality. In addition, individuals with slightly less dense bones, particularly males, appeared later than those with denser bones during the peak of the reproductive season. The high energy demands involved with aggression in males, often requires resource mobilization from various tissue compartments and could explain the slightly lower density in the latter half of the peak migration. Furthermore, Three-lips individuals that migrated earlier and later during the reproductive season may have more energy reserves than those that had been in the river for some time, hence the variable bone density between individuals. This study serves as a foundation for future studies on the effects of migration, changes in physiology and age on bone density analysis of Three-lips fish and other species in various ecosystems.
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