Non-Invasive Sex Determination of Nestlings and Adult Bonelli’s Eagles Using Morphometrics

SIMPLE SUMMARY: This study explores the use of biometric analysis to identify the sex of Bonelli’s eagles, a bird species with reverse sexual dimorphism where females are usually larger than males. By using linear discriminant analysis of biometric variables, we were able to obtain equations that accurately distinguished between male and female eagles, using just two measurements. The study included 137 Bonelli’s eagles, 82 nestlings and 55 adults, sampled in eastern Spain from 2015 to 2022. The validation procedure reduced the number of variables used, increasing the accuracy of sexing and saving time. Results showed that the lateral tarsus length and dorso-ventral tarsus length measurements were the most effective in distinguishing between male and female eagles of all age classes, while other variables showed some overlap between sexes. This non-invasive method has multiple applications, including estimation of sex ratio for further studies on population dynamics and extinction risk assessments, which could contribute to the conservation of this endangered species. ABSTRACT: Biometric analysis allows the sexing of most vertebrates, particularly birds. Birds of prey, and, especially, the Bonelli’s eagle (Aquila fasciata), show reverse sexual dimorphism (i.e., females are usually larger than males). In contrast to blood sampling, the use of morphometrics allows sex determination using a non-invasive method, and, therefore, it facilitates fieldwork. By means of a linear discriminant analysis of biometric variables, we obtained different equations that allow the sexing of nestlings and adult Bonelli’s eagles. We sampled 137 Bonelli’s eagles, 82 nestlings and 55 adults in eastern Spain during the period 2015–2022. The sexes obtained after linear discriminant analysis were compared with their molecular sexing. The validation procedure of the linear discriminant equations facilitated the reduction of the number of variables used and, consequently, optimised working time and sexing accuracy. After validation, some equations showed a 100% sexing efficiency for Bonelli’s eagles, particularly for adults. Our results showed that the variables with smaller overlap between the sexes were the lateral tarsus length and dorso-ventral tarsus length, particularly in nestlings. The rest of the variables showed some overlap between the sexes in both age classes. The results we obtained enable the sexing of juvenile and adult Bonelli’s eagles in the field using just these two measurements. Hence, this is an easy, accurate, quick and non-invasive method with multiple applications, including in studies on population dynamics, survival analysis or extinction risk assessments, which, ultimately, could contribute to the improvement of the conservation status of this endangered species.