Dear MARMAM Community,

On behalf of my co-authors at the Florida Fish and Wildlife Conservation 
Commission, I am pleased to announce two publications related to age estimation 
of Florida manatees using growth layers in their earbones:

Lonati, G. L., Howell, A. R., Hostetler, J. A., Schueller, P., de Wit, M., 
Bassett, B. L., Deutsch, C. J., and Ward-Geiger, L. I. (2019) Accuracy, 
precision, and error in age estimation of Florida manatees using growth layer 
groups in earbones. Journal of Mammalogy, 10(4): 1350-1363. 
https://doi.org/10.1093/jmammal/gyz079 (open access)

Ages of Florida manatees (Trichechus manatus latirostris) can be estimated by 
counting annual growth layer groups (GLGs) in the periotic dome portion of the 
tympanoperiotic complex of their earbones. The Florida Fish and Wildlife 
Conservation Commission manages an archive of more than 8,700 Florida manatee 
earbones collected from salvaged carcasses from 1989 to 2017. Our goal was to 
comprehensively evaluate techniques used to estimate age, given this large 
sample size and changes to processing protocols and earbone readers over time. 
We developed new standards for estimating ages from earbones, involving two 
independent readers to obtain measurements of within- and between-reader 
precision. To quantify accuracy, precision, and error, 111 earbones from 
manatees with approximately known ages (first known as calves: “KAC”) and 69 
earbones from manatees with minimum known ages (“MKA,” based on 
photo-identification sighting histories) were processed, and their ages were 
estimated. There was greater precision within readers (coefficient of 
variation, CV: 2.4–8.5%) than between readers (CV: 13.1–13.3%). The median of 
age estimates fell within the true age range for 63.1% of KAC cases and was at 
least the sighting duration for 75.0% of MKA cases. Age estimates were 
generally unbiased, as indicated by an average raw error ± SD of −0.05 ± 3.05 
years for the KAC group. The absolute error (i.e., absolute value of raw error) 
of the KAC data set averaged 1.75 ± 2.50 years. Accuracy decreased and error 
increased with increasing known age, especially for animals over 15 years old, 
whose ages were mostly underestimated due to increasing levels of resorption 
(the process of bone turnover that obscures GLGs). Understanding the degree of 
uncertainty in age estimates will help us assess the utility of age data in 
manatee population models. We emphasize the importance of standardizing and 
routinely reviewing age estimation and processing protocols to ensure that age 
data remain consistent and reliable.

--------------------

Lonati, G. L., Howell, A. R., Schueller, P., and Deutsch, C. J. (2021) Using 
tetracycline to evaluate age estimation in a long-lived aquatic mammal. 
Wildlife Society Bulletin, 45(2): 340-350. https://doi.org/10.1002/wsb.1192 
(available upon request - [email protected])

Age estimation is useful for understanding population parameters and, in many 
vertebrates, relies on the principle that growth layer groups (GLGs) are 
deposited annually in specific tissues. In Florida manatees (Trichechus manatus 
latirostris), GLGs in earbones are used to estimate age at death. Opportunities 
to validate the rate of GLG deposition in earbones from manatees >15 years old 
are rare, yet important for ensuring accurate age estimation across the 
species’ lifespan. Tetracycline injection is a useful method for validating GLG 
interpretations, particularly when the exact age of an animal is unknown. Since 
1997, we collected earbones from 10 manatees that were ≥13–69 years old at 
death and had been injected with tetracycline 9–37 years before death. The 
number of years since injection (YSI) was estimated by photographing earbone 
cross-sections under ultraviolet light, measuring the distance between the 
fluorescent tetracycline mark and earbone edge, processing the earbones to 
visualize GLGs, superimposing the distance to evaluate the marks’ locations 
relative to GLGs, and counting the GLGs between the mark and earbone edge. 
Seven earbones had tetracycline marks, although 2 of the marks were dull or 
discontinuous. On average, estimated YSI was 5.6 (SD = 7.5) years less than the 
known YSI; however, the error was nearly always ≤2 years for manatees that had 
been injected <20 years before death, consistent with an annual rate of GLG 
deposition at younger ages. Resorption (i.e., bone turnover) that obliterated 
GLGs was likely why YSI was underestimated in old manatees with longer 
post-injection intervals, although we cannot exclude the possibility that GLG 
deposition rate may slow in old age. We discuss how age, extrinsic stressors, 
life history events, and laboratory processing may affect tetracycline 
visibility, earbone growth, and GLG interpretation. Our study reinforces the 
challenges with accurately estimating the age of old individuals in long-lived 
mammalian species.

Please reach out with any questions.  Thank you!
Gina & colleagues

Gina Lonati
PhD Student (she/her)
University of New Brunswick Saint John
https://glonati2222.wixsite.com/research
[email protected]
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