*4.2. Sample Collection*

A total of 35 captive giraffes (*Gira*ff*a camelopardalis*) were involved in the current study. Based on physical examinations, giraffes did not show symptoms of diseases both before and during the urine sampling period. The giraffes were housed in five Italian zoos: Zoosafari Fasanolandia (FA) (N = 11), Safari Ravenna (RA) (N = 4), Giardino Zoologico di Pistoia (PT) (N = 1), Parco Natura Viva (VR) (N = 4), and Parco Faunistico Le Cornelle (BG) (N = 15).

The details for each giraffe are reported in Table 3. Their age ranged from a minimum of 6 months to a maximum of 20 years. The giraffes were categorized in 3 age classes: Young (from 6 months to 6 years old, N = 14), Adult (from 6 to 15 years old, N = 16), and Old (older than 15, N = 9), according to the following information. In female giraffes the first birth is at about 6.4 years old, even if sexual maturity is reached at 3–4 years [42,43]. Giraffe males are considered as adults when older than 6 years old, according to Lee et al. [44].


**Table 3.** Animal information.


**Table 3.** *Cont.*

The samples were collected between 10:00 a.m. and 2:00 p.m., in connection to the daily activities of the keepers. Urine samples were collected with a syringe from the ground. To limit the soil contaminants, only the upper part of the urine was collected, immediately after the spontaneous voiding, before it was absorbed by the soil. A sample from one male was also collected directly into a sterile beaker, preventing the sample from touching the ground. Four urine samples were collected from two females during and after pregnancy. After collection, the urine samples were centrifuged at 1500× *g* for 10 min, to further remove potential ground contaminants, and the supernatants were frozen at −80 ◦C.
