Antimicrobial resistance (AMR), encoded by plasmid‐mediated AMR genes (ARGs), is an increasing global public health threat. Wildlife play a fundamental role as sentinels, reservoirs and potential vectors of ARGs. For the first time in Galapagos, we have identified and quantified the presence of ARGs in free‐living giant tortoises (Chelonoidis porteri). We performed ARG analyses by quantitative PCR of faeces collected from the cloaca of 30 tortoises widely distributed across Santa Cruz Island. Validated samples (n = 28) were analysed by a panel of up to 21 different ARGs and all 28 tortoise samples were positive to one or more genes encoding resistance. Thirteen of 21 tested ARGs were present in at least one sample, and 10 tortoises (35.7%) had a multi‐resistant pattern. We recommend additional research so we may more fully understand resistance patterns across taxa and geographical locations throughout the Galapagos archipelago, and the implications of ARGs for the health of wildlife, domestic animals, and humans. In this study, we found 100% of sampled giant tortoises had ARGs present in their faeces, suggesting a large‐scale distribution of these genes within the archipelago.

Reference intervals (RIs) are an increasingly valuable tool for monitoring captive and free-living wildlife populations. Galapagos tortoises are one of the most emblematic species on Earth with 9 of the 12 extant species considered endangered due to human activities. As part of a long-term health assessment within the Galapagos Tortoise Movement Ecology Programme, we sampled a total of 210 free-living Santa Cruz Galapagos tortoises ( Chelonoidis porteri ). We collected blood from the brachial vein and performed packed cell volume (PCV), total solids (TS), morphological evaluation, white blood cell (WBC) count estimates and differentials and a VetScan biochemistry panel for each individual. We calculated 95% RIs and 90% confidence intervals (CIs) using the Reference Value Advisor tool and following international standard guidelines. Tortoises were categorized by estimated age and sex, with RI and CI reported here for 164 adult tortoises and 46 sub-adult tortoises. We found significant differences between sexes, with adult females presenting a lower PCV and higher values for both calcium and potassium. Among age groups, adult tortoises presented higher PCV, TS and albumin and lower WBC counts, aspartate aminotransferase and creatine kinase than sub-adult tortoises. We also found that tortoises presented higher numbers of lymphocytes during the dry season, but higher basophils, eosinophils, phosphorus, potassium and TS during the humid season. The heterophil:lymphocyte ratio did not differ between groups. To the authors’ knowledge, this is the first report of formal plasma biochemistry and haematology RI for free-living Galapagos tortoises. With the present study we provide an important diagnostic tool for captive-breeding programs in the Galapagos and zoological institutions globally that care for giant tortoises. The ultimate goal of this study is to contribute to the understanding of giant tortoise baseline health parameters and to inform local management decisions that help to conserve these emblematic species.