Background Reference intervals for haematology and serum biochemistry parameters were developed for free-ranging Lumholtz's tree-kangaroo (Dendrolagus lumholtzi) using 35 samples from 12 female and 15 male free-ranging animals. Captive tree-kangaroos (n = 12) were also sampled for comparison. Differences were found between free-ranging and captive animals in white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase. These differences may be attributed to diet, activity, capture methods or age group. Reference intervals generated may be used for both free-ranging and captive Lumholtz's tree-kangaroos. This study provides a valuable tool for the assessment of health in rescued and captive treekangaroos and will aid in investigations into population health and disease in free-ranging Lumholtz's tree-kangaroos.Objective To develop reference intervals (RIs) for haematology and serum biochemistry parameters in Lumholtz's tree-kangaroos.Methods Haematological and serum biochemical RIs were determined using 35 samples from 27 clinically healthy Lumholtz's tree-kangaroos from the Atherton Tablelands region of Queensland examined between 2014 and 2019. Haematology and serum biochemistry parameters were measured from 16 samples from 12 captive animals for comparison.Results Reference intervals based on 35 samples from freeranging animals showed higher mean and standard deviation values for white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase than results for 16 samples from captive animals. Captive individuals showed higher mean values than free-ranging individuals for albumin, protein, creatinine as well as Hb, MCV, MCH and MCHC. ConclusionThe haematological and serum biochemistry RIs developed for Lumholtz's tree-kangaroos in this study will provide a valuable tool during clinical examination and investigations into disease and population health by veterinarians and researchers. The differences in parameters between free-ranging and captive animals are consistent with differences in diet, age cohort, activity or capture methods. Reference intervals generated from free-ranging animals should also be valid for captive Lumholtz's tree-kangaroos.