BACKGROUND: As bacterial infections pose a major health risk to captive populations, disease prevention and management play a crucial role in the ex situ conservation of giraffes (Giraffa camelopardalis). This study describes the case of a giraffe that developed septicemia after an umbilical cord infection caused by Escherichia coli. To our knowledge, pathological changes in diseased giraffes caused by E. coli, which is an opportunistic pathogenic organism, have not been reported. This is the first report presenting an analysis of necropsies and subsequent microbiological investigations. CASE PRESENTATION: The baby giraffe's mother died shortly after birth, so it had to be fed milk powder. The giraffe was healthy at first but developed symptoms like depression, loss of appetite, and lameness at 8 days old. At 14 days of age, the juvenile giraffe showed astasia and gradually died, with a disease course of 7 days. Postmortem examination revealed opisthotonus and navel swelling. Serofibrinous arthritis, serofibrinous necrotizing inflammation of periarticular soft tissue, serous omphalitis, and severe adventitia hemorrhage of the umbilical artery were observed. Severe serofibrinous pericarditis, pleuritis, and peritonitis were also observed. The interstitium of the pulmonary lobule widened because it was filled with a pale yellow translucent gelatinous exudate. Histopathologically, the calf had diffuse serous interstitial pneumonia, serous necrotizing umbilical arteritis, degenerative hepatitis with mild fibrosis, degenerative nephritis, hemorrhagic lymphadenitis, necrotizing enteritis, and necrotizing thyroiditis. Blue-stained clumps of bacteria of varying sizes and neutrophil infiltration were scattered or diffused in the interstitial connective tissue and edematous serosa of all tissues and organs, as well as in small vessels and lymphatic vessels, which were filled with many neutrophils (lymphatic spread). Single gram-negative Escherichia coli were cultured from all tissues of the animal. Polymerase chain reaction results of 16S rRNA of the isolated Escherichia coli had 99.79% homology to KJB03889.1. CONCLUSIONS: The gross, histopathologic, microscopic, and polymerase chain reaction sequencing features reported in a juvenile giraffe were consistent with colibacillosis, which is a rare disease of giraffes. The gross, histopathologic, microscopic, and polymerase chain reaction sequencing features reported in a juvenile giraffe. This case serves as a paradigmatic illustration of a giraffe suffering from neglect and inadequate treatment, leading to severe consequences. In instances of giraffe Escherichia coli septicemia, it is imperative to thoroughly assess for underlying diseases, particularly in the absence of obvious predisposing factors. The rise of multidrug resistant organisms has constrained the efficacy of empirical antibiotic treatment, highlighting the importance of promptly conducting culture and sensitivity testing and employing antibiotic therapy guided by susceptibility results.