Background Neutrophil extracellular traps (NETs) represent a novel cellular mechanism of antimicrobial defense activity. Intravascular neutrophils produce extracellular web-like structures composed of chromatin, histones, and cytoplasmic granule proteins to attack and kill microbes. They may impact both pathogen and host; NETs correlate strongly with disseminated intravascular coagulation and mortality in critically ill humans. The mechanism was first discovered in human neutrophils in 2004. Presumptive heterophil extracellular traps (HETs) in a non-avian reptile species were first described in blood films of a gopher tortoise with systemic inflammation. Objective While prior reports are limited to blood film review and in vitro studies, this descriptive case series highlights the cytological identification of presumptive HETs in nine reptile patients. Methods Subjects included six gopher tortoises, one blood python (Python curtus), one Burmese python (P. bivittatus), and one desert king snake (Lampropeltis getula splendida). All six gopher tortoises (Gopherus polyphemus) had upper respiratory disease with bacterial etiology (including Helicobacter sp. and/or Mycoplasma sp.), and snakes had upper respiratory tract infection confirmed with serpentovirus (n = 2) or bacterial dermatitis (n = 1). Results Cytology samples with identified HETs included tissue imprints (n = 4), nasal discharge (n = 3), an oral swab (n = 1), and a fine needle aspirate of a skin lesion (n = 1). The identification of specific bacterial (n = 6) and/or viral pathogens (n = 2) was notable. Clinical relevance To the authors’ knowledge, this is the first report of presumptive HETs recognized in reptile cytology specimens, suggesting an active cellular process in vivo in response to systemic inflammation in non-avian reptiles, and contributing to further understanding of extracellular traps in these species.