Foot-and-mouth disease virus (FMDV) remains an important pathogen of livestock more than 120 years after it was identified, with annual costs from production losses and vaccination estimated at €5.3–€17 billion (US$6.5–US$21 billion) in FMDV-endemic areas. Control and eradication are difficult because FMDV is highly contagious, genetically and antigenically diverse, infectious for a wide variety of species, able to establish subclinical carriers in ruminants, and widely geographically distributed. For early detection, sustained control, or eradication, sensitive and specific FMDV surveillance procedures compatible with high through-put testing platforms are required. At present, surveillance relies on the detection of FMDV-specific antibody or virus, most commonly in individual animal serum, vesicular fluid, or epithelial specimens. However, FMDV or antibody are also detectable in other body secretions and specimens, e.g., buccal and nasal secretions, respiratory exhalations (aerosols), mammary secretions, urine, feces, and environmental samples. These alternative specimens offer non-invasive diagnostic alternatives to individual animal sampling and the potential for more efficient, responsive, and cost-effective surveillance. Herein we review FMDV testing methods for contemporary and alternative diagnostic specimens and their application to FMDV surveillance in livestock (cattle, swine, sheep, and goats).