Variability within the genus Phrynosoma in the occurrence of ocular-sinus blood-squirting, reportedly a defense used in canid encounters, is reviewed from the literature. Six species have been reported to squirt blood, and seven species remain unreported. Five of the latter species were tested in dog trials; one exhibited blood-squirting (Phrynosoma hernandesi), one exhibited precursor behaviors but failed to squirt blood (Phrynosoma ditmarsi), and three yielded negative results (Phrynosoma mcallii, Phrynosoma modestum, and Phrynosoma platyrhinos). Instances of blood-squirting in response to human encounters were collected and largely support the negative results for the three species P. mcallii, P. modestum, and P. platyrhinos. A phylogeny of blood-squirting and nonblood-squirting species is presented with blood-squirting being plesiomorphic in the genus and the synapomorphic condition of nonsquirting species being restricted to a single clade of P. mcallii-modestum-platyrhinos. The possibility of P. douglasii independently evolving an autapomorphic condition remains unresolved. Dog trials with 40 adult Phrynosoma cornutum were conducted to determine influences of body size and sex on squirt frequency and blood mass expelled, as well as to examine aspects of the potential physiological cost of the defense. In 153 trials, 85% of all lizards squirted in at least one trial, 82% squirted in more than one trial, and two lizards squirted daily over the seven-day trial period. Initial body mass positively correlated with the total number of squirts/individual (r2 = 0.28; P < 0.001) and the number of days a lizard continued squirting (r2 = 0.63; P < 0.01). Number of squirts/individual/day declined over the seven-day trial period (r2 = 0.20; P < 0.05). Cumulative mass loss for individual lizards attributable to blood-squirting averaged 0.7 ± 0.8 g (2.0 ± 2.0% body mass), with a high of 2.8 g (6.8% body mass). In addition, juvenile P. cornutum and P. hernandesi were shown to squirt blood in dog trials, illustrating the early developmental onset of the behavior.