A small-animal model of genital herpes simplex virus (HSV) infection has not been available for investigating the pathogenesis of disease in males. Male Hartley guinea pigs were inoculated on the medial thigh with 10(5)-10(6) plaque-forming units of HSV type 2. Animals developed vesicular lesions in the genital area, including the penis. Transient urinary retention, hindlimb paralysis, and acute urethral shedding of HSV also occurred. During acute infection, virus was detected in neural and genital tissue homogenates, including the peripheral nerve, lumbosacral dorsal root ganglia (DRG), spinal cord, brain, vas deferens, epididymis, testes, seminal vesicle, and prostate. Recurrent disease developed in all animals. Persistent virus was detected in DRG but rarely in the vas deferens, seminal vesicle, epididymis, and testes. This model produces disease similar to that seen in men and should prove useful for studying the pathogenesis and immunology of male genital HSV type 2 infections.