Mating behavior and early development of Cobitis sp. (BIWAE type D, TosaShima-dojo) were observed in the laboratory by inducing spawning of females with human chorionic gonadotropin (HCG). Subsequently, laboratory bred individuals were compared with spawned eggs and larvae obtained from the field. Mature adults, naturally spawned eggs, and larvae were collected from a river in eastern Kochi Prefecture, Shikoku Island, Japan, and a natural spawning ground adjacent to the river. Mating behavior was observed 15 times at night, and distinguished into four stages: phase 1, tracking; phase 2, approaching; phase 3, amplexus; and phase 4, spawning. Egg diameters after water absorption were 2.1–2.3 mm, the spherical, demersal eggs having a light-yellow yolk, no oil droplets, and slight viscosity. Newly hatched larvae [3.3–4.9 mm in total length (TL)] had 46 (32 + 14) myomeres, two pairs of outer gill filaments on the cheek, and melanophores on the head. Notochord flexion started at 6.0–7.2 mm TL (wild individuals at 6.0–6.3 mm TL) and was completed at 8.8 mm TL (6.6 mm TL). The formation of membranous fins and full fin-ray complements were attained in the fin order pectoral, caudal, dorsal, anal, and pelvic, and caudal, dorsal, anal, pectoral, and pelvic, respectively. Some individuals (7.0–8.0 mm TL at the flexion stage) had free neuromasts, each with a short cupula, laterally on the caudal region. In addition, following the postflexion stage, some morphological differences between artificially bred and wild individuals were observed but could not be quantified due to the small sample size.