An investigation into the developmental and morphological differences among the hatching larvae of three Chinese Oryzias curvinotus, was conducted. All embryos and larvae were cultured in a water bath with a temperature of (27.0±1.0)℃. The results showed that there are significant differences in morphology among the hatching larvae; specifically, the body length of O. sinensis[(4.04±0.19) mm], and [(3.88±0.17) mm]. One-way analysis of variance (ANOVA) revealed that 90.9% of measurements were significantly different among the three species' hatching larvae (20 of the 22 measurements). Further, Principal Component Analysis (PCA) showed that total body length (TL), head length (HL), HL vs. body depth (BD), TL/BD, anal length (AL), mouth width (MW), and yolk length (YL) were the main variables contributing to the morphological differences among the hatching larvae. Additionally, discriminant analysis confirmed the differences among the larvae, and the larvae were separated from one another in the combined plot for the discriminant analysis. The above results were also confirmed by analysis of similarity (ANOSIM). ANOSIM showed that the differences among the larvae had statistical significance (O. pectoralis and O. sinensis larvae, and larvae were isolated in the dendrogram constructed from the cluster analysis. This result correlates to their geographical distributions, because populations are found closer to one another than to populations in the wild. Finally, the chromatophore pigment pattern on the body could be used to discriminate the O. sinensis larvae than on the larvae of the other species, and the chromatophore pigment on the petrol fins of larvae pectoral fins. Also, many morphological measurements could be used to discriminate between larvae.