Picosecond time-resolved coherent anti-Stokes Raman spectroscopy (PTR/CARS) is used to generate high signal to noise (S/N) vibrational spectra of bathorhodopsin (batho) formed in the photoreaction of room-temperature rhodopsin (Rh-RT). These PTR/CARS spectra of batho(RT), measured as a function of only the time following 3-ps (full width at half maximum), 500-nm excitation of Rh-RT, demonstrate that the vibrational structure of batho(RT) in the 700-1700-cm(-1) region is distinct from that of Rh-RT and remains unchanged over at least the 10 ps [8 ps cross correlation time (CCT)] to 100 ns interval of the Rh-RT photoreaction. Given the experimental difficulties associated with the irreversibility of the Rh-RT photoreaction, these are the first time-resolved vibrational spectra of batho(RT) over the full 700-1700-cm(-1) region to be reported. The PTR/CARS spectra taken after 100 ns contain vibrational features other than those assignable to either Rh-RT or batho(RT) (potentially assignable to the blue-shifted intermediate, BSI). Excellent agreement is found between the major features of the Rh-RT and batho(RT) vibrational spectra measured via PTR/CARS and earlier resonance Raman (RR) spectra taken at low temperatures (LT) selected to thermally stabilize (freeze) bathe. Comparisons of the C=C stretching mode region reveal a 12-cm(-1) shift upon batho(RT) formation (PTR/CARS data), which agrees well with the 13-cm(-1) shift found for batho trapped at LT (RR data), These vibrational frequency changes also correlate well with the corresponding 38-nm (LT) and 31-nm (RT) shifts observed in the absorption maxima upon the formation of baths, thereby supporting an inverse relationship between C=C frequencies and absorption maxima proposed for retinal proteins, Comparisons of the C=C stretching frequencies at LT and RT reveal a temperature dependence characterized by red shifts of 4 cm(-1) in Rh and 3 cm(-1) in bathe, the direction of which is opposite to the blue shifts observed in the visible absorption maxima of Rh (7 nm) and bathe (14 nm). This latter observation suggests a stronger interaction of the protein (likely with the counterion Glu-113) with the all-trans-retinal in the batho(RT) structure than in the corresponding static batho(LT) structure.