Feline corneal sequestrum is a common ocular condition typified by brown to black discoloration of the cornea. The nature of the discoloration has not been identified. The purpose of this study was to perform a laboratory investigation of ocular samples from 12 clinical cases of feline corneal sequestrum in an attempt to characterize the nature of the discoloration. The 12 cases were referred to the Ophthalmology Unit at the Animal Health Trust between April and September 2000, and were also part of a clinical review of 64 cases of feline corneal sequestrum described separately. Five laboratory techniques that are routinely performed at the Biomaterials Unit, Aston University were employed for analysis of the ocular samples. Ocular material included corneal sequestrum, tear samples, meibomian gland secretions, and bandage contact lenses from the 12 clinical cases. High-performance liquid chromatography data showed that total tear lipid in affected eyes was significantly lower than in control eyes (P = 0.016); total tear lipid in affected eyes was lower than in the unaffected, contralateral eyes of the same cat but the difference was not significant (P = 0.29). The presence of an unknown lipid class was observed in tears and meibomian secretions of affected, contralateral and control eyes. Scanning electron microscopy showed that the discoloration in affected corneas was not due to the presence of iron. Fluorescence spectroscopic analysis of sequestra, unaffected corneas and contact lenses (from affected and contralateral/ unaffected eyes) showed that lipid and protein were present but did not play an important role in sequestra. Ultraviolet-visible light absorbance spectroscopy revealed a peak at 385 nm in unaffected corneas that was absent in sequestra and the difference was significant (P < 0.0001); this peak may be a characteristic feature of the normal feline cornea. The absorbance spectra displayed a peak at 280 nm in two sequestra suggesting that chromophore groups (e.g. melanin) were present. Optical microscopy performed on 10 sequestra revealed the presence of particles, which were consistent with the appearance of melanin particles, providing laboratory evidence that characterized the nature of the discoloration as melanin for the first time.
- Fluorescence spectrophotofluorimetry
- Optical microscopy
- Ultraviolet absorbance spectroscopy