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This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2-3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p < .01) thawed spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60-85°C min ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p < .05) percentage of motile spermatozoa and live spermatozoa with an intact acrosome. Vitrified-warmed sperm variables were at their best when the spermatozoa was diluted in TCG-6% egg yolk + 100 mm sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced (p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing.

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Mucoperiosteal exostoses (MpEs) of the tympanic bulla (TB), also referred as middle-ear otoliths, have been occasionally described in dogs and cats in association with clinical signs of otitis media or as an incidental finding, but they have not been recorded in other species. In this report, we describe the radiographic, gross, and histopathologic features of MpEs in 8 African lions (Panthera leo). All animals (5 males and 3 females) were adults that had been kept in captivity and had their skeletons conserved as part of an anatomic academic collection. A radiographic study revealed mineralized structures in the TB consistent with MpEs in 7 of the 16 examined TB; a computed tomography study identified MpEs in 12 of the 16 TB. Six TB from 4 lions were sectioned, and several MpEs were demineralized for histopathologic analysis. Grossly, MpEs appeared variable in number and shape. Some were globular structures that were loosely attached to the mucosal surface of the TB; others were isolated to coalescent bone spicules extending from the mucoperiosteum. Position was also variable, but MpEs frequently developed in the hypotympanum, especially on the ventromedial aspect of the TB wall. Microscopically, MpEs were composed of osteonal bone growing from the periosteum and not by dystrophic calcification of necrotic tissue debris, as is hypothesized in dogs. KeywordsAfrican lion, computed tomography, mucoperiosteal exostoses, middle ear, Panthera leo, pathology, tympanic bullaThe middle ear structure is well known in domestic animals, but it has only been occasionally studied in lions.6 Threedimensional anatomic reconstructions show that the middle ear of the lion and cat are similarly arranged but have large differences in absolute and relative size. 6,12 In mammals, the middle ear is lined by respiratory epithelium that is contiguous with the nasopharynx via the auditory tube; however, portions of the middle ear are lined by squamous to cuboidal epithelium, especially the petrous portion of the temporal and tympanic membrane. 1,12 In a recent exhaustive study on the histology and histopathology of the cat middle ear, Sula et al 12 introduced the term auricular mucoperiosteum to name this unique mucosal membrane lining the middle ear, which is characterized by the direct apposition of epithelia and submucosa with periosteal connective tissues.Little is known about the pathology of the middle ear in the majority of animal species. Infectious (bacterial) otitis media has been more frequently described in dogs, cats, and cows.7,12,13 Sula et al 12 recently described the histologic characteristics of diseased ears in a series of 50 cats and concluded that middle ear diseases in cats are far more common than gross lesions or clinical literature suggests.