Effects of Water Loss on New Mexico Spadefoot Toad (Spea multiplicata) Development, Spleen Cellularity, and Corticosterone Levels.

Amphibian metamorphosis is complex and larval morphology and physiology are completely restructured during this time. Amphibians that live in unpredictable environments are often exposed to stressors that can directly and indirectly alter physiological systems during development, with subsequent consequences (carryover effects) later in life. In this study, we investigated the effects of water level reduction on development rate, spleen size and cellularity, and examined the role of corticosterone levels in premetamorphic, metamorphic, and postmetamorphic New Mexico spadefoot toads (Spea multiplicata). Based on previous studies, we hypothesized that declining water level would increase tadpole developmental rate, but with the trade-off of increasing corticosterone to a level that would subsequently affect spleen size and cellularity, thus prolonging potential immunological suppression. Declining water levels increased developmental rate by 3 days; however, there were no significant body size effects. Corticosterone (CORT) was negatively correlated with total length, snout vent length, body weight, and spleen weight at metamorphosis, suggesting that size at metamorphosis and the immune system may be affected by excessive CORT levels. When compared to other studies, our results support the view that multiple factors may be acting as stressors in the field affecting amphibian responses, and simple pathways as tested in this study may not adequately represent field conditions.

Seasonal variation of the oviduct of the American alligator, Alligator mississippiensis (Reptilia: Crocodylia).

The annual oviductal cycle of the American alligator, Alligator mississippiensis, is described using light and electron microscopy. Previous work done by Palmer and Guillette ([1992] Biol Reprod 46:39-47) shed some light on the reproductive morphology of the female alligator oviduct; however, their study was limited and did not report details relating to variation across the reproductive season. We recognize six variable regions of the oviduct: infundibulum, tube, isthmus, anterior uterus, posterior uterus, and vagina. Each area shows variation, to some degree, in the histochemistry and ultrastructure of oviductal secretions. Peak secretory activity occurs during the months of May and June, with the greatest variation occurring in the tube and anterior uterus. During the month of May, high densities of neutral carbohydrates and proteins are found within the tubal and anterior uterine glands. The epithelium of the entire oviduct secretes neutral carbohydrates throughout the year, but many regions lack protein secretions, and the posterior uterine glands show little secretory activity of any type throughout the year. After oviposition, secretory activity decreases drastically, andthe oviduct resembles that of the premating season. This study also provides evidence to support the homology between alligator and bird oviducts. Sperm were observed in glands at the tubal-isthmus and utero-vaginal junctions in preovulatory, postovulatory and postovipository females.

Sperm storage in the oviduct of the American alligator.

Oviducts of the American alligator (Alligator mississippiensis) were examined histologically for the presence of stored sperm. Two regions containing sperm were identified, one at the junction of the posterior uterus and the vagina (UVJ) and the other at the junction of the tube and isthmus (TIJ). In these areas, sperm were found in the lumina of oviductal glands. The glands in these areas of the oviduct are diffuse and shallow and appear to allow better access to sperm than glands located elsewhere. Histochemically, the glands of the UVJ reacted weakly for carbohydrates and proteins, whereas those of the TIJ reacted strongly for these same two components, secretions of which are associated with sperm storage structures in other reptiles. Sperm were not in contact with the glandular epithelium, and glands at the UVJ contained more sperm than those at the TIJ. Oviductal sperm storage was observed not only in recently mated females but in all females possessing uterine eggs as well as all females known to be associated with a nest. We conclude that female alligators are capable of storing sperm in their oviductal glands, but not from one year to the next.

Renal sexual segment of the Cottonmouth snake, Agkistrodon piscivorous (Reptilia, Squamata, Viperidae).

The seasonal variation of the renal sexual segment (RSS) of males of the Cottonmouth snake, Agkistrodon piscivorous, is described using light and electron microscopy. This study is the first to describe the ultrastructure of the RSS of a viper (Viperidae) and only the fourth on a snake. Renal sexual segments from males collected February to May and from August to November are similar in appearance. The cells are eosinophilic and react with periodic acid/Schiff procedure (PAS) for neutral carbohydrates and bromphenol blue (BB) for proteins. At the ultrastructure level, the cells contain large (2 microm diameter), electron-dense secretory granules and smaller vesicles with a diffuse material, and these structures abut against the luminal border and upon clear vacuoles continuous with intercellular canaliculi. Evidence was found for both apocrine and merocrine processes of product release. In June and July, the RSS are significantly smaller in diameter, largely basophilic, and have only scattered granules that are PAS+ and BB+. Cytologically, the RSS from June to July lack electron-dense secretory granules and the smaller vesicles with diffuse material. Numerous condensing vacuoles and abundant rough endoplasmic reticulum, however, indicate that active product synthesis is occurring. This is the first report of significant seasonal variation in the histology and ultrastructure of the RSS of a snake, although such reports exist for lizards. The seasons when the RSS is most highly hypertrophied correspond to the fall and spring mating seasons of A. piscivorous, as determined by other studies.