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Hibernating ground squirrels rely on a short active period for breeding and mass accrual, and are thus vulnerable to extreme climate events that affect key periods in their annual cycle. Here, we observed how a heatwave in March 2012 led to a phenological mismatch between sexes in Richardson’s ground squirrels (Urocitellus richardsonii). Females emerged from hibernation and commenced breeding earlier in 2012 relative to average female emergence. Despite external indicators suggesting that males were prepared for breeding, it appeared that not all males were physiologically prepared since 58.6% of males had non-motile sperm when breeding commenced. We found that males with non-motile sperm had smaller accessory glands than males with motile sperm. Body condition, relative testes size, and the relative size of accessory glands were significant predictors of sperm motility. There was no difference in litter size among years, nor a decrease in the number of juveniles emerged in 2012 or female yearlings recruited in 2013. The impact of this heatwave on male ground squirrels emphasizes the importance of assessing the consequences of climate change on breeding success of hibernating species in both sexes, since the different cues for emergence led to a mismatch in timing under this event.

We validated a radioimmunoassay-based method quantifying fecal glucocorticoid metabolites (FGMs) from captive male and female Richardson’s ground squirrels Urocitellus richardsonii. Blood samples were drawn to explore the correlation between plasma cortisol and FGM concentrations. We also injected groups of squirrels with normal saline (CTL; control), adre-nocorticotropic hormone (ACTH; stimulating adrenal activity), or dexamethasone (DEX; suppressing adrenal activity). Potential correlations between stress and behaviour were explored through quantification of fecal pellet production and the intervention necessary to elicit defecation, as well as the behaviour of subjects in the context of handling. Changes in plasma cortisol concentration between capture (baseline), and following handling (stress-induced) were also quantified for free-living squirrels. While glucocorticoid concentrations recovered from feces during our captive-animal study were not well correlated with plasma cortisol concentrations, and uncorrelated with defecation or behaviour, FGM concentrations did reflect the activation of the hypothalamic-pituitary-adrenal (HPA) axis. FGM concentrations increased significantly during initial captivity, but declined to baseline level as individuals acclimated to the novel environment. Injection of subjects with ACTH increased FGMs above baseline, confirming activation of the HPA axis. Plasma cortisol concentrations increased significantly with induced stress, indicating that capture and handling activated the glucocorticoid stress response even among previously handled, free-living subjects. Our findings validate a non-invasive tool that will afford new insight into the physiological processes underlying social, reproductive and antipredator behaviour of Richardson’s ground squirrels.

The endangered Puerto Rican crested toad (Peltophryne [Bufo] lemur) has been held and bred in zoos for release into protected areas in Puerto Rico since 1982. In 2004, several cases of a novel syndrome of skin changes in toads were noticed at the Toronto Zoo. A total of 21 toads were found to have similar lesions and the condition has been seen in several other groups of toads in subsequent years. Affected toads show an uncharacteristic sheen of dark-brown leathery skin, followed by recurring dysecdysis, reduced appetite, weight loss, and death from secondary causes. Histologically the condition is characterized by epithelial hyperplasia and hyperkeratosis, ulceration, and the presence of superficial mats of bacterial and fungal agents. No etiology has been identified and to date toads have not permanently responded to treatment with various pharmaceutical and nutritional therapies.

The captive breeding program for the endangered Puerto Rican crested toad (Peltophryne [Bufo] lemur) has been hampered by an undiagnosed condition called “Brown Skin Disease” (BSD). Toads develop widespread skin darkening, skin thickening and abnormal shedding and eventually succumb to a chronic loss of viability. This project evaluated the marine toad (Bufo marinus) as a model for the PRCT, examining vitamin A deficiency as a potential cause of BSD. Wild caught marine toads had significantly higher liver vitamin A concentrations (61.89 ± 63.49 µg/g) than captive born marine toads (0.58 ± 0.59 µg/g); P

"Brown skin disease" (BSD) is a clinical syndrome of dysecdysis, chronic weight loss and death, previously reported in Puerto Rican crested toads (Peltophryne lemur). Although vitamin A deficiency has been suggested, its cause remains unknown and multiple treatments have failed to prevent or reverse the condition. This study compared the efficacy of vitamin A supplementation, administered in different forms and by different routes, in 48 captive born Puerto Rican crested toads fed from metamorphosis on gut-loaded, dusted, commercially raised crickets. Forty-five toads started to show clinical signs of BSD at 9 months of age; all toads were treated orally with an oil-based vitamin A formulation twice weekly for 2 months but continued to deteriorate. Two treatment groups were then compared: Animals in one group (n=19) received 2 IU injectable vitamin A (Aquasol-A) per gram bodyweight subcutaneously twice weekly for 3 months with no change in diet. Toads in the other group (n=22) received a single oral dose of vitamins A, D3 , and E, and were fed on earthworms and crickets gut-loaded with produce and a finely-ground alfalfa-based pellet, dusted with the same vitamin/mineral supplement. All affected animals developed severe BSD equally and died during, or were euthanized at the end of, the treatment regimen, with no clinical improvement. Animals supplemented with Aquasol-A had significantly higher liver vitamin A concentrations compared with the other treatment group, whereas serum retinol concentrations showed no significant difference. Vitamin A supplementation does not appear a successful treatment once BSD symptoms have developed.

Cerebral xanthomatosis was diagnosed in three female green water dragons (Physignathus cocincinus), all of which presented with progressive neurologic signs. No antemortem evidence for xanthomatosis was identified, but on postmortem examination cholesterol granulomas, composed of cholesterol clefts surrounded by macrophages and multinucleated giant cells, were found in the forebrain of each animal and were associated with significant displacement and pressure on the adjacent brain. Although the cause of xanthomatosis in these animals is unknown, nutrition and trauma may be involved in the pathogenesis of this condition. Cerebrum, cholesterol, green water dragon, Physignathus cocincinus, xanthoma.

The adaptive manipulation of offspring sex and number has been of considerable interest to ecologists and evolutionary biologists. The physiological mechanisms that translate maternal condition and environmental cues into adaptive responses in offspring sex and number, however, remain obscure. In mammals, research into the mechanisms responsible for adaptive sex allocation has focused on two major endocrine axes: the hypothalamic pituitary adrenal (HPA) axis and glucocorticoids, and the hypothalamic pituitary gonadal (HPG) axis and sex steroids, particularly testosterone. While stress-induced activation of the HPA axis provides an intuitive model for sex ratio and litter size adjustment, plasma glucocorticoids exist in both bound and free fractions, and may be acting indirectly, for example by affecting plasma glucose levels. Furthermore, in female mammals, activation of the HPA axis stimulates the secretion of adrenal testosterone in addition to glucocorticoids (GCs). To begin to untangle these physiological mechanisms influencing offspring sex and number, we simultaneously examined fecal glucocorticoid metabolites, free and bound plasma cortisol, free testosterone, and plasma glucose concentration during both gestation and lactation in a free-living rodent (Urocitellus richardsonii). We also collected data on offspring sex and litter size from focal females and from a larger study population. Consistent with previous work in this population, we found evidence for a trade-off between offspring sex and number, as well as positive and negative correlations between glucocorticoids and sex ratio and litter size, respectively, during gestation (but not lactation). We also observed a negative relationship between testosterone and litter size during gestation (but not lactation), but no effect of glucose on either sex ratio or litter size. Our findings highlight the importance of binding proteins, cross-talk between endocrine systems, and temporal windows in the regulation of trade-offs in offspring sex and number.

The captive breeding program for the endangered Puerto Rican crested toad (Peltophryne [Bufo] lemur) has been hampered by an undiagnosed condition called "Brown Skin Disease" (BSD). Toads develop widespread skin darkening, skin thickening and abnormal shedding and eventually succumb to a chronic loss of viability. This project evaluated the marine toad (Bufo marinus) as a model for the PRCT, examining vitamin A deficiency as a potential cause of BSD. Wild caught marine toads had significantly higher liver vitamin A concentrations (61.89 ± 63.49 µg/g) than captive born marine toads (0.58 ± 0.59 µg/g); P0.001). A significant difference in serum vitamin A concentration was found between the captive and wild caught toads (P=0.013) and between the low vitamin A-fed and wild caught toads (P=0.004), when controlling for liver vitamin A concentrations. After captive toads were treated with topical and/or oral vitamin A, their hepatic vitamin A concentrations were similar to those of the wild toads, averaging 48.41 ± 37.03 µg/g. However, plasma vitamin A concentrations pre- and post-vitamin A supplementation did not differ statistically. We concluded that plasma vitamin A concentrations do not provide a linear indication of liver/body vitamin A status, and that both topical and oral supplementation with an oil-based vitamin A formulation can increase liver stores in amphibians. No evidence of BSD or other signs of deficiency were noted in the marine toads, although this feeding trial was relatively short (127 days). To date, clinical, pathological and research findings do not support vitamin A deficiency as a primary factor underlying BSD.

This study investigates the use of an enzyme immunoassay to measure keratin glucocorticoid concentrations in reptilian shed skins. Keratin glucocorticoid concentrations were compared to fecal glucocorticoid concentrations during the period of keratin growth in the African House Snake (Lamprophis fuliginosus) and the Eastern Massasauga Rattlesnake (Sistrurus catenatus catenatus). Biochemical validation was performed for the shed skin and fecal corticosterone enzyme immunoassays in the African House Snake. Biological and physiological validations were attempted in the African House Snake. A statistically significant positive association was detected between shed skin corticosterone and the mean fecal corticosterone metabolites from 3 weeks before to 1 week after the previous ecdysis in the African House Snake. A statistically significant difference was not detected between the shed skin corticosterone concentrations of the minimally handled control and the weekly handled (or experimentally stressed) African House Snakes. Adrenocorticotropic hormone stimulation did not result in the physiological validation anticipated for shed skin corticosterone concentrations in the African House Snake.

AVIAN bornavirus (ABV), a newly discovered agent, has been identified as the causal agent of proventricular dilation disease (PDD) in psittacine birds (Honkavuori and others 2008, Kistler and others 2008). Subsequent research, including bird inoculation studies (Gancz and others 2009, Gray and others 2010) and outbreak investigations (Kistler and others 2010) have provided strong supporting evidence. PDD is a significant pathological syndrome, with high mortality affecting primarily psittacine birds, that has been reported worldwide since the late 1970s. Characteristic pathological findings of PDD …