The emergence of disease as a significant global threat to amphibian diversity has generated considerable interest in amphibian defenses against cutaneous microbial infection and disease. To date, however, the influence of sloughing on the susceptibility of amphibians to infection and disease has been largely overlooked. To investigate the potential for sloughing to regulate topical microbial loads, the abundance of cultivable cutaneous bacteria and fungi in the cane toad Rhinella marina were compared before and after sloughing. Toads were also exposed to fluctuating thermal regimes (10−20 and 20−30°C) and variable photoperiods to investigate possible effects of season and climate on sloughing periodicity. Sloughing substantially reduced the abundance of cultivable cutaneous bacteria and fungi by up to 100%. The intermoult interval of toads maintained at 10−20°C was twice that of animals at 20−30°C and did not appear to thermally acclimate. Photoperiod had no discernable influence on sloughing periodicity. Results of this study suggest that normal sloughing cycles could play a significant role in controlling the persistence and build-up of cutaneous microbes, including pathogens. The loss of non-pathogenic commensal and protective skin microbiota after sloughing may also influence host susceptibility to cutaneous pathogens. We suggest that the spatio-temporal dynamics of chytridiomycosis, the widespread and often fatal disease caused by the fungal pathogen Batrachochytrium dendrobatidis, are related to temperature not only because of its effect on the growth of the fungus, but also because of its effect on the frequency of host sloughing.KEY WORDS: Skin · Shedding · Temperature · Rhinella marina · Pathogen · Batrachochytrium dendrobatidis · Commensal microbe · Epidermis · Cutaneous Resale or republication not permitted without written consent of the publisherDis Aquat Org 101: [235][236][237][238][239][240][241][242] 2012 and microbes adherent to the most peripheral skin layers can be shed with the shed keratinocytes (Dahl 1993). In amphibians, the stratum corneum is shed on a regular basis (Larsen 1976, Withers 1995; moreover, unlike mammals, sloughing of the stratum corneum in amphibians is usually entire (i.e. all of the stratum corneum is shed at once) (Taylor & Ewer 1956, Ling 1972, Larsen 1976, Withers 1995. As such, sloughing could play a significant role in regulating the abundance and persistence of microbes, both beneficial and pathogenic, at the body's surface.To investigate the influence of sloughing on microbial abundances on amphibian skin, we examined changes in the abundance of cultivable cutaneous microbes post-sloughing in cane toads Rhinella marina (Linnaeus, 1758). As ectotherms, physiological processes such as sloughing are likely to be affected by environmental factors such as temperature and day length. Seasonal differences in temperature and photoperiod may, therefore, affect sloughing periodicity and in turn contribute to the seasonality of some amphibian diseases (Berger et...

Many of the recent global amphibian mass mortalities, declines and extinctions have been attributed to the emerging infectious disease chytridiomycosis. There have been mass mortalities due to ranaviral disease but no major declines or extinctions. Controlling the transmission and spread of disease is of utmost importance, especially where there is the potential for human involvement. We have reviewed current hygiene guidelines for working with wild frogs, identified potential flaws and recommended those most suitable and effective for the field environment. Our within-site hygiene measures aim to reduce the risk of transmission among individuals. These measures encompass the capture, handling and holding of amphibians, skin disinfection before and after invasive procedures, marking frogs, sealing open wounds and treatment of accessory equipment. Our between-site hygiene measures aim to mitigate the risk of pathogen spread among populations. We have designed a risk calculator to help simplify and standardise the decision-making process for determining the level of risk and appropriate risk mitigation strategies to reduce the risk of increasing pathogen spread above background levels. Calculation of an overall risk score for pathogen spread takes into account the prior activity of field workers, the proposed activity, remoteness of the site, presence of known pathogens and the consequences of increased pathogen spread for amphibians in a given area.

The skin is the first line of defence in preventing the establishment of pathogens and associated infections in frogs. Regular sloughing of the outer layer can reduce the abundance of cultivable cutaneous microbes in green tree frogs which has ramifications for our understanding of cutaneous pathogens like the amphibian chytrid fungus.

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A review of the current conservation status of Australian amphibians was recently completed as part of a World Conservation Union (IUCN) sponsored Global Amphibian Assessment (GAA). Fifty of 216 amphibian species (23%) in Australia are now recognized as threatened or extinct in accord with IUCN Red List Categories and Criteria. Here we report on the categories and criteria under which individual species qualified for listing and provide a summary of supporting information pertaining to population and distribution declines. Major threatening processes contributing to listing of species are also reviewed.

SUMMARYThe estuarine crocodile, Crocodylus porosus, inhabits both freshwater and hypersaline waterways and maintains ionic homeostasis by excreting excess sodium and chloride ions via lingual salt glands. In the present study, we sought to investigate the phenotypic plasticity, both morphological and functional, in the lingual salt glands of the estuarine crocodile associated with chronic exposure to freshwater (FW) and saltwater (SW) environments. Examination of haematological parameters indicated that there were no long-term disruptions to ionic homeostasis with prolonged exposure to SW. Maximal secretory rates from the salt glands of SW-acclimated animals (100.8±14.7·mol·100·g ). There were no differences in the mass-specific metabolic rate of salt gland tissue slices from FW-and SW-acclimated animals (558.9±49.6 and 527.3±142.8·l·O 2 ·g -1 ·h -1 , respectively). Stimulation of the tissue slices from SW-acclimated animals by methacholine resulted in a 33% increase in oxygen consumption rate. There was no significant increase in the metabolic rate of tissues from FW-acclimated animals in response to methacholine. Morphologically, the secretory cells from the salt glands of SW-acclimated animals were larger than those of FW-acclimated animals. In addition, there were significantly more mitochondria per unit volume in secretory tissue from SW-acclimated animals. The results from this study demonstrate that the salt glands of C. porosus are phenotypically plastic, both morphologically and functionally and acclimate to changes in environmental salinity.

 AbstractCramp, R.L., Franklin, C.E. and Meyer, E.A. 2004. The impact of prolonged fasting during aestivation on the structure of the small intestine in the greenstriped burrowing frog, Cyclorana alboguttata . -Acta Zoologica (Stockholm) 86 : 13-24The effects of short-term fasting and prolonged fasting during aestivation on the morphology of the proximal small intestine and associated organs were investigated in the green-striped burrowing frog, Cyclorana alboguttata (Anura: Hylidae). Animals were fasted for 1 week while active or for 3-9 months during aestivation. Short-duration fasting (1 week) had little effect on the morphology of the small intestine, whilst prolonged fasting during aestivation induced marked enteropathy including reductions in intestinal mass, length and diameter, longitudinal fold height and tunica muscularis thickness. Enterocyte morphology was also affected markedly by prolonged fasting: enterocyte cross-sectional area and microvillous height were reduced during aestivation, intercellular spaces were visibly reduced and the prevalence of lymphocytes amongst enterocytes was increased. Mitochondria and nuclei were also affected by 9 months of aestivation with major disruptions to mitochondrial cristae and increased clumping of nuclear material and increased infolding of the nuclear envelope. The present study demonstrates that the intestine of an aestivating frog responds to prolonged food deprivation during aestivation by reducing in size, presumably to reduce the energy expenditure of the organ.

SUMMARY Although green striped burrowing frogs (Cyclorana alboguttata)experience large reductions in the mass and absorptive surface area of the small intestine (SI) during aestivation, little is known about how this may affect the functional capacity of the SI. We examined changes in the function(l-proline uptake rate and capacity) and metabolism of the SI(in vitro oxygen consumption, Na+/K+-ATPase activity and abundance) of C. alboguttata following 6 months of aestivation. l-Proline uptake rate was significantly higher in aestivating frogs, but overall uptake capacity was lower than in active frogs. Total SI oxygen consumption rate (VO2) was also lower in aestivating frogs, despite no difference in mass-specific V̇O2. The proportion of intestinal V̇O2 associated with Na+/K+-ATPase activity and protein synthesis was equivalent between active and aestivating frogs, suggesting these processes were unaffected by aestivation. Indeed, the activity of Na+/K+-ATPase transporters in the SI of aestivating frogs was not different from that of active animals. Aestivating frogs maintained Na+/K+-ATPase activity, despite experiencing a reduction in the density of Na+/K+-ATPase transporters, by increasing the molecular activity of the remaining pumps to 2–3 times that of active frogs. These results show that functionality of the SI is maintained at the cellular level, potentially facilitating the reclamation of nutrients from the intestinal lumen while in aestivation. Despite this, the functional capacity of the SI in aestivating C. alboguttata is significantly reduced due to a reduction in tissue mass,helping frogs to conserve energy while in aestivation.