Steven J. Cork

The effects of elevated CO2 atmospheres on the nutritional quality of Eucalyptus foliage and its interaction with soil nutrient and light availability

Abstract Seedlings of Eucalyptus tereticornis (Smith) were grown under two levels of availability each of CO2 (352 and 793 µmol mol−1), soil nutrients (1/24 and 1/4 Hoagland’s solution) and light (full and 30% sunlight). Low soil nutrient availability or high light increased the C:N ratio of leaves, leading to lower leaf nitrogen concentrations, higher leaf specific weights and higher levels of both total phenolics and condensed tannins. These results were consistent with other studies of the effect of environmental resource availability on foliage composition. Similar results were observed when the C:N ratio of leaves was increased under elevated CO2. The changes in leaf chemistry induced by the treatments affected the performance of 4th-instar larvae of Chrysophtharta flaveola (Chapuis) fed on the leaves. Increased C:N ratios of leaves reduced digestive efficiencies and pupal body sizes and increased mortality. Below a threshold nitrogen concentration of approximately 1% dry mass, severe reductions in the performance of larvae were recorded. Such changes may have significant consequences for herbivores of Eucalyptus, particularly in view of projected increases in atmospheric CO2.

CONSEQUENCES OF BIOTRANSFORMATION OF PLANT SECONDARY METABOLITES ON ACID-BASE METABOLISM IN MAMMALS--A FINAL COMMON PATHWAY?

Regulation of acid-base homeostasis is essential for mammals and birds. Biotransformation and metabolism of absorbed plant secondary metabolites (PSMs) results in the production of organic acids that threaten acid-base homeostasis. Consequently these acids must be buffered and excreted from the body. The production of an acid load from detoxified PSMs should occur in herbivorous mammals and birds and with most PSMs and so may provide a unifying theme to explain many effects of PSMs on animal metabolism. Since the organic acids will be largely ionized at physiological pH, disposal of the hydrogen ion and the organic anion may proceed independently. Most hydrogen ions (H+) from organic acids are eliminated by one or more of three ways: (1) when they react with bicarbonate in the extracellular fluid to form carbon dioxide and the carbon dioxide is exhaled, (2) when they bind to dibasic phosphate and are excreted by the kidney as monobasic phosphate, and (3) when they are buffered and retained in the skeletal system. The secretion of phosphate ions and ammonium excretion are two ways in which the kidney replaces bicarbonate ions that have been eliminated as carbon dioxide. Secretion in the kidney tubule is an important means of excreting excessive organic anions rapidly. This process is saturable and may be subject to competition from a variety of different metabolites. Lagomorphs have limited capacity to form new bicarbonate from ammonium excretion and may therefore be obliged to excrete other cations such as sodium to balance the excretion of organic anions from PSMs. Acidemia has wide-ranging impacts on animals but browsing mammals and birds may have to break down muscle tissues to provide for urinary ammonium in order to generate bicarbonate for buffering. Acidemia also can affect the extent of urea recycling. Animals consuming browse diets may have to regulate feeding so that the rate of formation of hydrogen ions does not exceed the rate of disposal. The mechanisms by which this could occur are unknown.

Methods and pitfalls of extracting condensed tannins and other phenolics from plants: Insights from investigations onEucalyptus leaves.

Optimal conditions for extraction of tannins and other phenolics from tree foliage and their subsequent storage rarely have been investigated. We investigated methods of drying leaves, optimal solvents, and the effects of light and temperature on the extractability and stability of condensed tannins (proanthocyanidins) and total phenolics from leaves ofEucalyptus trees. Aqueous acetone was a better solvent than aqueous methanol for condensed tannins and total phenolics, but condensed tannins were less stable in aqueous acetone than aqueous methanol. Stability of condensed tannins also was decreased substantially by room temperature versus 4°C and by exposure to indirect sunlight, although the assay for total phenolics was unaffected. For quantitative estimation of condensed tannins, extraction with 50% acetone was better than methods of direct analysis of leaf tissue. The highest estimates of total condensed tannins were obtained by exhaustive extraction with 50% acetone followed by direct analysis of the residue. Lyophilization of fresh leaf increased yield of condensed tannin (although usually by less than 10%). Lyophilization and subsequent storage of extracts had little effect on assays for condensed tannins or total phenolics.

Use of fibrous diets by small herbivores: How far can the rules be 'bent'?

Allometric considerations suggest that small mammals should be unable to eat highly fibrous diets. A combination of the selective and more rapid passage of fibrous material through the gut, together with changes in gut capacity when energy requirements increase, may allow small mammals to escape these allometric constraints. Recent evidence that birds can absorb essential amino acids from the caecum (which has hitherto been considered insignificant in mammals) suggests that birds and mammals have evolved very different ways to be a herbivore.

ROLE OF DETOXIFICATION OF PLANT SECONDARY COMPOUNDS ON DIET BREADTH IN A MAMMALIAN HERBIVORE, Trichosurus- vulpecula

Theory predicts that mammalian herbivores detoxify different classes of plant secondary compounds via separate metabolic pathways and that generalist herbivores maintain broad diet breadth to avoid overloading individual detoxification pathways. We tested the hypothesis that a generalist marsupial herbivore, the common brushtail possum, Trichosurus vulpecula, can maintain a higher intake of food when allowed to select from two diets containing different profiles of secondary compounds (phenolics and terpenes) than when given access to the diets individually. Diets consisted of a fruit and vegetable mash to which was added ground leaves of either Eucalyptus melliodora or E. radiata. E. melliodora and E. radiata differ in their concentrations and types of secondary compounds. Brushtail possums include these eucalypt species as part of their natural diet. We measured food consumption and detoxification metabolites of possums on these diets. Consistent with the hypothesis, animals presented with a choice of both diets consumed more food than animals given diets singly. One of the two indicators of detoxification, acid load in urine, differed significantly between diets while the other, glucuronic acid, did not. These results provide partial support for the hypothesis that diet breadth is governed by detoxification abilities.

Diversity and habitat relationships of hypogeous fungi. II. Factors influencing the occurrence and number of taxa

Fruit-bodies of hypogeous fungi were sampled over two seasons across 136 forested study sites representing a stratified sample of the climatic, geological and topographic features of far south-eastern mainland Australia. Two hundred and nine species, over three-quarters being undescribed, were recorded. Statistical models based on various environmental attributes measured for each site were developed for the occurrence of several common taxa. At a landscape scale, climatic factors such as mean minimum temperature of the coldest month and annual mean moisture index were important explanatory variables for most taxa examined, but the type of response varied. More locally, topographic position, soil fertility, time since last fire and micro-habitat structures such as the leaf litter layer and number of large fallen trees also influenced the distribution of taxa in different ways. A model was then developed for the number of fungal species occurring at each site. Important explanatory variables were type of substrate, topography and diversity of potential host eucalypt species. The utility of each model constructed needs evaluation by further sampling of hypogeous fungi. Possible implications of our findings for forest management are discussed. Further analyses of our existing data are also identified.

Diversity and habitat relationships of hypogeous fungi. I. Study design, sampling techniques and general survey results

Hypogeous fungi are a large yet unknown component of biodiversity in forests of south-eastern mainland Australia. To better define their diversity and habitat relationships, we identified and counted fruit-bodies at 136 study sites sampling the climatic, geological and topographic features of the region. In one year 7451 fruit-bodies representing 209 species were collected in an autumn and spring sampling period. Only 57 of these species were previously described. Within genera, the number of species ranged from 1 to 21. Sites sampled in autumn averaged higher diversity of species and greater number of fruit-bodies than the same sites sampled in spring. Most major taxa occurred at more sites in autumn than in spring, whereas a few occurred more frequently in spring than in autumn. These patterns are consistent with those identified in previous smaller studies and likely reflect seasonal changes in soil moisture and temperature levels. Subsequent papers will explore factors influencing the occurrence, relative abundance and numbers of species of hypogeous fungi at the study sites and their community structure and possible host–plant relationships.

Mycophagy by small mammals in the coniferous forests of North America: nutritional value of sporocarps of Rhizopogon vinicolor, a common hypogeous fungus

Abstract We evaluated the nutritional value of sporocarps of Rhizopogon vinicolor, a common hypogeous fungus in the coniferous forests of North America, for two small mammal species: the Californian red-backed vole (Clethrionomys californicus) and the northern flying squirrel (Glaucomys sabrinus). Although the nitrogen concentration of sporocarps was high, much of it was in non-protein form or associated with cell walls, suggesting that it may be of low nutritional value or protected from mammalian digestive enzymes. Sporocarps also had high concentrations of cell wall constituents, indicating low availability of digestible energy. When fed a diet of this fungus alone in a controlled feeding experiment both mammal species lost a small amount of body mass. Digestibilities of dry matter, nitrogen, cell wall constituents and energy from sporocarps by both species were lower than the digestibilities of other food types by other similarly sized small mammals. Red-backed voles digested the various components of sporocarps at least as well as the flying squirrels, even though they were almost six-fold smaller in body mass. This observation supports the notion that red-backed voles, like other microtine rodents, have morphological and physiological adaptations of the digestive system that are postulated to permit greater digestion of fibrous diets than predicted on the basis of body size. Despite this, our results re-affirm previous conclusions that hypogeous fungi are only of moderate nutritional value for most small, hindgut-fermenting mammals. Future studies should focus on the importance of mixed-species of fungi in the diet of small mammalian mycophagists.

Production of Milk and Nutrition of the Dependent Young of Free‐Ranging Koalas (Phascolarctos cinereus)

The production of milk by lactating females, and energy expenditure and foliage intake of their dependent young, were investigated in free‐ranging koalas. Koalas had the lowest mass‐specific daily milk‐energy production at peak lactation so far recorded in a mammal, but the duration of reproduction was 58% longer than the combined marsupial and eutherian average. As a consequence, the total energy input to reproduction in koalas was similar to that in other mammals. We propose that the prolonged lactation and low daily rate of energy transfer to the young by female koalas is an adaptation to the low energy availability from their diet of Eucalyptus foliage. Energy requirements (field metabolic rates) of young koalas were lower than those expected for typical marsupials (only 60% at permanent pouch exit), which may be a necessary preadaptation that allows the low rate of maternal energy transfer. However, the energy requirements of the adult females were no lower than expected for marsupials. This pattern of energy requirements and age resulted in a linear relationship between field metabolic rate and mass for the koalas in this population. Differences in milk production between the years of the study coincided with fluctuations in the availability of preferred young foliage, which suggests that lactational output by koalas may be flexible and affected by diet quality. Despite the interannual differences in milk production, growth of the young was similar in the two years.

Metabolism of tannin-protein complex by facultatively anaerobic bacteria isolated from koala feces

The metabolic pathways involved in degradation of tannin-protein complex (T-PC) were investigated in various facultatively anaerobic bacteria, with specific reference to fecal isolates from the koala including T-PC-degrading enterobacteria (T-PCDE),Streptococcus bovis, Klebsiella pneumoniae, andK. oxytoca. It was demonstrated that T-PCDE andS. bovis biotype I were capable of degrading protein complexed with gallotannin (a hydrolyzable tannin), but not that complexed with quebracho (a condensed tannin). Subsequent studies showed that these strains metabolized gallic acid to pyrogallol. Strains ofKlebsiella pneumoniae andK. oxytoca, which did not degrade T-PC, also metabolized gallic acid into pyrogallol. Pyrogallol was not degraded by any strains studied, but it was not detected in fresh feces of the koalas. The majority of strains isolated from feces could degrade phloroglucinol. Based on these findings, we propose that members of the gut microflora of the koala cooperate in the degradation of T-PC.