Methyl jasmonate does not induce changes in Eucalyptus grandis leaves that alter the effect of constitutive defences on larvae of a specialist herbivore.

The up-regulation of secondary metabolic pathways following herbivore attack and the subsequent reduction in herbivore performance have been identified in numerous woody plant species. Eucalypts constitutively express many secondary metabolites in the leaves, including terpenes and formylated phloroglucinol compounds (FPCs). We used clonal ramets from six clones of Eucalyptus grandis and two clones of E. grandis x camaldulensis to determine if methyl jasmonate (MeJA) treatment could induce changes in the foliar concentrations of either of these groups of compounds. We also used bioassays to determine if any changes in the performance of larvae of Paropsis atomaria, a chrysomelid leaf beetle, could be detected in treated ramets versus the untreated controls, thus indicating whether MeJA induced the up-regulation of defences other than terpenes or FPCs. We found no significant effects of MeJA treatment on either the foliar concentrations of terpenes and FPCs or on herbivore performance. We did, however, detect dramatic differences in larval performance between Eucalyptus clones, thereby demonstrating large variations in the levels of constitutive defence. Larval feeding on clones resistant to P. atomaria resulted in high first instar mortality and disruption of normal gregarious feeding behaviour in surviving larvae. Histological examination of larvae feeding on a resistant clone revealed damage to the midgut consistent with the action of a toxin. These findings concur with mounting evidence that most evergreen perennial plants lack foliar-induced defences and suggest that constitutively expressed secondary metabolites other than those commonly examined in studies of interactions between insect herbivores and Eucalyptus may be important in plant defence.

Marker-based quantitative genetics in the wild?: the heritability and genetic correlation of chemical defenses in eucalyptus.

Marker-based methods for estimating heritability and genetic correlation in the wild have attracted interest because traditional methods may be impractical or introduce bias via G x E effects, mating system variation, and sampling effects. However, they have not been widely used, especially in plants. A regression-based approach, which uses a continuous measure of genetic relatedness, promises to be particularly appropriate for use in plants with mixed-mating systems and overlapping generations. Using this method, we found significant narrow-sense heritability of foliar defense chemicals in a natural population of Eucalyptus melliodora. We also demonstrated a genetic basis for the phenotypic correlation underlying an ecological example of conditioned flavor aversion involving different biosynthetic pathways. Our results revealed that heritability estimates depend on the spatial scale of the analysis in a way that offers insight into the distribution of genetic and environmental variance. This study is the first to successfully use a marker-based method to measure quantitative genetic parameters in a tree. We suggest that this method will prove to be a useful tool in other studies and offer some recommendations for future applications of the method.

Differential susceptibility to Eucalyptus secondary compounds explains feeding by the common ringtail (Pseudocheirus peregrinus) and common brushtail possum (Trichosurus vulpecula).

The effect of two plant secondary metabolites, tannins and formylated phloroglucinol compounds (FPCs), on the intake of Eucalyptus foliage by common ringtail (Pseudocheirus peregrinus) and common brushtail possums (Trichosurus vulpecula) was studied. We manipulated the amount of tannin that was free to bind with protein by coating foliage with polyethylene glycol 4000 (PEG) and relied on natural intraspecific variation in FPC concentrations. In contrast to ringtail possums, brushtail possums showed a greater tolerance to FPCs and ate more foliage when it was coated with PEG, suggesting that tannins limited their food intake. Brushtails detected the effects of tannins through immediate oral sensations rather than through systemic effects. Ringtail possums appeared highly tolerant of foliar tannins yet susceptible to low concentrations of FPCs. We could not detect any interaction between tannins and FPCs that affected the intake of Eucalyptus foliage by either species of possum. Although ringtail and brushtail possums are widely regarded as specialist and generalist folivores, respectively, their differential susceptibility to co-occurring secondary metabolites suggests greater complexity. Each possum species appears to be a specialist in its own right, which leads to a partitioning of available foliage. Brushtails avoid tannins and ringtails avoid FPCs.

The effect of excesses and deficiencies in amino acids on the feeding behaviour of the common brushtail possum (Trichosurus vulpecula).

In this study of the amino acid nutrition of a marsupial we tested three hypotheses: (a) that brushtail possums eat less when diets contain excesses or deficiencies in essential amino acids, (b) that brushtail possums choose diets that do not contain amino acid excesses, and (c) that amino acid consumption is mediated partly by the 5HT3 receptor. Possums ate less when 0.2-1.0% methionine (wet matter) was added to the diet, but similar concentrations of lysine and threonine had little effect. However, when given a choice, possums always selected the basal ration over one with added lysine, methionine or threonine at concentrations between 0.05% and 0.9%. In contrast to the experiments with excess amino acids, possums did not eat less of a diet almost devoid of an essential amino acid. Instead, the possums ate less when their diets contained synthetic amino acids rather than similar amounts and proportions of amino acids as casein. Contrary to the third hypothesis, the 5HT3 receptor antagonist, ondansetron, did not affect feeding by possums given a diet containing 0.8% methionine, suggesting that post-ingestive feedback, via the 5HT3 receptor, does not regulate amino acid intake when diets contain amino acid excesses.

Threonine requirements of broiler chickens: why do published values differ?

1. A statistical analysis of published data using restricted maximum likelihood showed that the age of the bird and the dietary crude protein concentration in test diets explained most of the variation in published threonine requirements. The sex of the bird and the main ingredients in the test diets were less important contributors of variation. The analysis resulted in a model that we tested in an accompanying paper. 2. Published data on threonine requirements are less variable than they first appear. There is no need for a plethora of studies on nutrient requirements other than those that test specific hypotheses, take a modelling approach and endeavour to explain requirements as units of nutrient per unit of production. 3. Models that use published data for predicting the responses of broilers to amino acids should first standardise the data by removing the variation due to factors such as age of bird, dietary crude protein content and the raw materials used in the ration.

Threonine requirements of broiler chickens: an experimental validation of a model using growth responses and carcase analysis.

1. This paper describes the experimental testing of a model derived from an analysis of published data on the threonine requirements of broiler chickens. The model, published in a separate paper, showed that the age of the bird and dietary crude protein were highly significant determinants of threonine requirements. 2. We tested this model by measuring the threonine requirements of male broilers aged 7 to 21 d and 21 to 42 d fed on wheat-peanut meal diets containing graded concentrations of threonine. The growth of those birds given adequate threonine was compared with that of others fed a typical wheat-soyabean diet to measure the relative value of peanut meal as a protein concentrate. Finally, we measured the dry matter, nitrogen and fat concentrations in the carcase using near infrared reflectance spectroscopy (NIRS) to test the hypothesis that birds given diets deficient in an amino acid produce carcases with more fat. 3. Increasing the threonine concentration of the diet from 5.7 to 7.2 g/kg improved the growth and feed conversion ratio (FCR) of broilers. However, a further increase in dietary threonine to 7.7 g/kg had an adverse effect. The determined threonine requirement agreed with our prediction model. 4. From 7 to 21 d birds given the diet containing 7.2 g/kg threonine ate more, weighed more, had an improved FCR and, in absolute terms, had carcases with more dry matter, fat and protein than did birds given the wheat-soyabean diet. However, after correction for treatment differences in body mass there were no differences in the carcase parameters. Likewise, there were no differences in any measurements between birds given these two diets between 21 and 42 d.

Can betaine partially replace or enhance the effect of methionine by improving broiler growth and carcase characteristics?

1. Growth rates and carcase characteristics were measured in male broiler chickens fed on a control diet deficient in methionine (c. 2.8 g/kg methionine) or a series of diets containing graded levels of betaine or DL-methionine or both additives. 2. We aimed to answer 2 main questions. First, can betaine replace part of the methionine in a broiler ration? Secondly is there a synergism between methionine and betaine? 3. Birds given the control diet or that supplemented only with betaine ate less, grew more slowly, had higher food convension ratio (FCR) and varied more in mass at 42 d than birds fed diets with DL-methionine. Adding 1.2 g/kg DL-methionine to the control ration produced the heaviest birds at 42 d (2500 g) with the 2nd heaviest breast muscle (366 g). 4. After correcting for treatment differences in body mass (analysis of convariance), birds fed on the control diet and the diet supplemented with betaine only, had relatively lighter breast muscles but relatively heavier abdominal fat pads than those of birds given diets supplemented with DL-methionine. However, adding betaine to diets containing added methionine further improved the relative breast muscle yield. 5. After correcting for differences in body mass between treatments, birds fed on diets containing most methionine had lighter viscera than birds fed diets deficient in methionine. This demonstrated gut plasticity, suggesting that the viscera enlarged to sequester methionine from low-methionine diets. 6. Our data refute the hypothesis that betaine can substitute for methionine in broilers fed diets that are marginally deficient in methionine plus cystine. However, betaine may improve carcase composition, especially breast meat yield.

Metabolic and organ mass responses to selection for high growth rates in the domestic chicken (Gallus domesticus).

Evolutionary hypotheses suggest that higher rates of postembryonic development in birds should either lower the resting metabolic rate (RMR) in a trade-off between the costs of growth and maintenance or increase RMR because of a buildup of metabolic machinery. Furthermore, some suggest that higher rates of postembryonic development in birds should reduce peak metabolic rate (PMR) through delayed tissue maturation and/or an increased energy allocation to organ growth. We studied this by comparing metabolic rates and organ sizes of fast-growing meat-type chickens (broilers) with those of birds from a laying strain, which grow much slower. During the first week of life, despite growing six times faster, the RMR of the broiler chickens was lower than that of birds of the laying strain. The difference between strains in RMR disappeared thereafter, even though broilers continued to grow twice as fast as layers. The differences between strains in growth rate during the first week after hatching were not reflected in similar differences in the relative masses of the heart, liver, and small intestine. However, broilers had heavier intestines once they reached a body mass of 80 g. In contrast, broilers had relatively smaller brains than did layers. There was a positive correlation, over both strains, between RMR and the masses of leg muscles, intestine, and liver. Furthermore, despite delayed maturation of muscle tissue, broilers exhibited significantly higher PMR. We hypothesize that a balance between the larger relative muscle mass but lower muscle maturation level explains this high PMR. Another correlation, between leg muscle mass and PMR, partly explained the positive correlation between RMR and PMR.

Reference intervals and physiologic alterations in hematologic and biochemical values of free-ranging desert tortoises in the Mojave Desert.

Desert tortoise (Gopherus agassizii) populations have experienced precipitous declines resulting from the cumulative impact of habitat loss, and human and disease-related mortality. Evaluation of hematologic and biochemical responses of desert tortoises to physiologic and environmental factors can facilitate the assessment of stress and disease in tortoises and contribute to management decisions and population recovery. The goal of this study was to obtain and analyze clinical laboratory data from free-ranging desert tortoises at three sites in the Mojave Desert (California, USA) between October 1990 and October 1995, to establish reference intervals, and to develop guidelines for the interpretation of laboratory data under a variety of environmental and physiologic conditions. Body weight, carapace length, and venous blood samples for a complete blood count and clinical chemistry profile were obtained from 98 clinically healthy adult desert tortoises of both sexes at the Desert Tortoise Research Natural area (western Mojave), Goffs (eastern Mojave) and Ivanpah Valley (northeastern Mojave). Samples were obtained four times per year, in winter (February/March), spring (May/June), summer (July/August), and fall (October). Years of near-, above- and below-average rainfall were represented in the 5 yr period. Minimum, maximum and median values, and central 95 percentiles were used as reference intervals and measures of central tendency for tortoises at each site and/or season. Data were analyzed using repeated measures analysis of variance for significant (P < 0.01) variation on the basis of sex, site, season, and interactions between these variables. Significant sex differences were observed for packed cell volume, hemoglobin concentration, aspartate transaminase activity, and cholesterol, triglyceride, calcium, and phosphorus concentrations. Marked seasonal variation was observed in most parameters in conjunction with reproductive cycle, hibernation, or seasonal rainfall. Year-to-year differences and long-term alterations primarily reflected winter rainfall amounts. Site differences were minimal, and largely reflected geographic differences in precipitation patterns, such that results from these studies can be applied to other tortoise populations in environments with known rainfall and forage availability patterns.

Energy metabolism in potoroine marsupials.

Fasting and fed metabolic rates were measured in three species of potoroine marsupials, the rufous rat-kangaroo (Aepyprymnus rufescens), the long-nosed potoroo (Potorous tridactylus) and the brush-tailed bettong (Bettongia penicillata). There were no significant differences among potoroine species in fasting metabolic rate. The lowest fasting heat production for each species was 11-20% less than the interspecific value of 295 kJ.kg-0.75.day-1 for basal metabolism of mature, non-reproductive eutherian homeotherms. The respiratory quotient of all species was reduced significantly as starvation proceeded, but only for B. penicillata was there a significant effect of starvation duration on fasting heat production. The night-time activity of P. tridactylus and B. penicillata doubled their daytime fasting heat production; the corresponding increase for A. rufescens was only 25%. The calorimetric measurement of fed animals showed no differences in digestible energy or metabolisable energy between species. Nevertheless, P. tridactylus and B. penicillata produced more heat per unit metabolic body mass. The maintenance energy requirements (kJ.kg-0.75.day-1) were 479, 494 and 345 for P. tridactylus, B. penicillata and A. rufescens, respectively. The net availability of metabolisable energy was about 0.70 in the three species. The combined heat production of fed female A. rufescens and their pouch young stayed relatively constant for the first two-thirds of pouch life, after which it rose sharply (20%) in response to the rapid growth of the young. Only during the last week of pouch life did the female enter negative energy balance. There was no indication that the metabolism of the female increased in response to the presence of a pouch young. The presence of pouch young did not alter the efficiency of utilisation of metabolisable energy. The daily energy requirement for maintenance was 0.83 MJ.day-1 or 0.36 MJ.kg-0.75.day-1.

Interactions between wheat and other dietary cereals with respect to metabolisable energy and digestible amino acids.

Small, varying improvements in apparent ileal amino acid digestibilities were obtained when wheat was combined with certain other cereals in broiler diets. Synergistic interactions, sometimes resulting in large improvements in the apparent metabolisable energy (AME) contents of the diets, were also observed with certain cereal combinations. Combining wheat with maize or oats enhanced the utilisation of dietary energy and amino acids.

The influence of environmental temperature, age and sex on the digestibility of amino acids in growing broiler chickens.

The ability of the broiler chicken to metabolise energy and to digest and absorb amino acids increased from 30 to 50 d of age. Although sex had no major effect on metabolisable energy or amino acid digestibilities at these ages, the influence of environmental temperature on amino acid digestibilities appeared to be sex-related, there being decreased digestibilities of most amino acids at higher temperatures in female but not male birds.