Temperature regulation in the echidna (Tachyglossus aculeatus)

The echidna (Tachyglossus aculeatus) maintained a body temperature of 30.7°C ± 1.03 s.d. (N = 23) at ambient temperatures (T_A) between 0 and 25°C. It may, however, also become hypothermic at low T_A. At T_A = 30°C or above the echidna became hyperthermic. The thermoneutral range was about 20–30°C. At low T_A the metabolic rate might be increased several fold. The thermal conductance was at a minimum at T_A = 20°C, and was not further reduced at lower T_A. At higher T_A the thermal conductance increased up to five-fold. The evaporation showed little change with increasing T_A. At the highest T_A we used (33°C) the evaporation on the average accounted for the dissipation of only about one-third of the metabolic heat produced. These findings suggest that the echidna, although it can maintain its body temperature at low ambient temperature, cannot rely upon evaporation as the major avenue for heat loss at high ambient temperature.     

Control of breathing in the echidna (Tachyglossus aculeatus) during hibernation

Resting non-hibernating echidnas are characterised by low metabolic rates, but also have a very low respiratory frequency and a variable respiratory minute volume, often resulting in low levels of arterial O(2) and high CO(2). As the echidna lies at one physiological extreme among the hibernators, in terms of its large size and low metabolism and ventilatory requirement when not hibernating, a study of control of breathing during hibernation in echidnas should provide a useful test of the generality of various models. We used non-invasive techniques to study breathing patterns and the control of ventilation in 6 echidnas. Hibernating echidnas (T(b) range 7-10 degrees C) showed episodic breathing with bursts of breaths (average 36+/-16 breaths in 24+/-5 min) followed by a period of apnea (76+/-17 min) then a series (8+/-4) of slow breaths at 14+/-1 min intervals leading up to the next burst. Increasing CO(2) levels in the inspired air increased the number of breaths in a burst, eventually leading to continuous breathing. Inter burst breaths were controlled by O(2): hypoxia increased inter burst breaths, and decreased burst length, while hyperoxia abolished inter burst breaths and increased the apneic period. Overall, while CO(2) was a strong respiratory stimulus in hibernating echidnas, O(2) had little effect on total ventilation, but did have a strong effect on the breathing pattern.     

The anatomy of the cerebral cortex of the echidna (Tachyglossus aculeatus)

The cerebral cortex of the echidna is notable for its extensive folding and the positioning of major functional areas towards its caudal extremity. The gyrification of the echidna cortex is comparable in magnitude to prosimians and cortical thickness and neuronal density are similar to that seen in rodents and carnivores. On the other hand, many pyramidal neurons in the cerebral cortex of the echidna are atypical with inverted somata and short or branching apical dendrites. All other broad classes of neurons noted in therian cortex are also present in the echidna, suggesting that the major classes of cortical neurons evolved prior to the divergence of proto- and eutherian lineages. Dendritic spine density on dendrites of echidna pyramidal neurons in somatosensory cortex and apical dendrites of motor cortex pyramidal neurons is lower than that found in eutheria. On the other hand, synaptic morphology, density and distribution in somatosensory cortex are similar to that in eutheria. In summary, although the echidna cerebral cortex displays some structural features, which may limit its functional capacities (e.g. lower spine density on pyramidal neurons), in most structural parameters (e.g. gyrification, cortical area and thickness, neuronal density and types, synaptic morphology and density), it is comparable to eutheria.     

Immunoglobulin genetics of Ornithorhynchus anatinus (platypus) and Tachyglossus aculeatus (short-beaked echidna)

In this paper, we review data on the monotreme immune system focusing on the characterisation of lymphoid tissue and of antibody responses, as well the recent cloning of immunoglobulin genes. It is now known that monotremes utilise immunoglobulin isotypes that are structurally identical to those found in marsupials and eutherians, but which differ to those found in birds and reptiles. Monotremes utilise IgM, IgG, IgA and IgE. They do not use IgY. Their IgG and IgA constant regions contain three domains plus a hinge region. Preliminary analysis of monotreme heavy chain variable region diversity suggests that the platypus primarily uses a single VH clan, while the short-beaked echidna utilises at least 4 distinct VH families which segregate into all three mammalian VH clans. Phylogenetic analysis of the immunoglobulin heavy chain constant region gene sequences provides strong support for the Theria hypothesis. The constant region of IgM has proven to be a useful marker for estimating the time of divergence of mammalian lineages.     

Iron(III) binding proteins of echidna (Tachyglossus aculeatus) and platypus (Ornithorhynchus anatinus)

Iron (III) binding proteins are isolated from echidna (Tachyglossus aculeatus multiaculeatus) and platypus (Ornithorhynchus anatinus) milk and blood. On the basis of several criteria it is shown that the milk proteins are not lactoferrins, but are transferrins similar to the corresponding transferrins from the blood. The heterogeneity of the proteins, particularly the echidna milk transferrin, is, at least in part, due to different levels of sialic acid. Their N-terminal sequences (30 residues) are determined and compared with those of other transferrins and lactoferrins. The role of the proteins is discussed.     

Type I interferon genes from the egg-laying mammal, Tachyglossus aculeatus (short-beaked echidna)

The type I IFN are an important group of multifunctional cytokines that have, for whatever reason, evolved to a high level of complexity in eutherian mammals such as humans and mice. However, until recently, little was known about the type I IFN systems of the other two groups of extant mammals, the marsupials and the egg-laying monotremes. Preliminary partial type I IFN sequences from the short-beaked echidna were previously found to cluster only with the IFN-beta subtype in phylogenetic analyses, but a lack of sequence information made interpretation of these results tenuous. Here, we report cloning of the full-length genes of representatives from the two previously defined groups of echidna type I IFN by genomic walking PCR. Along with analysis of conserved cysteine placement and promoter elements, phylogenetic analysis incorporating these sequences strongly suggest that the two groups of echidna type I IFN genes are in fact homologous to IFN-alpha and IFN-beta, confirming that the duplication leading to these two major classes of type I IFN occurred prior to the divergence of eutherians and monotremes some 180 million years ago. Thus, even though there are major differences in gene copy number and heterogeneity, separate IFN-alpha and IFN-beta gene families are a feature of the cytokine networks of all three groups of living mammals.     

The isolation and amino acid sequences of echidna (Tachyglossus aculeatus) milk lysozyme I and II.

Comparative studies of monotreme proteins are of particular value in gaining an understanding of the origin of mammals and their interrelationships. The presence of two lysozyme variants, echidna lysozyme I and II, has been confirmed in mature milk samples of Tachyglossus aculeatus multiaculeatus and Tachyglossus aculeatus aculeatus respectively. A simplified procedure is described for their isolation. Their amino acid sequences, the first determined for a monotreme secretory protein, are unusual. They are shown to be c-type lysozymes, each consisting of a single chain of 125 residues (terminating at Cys 125). The only other known c-type lysozyme with this termination is that of pigeon eggwhite. Echidna lysozyme is unique in having no Cys at position 6, but at position 9. It has precisely the residues relevant to the binding of Ca(II), and most of the residues implicated in the galactosyl transferase modifier action of alpha-lactalbumin. However, the weak modifier action previously observed for variant I, prepared by a different method, was not found for the present preparation. The evolutionary significance of the results is discussed.     

Morphology,histochemistry and physiology of the major salivary glands in the echidna Tachyglossus aculeatus (Monotremata)

The three major salivary glands of the monotreme echidna are described. The parotid is a typical serous gland with tubulo-acinar secretory endpieces and a well-developed system of striated ducts. The mandibular gland, although light microscopically resembling a mucous gland, secretes very little glycoprotein. Its cells are packed instead with serous granules, resembling in fine structure the “bull's eye” granules in the mandibular gland of the European hedgehog Erinaceus europaeus. The sublingual glands secrete an extremely viscous mucous saliva. Expulsion of this saliva through the narrow ducts is probably aided by contraction of the extensive myoepithelial sheaths surrounding the secretory tubules. Application of the glyoxylic acid induced fluorescence method failed to demonstrate adrenergic innervation in any of the glands.     

Topography and chemoarchitecture of the striatum and pallidum in a monotreme, the short-beaked echidna (Tachyglossus aculeatus)

The topography and chemoarchitecture of the striatum and pallidum in a monotreme, the short-beaked echidna (Tachyglossus aculeatus) have been studied using Nissl staining in conjunction with myelin staining, enzyme reactivity to acetylcholinesterase and NADPH diaphorase, and immunoreactivity to parvalbumin, calbindin, calretinin, tyrosine hydroxylase, neuropeptide Y, and neurofilament protein (SMI-32 antibody). All those components of the striatum and pallidum found in eutherian mammals could also be identified in the echidna's brain, with broad chemoarchitectural similarities to those regions in eutherian brains also apparent. There was a clear chemoarchitectural gradient visible with parvalbumin immunoreactivity of neurons and fibers, suggesting a subdivision of the echidna caudatoputamen into weakly reactive rostrodorsomedial and strongly reactive caudoventrolateral components. This may, in turn, relate to subdivision into associative versus sensorimotor CPu and reflect homology to the caudate and putamen of primates. Moreover, the chemoarchitecture of the echidna striatum suggested the presence of striosome-matrix architecture. The morphology of identified neuronal groups (i.e., parvalbumin, calbindin, and neuropeptide Y immunoreactive) in the echidna striatum and pallidum showed many similarities to those seen in eutherians, although the pattern of distribution of calbindin immunoreactive neurons was more uniform in the caudatoputamen of the echidna than in therians. These observations indicate that the same broad features of striatal and pallidal organization apply across all mammals and suggest that these common features may have arisen before the divergence of the monotreme and therian lineages.     

Body temperature as an indicator of egg-laying in the echidna,Tachyglossus aculeatus

We investigated the usefulness of body temperature (T_b) as a guide to egg-laying in a monotreme mammal, the echidna, Tachyglossus aculeatus, and attempted to quantify changes in T_b and relate them to specific reproductive events. Six female echidnas were implanted with temperature loggers and then radio-tracked in the wild for up to 6 years. In reproductive years there was a significant reduction in T_b variability 21.7±2.5 days after final arousal from hibernation, which coincided with the time at which the female entered the nursery burrow. Egg-laying occurred within 2 days of this T_b change which lasted an average of 43±4 days.