Haematological characteristics of captive Parma wallabies (Macropus parma)

 Blood was collected from 31 Parma wallabies from five different captive populations. The morphology of erythrocytes, leucocytes and platelets was similar to that previously described for macropodids. The effects of location (population), anaesthesia and analyser on the haematological characteristics of the wallabies were considered. Location had an affect on many of the values, anaesthesia affected some of the leucocyte values, and analyser had little effect. Received: 22 November 2002 / Accepted 13 February 2003 Acknowledgements We thank Mrs R. Power, Ms S. Bayliss, Ms T. Smith and Mrs P. Slack for technical assistance, Mr D. Hopcroft for assistance with electron microscopy and Mr D. Hedderly for performing the statistical analyses.     

Construction of a highly enriched marsupial Y chromosome-specific BAC sub-library using isolated Y chromosomes

The Y chromosome is perhaps the most interesting element of the mammalian genome but comparative analysis of the Y chromosome has been impeded by the difficulty of assembling a shotgun sequence of the Y. BAC-based sequencing has been successful for the human and chimpanzee Y but is difficult to do efficiently for an atypical mammalian model species (Skaletsky et al. 2003, Kuroki et al. 2006). We show how Y-specific sub-libraries can be efficiently constructed using DNA amplified from microdissected or flow-sorted Y chromosomes. A Bacterial Artificial Chromosome (BAC) library was constructed from the model marsupial, the tammar wallaby (Macropus eugenii). We screened this library for Y chromosome-derived BAC clones using DNA from both a microdissected Y chromosome and a flow-sorted Y chromosome in order to create a Y chromosome-specific sub-library. We expected that the tammar wallaby Y chromosome should detect ∼100 clones from the 2.2 times redundant library. The microdissected Y DNA detected 85 clones, 82% of which mapped to the Y chromosome and the flow-sorted Y DNA detected 71 clones, 48% of which mapped to the Y chromosome. Overall, this represented a ∼330-fold enrichment for Y chromosome clones. This presents an ideal method for the creation of highly enriched chromosome-specific sub-libraries suitable for BAC-based sequencing of the Y chromosome of any mammalian species.     

Reproduction in male swamp wallabies (Wallabia bicolor): puberty and the effects of season

This study describes pubertal changes in testes and epididymides and seasonal changes in the adult male reproductive organs and plasma androgen concentrations of the swamp wallaby (Wallabia bicolor). Pre-pubescent males had testes with solid seminiferous cords and spermatogenesis only to the stage of gonocytes. Their epididymides had empty lumina along their entire length. The testes of three males undergoing puberty had some lumen formation and mitotic activity. Their epididymides were similar in appearance to those of adult males but were entirely devoid of any cells within the lumen of the duct. Three other pubescent males showed full lumen formation in the testes and spermatogenesis up to the elongating spermatid stage. Their epididymides were similar in appearance to those of adult males but with no spermatozoa in the duct. However, cells of testicular origin were found in the lumen of the duct in all regions suggesting that testicular fluids and immature germ cells shed into the rete testes flow through the seminiferous tubules into the epididymis before the release of mature testicular spermatozoa. The weights of testes and epididymides of adult males showed no change throughout the year but prostate weight and plasma androgen concentrations varied significantly with season, with maximums in spring and summer and minimums in winter. The volume fraction of Leydig cells and seminiferous tubules was significantly lower in winter than in summer; but, despite this, maturing spermatozoa were found in the testes throughout the year. Females in the area conceived year-round, suggesting that seasonal changes in the male reproductive tract did not prevent at least some males from breeding throughout the year.     

Brain-derived neurotrophic factor is expressed in a gradient in the superior colliculus during development of the retinocollicular projection

Abstract Theoretical models of topographic map formation have postulated a gradient of attractant in addition to a gradient of repulsion in the target. In species where many axons grow past their correct positions initially, it has also been argued that a parallel gradient of attractant or branching signal is required to ensure collateral formation at the correct position (O'Leary et al., 1999). Brain-derived neurotrophic factor (BDNF) is a known attractant and promotes branching of retinal axons. We have examined its distribution in the superior colliculus and that of its receptor, trkB, in the retina, using immunohistochemistry and in situ hybridization, respectively, during the development of the topographic retinocollicular projection in the wallaby, a marsupial mammal. The number of glial endfeet expressing BDNF at the surface of the colliculus was found to be in a high caudal-to-low rostral gradient during the time when the retinocollicular projection was developing. When the projection was mature the rostrocaudal gradient had disappeared and the number of detectable endfeet expressing BDNF was very low. Messenger RNA for TrkB was expressed in the retinal ganglion cell layer throughout the time when the retinocollicular projection was developing, with no difference in expression across the nasotemporal axis of the retina. The low rostral to high caudal distribution of BDNF in glial endfeet supports the idea that it is providing a parallel gradient of attractant or branching signal in the colliculus.     

The development of the gubernaculum and inguinal closure in the marsupial Macropus eugenii

This study reports the developmental anatomy of testicular descent and inguinal closure of the tammar wallaby (Macropus eugenii) from birth to maturity. In females the ovary migrated caudally between days 10 and 20 after birth. The gubernaculum differentiates into the round ligament in the abdomen and extra-abdominally as the ilio-marsupialis muscle of the mammary glands. In males the testes migrated to the internal inguinal ring by day 20 post partum (pp), coinciding with the enlargement of the gubernaculum, and from the internal inguinal ring to the scrotum between days 20 and 65 pp. During descent there was an increase in the hyaluronic acid concentration in cells of the gubernaculum and scrotum. Development of the cremaster muscle began by day 10 pp on the periphery of the gubernaculum and its basic structure was completed by day 60 pp. After descent the inguinal canal closed between days 50 and 60 pp, but a small irregular lumen persisted, somewhat similar to that seen in the congenital scrotal hydrocoele of humans. Tammars have a hopping mode of locomotion and, like humans, are essentially bipedal. We suggest that inguinal closure evolved in these two species because their upright posture may otherwise lead to a high incidence of inguinal hernias.     

Erythrocyte metabolism in the Koala, the common brushtail possum and the whiptail wallaby

Erythrocyte metabolism was studied in the koala (Phascolarctos cinereus), the common brushtail possum (Trichosurus vulpecula) and the whiptail wallaby (Macropus parryi). Aspects of metabolism studied were the levels of ATP, 2,3-diphosphoglycerate (2,3-DPG) and reduced glutathione (GSH), activities of erythrocyte enzymes, glycolytic rate, the rate of glutathione S-conjugate transport (GCT), susceptibility of erythrocytes to an oxidising agent and the effect of eucalyptus oil on the red cells. Some of the major findings were that compared to common brushtail possum and the whiptail wallaby, the koala red cells (a) produced significantly greater amounts of lactate; (b) had significantly lower activity of glutathione S-transferase, (c) had significantly lower rate of GCT, (d) were more susceptible to oxidant stress induced by acetylphenylhydrazine (APH) and (e) exhibited a greater degree of haemolysis when incubated with eucalyptus oil.     

Interactions between herbivores,vegetation and eucalypt tree seedlings in a plantation forestry environment

Non-lethal management methods for reducing mammal browsing damage to Eucalyptus seedlings during plantation establishment are currently being investigated in Tasmania, Australia. One method is use of selective retention of non-seedling vegetation in plantations. To assess its potential, information is needed on how herbivores use such vegetation and whether it has positive or negative effects on browsing and growth of commercial tree seedlings. We quantified the populations of five mammalian herbivore species in and around a spot-herbicided E. nitens plantation during its establishment. We also investigated several aspects of the interactions between those mammalian herbivores, insect herbivores, tree seedlings and non-seedling vegetation. We found that overall population densities were high for red-bellied pademelons (Thylogale billardierii) and red-necked wallabies (Macropus rufogriseus rufogriseus), but very low for common brushtail possums (Trichosurus vulpecula), common wombats (Vombatus ursinus) and the introduced European rabbit (Oryctolagus cuniculus). Population densities of the five mammalian species did not change in response to planting E. nitens seedlings. Dominant vegetation in the plantation was grass, then forbs and non-grass monocots. Herbivores ate a large proportion of available plantation vegetation in unfenced plots. Mammal browsing had little effect on survival of E. nitens seedlings 15 weeks after planting, but significantly reduced their growth. In unfenced plots, insects, mainly Heteronyx spp., caused twice as much damage to seedlings as browsing mammals. The absence of mammalian browsing in fenced plots, which resulted in high grass cover, was associated with reduced insect damage to seedlings.     

The vomeronasal organ of the tammar wallaby

The vomeronasal organ is the primary olfactory organ that detects sexual pheromones in mammals. We investigated the anatomy of the vomeronasal organ of the tammar wallaby ( Macropus eugenii ), a small macropodid marsupial. Pheromones may be important for activation of the hypothalamo-pituitary axis of tammar males at the start of the breeding season because plasma testosterone and luteinizing hormone concentration in males rise concurrently with pregnancy and the post-partum ovulation in females. The gross anatomy and the connection to the brain of the vomeronasal organ were examined by light and electron microscopy in adult male and female tammars. The vomeronasal organ was well developed in both sexes. The vomeronasal organ is a tubular organ connected at the rostral end via the nasopalatine duct (incisive duct) to the mouth and nasal cavity. At the rostral end the lumen of the vomeronasal organ was crescent shaped, changing to a narrow oval shape caudally. Glandular tissue associated with the vomeronasal organ increased towards the blind end of the organ. The tammar has the typical pattern of mammalian vomeronasal organs with electron-dense supporting cells and electron-lucent receptor cells. Microvilli were present on the surface of both epithelia while cilia were only found on the surface of the non-receptor epithelium. Some non-receptor epithelial cells appeared to secrete mucus into the vomeronasal organ lumen. The vomeronasal organ shows a high degree of structural conservation compared with eutherian mammals. The degree of vomeronasal organ development makes it likely that, as in other mammals, pheromones are important in the reproduction of the tammar.     

Serological relationships among maize rough dwarf-like viruses.

The serological relationships of most of the known maize rough dwarf-like viruses were investigated using immune electron microscopy and gel-diffusion tests to detect the reactions of the inner capsids and the B spikes of the virus particles. The antigens were those of maize rough dwarf virus (MRDV), pangola stunt virus (PSV), cereal tillering disease virus (CTDV), oat sterile dwarf virus (OSDV), Arrhenatherum blue dwarf virus (ABDV) and Lolium enation virus (LEV). The sera were those against MRDV, rice black streaked dwarf virus (RBSDV), and Fiji diseases virus (FDV). These were absorbed with healthy material and dsRNA if necessary. The results showed that MRDV, RBSDV, PSV, and CTDV were serologically related in both the inner capsid and the B spike. However, MRDV, PSV, and CTDV did not react with FDV antiserum (RBSDV was not available). OSDV, ABDV, and LEV did not react with sera against MRDV, RBSDV, or FDV. We conclude that the MRDV-like viruses fall into at least three serological groups: (I) MRDV, RBSDV, PSV, and CTDV (interrelated, but not related to II and III); (II) FDV (not related to I and III); (III) OSDV, ABDV, and LEV (not related to I and II, but interrelationships unknown).     

Immunohistochemistry of the lymphoid tissues of the tammar wallaby,Macropus eugenii

The lymphoid tissues of the metatherian mammal, the adult tammar wallaby, Macropus eugenii, were investigated using immunohistochemical techniques. Five cross-reactive antibodies previously shown to recognize surface markers in marsupial tissues and five previously untested antibodies were used. The distribution of T-cells in the tissue beds of spleen, lymph node, thymus, gut-associated lymphoid tissue (GALT) and bronchus-associated lymphoid tissue (BALT) was documented using antibodies to CD3 and CD5. Similarly, B-cells were identified in the same tissues using anti-CD79b. Antibodies to CD8, CD31, CD79a and CD68 failed to recognize cells in these tissue beds. In general the pattern of cellular distribution identified using these antibodies was similar to that observed in other marsupial and eutherian lymphoid tissues. This study provides further information on the commonality of lymphoid tissue structure in the two major groups of extant mammals, metatherians and eutherians.