Polymorphic microsatellites identified in an endangered dasyurid marsupial,the dibbler (Parantechinus apicalis)

Dibblers are carnivorous marsupials that were considered extinct for over 60 years. They now persist in small, highly fragmented populations. In this work we describe the characterization of seven polymorphic microsatellite loci that are being used to investigate population parameters. For 22 individuals, from the mainland population, these markers had allelic diversities from three to 11 (alleles per locus) and expected heterozygosities ranging from 0.53 to 0.89. Primers were also tested on a number of closely related dasyurids and in other marsupials, suggesting that their usefulness in population studies may go beyond investigations of the dibbler.     

Molecular Evidence for a Deep Clade of Dunnarts (Marsupialia: Dasyuridae: Sminthopsis)

Sminthopsis is the most speciose genus of living dasyurid marsupials and, along with its close relatives Antechinomys and Ningaui, constitutes the clade Sminthopsini. Phylogenetic relationships among the 23 species in this clade have been the subject of much morphological and molecular investigation, including a recent integration of penis morphology (in Sminthopsis) with molecular systematics. Several phylogenetic issues remain open, however, including the monophyly of Sminthopsis and branching order among early sminthopsin lineages. In this study, we revisit sminthopsin systematics with an expanded molecular data set, including new DNA sequences from mitochondrial (valine transfer-RNA and 16S ribosomal RNA) and nuclear (interphotoreceptor retinoid binding protein and beta-fibrinogen) loci, along with previously published sequences of cytochrome b, 12S ribosomal RNA, control region, and protamine P1. Our results again fail to establish the monophyly of Sminthopsis, but do provide a clearer resolution of early sminthopsin branching. Specifically, our phylogeny suggests three major groups of Sminthopsis species: S. longicaudata (perhaps the sister of Antechinomys); the Macroura species group of previous authors (S. crassicaudata, S. macroura, S. virginiae, S. douglasi, and S. bindi); and the remaining 13 species allied with the Murina species group. Our results depart from previous molecular findings by reuniting S. ooldea with the Murina group, while resolving S. psammophila as sister to the hairy-footed dunnarts (S. hirtipes and S. youngsoni). We suggest that this conflict traces to anomalous phylogenetic signal in previously published cytochrome b sequences. Penis morphology maps reasonably well onto our phylogeny, requiring parallel origination of only one of the ten morphotypes described for Sminthopsis.     

Milk composition in the eastern quoll, Dasyurus viverrinus (Marsupialia:Dasyuridae)

The milk constituents of Dasyurus viverrinus, a carnivorous marsupial, exhibited major quantitative and qualitative changes during the course of lactation. The milk produced in the early stages of lactation was dilute, about 13-16% (w/w) solids before 3 weeks with carbohydrate representing the major fraction. In the latter stages of lactation the milk was concentrated, around 30% solids, and lipid was the predominant fraction. Palmitic acid was the major fatty acid present in early-stage milk but oleic acid became predominant in milk after 10 weeks post-partum. The changes in milk composition in D. viverrinus were similar to those described for the milks of herbivorous marsupials which therefore suggests that this pattern may be uniform throughout the Marsupialia.     

Molecular Relationships of New Guinean Three-Striped Dasyures, (Myoictis, Marsupialia: Dasyuridae)

Complete nucleotide sequences of the cytochrome b and 12S rRNA genes and partial sequences of the mitochondrial 16S rRNA gene and the nuclear ɛ-globin gene were obtained from multiple exemplars of the New Guinean dasyurid, Myoictis. Allozyme data were also obtained from most of the same animals. The molecular data show that the genus comprises a number of genetically distinct lineages which correspond with groups proposed by Woolley (2005) on the basis of a number of morphological traits, including the form of the tail i.e. Myoictis leucura (sp. nov.), M. melas, M. wallacei and M. wavicus (new status). Divergence dates estimated from the weighted-average distances for the combined cytochrome b and 12S rRNA data, calibrated with a dasyurid-thylacine divergence 25 million years ago, suggest that the early cladogenic events separating Myoictis took place in the late Miocene. Subsequent separation of M. wavicus and M. leucura from a common ancestor as well as some genetic differentiation within M. melas, took place in the medial Pliocene.     

Seasonal changes in the reproductive anatomy of male Antechinus stuartii (Marsupialia: Dasyuridae)

Male Antechinus stuartii (Marsupialia: Dasyuridae) undergo significant endocrinological changes during their life history. The endocrine changes are associated with complete male mortality. The accessory reproductive tract of males is important for seminal plasma production, and changes associated with the reproductive cycle were not described in detail by earlier studies. The present study sought to describe some of the seasonal changes in structure of the male accessory reproductive tract in relation to the known hormonal changes. The epididymis, prostate, and bulbourethral glands are relatively undifferentiated in February and May, a time when plasma concentrations of testosterone are known to be low. By July, considerable hypertrophy and differentiation of the accessory reproductive tract are observed. This is most obvious in the bulbourethral glands, which change from being indistinguishable from one another in February and May to being three large morphologically and histologically distinct glands in July. The hypertrophy and secretory activity continue into August, the breeding season. These findings correlate with the hormonal profiles found in other studies of A. stuartii. J Morphol 231:261–275, 1997. © 1997 Wiley-Liss, Inc.     

Plasma prolactin concentrations throughout lactation in the eastern quoll, Dasyurus viverrinus (Marsupialia:Dasyuridae)

Blood samples were collected throughout lactation from eight D. viverrinus, and the concentrations of prolactin in the plasma determined by an heterologous, double-antibody radioimmunoassay. For the first 45-50 days of lactation concentrations fluctuated between 2 and 80 ng/ml but were generally less than 60 ng/ml. They then increased sharply to over 120 ng/ml and greater than 200 ng/ml in some animals. Levels began to decline after 120-130 days of lactation and were less than 40 ng/ml in all animals by the time lactation had ceased, between 160 and 185 days, when the young became independent. The timing of these changes correlates well with a rapid increase in the growth rate of the young, the time at which they first relinquish the teat and remain in the nest, their first intake of solid food shortly after eruption of the first molars and specific changes in milk composition. The pattern of prolactin concentrations throughout lactation in the quoll is therefore very similar to that previously described for the tammar, Macropus eugenii, and the possum, Trichosurus vulpecula.     

Social and foraging components of the home range in Antechinus stuartii (Dasyuridae: Marsupialia)

The home range of Antechinus stuartii has two components. A small area (0.94 ha for males, 0.38 ha for females) is used for foraging, while a much greater social range (mean for males in a given year may exeed 5 ha; the mean for females never exceeded 3 ha) encompasses communal nests, which are used by both males and females. The foraging range of females is stable throughout the year. The foraging range may not coincide at all, or only partially coincide with her social range. By contrast, male foraging ranges drift through the year, and generally lie within their social range. Intersexual differences reflect the different selection pressures on the sexes. Females require access to resources to sustain a costly lactation, while males require a knowledge of many possible sites for mating and early resolution of agonistic interactions with other males with which they cohabit during mating.     

Habitat of the white-footed dunnart Sminthopsis leucopus (Gray) (Dasyuridae: Marsupialia) in a logged,burnt forest near Bega,New South Wales

In the winter of 1981, 9 months after an intense fire in November 1980 in the recently logged Mumbulla State Forest, the numbers of a small population of Sminthopsis leucopus had slightly increased, while the common species of small mammals had declined sharply from the previous winter. The study reported in this paper aimed to identify the habitat of S. leucopus. Ninety study plots were divided equally among three classes of forest (unlogged forest, and forest logged in 1979 and 1980), between two aspects (north- and south-facing slopes), and among three topographic positions (ridge, midslope and gully). Plots were assessed on the basis of both floristic data and the structural components of ground, shrub and tree cover. The primary finding was that the habitat of S. leucopus was the treeless ridges and mid-slopes with less than 51% ground cover in the logged areas of burnt forest. The floristic analysis showed that the vegetation subcommunities restricted to gullies, or carrying a ferny ground layer of vegetation, were not selected by S. leucopus. An analysis of movements of male S. leucopus identified two classes. ‘Resident’ males occurred on logged ridges and midslopes characterized by a floristic subcommunity of open forest with a grassy understorey.‘Explorer’ males moved through a variety of ridge habitats, including unlogged forest, and were spread more widely through the floristic communities. The pattern of habitat selection identified in this study shows S. leucopus to be ecologically distinct from the other species of small mammals in the forest and thus in need of special consideration in management programmes if it is to be conserved. Under the current regime, the uniform treatment of the forest leads to widespread stands of dense regrowth to the exclusion of the species and is inimical to its survival.     

Sperm morphology and storage in the female reproductive tract of the fat-tailed dunnart,Sminthopsis crassicaudata (Marsupialia: Dasyuridae)

A study of spermatozoa in the isthmus of the oviduct and of the surrounding epithelial cells in the dasyurid marsupial, Sminthopsis crassicaudata, was carried out. At least 10% of the ejaculated spermatozoa probably populate the isthmus region, where many come to reside in crypts until around the time of ovulation. Ultrastructural observations of spermatozoa in this region indicated that they had intact acrosomes and were identical in their morphology with those in the cauda epididymidis. After ovulation spermatozoa rapidly disappeared, some of which may be phagocytosed by the cells lining the crypts. These epithelial cells were also found to have many large, electron-dense granules at the time of sperm storage, but the contents did not appear to be released until the zygotes passed along the tract. The secretory activity of these cells may thus relate more to the production of the shell membrane that comes to surround the zygote than to the cells performing a nutritive or protective function for the spermatozoa during their period of storage within the female reproductive tract.     

Demographic responses of Antechinus stuartii (Marsupialia) to supplementary food

This study investigates the effects of supplementary feeding on populations of Antechinus stuartii, an insectivorous, semi-arboreal species of dasyurid marsupial. Animals were live-trapped, marked and released in three study areas in forest near Canberra, from September 1980 to November 1981. Supplementary food was supplied above ground in trees in one study area from January to November 1981, and in the second study area from January to May 1981, while the third study area was left unmanipulated as a control. The supplemented populations increased in numbers, mean body weights and survival, and showed decreased movements between traps and smaller overlaps in home ranges. Discontinuation of food in the second study area in May 1981 produced reciprocal shifts in these parameters. Reproduction, home-range area, mean head-body length, use of the terrestrial and arboreal parts of the habitat were largely unaffected by provision of supplementary food. In contrast to rodents, where fed populations usually increase rapidly due to immigration or increased reproduction, the supplemented populations of A. stuartii responded relatively slowly via increased survival of young. Population responses were probably retarded at the beginning of the experiments in summer due to the abundance of natural invertebrate prey, but became more obvious in winter when natural food was scarce. The added food probably contributed directly to increased numbers, survival and body weights, but may also have enhanced these parameters indirectly by allowing foraging animals to space out and avoid intraspecific intolerant contacts. These responses in A. stuartii suggest strongly that populations are limited by the food supply.     

A Reexamination of Proposed Morphology-Based Synapomorphies for the Families of Dasyuromorphia (Marsupialia). I. Dasyuridae

The validity of eight morphological features previously advanced as synapomorphic for Dasyuridae is investigated in the light of new fossil and molecular data. Results indicate that one of these features (alisphenoid–periotic enclosure of the foramen ovale) is common to outgroups for Dasyuromorphia. Another feature (loss of intestinal cecum) is a likely synapomorphy for Dasyuromorphia. Two features (development of a hypoconulid notch, enlargement of stylar cusp D) may represent shared–derived characters within Dasyuromorphia but not at the family level for Dasyuridae (i.e., probably unite Dasyuridae–Thylacinidae). Another two features (loss of posterolateral palatine foramina, reduction of P₃) are also apomorphic within Dasyuromorphia but unite specialized clades within Dasyuridae. Only two previously treated features are probable synapomorphies for the family (enlargement of the alisphenoid tympanic wing and development of a distinct periotic hypotympanic sinus). An additional feature is identified as a dasyurid synapomorphy (presence of a distinct tubal foramen). Of all putative synapomorphies proposed to date, only the presence of a periotic hypotympanic sinus and tubal foramen are unique for Dasyuridae among dasyuromorphians. Results suggest considerable homoplasy for basicranial features within Dasyuromorphia. Independent acquisition for alisphenoid enclosure of the foramen ovale, development of secondary foramina ovale and loss of posterolateral palatal foramina has occurred in derived thylacinid and dasyurid clades. Convergence is also indicated for hypertrophy of the alisphenoid tympanic wing shown for dasyurids and myrmecobiids, and the development of a squamosal epitympanic sinus in Thylacinidae, Dasyuridae, and Myrmecobiidae. The finding of plesiomorphy for alisphenoid–periotic enclosure of the foramen ovale within Dasyuromorphia undermines the strongest morphology-based synapomorphy uniting a monophyletic Dasyuridae–Myrmecobiidae. Phylogenetic placement for some plesiomorphic fossil dasyuromorphians, known only from dental material, within Dasyuridae is currently untenable, with no dental synapomorphies uniting the family. The value of identifying morphoclines within clades known from robust phylogenetic data for consideration in character analysis is stressed, as is the importance of form–function and ontogenetic data.