Frank N. Carrick

Phylogeographic differentiation in the mitochondrial control region in the koala, Phascolarctos cinereus (Goldfuss 1817)

The koala, Phascolarctos cinereus, is a geographically widespread species endemic to Australia, with three currently recognized subspecies: P.c. adustus, P.c. cinereus, and P.c. victor. Intraspecific variation in the mitochondrial DNA (mtDNA) control region was examined in over 200 animals from 16 representative populations throughout the species’ range. Eighteen different haplotypes were defined in the ≈ 860 bp mtDNA control region, as determined by heteroduplex analysis/temperature gradient gel electrophoresis (HDA/TGGE). Any single population typically possessed only one or two haplotypes yielding an average within‐population haplotypic diversity of 0.180 ± 0.003, and nucleotide diversity of 0.16%. Overall, mtDNA control region sequence diversity between populations averaged 0.67%, and ranged from 0% to 1.56%. Nucleotide divergence between populations averaged 0.51%, and ranged from 0% to 1.53%. Neighbour‐joining methods revealed limited phylogenetic distinction between geographically distant populations of koalas, and tentative support for a single evolutionarily significant unit (ESU). This is consistent with previous suggestions that the morphological differences formalized by subspecific taxonomy may be interpreted as clinal variation. Significant differentiation in mtDNA‐haplotype frequencies between localities suggested that little gene flow currently exists among populations. When combined with microsatellite analysis, which has revealed substantial differentiation among koala populations, we conclude that the appropriate short‐term management unit (MU) for koalas is the local population.

Tree use, diet and home range of the koala (Phascolarctos cinereus) at Blair Athol, central Queensland

Free-ranging koalas (Phascolarctos cinereus) were monitored by means of radio-relocation in the area of Blair Athol Coal Mine and surrounding properties. Daytime tree use, home range and diet of these koalas was determined in spring and autumn, as was the leaf moisture composition of potential fodder species. Koalas used on average 93 (male koalas) and 56 (female koalas) trees during the period of observation, occupying home ranges of 135 and 101 ha respectively. Mean sightings per tree were 1.19 for both males and females and home-range sizes were not significantly different between sexes or seasons. Koalas were observed returning to previously used daytime roosting trees infrequently (<12%). Although koalas were observed roosting in trees of the species that they ate, proportional species representation in the diet of these koalas during spring and autumn did not accurately reflect concurrent observations of their daytime tree-roosting behaviour. Koalas were observed to utilise non-fodder species for daytime roosting, and patterns of daytime tree use and diet selection varied between individuals inhabiting adjacent communities within the site. Leaf moisture of tree species represented in koala diets was greater in autumn than spring.

Dispersal patterns in a regional koala population in south-east Queensland

Koala dispersal was investigated as part of a detailed ecological study of a nationally significant koala population located 20 km south-east of Brisbane, Queensland. From 1996 to 2000, 195 koalas from three sites were captured and fitted with radio-collars. A total of 40 koalas (23 males and 17 females) dispersed from these sites. Most (93%) dispersing individuals were 20–36 months of age. Three adult females (more than 36 months old) dispersed and no adult males dispersed during the study. A significantly higher proportion of young males dispersed than females. Dispersal occurred between June and December, with most dispersal of males commencing in July and August and that of females commencing between September and November prior to, and early in, the annual breeding season. The mean straight-line distance between the natal and breeding home ranges for males and females was similar and was measured at 3.5 km (range 1.1–9.7 km) and 3.4 km (range 0.3–10.6 km) respectively. Dispersing males and females tended to successfully disperse south and west of their natal home ranges and were generally unable to successfully disperse to urban areas within the study area, as a high proportion of the mortality of dispersing koalas was associated with attacks by domestic dogs and with collisions with vehicles on roads. Information from other studies indicates that most young koalas disperse from their natal areas. It is likely that the social behaviour and mating systems of koala populations provide mechanisms for young koalas to disperse. The potential role of dispersal in the dynamics of regional koala populations is discussed.

Genetic variation and structuring in the threatened koala populations of Southeast Queensland

Habitat fragmentation can act to cause reproductive isolation between conspecifics and undermine species’ persistence, though most studies have reported the genetic condition of populations that have already declined to a very small size. We examined genetic diversity within the vulnerable, declining koala (Phascolarctos cinereus) population in Southeast Queensland, Australia to determine the genetic impact of ongoing threatening processes. Five hundred and twelve koalas from ten Southeast Queensland Local Government Areas on the mainland and one island were genotyped at six polymorphic microsatellite loci. Based on Bayesian cluster analysis incorporating spatial data, the regional koala population was subdivided into six clusters, with location of major roads and rivers appearing to be consistent with being barriers to gene flow. The distribution of mtDNA control region haplotypes identified distinct coastal and inland clades suggesting that historically there was gene flow between koalas along the coast (though little interchange between coastal and inland animals). In contrast, koalas from the Koala Coast (Brisbane City, Logan City and Redland Shire) were shown by microsatellite analysis to be genetically distinct from adjacent areas. It is likely, therefore, that more recent reductions in population size and restricted gene flow through urbanisation have contributed to the genetic differentiation of koalas in the Koala Coast region.

Seasonal-Variation in Water Flux, Field Metabolic-Rate and Food-Consumption of Free-Ranging Koalas (Phascolarctos-Cinereus)

Mass-corrected field metabolic rates of free-ranging male koalas in central Queensland, Australia, varied between 0.329 MJ kg0.75 day-1 in summer and 0.382 MJ kg0.75 day-1 in winter. Field water influx measured 50.8 mL kg-0.8 day-1 in winter, increasing to 59.9 mL kg0.8 day-1 in summer for the same koalas, and was positively correlated with values for leaf moisture of food. Winter rates of water influx for koalas from Springsure were lower than those recorded for koalas from Victoria for the same period of the year. Mass-corrected feeding rates were lower in summer than winter; wet food intake was significantly lower than reported for similar sized female koalas from Victoria. The preferred browse was Eucalyptus crebra in winter and E. tereticornis in summer. Our study indicates that in central Queensland seasonal changes in diet selection by male koalas reflect increased energy requirements in winter and increased water requirements in summer.

Aspects of the epidemiology of Chlamydia psittaci infection in a population of koalas (Phascolarctos cinereus) in southeastern Queensland, Australia.

A population of free-ranging koalas in southeastern Queensland was examined to determine the prevalence of Chlamydia psittaci infections. Although C. psittaci was isolated from 46 of 65 (71%) koalas studied, only six (9%) of these had clinical signs of disease. Most adult females (82%) had back or pouch young present even though 67% of them were infected. There were no significant correlations between age, sex or site of sampling (urogenital versus conjunctival tissues) and the isolation of C. psittaci. No other important bacterial or fungal pathogens were isolated. The complement fixation test had a sensitivity of 7% and a specificity of 94% in detecting chlamydial infections, suggesting that it is unsuitable for use as a screening test. Chlamydia psittaci infection within this population appeared to represent a generally well-balanced host-parasite relationship and few animals had clinical signs of disease. Only four of 27 (15%) healthy koalas infected with C. psittaci followed for 24 wk after sampling developed eye disease or “dirty tail.” Two koalas with keratoconjunctivitis recovered without treatment during the study period. Additional factors, including the stresses imposed by loss of habitat, may act to produce overt disease in koalas with latent C. psittaci infections.

Low genetic diversity and inbreeding depression in Queensland koalas

The amount of genetic variation in two natural populations of Queensland koalas (Phascolarctos cinereus adustus) was assessed by analysis of mitochondrial DNA. Levels and any adverse effects of inbreeding (inbreeding depression) were estimated from the pedigree of a well-characterised captive colony. Genetic diversity of mitochondrial DNA was found to be exceedingly low both within and between the two populations, but the variation detected was found to be strongly structured geographically. Inbreeding levels in the captive colony were moderate to high yet the only apparent evidence of inbreeding depression was a male-biased sex ratio. There was no evidence for decreased juvenile survivorship or growth rate with inbreeding. Because of the limited data it would be premature to conclude that koalas are relatively resistant to the effects of inbreeding. However, we suggest the hypothesis that koalas have a history of small population size, resulting in reduced susceptibility to inbreeding depression.

Cytochrome P450 aromatase in grey mullet: cDNA and promoter isolation; brain, pituitary and ovarian expression during puberty.

In a study towards elucidating the role of aromatases during puberty in female grey mullet, the cDNAs of the brain (muCyp19b) and ovarian (muCyp19a) aromatase were isolated by RT-PCR and their relative expression levels were determined by quantitative real-time RT-PCR. The muCyp19a ORF of 1515bp encoded 505 predicted amino acid residues, while that of muCyp19b was 1485 bp and encoded 495 predicted amino acid residues. The expression level of muCyp19b significantly increased in the brain as puberty advanced; however, its expression level in the pituitary increased only slightly with pubertal development. In the ovary, the muCyp19a expression level markedly increased as puberty progressed. The promoter regions of the two genes were also isolated and their functionality evaluated in vitro using luciferase as the reporter gene. The muCyp19a promoter sequence (650 bp) contained a consensus TATA box and putative transcription factor binding sites, including two half EREs, an SF-1, an AhR/Arnt, a PR and two GATA-3 s. The muCyp19b promoter sequence (2500 bp) showed consensus TATA and CCAAT boxes and putative transcription binding sites, namely: a PR, an ERE, a half ERE, a SP-1, two GATA-binding factor, one half GATA-1, two C/EBPs, a GRE, a NFkappaB, three STATs, a PPAR/RXR, an Ahr/Arnt and a CRE. Basal activity of serially deleted promoter constructs transiently transfected into COS-7, alphaT3 and TE671 cells demonstrated the enhancing and silencing roles of the putative transcription factor binding sites. Quinpirole, a dopamine agonist, significantly reduced the promoter activity of muCyp19b in TE671. The results suggest tissue-specific regulation of the muCyp19 genes and a putative alternative promoter for muCyp19b.

Climate change and the koala Phascolarctos cinereus: water and energy

We studied two groups of koalas during a drought in central Queensland to investigate potential impacts of climatic variability on the physiology and behaviour of this species. The tree use, water turnover, field metabolic rate and diet of koalas during autumn and spring were compared to a similar study of koalas in summer and winter, also in central Queensland, to generate a seasonal picture of the response of koalas to climatic variation. We also compared the microclimate temperature of a range of food and non-food tree species against daily ambient temperatures, to examine the benefit to koalas of using of non-food species. Field metabolic rate, adjusted for body mass, was significantly higher in spring than autumn and there was no difference between males and females. Neither females with pouch young nor those with back young had significantly different FMR to that of females without young, confirming that koalas may compartmentalize energy demands during lactation. Estimations of theoretical water in...

Breeding dynamics of koalas in open woodlands

The spatial and breeding dynamics of koalas in sub-tropical woodlands at Blair Athol in central Queensland were intensively monitored between 1993 and 1998. Genetic relationships among koalas at this locality were studied to determine the breeding dynamics of males, including whether ‘resident’ or ‘transient’ males dominate as sires. Males and females were radio-collared and tracked periodically throughout each year of the study. Genotypes from hypervariable microsatellite loci identified uniquely all individuals and were used to analyse parentage as well as to determine population genetic parameters when compared with other regional localities. Koalas at Blair Athol comprise a population in genetic equilibrium. Gene diversity estimates show the population to be similar to other populations found in similar habitat in the region, and estimates of genetic differentiation among four regional populations showed that gene flow conforms to a model of isolation by distance. Analysis of parentage found that both resident and transient males sired about equal numbers of offspring. Familial DNA analysis revealed multiple paternities of successive young in this population. The conclusion from this study is that ‘resident’ status among males does not confer any advantage for parentage.