Activity Patterns of Free-Ranging Koalas (Phascolarctos cinereus) Revealed by Accelerometry

An understanding of koala activity patterns is important for measuring the behavioral response of this species to environmental change, but to date has been limited by the logistical challenges of traditional field methodologies. We addressed this knowledge gap by using tri-axial accelerometer data loggers attached to VHF radio collars to examine activity patterns of adult male and female koalas in a high-density population at Cape Otway, Victoria, Australia. Data were obtained from 27 adult koalas over two 7-d periods during the breeding season: 12 in the early-breeding season in November 2010, and 15 in the late-breeding season in January 2011. Multiple 15 minute observation blocks on each animal were used for validation of activity patterns determined from the accelerometer data loggers. Accelerometry was effective in distinguishing between inactive (sleeping, resting) and active (grooming, feeding and moving) behaviors. Koalas were more active during the early-breeding season with a higher index of movement (overall dynamic body acceleration [ODBA]) for both males and females. Koalas showed a distinct temporal pattern of behavior, with most activity occurring from mid-afternoon to early morning. Accelerometry has potential for examining fine-scale behavior of a wide range of arboreal and terrestrial species.     

Genetic Variation in Captive Koalas (Phascolarctos cinereus): Parentage Determination and Individual Identification

Highly repeatable randomly amplified polymorphicDNA (RAPD) markers were developed for parentage studiesin the koala (Phascolarctos cinereus). Of the 25 RAPDprimers screened, 5 (20.0%) produced 32 repeatable polymorphic RAPD bands (average/primer = 6.4± 4.2). A high level of polymorphism was observedfor each group of koalas (Featherdale, 71.9%; Lone Pine,84.4%). All 25 koalas could be uniquely identified using either RAPD or microsatellite markers. Of the32 RAPD markers generated in koalas, 25 were informativefor parentage analyses. These RAPD markers successfullydetermined both parents to three offspring and a male parent to a fourth offspring.Paternity analysis (where the female parent is known)succeeded in assigning the correct male parent to sevenoffspring. Our RAPD–PCR method generatesinformative genetic markers that are useful for parentagedetermination and individual identification of captivekoalas. This would provide genetic analysis to zoos andwildlife parks as a low-cost alternative to the more expensive microsatellite markers.     

Perception of male caller identity in Koalas (Phascolarctos cinereus): acoustic analysis and playback experiments

The ability to signal individual identity using vocal signals and distinguish between conspecifics based on vocal cues is important in several mammal species. Furthermore, it can be important for receivers to differentiate between callers in reproductive contexts. In this study, we used acoustic analyses to determine whether male koala bellows are individually distinctive and to investigate the relative importance of different acoustic features for coding individuality. We then used a habituation-discrimination paradigm to investigate whether koalas discriminate between the bellow vocalisations of different male callers. Our results show that male koala bellows are highly individualized, and indicate that cues related to vocal tract filtering contribute the most to vocal identity. In addition, we found that male and female koalas habituated to the bellows of a specific male showed a significant dishabituation when they were presented with bellows from a novel male. The significant reduction in behavioural response to a final rehabituation playback shows this was not a chance rebound in response levels. Our findings indicate that male koala bellows are highly individually distinctive and that the identity of male callers is functionally relevant to male and female koalas during the breeding season. We go on to discuss the biological relevance of signalling identity in this species' sexual communication and the potential practical implications of our findings for acoustic monitoring of male population levels.     

Keratitis in free-ranging koalas (Phascolarctos cinereus) on Magnetic Island, Townsville.

Seventy free-ranging koalas (Phascolarctos cinereus) from Magnetic Island (Queensland, Australia) underwent an ocular examination, blood collection and serological examination for Chlamydia psittaci antibodies, and an examination of their teeth and genitalia. In 12 koalas long-standing unilateral keratitis was noted and in another 10 animals long-standing bilateral keratitis was observed. All animals were seronegative for Chlamydia psittaci and apart from some nasal discharge and a variety of assorted medical findings there was no sign of chlamydial infection. These ocular findings probably represent a new disease of unknown etiology.     

A Prototype Recombinant-Protein Based Chlamydia pecorum Vaccine Results in Reduced Chlamydial Burden and Less Clinical Disease in Free-Ranging Koalas (Phascolarctos cinereus)

Diseases associated with Chlamydia pecorum infection are a major cause of decline in koala populations in Australia. While koalas in care can generally be treated, a vaccine is considered the only option to effectively reduce the threat of infection and disease at the population level. In the current study, we vaccinated 30 free-ranging koalas with a prototype Chlamydia pecorum vaccine consisting of a recombinant chlamydial MOMP adjuvanted with an immune stimulating complex. An additional cohort of 30 animals did not receive any vaccine and acted as comparison controls. Animals accepted into this study were either uninfected (Chlamydia PCR negative) at time of initial vaccination, or infected (C. pecorum positive) at either urogenital (UGT) and/or ocular sites (Oc), but with no clinical signs of chlamydial disease. All koalas were vaccinated / sampled and then re-released into their natural habitat before re-capturing and re-sampling at 6 and 12 months. All vaccinated koalas produced a strong immune response to the vaccine, as indicated by high titres of specific plasma antibodies. The incidence of new infections in vaccinated koalas over the 12-month period post-vaccination was slightly less than koalas in the control group, however, this was not statistically significant. Importantly though, the vaccine was able to significantly reduce the infectious load in animals that were Chlamydia positive at the time of vaccination. This effect was evident at both the Oc and UGT sites and was stronger at 6 months than at 12 months post-vaccination. Finally, the vaccine was also able to reduce the number of animals that progressed to disease during the 12-month period. While the sample sizes were small (statistically speaking), results were nonetheless striking. This study highlights the potential for successful development of a Chlamydia vaccine for koalas in a wild setting.     

Epidemiology of Mycobacterium ulcerans infection in koalas (Phascolarctos cinereus) on Raymond Island, southeastern Australia

Mycobacterium ulcerans infections were found in 11 koalas (Phascolarctos cinereus) between 1980 and 1985, in a population of approximately 200 koalas on Raymond Island in southeastern Australia. Ulcers caused by the infection occurred on the face, forearm, rump, groin and footpads. Seven koalas had multiple ulcers. All the infected animals were mature (age classes 4, 5 and 6), and eight were male. The distribution of ulcers corresponded with the distribution of wounds in a sample of 87 koalas. Many of these wounds were associated with social behaviour.     

Maternal effort and joey growth in koalas (Phascolarctos cinereus)

The extent to which sex-biased maternal investment characterizes mammals is controversial, with less information available for evaluating patterns of maternal effort in marsupials than in placentals. Koalas Phascolarctos cinereus are size-dimorphic animals with a lengthy period of dependency and they reside in mating systems that might favour sex-biased maternal investment. We examined 18 years of data recorded from koalas living at the San Diego Zoo in order to examine how joey development and maternal condition might be connected. Koalas are pregnant for only 1 month, but joey emergence from the pouch does not occur until 32 weeks of age. Neither maternal condition nor age affected sex ratio at joey emergence, and both sexes had the same survivorship prospects. Koala dams transport and nurse joeys for close to 1 year, at which time the two sexes are size dimorphic. Given the poor-quality diet of koalas, combined with maternal transport of infants who are at least 25% of maternal mass, we suggest that infant rearing poses high energetic costs on koala females. We suggest that ecological and energetic constraints have moulded koala maternal strategies such that females maximize allocation of resources to offspring, regardless of sex, in order to increase prospects for joey survivorship.     

Detection of a novel gammaherpesvirus in koalas (Phascolarctos cinereus)

A novel gammaherpesvirus was detected in wild koalas (Phascolarctos cinereus) captured at different locations during 2010. Sequence analysis of the DNA polymerase gene revealed that the virus was genetically distinct from all known gammaherpesviruses. This is the first herpesvirus to be definitively identified in the Vombatiforme suborder (koalas and wombats).     

Dietary aluminium and renal failure in the koala (Phascolarctos cinereus)

The study investigated the link between the potentially nephrotoxic levels of aluminium ingested in the natural diet of eucalypt leaves by koalas in the Adelaide Hills, South Australia, and the high incidence of renal failure in koalas within this habitat. Routine histology of kidney specimens revealed no pathologies at the light microscopic level and contrasted sharply with the clinical signs of renal failure. However staining with solochrome azurine and Perl's Prussian blue showed aluminium was present in some proximal convoluted tubules in all specimens. Aluminium was also found in bone samples. The presence of aluminium in bone and kidney tissues was confirmed using electron dispersive x-ray analysis with transmission and scanning electron microscopy. Ultrastructural changes, including a decrease in lysosomal numbers, were seen in proximal convoluted tubules and these changes were shown to coincide with the presence of aluminium. No aluminium was found in koalas that died from causes other than renal failure. It was concluded that renal failure in the koalas of the Adelaide Hills is characterised by the presence of aluminium in the kidneys and bone and it is probably related to the high levels of aluminium in their restricted diet of eucalypt leaves. However, it is not known if the presence of aluminium is the cause or effect of the renal failure. The study is the first account where aluminium ingested as part of the natural diet of mammals has been shown to accumulate in the animal and be implicated with nephrotoxicity.     

Nitrogen and the koala (Phascolarctos cinereus): some indirect evidence

Indirect evidence provides additional support for the hypothesis that the level of available nitrogenous food is the major limiting influence in the abundance of the koala. It appears that koalas may select for low fibre content which is usually correlated with high nitrogen levels of leaves.     

Chemical characterization of milk oligosaccharides of the koala (Phascolarctos cinereus)

Previous structural characterizations of marsupial milk oligosaccharides had been performed in only two macropod species, the tammar wallaby and the red kangaroo. To clarify the homology and heterogeneity of milk oligosaccharides among marsupial species, which could provide information on their evolution, the oligosaccharides of the koala milk carbohydrate fraction were characterized in this study. Neutral and acidic oligosaccharides were separated from the carbohydrate fraction of milk of the koala, a non-macropod marsupial, and characterized by ¹H-nuclear magnetic resonance spectroscopy. The structures of the neutral saccharides were found to be Gal(β1-4)Glc (lactose), Gal(β1-3)Gal(β1-4)Glc (3′-galactosyllactose), Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc (3′,3″-digalactosyllactose), Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (lacto-N-novopentaose I) and Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl lacto-N-novopentaose I), while those of the acidic saccharides were Neu5Ac(α2-3)Gal(β1-4)Glc (3′-SL), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-4)Gal (sialyl 3′-galactosyllactose), Neu5Ac(α2-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose a), Gal(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose b), Gal(β1-3)[Neu5Ac(α2-3)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose c), and Neu5Ac(α2-3)Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl sialyl lacto-N-novopentaose a). The neutral oligosaccharides, other than fucosyl lacto-N-novopentaose I, a novel hexasaccharide, had been found in milk of the tammar wallaby, a macropod marsupial, while the acidic oligosaccharides, other than fucosyl sialyl lacto-N-novopentaose a had been identified in milk carbohydrate of the red kangaroo. The presence of fucosyl oligosaccharides is a significant feature of koala milk, in which it differs from milk of the tammar wallaby and the red kangaroo.     

Metabolites of dietary 1,8-cineole in the male koala (Phascolarctos cinereus)

The in vivo metabolic fate of 1,8-cineole was investigated in six male koalas. Koalas were fed ad lib a diet of Eucalyptus cephalocarpa leaf with a 1,8-cineole concentration of 2.53+/-0.70% dry mass of leaf, corresponding to a 1,8-cineole intake of 2.4+/-1.1 mmol/kg (3.1+/-1.3 g). Urine and faeces were collected for 24 h and metabolites identified by GC-MS and LC-MS. Metabolites were quantified before and after hydrolysis with beta-glucuronidase to give free and total levels, respectively. Fractional recovery of ingested 1,8-cineole was 1.3+/-0.4 and 1.4+/-0.4 (mean+/-S.D.) for free and total measurements, respectively. Seven metabolites were identified and quantified: 9- and 7-hydroxycineole, 9- and 7-cineolic acid, 7-hydroxy-9-cineolic acid, 9-hydroxy-7-cineolic acid and 7,9-dicineolic acid. The hydroxycineolic acids dominated the metabolite profile (85%). 7,9-Dicineolic acid, a novel metabolite of 1,8-cineole, accounted for almost 10% of the recovered dose making it the second most abundant metabolite after 7-hydroxy-9-cineolic acid (77%). Together, the less oxidised metabolites, the hydroxycineoles and cineolic acid, accounted for only 5% of the cineole consumed. Significant conjugation only occurred with four minor, less oxidised, alcohol and carboxylic acid metabolites. We have shown that the koala detoxifies and eliminates 1,8-cineole primarily by extensive oxidation without utilising conjugation pathways.