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Oral Diseasein Animals: The Australian Perspective. Isolation and Characterisation of Black-Pigmented Bacteria from the Oral Cavity of Marsupials

Philip S. Bird1,*, Sharnan C. Huynh1,2, Deirdre Davis1,2, Daria N. Love3,w, Linda L. Blackall2 and GregoryJ. Seymour1

1Oral Biology and Pathology, School of Dentistry, The University of Queensland, St Lucia, Queensland 4072 Australia

 2 Department of Microbiology Parasitology, School of Molecular and Microbial Sciences, The University of Queensland, St Lucia, Queensland 4072 Australia

3 Faculty of Veterinary Science, The University of Sydney, New South Wales 2006 Australia

ABSTRACT

A wide range of animals suffer from periodontal disease. However, there is very little reported on disease and oral micro-biota of Australian animals. Therefore, the oral cavity of 90 marsupials was examined for oral health. Plaque samples were collected from the subgingival margins using curettes or swabs. Plaque samples were plated onto non-selective trypticase soy agar plates, selective trypticase soy agar, non-selective and selective Wilkens Chalgrens Agar. Plates were incubated in an anaerobic atmosphere and examined after 7–14 days for the presence of black–brown-pigmented colonies. A combination of morphological and biochemical tests were used (colonial morphology, pigmentation, aerobic growth, Gram reaction, fluorescence under long-wave UV light (360 nm), production of catalase, enzymatic activity with fluorogenic substrates and haemagglutination of sheep red cells) to identify these organisms. Black-pigmented bacteria were cultivated from the plaque of 32 animals including six eastern grey kangaroos, a musky rat kangaroo, a whiptail and a red-necked wallaby, 18 koalas, a bandicoot and five brushtail possums. No black-pigmented colonies were cultivated from squirrel or sugar gliders or quokkas or from marsupial mice. The majority of isolates were identified as Porphyromonas gingivalis-like species with the higher prevalence of isolation from the oral cavity of macropods (the kangaroos and wallabies). Oral diseases, such as gingivitis can be found in native Australian animals with older koalas having an increase in disease indicators and black-pigmented bacteria. Non-selective Wilkens Chalgren Agar was the medium of choice for the isolation of black-pigmented bacteria.