Pulmonary Actinomycosis in South Australian Koalas (Phascolarctos cinereus).

Pneumonia has been reported in both free-ranging and captive koalas and a number of causative agents have been described. Between 2016 and 2019, 16 free-ranging and 1 captive koala (Phascolarctos cinereus) from the Mount Lofty Ranges of South Australia were identified with pyogranulomatous lobar pneumonia, which involved the left caudal lobe in 14/17 (82%) cases. Within lesions, numerous gram-positive or gram-variable, non-acid-fast filamentous bacteria were observed in association with Splendore-Hoeppli phenomenon. Culture yielded growth of anaerobic bacteria, which were unidentifiable by MALDI-TOF-MS (matrix-assisted laser desorption ionization-time of flight mass spectrometry) analysis in 5/5 cases. Sequencing of the bacterial 16S rRNA gene identified a novel Actinomyces species in 4 samples, confirming a diagnosis of pulmonary actinomycosis. Concurrent examination of resin lung casts from healthy koalas suggested greater laminar flow of air to the left caudal lung lobe in koalas. Actinomyces spp. have been reported as commensals of the oral microbiome in other species, and an association with similar pulmonary lesions in other species. Considering the predilection for involvement of the left caudal lung lobe, aspiration is suggested as the likely cause in some cases of pulmonary actinomycosis in koalas. Pulmonary actinomycosis has not been previously described in koalas and further work needs to be undertaken in order to classify this organism within the Actinomyces genus.

Intravenous 15% isoflurane lipid nanoemulsion for general anesthesia in dogs.

OBJECTIVE: To investigate the efficacy of a new intravenous (IV) nanoemulsified isoflurane formulation for maintenance of general anesthesia in dogs. STUDY DESIGN: Prospective, crossover, experimental study. ANIMALS: Seven healthy, mature, mixed-breed dogs, three male and four female, weighing 11.5 ± 1.5 kg. METHODS: Anesthesia was induced with propofol for instrumentation. Measurements were obtained before administration of either inhaled isoflurane (Iso-I) or IV 15% isoflurane-loaded lipid nanoemulsion (Iso-nano). The minimum alveolar concentration (MAC) of isoflurane was determined using the 'up-and-down' technique. A tail clamp was applied every 15 minutes for a total time of 90 minutes and isoflurane administration was adjusted according to the response. Data were recorded at 30, 60 and 90 minutes for end-tidal isoflurane concentration (Fe´Iso), end-tidal carbon dioxide partial pressure (Pe'CO2), inspired isoflurane concentration (FIIso), arterial hemoglobin oxygen saturation (SaO2), peripheral hemoglobin oxygen saturation (SpO2), respiratory rate (fR), heart rate (HR), arterial blood pH, PaCO2, PaO2, base excess (BE), bicarbonate (HCO3-), systemic arterial pressure (sAP), and biochemical variables of blood urea nitrogen, alanine aminotransferase, creatine kinase and creatinine. RESULTS: No significant differences between treatments were detected for HR, fR, SaO2 or any biochemical variables (p > 0.05). In the Iso-nano treatment, sAP was significantly decreased throughout the study. Significant decreases in pH, Pe'CO2, BE and HCO3- were measured in the Iso-nano treatment. Isoflurane MAC was significantly lower in the Iso-nano than the Iso-I treatment. The dose of isoflurane (g hour-1) required to maintain general anesthesia did not differ significantly between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of 15% isoflurane-loaded lipid nanoemulsion IV was effective in maintaining general anesthesia in dogs but did not reduce the amount of isoflurane necessary to maintain general anesthesia. Significant hypotension and nonrespiratory acidosis occurred with the injectable form.

General anesthesia with an injectable 8% v/v sevoflurane lipid emulsion administered intravenously to dogs.

OBJECTIVE: To evaluate the potential of an intravenous (IV) sevoflurane formulation for maintenance of general anesthesia in dogs. STUDY DESIGN: Prospective crossover design. ANIMALS: Six healthy, mature, mixed-breed dogs, four males and two females, weighing 11.7 ± 3.4 kg. METHODS: Anesthesia was induced and maintained with propofol IV for instrumentation. Baseline measurements were recorded before administration of either sevoflurane in oxygen (Sevo-Inh) or lipid-emulsified sevoflurane 8% v/v in 30% Intralipid IV (Sevo-E), 0.5 mL kg(-1) over 5 minutes followed by an infusion at 0.1-0.3 mL kg(-1)  minute(-1) . Dogs were breathing spontaneously. The 'up-and-down' technique was used to determine the minimum alveolar concentration (MAC) of sevoflurane. Over 120 minutes, a tail clamp was applied every 15 minutes and sevoflurane administration was adjusted depending on the response. End-tidal sevoflurane concentration and variables were recorded at 30, 60, 90, and 120 minutes: heart rate (HR), systemic arterial pressure (sAP), respiratory rate (fR ), end-tidal carbon dioxide tension, hemoglobin oxygen saturation (SaO2 ), arterial pH and blood gases, blood urea nitrogen, alanine aminotransferase, creatine kinase, gamma-glutamyl transferase, and aspartate aminotransferase. RESULTS: There were no significant differences between treatments for HR, sAP, fR , SaO2 , and biochemical variables (p > 0.05). pH and HCO3-were significantly decreased, and PaCO2 increased from baseline in Sevo-E (p < 0.05). MAC was significantly lower for Sevo-E than for Sevo-Inh, although the required dose of sevoflurane (g hour(-1) ) to maintain general anesthesia was not significantly different between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of 8% v/v sevoflurane lipid emulsion IV was effective in maintaining general anesthesia in dogs, but resulted in moderate cardiopulmonary depression, metabolic and respiratory acidosis. The amount of sevoflurane (g hour(-1) ) required to maintain general anesthesia was significantly lower for inhaled than for IV sevoflurane.

Identification of MHCII variants associated with chlamydial disease in the koala (Phascolarctos cinereus).

Chlamydiosis, the most common infectious disease in koalas, can cause chronic urogenital tract fibrosis and infertility. High titres of serum immunoglobulin G against 10 kDa and 60 kDa chlamydial heat-shock proteins (c-hsp10 and c-hsp60) are associated with fibrous occlusion of the koala uterus and uterine tube. Murine and human studies have identified associations between specific major histocompatibility complex class II (MHCII) alleles or genotypes, and higher c-hsp 60 antibody levels or chlamydia-associated disease and infertility. In this study, we characterised partial MHCII DAB and DBB genes in female koalas (n = 94) from a single geographic population, and investigated associations among antibody responses to c-hsp60 quantified by ELISA, susceptibility to chlamydial infection, or age. The identification of three candidate MHCII variants provides additional support for the functional role of MHCII in the koala, and will inform more focused future studies. This is the first study to investigate an association between MHC genes with chlamydial pathogenesis in a non-model, free-ranging species.

Expression profiles of the immune genes CD4, CD8ß, IFNγ, IL-4, IL-6 and IL-10 in mitogen-stimulated koala lymphocytes (Phascolarctos cinereus) by qRT-PCR.

Investigation of the immune response of the koala (Phascolarctos cinereus) is needed urgently, but has been limited by scarcity of species-specific reagents and methods for this unique and divergent marsupial. Infectious disease is an important threat to wild populations of koalas; the most widespread and important of these is Chlamydial disease, caused by Chlamydia pecorum and Chlamydia pneumoniae. In addition, koala retrovirus (KoRV), which is of 100% prevalence in northern Australia, has been proposed as an important agent of immune suppression that could explain the koala's susceptibility to disease. The correct balance of T regulatory, T helper 1 (Th1) and Th2 lymphocyte responses are important to an individual's susceptibility or resistance to chlamydial infection. The ability to study chlamydial or KoRV pathogenesis, effects of environmental stressors on immunity, and the response of koalas to vaccines under development, by examining the koala's adaptive response to natural infection or in-vitro stimulation, has been limited to date by a paucity of species- specific reagents. In this study we have used cytokine sequences from four marsupial genomes to identify mRNA sequences for key T regulatory, Th1 and Th2 cytokines interleukin 4 (IL-4), interleukin 6 (IL-6), interleukin 10 (IL-10) and interferon gamma (IFNγ) along with CD4 and CD8ß. The koala sequences used for primer design showed >58% homology with grey short-tailed opossum, >71% with tammar wallaby and 78% with Tasmanian devil amino acid sequences. We report the development of real-time RT-PCR assays to measure the expression of these genes in unstimulated cells and after three common mitogen stimulation protocols (phorbol myristate acetate/ionomycin, phorbol myristate acetate/phytohemagglutinin and concanavalin A). Phorbol myristate acetate/ionomycin was found to be the most effective mitogen to up-regulate the production of IL-4, IL-10 and IFNγ. IL-6 production was not consistently up-regulated by any of the protocols. Expression of CD4 and CD8ß was down-regulated by mitogen stimulation. We found that the reference genes GAPDH and 28s are valid for normalising cytokine expression by koala lymphocytes after mitogen stimulation.

A retrospective study of admission trends of koalas to a rehabilitation facility over 30 years.

To identify threats to the survival of koalas (Phascolarctos cinereus) in coastal New South Wales, Australia, we compared 3,781 admission records of koalas, admitted between 1 January 1975 and 31 December 2004 to a koala rehabilitation facility on the midnorthern coast of New South Wales, against local wild population demographics, with the use of multinomial logistic regression and chi-square analyses. Trauma, the most frequent reason for admission, affected young and male animals more frequently than other groups. Seasonal differences in the probability of males presenting as trauma cases suggest behavioral factors as an important risk factor for this group. An increasing probability of koalas presenting as a result of motor vehicle accident since 1985 strongly supports the enhanced action of local authorities to pursue traffic-calming strategies if urban koala populations are to be maintained in this area. Koalas with clinical signs of chlamydiosis made up the second most frequent admission group, and these animals were more likely to be aged. This study highlights the potential usefulness of wildlife rehabilitation centers in detailing threats to local wildlife populations, provided record keeping is efficient and focused, and the role of such studies in providing evidence for focusing threat-mitigation efforts. Continual community engagement by koala researchers is important to ensure that maximum benefit is obtained from activities of special interest groups.