Diana Mendez

The distribution and host range of the pandemic disease chytridiomycosis in Australia, spanning surveys from 1956–2007

Chytridiomycosis is the worst disease to affect vertebrate biodiversity on record. In Australia, it is thought to have caused the extinction of four frog species, and it threatens the survival of at least 10 more. We report the current distribution and host range of this invasive disease in Australia, which is essential knowledge for conservation management. We envisage that the data be used in a global and national context for predictive modeling, meta-analyses, and risk assessment. Our continent-wide data set comprises 821 sites in Australia and includes 10 183 records from >80 contributors spanning collection dates from 1956 to 2007. Sick and dead frogs from the field and apparently healthy frogs from museum collections were tested opportunistically for the presence of Batrachochytrium dendrobatidis, the fungal pathogen causing chytridiomycosis, and apparently healthy frogs and tadpoles found during surveys were tested purposively. The diagnostic tests used were histology of skin samples and quantitative PCR of skin swabs. Chytridiomycosis was found in all Australian states and the Australian Capital Territory, but not in the Northern Territory. Currently it appears to be confined to the relatively cool and wet areas of Australia, such as along the Great Dividing Range and adjacent coastal areas in the eastern mainland states of Queensland, New South Wales, and Victoria, eastern and central Tasmania, southern South Australia, and southwestern Western Australia. Batrachochytrium dendrobatidis may have been introduced into Australia via the port of Brisbane around 1978 and spread northward and southward. It did not appear to arrive in Western Australia until 1985. The earliest records from South Australia and Tasmania are from 1995 and 2004, respectively, although archival studies from these states are lacking. We also report negative findings showing that the disease does not currently occur in some areas that appear to be environmentally suitable, including Cape York Peninsula in Queensland and most of the World Heritage Area in western Tasmania. Infection with B. dendrobatidis has been recorded from 63 frog species in Australia to date, all belonging to the Hylidae, Limnodynastidae, and Myobatrachidae, with the exception of one individual of a species from the Microhylidae and the introduced cane toad of the family Bufonidae.

Validation of Diagnostic Tests in Wildlife: The Case of Chytridiomycosis in Wild Amphibians

Abstract We validated the diagnostic tests for the high-profile disease, chytridiomycosis, in wild amphibian populations. We compared histological samples with a Taqman real time quantitative PCR (qPCR) test in five species of frogs at different times of the year at six locations in the wet tropics of northern Queensland. The sensitivity and specificity of each test were estimated using prior estimates from previous laboratory studies and Bayesian methods. The qPCR test was almost three times as sensitive 72.9% (62.7–82.2%) than histology 26.5% (19.9–33.9%) but was less specific 94.2% (89.3–98.6%) than histology 99.5% (98.4–100%), which was likely caused by contamination. Monitoring of the negative control success rate of the qPCR test is potentially a good indicator of specificity. It is likely that using individual gloves for handling amphibians reduces cross-contamination and, therefore, improves specificity rather than cause inhibition of the qPCR. Classifying indeterminate results as positive will increase the qPCR test sensitivity but will lower specificity to a lesser degree depending on the likelihood of contamination. Although PCR is the preferred test for amphibian populations, histology is useful when wishing to confirm a diagnosis of infection and in situations where observing the severity of infection and pathology in skin is desired. In this study, we show that diagnostic test validation in wild animals is now relatively straight forward using modern computing power and can incorporate prior knowledge generated from laboratory studies using Bayesian approaches.

Presence of the amphibian chytrid fungus Batrachochytrium dendrobatidis in threatened corroboree frog populations in the Australian Alps

Since the early 1980s, the southern corroboree frog Pseudophryne corroboree and northern corroboree frog P. pengilleyi have been in a state of decline from their sub-alpine and high montane bog environments on the southern tablelands of New South Wales, Australia. To date, there has been no adequate explanation as to what is causing the decline of these species. We investigated the possibility that a pathogen associated with other recent frog declines in Australia, the amphibian chytrid fungus Batrachochytrium dendrobatidis, may have been implicated in the decline of the corroboree frogs. We used histology of toe material and real-time PCR of skin swabs to investigate the presence and infection rates with B. dendrobatidis in historic and extant populations of both corroboree frog species. Using histology, we did not detect any B. dendrobatidis infections in corroboree frog populations prior to their decline. However, using the same technique, high rates of infection were observed in populations of both species after the onset of substantial population declines. The real-time PCR screening of skin swabs identified high overall infection rates in extant populations of P. corroboree (between 44 and 59%), while significantly lower rates of infection were observed in low-altitude P. pengilleyi populations (14%). These results suggest that the initial and continued decline of the corroboree frogs may well be attributed to the emergence of B. dendrobatidis in populations of these species.

Unexpected Result of Hendra Virus Outbreaks for Veterinarians, Queensland, Australia

A qualitative study of equine veterinarians and allied staff from Queensland, Australia, showed that veterinarians are ceasing equine practice because of fears related to Hendra virus. Their decisions were motivated by personal safety and legal liability concerns.

Application of the survey protocol for chytridiomycosis to Queensland, Australia

Spread of the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd), which causes chytridiomycosis, has resulted in the extinction of frogs, but the distribution of Bd is incompletely known. We trialled the survey protocol for Bd by attempting to systematically map its distribution in Queensland, Australia. Bd was easily detected in known infected areas, such as the Wet Tropics and South East Queensland. It was not detected in bioregions adjacent to, but inland from or to the north of, infected regions: Einasleigh Uplands and Cape York adjacent to the infected Wet Tropics; and Brigalow Belt South adjacent to the infected South East Queensland bioregion. These regions where Bd was not detected have bordered infected regions for between 15 yr (in northern Queensland) and 30 yr (in southern Queensland), and so they define the geographical limits of Bd with regard to the long-term environmental conditions in Queensland. The Gulf Plains, a bioregion distant from infected bioregions, was also negative. Bd was confined to rainforest and bordering habitats, such as wet eucalypt forests. Infections were largely confined to permanent water-associated species, consistent with this being an important cause of this group having the greatest declines. Our data supports biogeographic climatic models that show much of inland and northern Australia to be too hot and dry to support Bd. As there is limited opportunity for Bd to spread further in Queensland, the priority for management is reducing the impact of Bd in affected populations and assisting frogs to disperse into their former distributions. Given that the survey protocol has been applied successfully in Australia it may be useful for mapping the distribution of Bd in other parts of the world.

Survival of the amphibian chytrid fungus Batrachochytrium dendrobatidis on bare hands and gloves: hygiene implications for amphibian handling

Hygiene protocols for handling amphibians in the field and in laboratories have been proposed to decrease the transmission of chytridiomycosis caused by infection with the amphibian chytrid fungus Batrachochytrium dendrobatidis, which is responsible for global amphibian declines. However, these protocols are mainly based on theoretical principles. The aim of this study was to develop an evidence-based approach to amphibian handling hygiene protocols by testing the survival of B. dendrobatidis on human hands and various gloves. Bare or gloved human fingers were exposed to cultured zoospores and zoosporangia of B. dendrobatidis. Survival of B. dendrobatidis on hands and gloves was tested for up to 10 min post-exposure by inoculation onto tryptone/gelatin hydrolysate/lactose (TGhL) agar plates. The effects of repeated hand washings with water and with 70% ethanol and of washing gloves with water were also tested. Bare human skin demonstrated a fungicidal effect on B. dendrobatidis by 2 min and killed 100% of cells by 6 min, but this killing effect was reduced by repeated washing with water and ethanol. Nitrile gloves killed all B. dendrobatidis on contact, but washing in water decreased this effect. Latex and polyethylene gloves had no killing effect, and B. dendrobatidis survived for over 6 min. The killing effect of vinyl gloves varied with brands and batches. These results support the use of an unused pair of gloves for each new amphibian handled in either the field or the laboratory, and if this is not possible, bare hands are a preferable, although imperfect, alternative to continual use of the same pair of gloves.

Testing for Hendra virus: difficulties experienced by veterinarians in Queensland prior to 2011.

OBJECTIVE To identify the perceived barriers to Hendra virus (HeV) management by private equine veterinarians in Queensland. DESIGN An exploratory qualitative study of private equine veterinarians registered and working in coastal Queensland. METHODS A questionnaire that included eight open-ended questions about the management of HeV was used in face-to-face in-depth interviews with 21 veterinary personnel working in equine or mixed private practice between Far North and South-East Queensland in 2009-10. The qualitative data was entered and analysed thematically using QSRs Internationals Nvivo 9 qualitative data analysis software. RESULTS This study revealed key issues associated with HeV testing: (1) inadequate knowledge of testing procedures and laboratory diagnostic pathways; (2) difficulty in accessing laboratory services; (3) responsibility for cost of collection and transport of specimen; and (4) the role of government. Participants perceived these issues as reducing potential HeV case management efficiency. CONCLUSION Although HeV management plans have been modified in part since 2009-10, this study highlights the importance of considering the perspectives of private veterinary practitioners in any biosecurity protocols.

Management of the slowly emerging zoonosis, Hendra virus, by private veterinarians in Queensland, Australia: a qualitative study

BackgroundVeterinary infection control for the management of Hendra virus (HeV), an emerging zoonosis in Australia, remained suboptimal until 2010 despite 71.4% (5/7) of humans infected with HeV being veterinary personnel or assisting a veterinarian, three of whom died before 2009. The aim of this study was to identify the perceived barriers to veterinary infection control and HeV management in private veterinary practice in Queensland, where the majority of HeV outbreaks have occurred in Australia.ResultsMost participants agreed that a number of key factors had contributed to the slow uptake of adequate infection control measures for the management of HeV amongst private veterinarians: a work culture characterised by suboptimal infection control standards and misconceptions about zoonotic risks; a lack of leadership and support from government authorities; the difficulties of managing biosecurity and public health issues from a private workforce perspective; and the slow pattern of emergence of HeV. By 2010, some infection control and HeV management changes had been implemented. Participants interviewed agreed that further improvements remained necessary; but also cautioned that this was a complex process which would require time.ConclusionPrivate veterinarians and government authorities prior to 2009 were unprepared to handle new slowly emerging zoonoses, which may explain their mismanagement of HeV. Slowly emerging zoonoses may be of low public health significance but of high significance for specialised groups such as veterinarians. Private veterinarians, who are expected to fulfil an active biosecurity and public health role in the frontline management of such emerging zoonoses, need government agencies to better recognise their contribution, to consult with the veterinary profession when devising guidelines for the management of zoonoses and to provide them with greater leadership and support. We propose that specific infection control guidelines for the management of slowly emerging zoonoses in private veterinary settings need to be developed.

Hendra virus in Queensland, Australia, during the winter of 2011: Veterinarians on the path to better management strategies

Abstract Following the emergence of Hendra virus (HeV), private veterinarians have had to adopt additional infection control strategies to manage this zoonosis. Between 1994 and 2010, seven people became infected with HeV, four fatally. All infected people were at a higher risk of exposure from contact with horses as they were either veterinary personnel, assisting veterinarians, or working in the horse industry. The management of emerging zoonoses is best approached from a One Health perspective as it benefits biosecurity as well as a public health, including the health of those most at risk, in this case private veterinarians. In 2011 we conducted a cross-sectional study of private veterinarians registered in Queensland and providing veterinary services to horses. The aim of this study was to gauge if participants had adopted recommendations for improved infection control, including the use of personal protective equipment (PPE), and the development of HeV specific management strategies during the winter of 2011. A majority of participants worked in practices that had a formal HeV management plan, mostly based on the perusal of official guidelines and an HeV field kit. The use of PPE increased as the health status of an equine patient decreased, demonstrating that many participants evaluated the risk of exposure to HeV appropriately; while others remained at risk of HeV infection by not using the appropriate PPE even when attending a sick horse. This study took place after Biosecurity Queensland had sent a comprehensive package about HeV management to all private veterinarians working in Queensland. However, those who had previous HeV experience through the management of suspected cases or had attended a HeV specific professional education programme in the previous 12 months were more likely to use PPE than those who had not. This may indicate that for private veterinarians in Queensland personal experience and face-to-face professional education sessions may be more effective in the improvement of HeV management than passive education via information packages. The role of different education pathways in the sustainable adoption of veterinary infection control measures should be further investigated.

Willingness to Consult a Veterinarian on Physician’s Advice for Zoonotic Diseases: A Formal Role for Veterinarians in Medicine?

Physicians appear to find zoonotic diseases a challenge and consider that this topic belongs more to the veterinary profession. However, veterinarians have no formal role in clinical medicine. Data were collected as part of the Queensland Social Survey 2014 to determine the willingness of the public, if diagnosed with a zoonotic disease, to consult a veterinarian on the advice of a physician. Self-reported willingness to consult with a veterinarian at the respondent’s own expense was 79.8% (95% CI: 81.96%-77.46%) (976/1223). If the cost was funded by Medicare, the Australian public health insurance scheme, 90.7% (95% CI: 92.18%-88.92%) (1109/1223) would be willing to consult a veterinarian. Therefore, a large majority of Australian residents would be willing to consult with a veterinarian on the advice of their physician if they had a zoonotic disease. Does this indicate a possible new role for veterinarians under Clinical One Health?