Chronic phalaris toxicity in macropods is widespread and peaks in July in Victoria, Australia.

Phalaris aquatica is pasture species introduced into Australia during early European settlement. Consumption of the plant can cause the neurological condition chronic phalaris toxicity (CPT) in sheep and cattle. In recent years, there has been an increase in reports of CPT in macropods, which has raised concerns regarding its impacts on their welfare. Currently, little is known about the distribution or seasonal patterns of this disease in wildlife, information pivotal in assessing its potential risks. Between 2021 and 2022, we conducted a survey targeting government bodies, veterinary businesses and wildlife organisations to investigate the locations and time of occurrence of CPT in macropods in the state of Victoria, Australia. We received 13 survey responses, 12 verbal reports, a full record of investigated cases from a university veterinary school and cases from a wildlife rescue organisation. Over the period of 11 years, Victoria had 918 cases of CPT recorded in macropods from 36 local government areas, with cases concentrated centrally just north of the state capital of Melbourne and July (midwinter) being the month with the highest case count (n = 220). There was a significant positive correlation between case count and both the abundance of kangaroos (Macropus giganteus and Macropus fuliginosus) (P < 0.01) and the abundance of P. aquatica (P = 0.009), and a significant negative correlation between annual case count and average rainfall of March (P = 0.016) and April (P = 0.02). Understanding these relationships will assist land and wildlife managers in predicting the risk and magnitude of disease outbreaks of CPT each in Victoria.

Embryo mortality in a captive-bred, Critically Endangered amphibian.

The Critically Endangered southern corroboree frog Pseudophryne corroboree is dependent upon captive assurance colonies for its continued survival. Although the captive breeding programme for this species has largely been successful, embryonic mortality remains high (40-90% per year). This study aimed to investigate the causes of mortality in P. corroboree embryos in the captive collection at Melbourne Zoo. During the 2021 breeding season, we investigated 108 abnormal embryos to determine the impact of infections and anatomical deformities on survival and used culture and molecular methods to identify microbes. Overall, 100% of abnormal embryos had fungal infections, and of these, 41.6% also had anatomical deformities. The mortality rate in abnormal embryos was 89.8%; however, we detected no difference in survival in any of the 3 observed fungal growth patterns or between deformed and non-deformed embryos. Sanger sequencing of the ITS region identified fungal isolates belonging to the genus Ilyonectria, the first record in a vertebrate host, and another as a Plectosphaerella sp., which is the first record of infection in an embryo. Dominant bacteria identified were of the genera Herbaspirillum and Flavobacterium; however, their role in the mortality is unknown. Fungal infection and deformities have a significant impact on embryo survival in captive-bred P. corroboree. In a species which relies on captive breeding, identifying and reducing the impacts of embryonic mortality can inform conservation efforts and improve reintroduction outcomes.

Haematological and serum biochemical reference intervals of free-ranging Lumholtz's tree-kangaroos (Dendrolagus lumholtzi).

BACKGROUND: Reference intervals for haematology and serum biochemistry parameters were developed for free-ranging Lumholtz's tree-kangaroo (Dendrolagus lumholtzi) using 35 samples from 12 female and 15 male free-ranging animals. Captive tree-kangaroos (n = 12) were also sampled for comparison. Differences were found between free-ranging and captive animals in white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase. These differences may be attributed to diet, activity, capture methods or age group. Reference intervals generated may be used for both free-ranging and captive Lumholtz's tree-kangaroos. This study provides a valuable tool for the assessment of health in rescued and captive tree-kangaroos and will aid in investigations into population health and disease in free-ranging Lumholtz's tree-kangaroos. OBJECTIVE: To develop reference intervals (RIs) for haematology and serum biochemistry parameters in Lumholtz's tree-kangaroos. METHODS: Haematological and serum biochemical RIs were determined using 35 samples from 27 clinically healthy Lumholtz's tree-kangaroos from the Atherton Tablelands region of Queensland examined between 2014 and 2019. Haematology and serum biochemistry parameters were measured from 16 samples from 12 captive animals for comparison. RESULTS: Reference intervals based on 35 samples from free-ranging animals showed higher mean and standard deviation values for white blood cell and neutrophil counts, and levels of aspartate aminotransferase, alkaline phosphatase, bilirubin, creatine kinase, phosphate, triglycerides and lipase than results for 16 samples from captive animals. Captive individuals showed higher mean values than free-ranging individuals for albumin, protein, creatinine as well as Hb, MCV, MCH and MCHC. CONCLUSION: The haematological and serum biochemistry RIs developed for Lumholtz's tree-kangaroos in this study will provide a valuable tool during clinical examination and investigations into disease and population health by veterinarians and researchers. The differences in parameters between free-ranging and captive animals are consistent with differences in diet, age cohort, activity or capture methods. Reference intervals generated from free-ranging animals should also be valid for captive Lumholtz's tree-kangaroos.

More bark than bite: Comparative studies are needed to determine the importance of canine zoonoses in Aboriginal communities. A critical review of published research.

The objective of this review was to identify and critique over forty years of peer-reviewed literature concerned with the transmission of canine zoonoses to Aboriginal people and determine the zoonotic organisms documented in dogs in Australian Aboriginal communities. A systematic literature search of public health, medical and veterinary databases identified 19 articles suitable for critical appraisal. Thirteen articles documented the occurrence of recognized zoonotic organisms in dogs in Aboriginal communities, including Toxocara canis, Dirofilaria immitis, Streptococcus dysgalactiae, Rickettsia felis, Sarcoptes scabiei and Giardia. Currently, there is definitive evidence indicating that dogs act as a reservoir for human scabies in Aboriginal communities. However, there is a need for large-scale, high-quality, comparative studies of dogs and humans from the same household to assess the occurrence and importance of transmission of S. scabiei and other diseases between dogs and humans. These studies should use current genetic and molecular techniques along with traditional techniques to identify and type organisms in order to better understand their epidemiology. This review has revealed that there is a lack of high-quality comparative studies to determine whether dogs are contributing to human disease by transmitting zoonoses. Our recommendations differ significantly from current public health policy and may have substantial implications for human and dog health.

Avian influenza in Australia: a summary of 5 years of wild bird surveillance.

BACKGROUND: Avian influenza viruses (AIVs) are found worldwide in numerous bird species, causing significant disease in gallinaceous poultry and occasionally other species. Surveillance of wild bird reservoirs provides an opportunity to add to the understanding of the epidemiology of AIVs. METHODS: This study examined key findings from the National Avian Influenza Wild Bird Surveillance Program over a 5-year period (July 2007-June 2012), the main source of information on AIVs circulating in Australia. RESULTS: The overall proportion of birds that tested positive for influenza A via PCR was 1.9 ± 0.1%, with evidence of widespread exposure of Australian wild birds to most low pathogenic avian influenza (LPAI) subtypes (H1-13, H16). LPAI H5 subtypes were found to be dominant and widespread during this 5-year period. CONCLUSION: Given Australia's isolation, both geographically and ecologically, it is important for Australia not to assume that the epidemiology of AIV from other geographic regions applies here. Despite all previous highly pathogenic avian influenza outbreaks in Australian poultry being attributed to H7 subtypes, widespread detection of H5 subtypes in wild birds may represent an ongoing risk to the Australian poultry industry.

In vitro sensitivity of the amphibian pathogen Batrachochytrium dendrobatidis to antifungal therapeutics.

Chytridiomycosis, a skin disease caused by Batrachochytrium dendrobatidis, has caused amphibian declines worldwide. Amphibians can be treated by percutaneous application of antimicrobials, but knowledge of in vitro susceptibility is lacking. Using a modified broth microdilution method, we describe the in vitro sensitivity of two Australian isolates of B. dendrobatidis to six antimicrobial agents. Growth inhibition was observed, by measurement of optical density, with all agents. Minimum inhibitory concentrations (µg/ml; isolate 1/2) were - voriconazole 0.016/0.008; itraconazole 0.032/0.016; terbinafine 0.063/0.063; fluconazole 0.31/0.31; chloramphenicol 12.5/12.5; amphotericin B 12.5/6.25. Killing effects on zoospores were assessed by observing motility. Amphotericin B and terbinafine killed zoospores within 5 and 30 min depending on concentration, but other antimicrobials were not effective at the highest concentrations tested (100 µg/ml). This knowledge will help in drug selection and treatment optimization. As terbinafine was potent and has rapid effects, study of its pharmacokinetics, safety and efficacy is recommended.

Causes of morbidity and mortality of wild aquatic birds at Billabong Sanctuary, Townsville, North Queensland, Australia.

Infectious diseases are common causes of significant morbidity and mortality events of wild aquatic birds (WABs) worldwide. Reports of Australian events are infrequent. A 3-yr passive surveillance program investigating the common causes of morbidity and mortality of WABs was conducted at Billabong Sanctuary near Townsville, North Queensland, from April 2007 to March 2010. Forty-two carcasses were obtained and evaluated by clinico-pathologic, histologic, bacteriologic, and virologic (molecular) examinations. Morbidity and mortality were sporadic and more commonly observed in chicks and juvenile birds in April than other months of the year. Morbid birds were frequently unable to walk. Hemorrhagic lesions and infiltration of lymphocytes in various organs were the most common findings in dead birds. Identified bacterial diseases that could cause bird mortality were colibacillosis, pasteurellosis, and salmonellosis. Salmonella serotypes Virchow and Hvittingfoss were isolated from an Australian white ibis (Threskiornis molucca) chick and two juvenile plumed whistling ducks (Dendrocygna eytoni) in April 2007. These strains have been previously isolated from humans in North Queensland. A multiplex real time reverse transcriptase-PCR (rRT-PCR) detected Newcastle disease viral RNA (class 2 type) in one adult Australian pelican (Pelecanus conspicillatus) and a juvenile plumed whistling duck. No avian influenza viral RNA was detected from any sampled birds by the rRT-PCR for avian influenza. This study identified the public health importance of Salmonella in WABs but did not detect the introduction of the high pathogenicity avian influenza H5N1 virus in the population. A successful network was established between the property owner and the James Cook University research team through which dead birds, with accompanying information, were readily obtained for analysis. There is an opportunity for establishing a long-term passive disease surveillance program for WABs in North Queensland, an important region in Australian biosecurity, thus potentially significantly benefitting public health in the region and the country.

Monitoring of wild birds for Newcastle disease virus in north Queensland, Australia.

Wild aquatic birds (WABs) are considered as reservoir hosts for Newcastle disease viruses (NDVs) and may act as vectors for transferring these viruses to poultry, causing outbreaks of disease. A 3-year epidemiological study was conducted on WABs of north Queensland from April 2007 to March 2010. Swab and fresh moist faecal samples of WABs were screened to detect Newcastle disease viral (NDV) RNA by one-step real time reverse transcriptase polymerase chain reaction (rRT-PCR) in multiplex primers, targeting the matrix gene. The potential reactor samples in rRT-PCR were processed for sequencing of the different NDV genes using conventional PCR. The overall NDV RNA prevalence was 3.5% for live bird samples (N=1461) and 0.4% for faecal samples (N=1157). Plumed whistling ducks (PWDs) had a higher prevalence (4.2%) than Pacific black ducks (PBDs) (0.9%) (χ(2) test, p=0.001). Univariate and multivariate logistic regression analyses were used to estimate the association between the proportion of reactor and non-reactor NDV RNA samples of PWDs and potential risk factors. The odds of reactor samples were 2.7 (95% Confidence Interval 1.5-4.9) times more likely in younger than older ducks (p=0.001) (data set B, multivariate analysis). Both NDV RNA class-one and class-two types were identified in samples of WABs (12 and 59, respectively) (Supplementary Table 1). Phylogenetic analysis of the matrix gene identified two reactor sequences of class-one type NDV RNA (PWD-48 and 55) which were closely related to the sequences of Australian Ibis and duck isolates (Fig. 2). Another reactor sample sequence was determined as class-two type NDV RNA (PWD-46, avirulent) based on analysis of the matrix and fusion genes which was more similar to the sequences of Australian I-2 progenitor virus and vaccine strain virus (Figs. 3 and 4). Our findings of higher prevalence in PWDs along with confirmation of class-one and class-two type NDV RNAs will significantly contribute to the design of surveillance programs for NDVs in northern Australia.

Adaptations of a competitive enzyme-linked immunosorbent assays for the detection of antibodies to influenza a virus in horse sera for use in wild aquatic birds.

We applied a competitive enzyme-linked immunosorbent assay for the detection of antibodies for influenza A in equine sera to their detection in sera from wild aquatic birds. Suboptimal results were obtained for the optical density (OD) of the monoclonal antibody (MAb) control and reproducibility between duplicate analyses in the initial assessment. It was therefore necessary to modify the assay to deliver increased reliability and reproducibility while maintaining adequate sensitivity. We optimized reagent concentrations to obtain optimal OD values (close to 2) for the monoclonal antibody control and used 2, 2'-Azino-bis: 3-Benzthiazoline-6-Sulphonic Acid as an alternative chromogen to potentially reduce variability in duplicate analyses. The original assay was compared with the optimized versions, with and without post coating, for the detection of avian influenza viral antibodies in 240 sera obtained from wild plumed whistling ducks. A separate analytical sensitivity study on diluted positive field sera of plumed whistling ducks and a test of antigen stability after post coating were also performed. Some quantitative differences were detected between the original and modified assays. The original assay recorded higher percentage inhibition results which were potentially indicative of increased sensitivity. However, when reagent concentrations were increased in the original assay to the same levels as used in the modified versions, there were no quantitative differences for practical purposes. The original assay produced a median (OD) value of 0.81 for the (MAb) controls that is at the limit of acceptability. By contrast, the modified assays always produced acceptable optical density values for MAb controls. Our overall results indicated the modified assays were potentially more reliable (OD values close to 2), and of adequate sensitivity compared to the original assay in the detection of avian influenza viral antibodies in wild bird sera. Although further optimization of antigen and MAb concentrations should also be considered to increase the sensitivity of a modified assay, while maintaining acceptable optical density values for the MAb control. Post coating had a minimal quantitative effect on the results and stabilized the plates for 214 days. We therefore recommend the incorporation of post coating.

Factors limiting the health of semi-scavenging ducks in Bangladesh.

Duck rearing is well suited to coastal and lowland areas in Bangladesh. It is an important component of sustainable livelihood strategies for poor rural communities as an additional source of household income. An epidemiological study was conducted during January 2005-June 2006 on 379 households in Chatkhil of the Noakhali District, Bangladesh which were using the recently devised "Bangladesh duck model". The overall objective of the study was to identify factors that significantly contributed to mortality and constrained productivity and to generate sufficient knowledge to enable establishment of a disease surveillance system for household ducks. The overall mortality was 15.0% in Chatkhil, with predation causing a significantly higher mortality compared with diseases (p < 0.001). Common diseases were duck plague and duck cholera. Morbid ducks frequently displayed signs associated with diseases affecting the nervous and digestive systems. Haemorrhagic lesions in various organs and white multiple foci on the liver were frequently observed in dead ducks. Epidemiological analysis with a shared frailty model that accounted for clustering of data by farm was used to estimate the association between survival time and risk factors. The overall mortality rate due to disease was significantly lower in vaccinated than in non-vaccinated ducks in all zones except zone 2 (p < 0.001). Only vaccinated ducks survived in zone 1. In conclusion, duck mortality and untimely sale of ducks appeared to be important constraints for household duck production in Chatkhil. Vaccination against duck plague appears to be an effective preventive strategy in reducing the level of associated duck mortality. A successful network was established amongst farmers and the surveillance team through which dead ducks, with accompanying information, were readily obtained for analysis. Therefore, there is an opportunity for establishing a long-term disease surveillance programme for rural ducks in Chatkhil of the Noakhali District of Bangladesh.

Monitoring the health and production of household Jinding ducks on Hatia Island of Bangladesh.

Duck rearing is an important component of sustainable living in poor rural communities, especially as a source of subsistence. A study was conducted on 118 households (N = 1,373 Jinding ducks, Anas platyrhynchus) from December 2002 to February 2004 on Hatia Island in Bangladesh with the aim of identifying the factors that limit the health and production of Jinding ducks. Overall duck mortality was 29.3%, with disease (19.7%) being a more significant factor than predation (9.6%; p = 0.001). Duck mortality also varied significantly among study zones (p < 0.001). Common diseases were duck plague (21.1%) and duck cholera (32.1%). Helminth infection was prevalent, with endemic trematode (Prosthogonimus spp., Trichobilharzia spp., Echinostoma spp.) and nematode (Cyathostoma bronchialis, Amidostomum anseris, Heterakis gallinarum, Capillaria spp., and Echinuria spp.) infections and epidemic cestode infections due to Hymenolepsis setigera. The median egg production rate per duck per household was 93 for a 6-month laying period. The odds of diminished egg production (average ≤ 93 eggs per duck per household for a 6-month laying period) was 25.4 times higher in ducks that were kept in traditional duck houses (p < 0.001) and 14.2 times higher in ducks that experienced delays in the onset of sexual maturity (days 191 and 280; p < 0.001). Ducks that were provided snails for a shorter period of time over the laying period were 18.2 times more likely to produce fewer eggs than their longer fed peers (p = 0.002). In conclusion, duck mortalities due to diseases and predation and parasitic infections appear to be common constraints on household duck production on Hatia Island. Additionally, improving duck housing and providing longer nutritional supplementation with snails increased the production capabilities of household-raised Jinding ducks on Hatia Island.

Meteorological effects on the daily activity patterns of tabanid biting flies in northern Queensland, Australia.

Information on the daily activity patterns of tabanid flies is important in the development of strategies that decrease the risk of pathogens transmitted by them. In addition, this information is useful to maximize numbers of tabanids trapped during short-term studies and to target feeding behavior studies of certain tabanid species to their times of peak activity. The current study examined the effects of various meteorological factors on the daily activity patterns of common tropical species of tabanids in north Queensland. Each species studied responded differently to weather factors. Tabanus townsvilli Ricardo (Diptera: Tabanidae) was most active during late morning and early afternoon, whereas Pseudotabanus silvester (Bergroth) and Tabanus pallipennis Macquart were most active in the late afternoon. Tabanus dorsobimaculatus Macquart was most active in the morning and early afternoon. Data on daily activity patterns of tabanid flies indicates that in an area such as Townsville, North Queensland, where several species of tabanid are present concurrently in high numbers, the overlapping periods of high activity for these species indicate a high risk of pathogen transmission for most of the day (10.00-19.00 hours). Similarly, because each species responds differently to weather variables, only extreme weather conditions are likely to inhibit activity of all species. These data also indicate that for maximal results, trapping and feeding behavior studies should be tailored to the preferred activity period of the species under investigation.

Host preferences of tabanid flies based on identification of blood meals by ELISA.

Tabanid flies in Australia are potential vectors of the parasite Trypanosoma evansi which causes the animal disease surra. It is endemic to most of south-east Asia and could enter Australia, but evaluation of the potential impact of a surra incursion requires identification of the major hosts of Australian tabanids. This study investigated the natural pattern of feeding and host preference by tabanid flies of Townsville, north Queensland by identification of ingested blood in trap-caught tabanids using ELISA. The assays were developed for identification of horse, cow, macropod and pig blood meals. Macropods were the most frequent food source for each of six major tabanid species in the area. This did not vary with location for one species, Tabanus pallipennis, despite macropod densities being lower than other hosts such as cattle and horses in some locations. Feeding patterns on other hosts generally depended on availability and density of animals. All tabanid species fed on at least three of the host species tested and mixed meals were also commonly encountered, suggesting a level of opportunistic feeding in addition to a preference for macropods. Some of the blood meals detected were possibly from previous gonotrophic cycles. The results indicate that all tabanid species examined could potentially transmit surra and all the host types investigated could be affected, but macropods face the highest transmission risk.

Factors limiting traditional household duck production in Bangladesh.

A cross sectional survey of duck production was carried out in 2002 on 771 traditional, semiscavenging household duck farms on the coastal Island of Hatia. We determined the socioeconomic characteristics of duck farmers and their management systems, identified the factors associated with egg production, and measured the level of selected duck diseases and current preventive strategies. Household family size varied from 1 to 14 individuals and women were the main caretakers of ducks. Around 34% of keepers were illiterate. Most duck products (eggs and meat; 85%) were sold at the local market. Duck houses were poorly ventilated and a variety of bedding materials were used. Feed was available in nearby scavenging areas; however, additional feed was frequently supplied by farmers. Almost all farmers (96%) ranked the rainy season as the best time for rearing ducks due to greater feed availability. The annual egg production was 79 eggs per layer with a weight of 48 g and a hatchability rate of 87%. Egg production varied by zone (p < 0.05). The odds of suboptimal egg production was 0.5 times lower in educated farmers (p = 0.001). The odds of suboptimal egg production was 2.5 times more likely in ducks that attained sexual maturity at >22 weeks (p<0.001). Most farmers ranked duck plague as the most important disease, followed by duck cholera, botulism, and duck viral hepatitis. Preventive vaccination was sporadic and used by few farmers (28%). There are significant opportunities for improved duck production on the Island of Hatia and in Bangladesh generally.

Feeding success and trappability of horse flies evaluated with electrocuting nets and odour-baited traps.

The relative capacity of different tabanid species to mechanically transmit infectious diseases was assessed by comparing their feeding success on a horse. Ten species were intercepted with electrocuting nets while approaching and leaving the horse: the most abundant were Tabanus pallipennis, Pseudotabanus silvester and Tabanus townsvilli. Of the tabanid species that managed to bite, the highest proportions of partially fed flies, which could potentially transmit pathogens to their next host, were for T. pallipennis (49%) and Dasybasis oculata (48%), while the lowest was in P. silvester (11%). T. townsvilli and T. dorsobimaculatus did not obtain blood meals, possibly because of their sensitivity to host defensive movements. The study also investigated whether catches from odour-baited traps could reliably predict tabanid activity around horses. The traps caught fewer flies, but captured the same species as the electrocuting nets surrounding a horse. A significant linear relationship between the two was observed for P. silvester, T. pallipennis and T. townsvilli but not for T. dorsobimaculatus. The results also demonstrate differences in trappability among tabanid species.

Alighting and feeding behaviour of tabanid flies on horses, kangaroos and pigs.

Successful mechanical transmission of surra between animals by tabanid flies (Diptera: Tabanidae) depends to a large extent on the blood-feeding behaviour of the tabanid species prevalent in the area. We studied tabanid-host interactions in Australia to better predict risk of surra transmission and design intervention strategies. At least six tabanid species were observed alighting on horses, pigs and kangaroos, but the most abundant were Tabanus pallipennis Macquart, Pseudotabanus silvester Bergroth and T. townsvilli Ricardo. The behaviour of tabanids in terms of landing location on the host body, duration of feeding and the proportion completing the blood-meal varied with fly species and host species. The findings predict that some species of tabanid such as T. pallipennis should be better vectors and some species of host such as pigs should be better reservoirs of surra based on the inability of flies to feed to repletion and longer feeding durations. This will result in multiple feeds and increased risk of exposure to the infectious agent, respectively, which increases the risk of transmission. Insecticide treatments should target preferred feeding sites on the host's body.

Minimising exposure of amphibians to pathogens during field studies.

Many of the recent global amphibian mass mortalities, declines and extinctions have been attributed to the emerging infectious disease chytridiomycosis. There have been mass mortalities due to ranaviral disease but no major declines or extinctions. Controlling the transmission and spread of disease is of utmost importance, especially where there is the potential for human involvement. We have reviewed current hygiene guidelines for working with wild frogs, identified potential flaws and recommended those most suitable and effective for the field environment. Our within-site hygiene measures aim to reduce the risk of transmission among individuals. These measures encompass the capture, handling and holding of amphibians, skin disinfection before and after invasive procedures, marking frogs, sealing open wounds and treatment of accessory equipment. Our between-site hygiene measures aim to mitigate the risk of pathogen spread among populations. We have designed a risk calculator to help simplify and standardise the decision-making process for determining the level of risk and appropriate risk mitigation strategies to reduce the risk of increasing pathogen spread above background levels. Calculation of an overall risk score for pathogen spread takes into account the prior activity of field workers, the proposed activity, remoteness of the site, presence of known pathogens and the consequences of increased pathogen spread for amphibians in a given area.

Determining causality and controlling disease is based on collaborative research involving multidisciplinary approaches.

Understanding the causes of infectious disease to facilitate better control requires observational and experimental studies. Often these must be conducted at many scales such as at the molecular, cellular, organism, and population level. Studies need to consider both intrinsic and extrinsic factors affecting the pathogen/host interaction. They also require a combination of study methods covered by disciplines such as pathology, epidemiology, microbiology, and ecology. Therefore, it is important that disciplines work together when designing and conducting studies. Finally, we need to integrate and interpret data across levels and disciplines to better formulate control strategies. This requires another group of specialists with broad cross-disciplinary training in epidemiology and an ability to readily work with others.

Storage of samples at high temperatures reduces the amount of amphibian chytrid fungus Batrachochytrium dendrobatidis DNA detectable by PCR assay.

Chytridiomycosis, caused by the amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd), is an emerging infectious disease responsible for amphibian declines on several continents. In laboratory conditions, optimal temperatures for Bd growth and survivorship are between 17 and 25 degrees C. We investigated the effect of different storage temperatures, both in field and laboratory conditions, on detection of Bd from swabs stored for 7 d. We sampled 52 wild Litoria wilcoxii males for Bd by simultaneously running 2 cotton swabs along the skin of the frog. One group of swabs was stored in a freezer within 2 h of sampling and the other was kept in a car in an exposed environment for 7 d before being stored in the freezer. In the laboratory experiment, swabs were inoculated with zoospores of Bd and underwent one of 4 treatments: immediate DNA extraction, or storage at 27, 38 or 45 degrees C for 7 d prior to DNA extraction. Swabs from all treatments were analyzed by quantitative (real-time) PCR test. Though prevalence of Bd did not differ significantly between swabs that were frozen and those that remained in a car for 7 d (19.2 vs. 17.3%, respectively), the number of Bd zoospores detected on car swabs taken from infected frogs was, on average, 67% less than that detected on the corresponding frozen swab. In the laboratory experiment, the number of zoospore equivalents varied significantly with treatment (F(3,35) = 4.769, p = 0.007), indicating that there was reduced recovery of Bd DNA from swabs stored at higher temperatures compared with those stored at lower temperatures or processed immediately. We conclude that failure to store swabs in cool conditions can result in a significant reduction in the amount of Bd DNA detected using the PCR assay. Our results have important implications for researchers conducting field sampling of amphibians for Bd.

A comparison of trapping methods for Tabanidae (Diptera) in North Queensland, Australia.

The ability to monitor the abundance and diversity of tabanid flies over wide areas requires effective and low-cost surveillance methods. Such monitoring activities help to quantify the risk of transmission of pathogens by tabanids. Here we examine the effectiveness and practicality of two types of trap (canopy traps and Nzi traps) and two types of attractant (octenol and carbon dioxide) for monitoring tabanid flies in tropical Australia. The Nzi trap consistently caught more tabanids and more species of tabanids than the canopy trap. It was also more robust and therefore required less maintenance in remote locations. The use of attractants substantially increased capture rates, both of individuals and species, and traps using both attractants were consistently the most effective. However, in remote locations, where it is not possible to check traps frequently, the use of attractants may not be feasible. When attractants were not used, the canopy trap caught very few tabanids, but the Nzi trap remained effective enough to be useful as a monitoring device. In addition, the number of tabanid species caught by the Nzi traps remained high, and included those that were most abundant. We therefore conclude that, in this region, Nzi traps are preferable for tabanid monitoring and that attractants greatly improve their effectiveness. However, for longterm monitoring, especially in remote locations, Nzi traps without attractants are a satisfactory option.