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.     

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.     

A zoospore inhibition technique to evaluate the activity of antifungal compounds against Batrachochytrium dendrobatidis and unsuccessful treatment of experimentally infected green tree frogs (Litoria caerulea) by fluconazole and benzalkonium chloride

Effective and safe treatments of chytridiomycosis in amphibians, caused by Batrachochytrium dendrobatidis, are needed to help prevent mortality in captive programs for threatened species, to reduce the risk of spread, and to better manage the disease in threatened populations. We describe a simple method to determine minimum inhibitory concentrations (MICs) of antifungal agents that involves adding zoospores to various drug concentrations in 96 well plates and microscopic observation after four days. We report results from testing 10 commercially available antifungal compounds: benzalkonium chloride (<0.78 μg/ml), povidone iodine (312.5 μg/ml), amphotericin B (3.125 μg/ml), fluconazole (<1.56 μg/ml), itraconazole (<1.56 μg/ml), enilconazole (<1.56 μg/ml), mercurochrome (6.25 μg/ml), sodium chloride (12.5 mg/ml), methylene blue (<1.56 μg/ml) and Virkon (3.125 μg/ml). For treatment trials of juvenile Litoria caerulea, baths of benzalkonium chloride at 1 mg/L and fluconazole at 25 mg/L were used on 18 experimentally infected frogs per treatment. Although these treatments resulted in longer survival times (mean 43.7 ± 11.3 days) than in the untreated controls (37.9 ± 9.3 days), the mortality rate was still 100%. Higher doses of fluconazole are suggested for further animal trials.     

Echinococcus granulosus in northern Queensland. 1. Prevalence in cattle

OBJECTIVE: To determine the prevalence and geographical distribution of hydatidosis and investigate factors that might be expected to influence the prevalence of hydatids in cattle in Queensland north of the Tropic of Capricorn. To determine the effect of natural levels of infection on carcase weight and subsequent economic loss. PROCEDURE: An abattoir survey conducted in 1981 provided information on the distribution, prevalence and viability of hydatid cysts in cattle from all shires north of the Tropic of Capricorn in Queensland. Livers, lungs and spleens from 10,382 cattle were palpated at abattoirs in Cairns, Townsville and Rockhampton to detect hydatid cysts. Prevalence of infection in cattle in each shire was estimated from results of the abattoir study together with reports of infection in a further 22,185 cattle obtained from abattoir records. Linear modelling was used to define the effect of geographical origin, age, breed and sex on prevalence of infection. Differences in the weights of carcases between infected and non-infected cattle of the same age, sex, breed and property of origin were examined. The economic loss to the beef industry in the region surveyed was estimated. RESULTS: Cattle infected with hydatids originated almost entirely from regions to the east of the Great Dividing Range. The mean prevalence inside this zone was 28% compared with 3% in other areas. Viable protoscoleces were found in 0.7% of cysts. Geographical origin and age of the cattle were the most significant factors influencing prevalence. Infection with hydatids had no effect on carcase weight. Economic loss was limited to that associated with condemnations of organs at meat inspection, estimated to be 0.5 million dollars per annum in 1981 and 6 million dollars in 2004. The distribution of hydatids in Queensland north of the Tropic of Capricorn corresponded most closely with the distribution of small wallabies such as Macropus dorsalis (black-striped wallaby), M parryi (whiptail wallaby) and M rufogriseus (red-necked wallaby). CONCLUSIONS: It was concluded that cattle are not an important part of maintaining the life-cycle of E granulosus in Queensland north of the Tropic of Capricorn. Within the endemic zone, which is almost all to the east of the Great Dividing Range, the local pattern of bovine echinococcosis is most likely to be determined by the presence or absence of small species of wallaby such as M dorsalis, M parryi and M rufogriseus.     

Echinococcus granulosus in northern Queensland. 2. Ecological determinants of infection in beef cattle

OBJECTIVE: To provide information on possible ecological determinants of infection with Echinococcus granulosus in a beef pastoral area of northern Queensland. PROCEDURE: An ecological study was carried out on the prevalence and viability of infection with Echinococcus granulosus in definitive and potential intermediate hosts, and their predator prey relationships. Seven adjacent extensive beef properties 100 km south of Townsville, that included areas of savannah, open woodland and dense closed scrub, were selected for the study. Infection with E granulosus in dingoes was determined at post mortem, and in domestic dogs by examining duodenal mucus after purging with arecoline hydrobromide. Cattle, wild pigs and macropods were examined at post mortem for viable hydatid cysts. The diet of dingoes was investigated by identifying the hair of prey species found in their stomach and colon, and that of domestic dogs by questioning their owners. RESULTS: Prevalence of hydatidosis in adult cattle ranged from 41% in animals from properties with large areas of dense closed scrub, to 3% on properties with little or no scrub. Hydatid cysts were found in 21.8% of black-striped wallabies (Macropus dorsalis), 9.4% of feral pigs, 1.5% of wallaroos (Macropus robustus), and 1.4% of eastern grey kangaroos (Macropus giganteus). No rufous rat kangaroos (Aepyprymnus rufescens) or swamp wallabies (Wallabia bicolor) were infected. Most cysts in macropods were viable, whereas in pigs about half were viable and in cattle only 0.7% contained viable protoscoleces. Infection with E granulosus was detected in 76% of dingoes, whereas no infection was detected in domestic dogs in the study area. CONCLUSIONS: It was concluded that the sylvatic cycle of E granulosus in the study area was maintained mainly through predation of black-striped wallabies by dingoes, and that the verges of dense scrub were the main nidus of infection.     

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.