R.P. Hobbs

Molecular and morphological characterization of Echinococcus granulosus of human and animal origin in Iran

Iran is an important endemic focus of cystic hydatid disease (CHD) where several species of intermediate host are commonly infected with Echinococcus granulosus. Isolates of E. granulosus were collected from humans and other animals from different geographical areas of Iran and characterized using both DNA (PCR-RFLP of ITS1) and morphological criteria (metacestode rostellar hook dimensions). The sheep and camel strains/genotypes were shown to occur in Iran. The sheep strain was shown to be the most common genotype of E. granulosus affecting sheep, cattle, goats and occasionally camels. The majority of camels were infected with the camel genotype as were 3 of 33 human cases. This is the first time that cases of CHD in humans have been identified in an area where a transmission cycle for the camel genotype exists. In addition, the camel genotype was found to cause infection in both sheep and cattle. Results also demonstrated that both sheep and camel strains can be readily differentiated on the basis of hook morphology alone.

Molecular and morphological characterization of Echinococcus in cervids from North America.

Many issues concerning the taxonomy of Echinococcus have been resolved in recent years with the application of molecular tools. However, the status of Echinococcus maintained in transmission cycles involving cervid intermediate hosts remains to be determined. The recent characterization of the parasite from cervids in Finland has highlighted the paucity of data available, particularly that from North America. In this study, we have characterized a large number of Echinococcus isolates from cervids from Western Canada on the basis of morphology and molecular genetic techniques. Our results support earlier studies suggesting that Echinococcus of cervid origin is phenotypically and genetically distinct to Echinococcus maintained in domestic host assemblages, and also confirms that Echinococcus of cervid origin does not constitute a genetically homogeneous group. However, our data do not support the existence of 2 distinct genotypes (strains/subspecies) with separate geographical distributions. Our data appear to support the existence of only 1 species in cervids, but additional isolates from cervids and wolves in other endemic regions should be characterized before a final decision is made on the taxonomic status of Echinococcus in cervids.

Rostellar hook morphology of Echinococcus granulosus (Batsch, 1786) from natural and experimental Australian hosts, and its implications for strain recognition

An analysis of the rostellar hooks of Australian isolates of Echinococcus granulosus revealed that there was less variation in larval (metacestode) than adult characters and that metacestode characters could be measured directly from adult worms. A factor analysis indicated that two factors, one representing a contrast between number of hooks and their length, and the other representing blade lengths, were sufficient to account for 87.5% of the variance in metacestode hook measurements. These results indicate that rostellar hook morphology is not useful for discriminating strains of E. granulosus in Australia. The Tasmanian and mainland domestic strains were found to be indistinguishable using rostellar morphology. Although many of the isolates from sylvatic hosts differed from those from domestic hosts, there was not a clear separation as would be expected if two distinct strains existed. Evidence was presented to show that the morphological differences seen in sylvatic hosts could be attributed to host-induced effects, and that the previously accepted existence of two mainland strains should be investigated further.

Efficacy of albendazole against Giardia and hookworm in a remote Aboriginal community in the north of Western Australia.

The parasitological, clinical efficacy and tolerability of albendazole in the treatment for both giardiasis and hookworm infection in a remote Aboriginal population was investigated. Albendazole at a dose rate of 400 mg daily for 5 days was highly effective in reducing hookworm egg numbers and both Giardia antigen and cysts. The 36.6% prevalence of Giardia prior to treatment fell to 12% between days 6 and 9, 15% for days 10-17 and rose to 28% between days 18 and 30. Tolerability and clinical efficacy were excellent. The effect of albendazole on hookworm was longer lasting than that on Giardia, reducing percent infection from over 76-2% on days 6-9 and zero by day 18-30 despite conditions highly conducive to rapid re-infection. We conclude that albendazole is highly efficacious against both parasites when used as described but that long term community benefit may require additional education programmes to avoid re-infection with Giardia although treatment strategies would seem appropriate for hookworm.

Genetic diversity and genetic differentiation in Echinococcus granulosus (Batsch, 1786) from domestic and sylvatic hosts on the mainland of Australia

Enzyme electrophoresis was used to examine genetic variation within and between populations of Echinococcus granulosus from domestic and sylvatic hosts in western and eastern Australia. Substantial genetic diversity was found within all populations. There was no evidence, however, of genetic differentiation between populations from different hosts or geographic areas. When isolates were grouped into previously described domestic or sylvatic strains on the basis of rostellar hook morphology, most (94%) of the genetic variation occurred within, rather than between strains. These results conflict with the currently accepted theory of separate domestic and sylvatic strains of E. granulosus on the mainland of Australia.

A case of mistaken identity - reappraisal of the species of canid and felid hookworms (Ancylostoma) present in Australia and India

This study serves to clarify the current status of canid and felid Ancylostoma species present in Australia. The morphological identification of A. ceylanicum from cats for the first time in Townsville, Australia, appears to be in error, together with the genetic markers provided for the species. Morphological and genetic data presented herein provide strong evidence that the hookworms from cats in Towsville are not A. ceylanicum as previously identified (i.e. the first report of this species in Australia), but are A. braziliense. Therefore the subsequent genetic markers established for A. ceylanicum in subsequent molecular studies based on these Townsville specimens should also be attributed to A. braziliense. Based on this information, a study of canine hookworm species present in northern India is also in error and it is apparent that the hookworms found in this region are those of A. ceylanicum. The distribution of A. braziliense and A. ceylanicum in the Americas and Asia Pacific region is discussed together with the importance of combining parasite morphology with genetic data for parasite diagnosis in epidemiological studies.

Comparative development of Echinococcus multilocularis in its definitive hosts

The comparative development of Echinococcus multilocularis was studied in its definitive hosts, the fox, dog, cat and raccoon dog, beyond the pre-patent period to 90 days post-infection. All host species, apart from cats were susceptible to infection and capable of supporting substantial worm burdens. Although worms in cats matured and produced thick-shelled eggs, their overall development was retarded compared to that in other species in which the parasite matured rapidly producing large populations of gravid worms. E. multilocularis matured rapidly in foxes and raccoon dogs and this was sustained in raccoon dogs but not in foxes in which maturation of worms declined during the later stages of infection, in contrast to that in both raccoon dogs and dogs. These populations were sustained for longer in raccoon dogs and dogs compared to foxes. Cats would appear to have only a minor role in the maintenance of E. multilocularis in endemic areas, and infections in cats may be of minimal public health significance. In contrast, foxes, dogs and the recently recognized definitive host the raccoon dog, are all capable of playing significant roles in the epidemiology of alveolar echinococcosis. This study also demonstrated that the developmental processes of growth, segmentation, proglottization and maturation in adult Echinococcus are independent and can be influenced by environmental factors thus confirming earlier in vitro observations.

Comparative studies on the growth dynamics of two genetically distinct isolates of Giardia duodenalis in vitro

The in vitro growth behaviour of the intestinal protozoan Giardia duodenalis was studied in detail and comparisons were made between two genetically and biologically distinct cloned isolates. Replicates of each clone were grown at six different initial cell concentrations and in culture media at four different pH values. Significant differences in in vitro growth were found between the two isolates, BAH12 and P1. BAH12 had a specific narrow pH requirement, with satisfactory growth only obtained at pH 6. The mean generation time of BAH12 at pH 6 between days 1 and 3 was 10.8 h, compared to an average of 6 h for the same period for P1, both at pH 6 and pH 7. Comparative health of cultures was assessed during both the pH and growth experiments using a suite of six variables. Consistent changes in the health of cultures over time were found to reflect growth behaviour over time. These results provide the first detailed evidence that genetically different isolates of Giardia may differ in such fundamental biological parameters as growth rate and pH requirements. These differences may have important epidemiological and taxonomic implications.

Pathogens of house mice on arid Boullanger Island and subantarctic Macquarie Island, Australia

Studies on island populations of house mice (Mus domesticus) and their viruses reveal insights into viral persistence in isolated communities. We surveyed the ectoparasites, endoparasites, and antiviral antibodies for 11 murine viruses and two bacteria of house mice inhabiting two islands off Australia. House mice on Boullanger Island were seropositive to two viruses, murine cytomegalovirus and epizootic diarrhea of infant mice. On subantarctic Macquarie Island, house mice were seropositive for five viruses: murine cytomegalovirus, lymphocytic choriomeningitis virus, mouse parvovirus, epizootic diarrhea of infant mice, and Theilers murine encephalomyelitis virus. The diversity of antiviral antibodies was lower among populations of house mice on islands than those inhabiting mainland Australia. The decreased diversity of viruses in island populations of house mice may be a function of which agent the founder mice transfer to the island and related to the low densities which the host population may periodically reach over time.

Larval anisakid infections of some tropical fish species from north-west Australia

Despite the commercial and zoonotic importance of larval anisakid infestations of teleosts, their distribution among Australias diverse marine fish fauna is poorly understood. A preliminary survey of Australias tropical north-west revealed a generally high prevalence of larval anisakids representing four genera (Anisakis, Terranova, Thynnascaris and Raphidascaris) among only seven fish species. The potential impact of high larval anisakid infections on both the health of recreational fishermen and aquaculture environments is discussed.