The life-cycle of the tapeworm Echinococcus multilocularis is dependent upon the predator - prey relationship between canids (e.g. Vulpes sp) and small mammals. Accidental ingestion of E. multilocularis eggs can lead to the rare but fatal zoonotic disease alveolar echinococcosis (AE) in humans. Detecting early asymptomatic infections greatly increases patients' life expectancy. Mass-screening detect unknown cases using ultrasonography. Reliable predictive methods could help medics target resources. Here epidemiology, landscape ecology, satellite remote sensing and spatial modelling are integrated for the purposes of analytical inference and spatial prediction.

Echinococcosis caused by E.granulosus or E.multilocularis is a chronic, debilitating and potentially fatal zoonotic cestode disease of humans. The natural primary definitive host for E.granulosus is the domestic dog and for E.multilocularis it is wild foxes. In areas of the eastern Tibetan plateau, it was suspected that the domestic dog played a vital role in the transmission of both species of parasite to man, contributing to some of the highest prevalence rates globally. A new species, E.shiquicus is distributed sympatrically in the same location but its zoonotic potential was unknown. In order to investigate the role of canids in transmission of E.shiquicus, E.granulosus and E.multilocularis, 197 faecal samples from Tibetan foxes (Vulpes ferrilata) in Sichuan, Qinghai and Tibet Autonmous Region (TAR) and more than 600 faecal samples from owned domestic dogs in Sichuan were collected and analysed by an Echinococcus genus specific coproantigen- ELISA and three species specific copro-PCR tests. The copro-ELISA was shown to have a sensitivity of 86.6% for E.granulosus and 72.7% for E.multilocularis. The specificity was 100% when compared with faecal samples from other taeniid tapeworm infections. Copro-PCR tests were considered the most specific for use in the eastern Tibetan plateau co-endemic communities. The primary objective endeavoured to understand the role of the domestic dog in maintaining transmission of E.multilocularis in Shiqu County, Sichuan. A cohort of 308 dogs were followed up for one year after a single treatment with praziquantel for a re-infection study at 2 month, 5 months and 12 months. This research was the first to confirm E.multilocularis is found in foxes across the plateau into central TAR. The prevalence ranged from 2.6% to 25% dependant on location. In foxes E.shiquicus was distributed ~350 miles west of Shiqu County (where it was first described) at a prevalence of 6.1% and the prevalence appeared to be increasing along a gradient from north to south of the plateau. No dog faecal samples were positive for E.shiquicus DNA. The prevalence of E.multilocularis in dogs reached 8.9% in one endemic foci (Shiqu County) whilst E.granulosus was distributed evenly across the study sites. There was no significant difference between the prevalence of E.multilocularis in the dog and fox populations. The re-infection study of dogs demonstrated they are Echinococcus copro-ELISA test positive at a prevalence of 8.4% after 2 months, 2.2% after 5 months and 9.5% after 12 months. No positive copro-PCR results were obtained at 5 months and 12 month post treatment however knowledge of the parasite biology and host availability/behaviour meant that some assumptions could be made. It was considered that the infection pressure to dogs from small mammals infected with E.multilocularis is at a peak in the late spring to early summer whilst the infection pressure from livestock infected with E.granulosus to dogs is at a peak in late autumn to early winter. Furthermore, the data indicated that dogs may have the ability to maintain E.multilocularis transmission without the input of a fox definitive host. This was based on the significant reduction in copro-prevalence 12 months post treatment and the probable effect the dosing had on transmission of E.multilocularis to small mammals in the research area. The only significant risk factor for dog echinococcosis in the current study was the release of dogs at night by owners which allows them to roam in the villages. It was thought that these dogs have more access to small mammals or livestock carcasses infected with Echinococcus spp. Identification of peak Echinococcus transmission periods are discussed with a view to control via dog dosing schemes on the plateau.

Echinococcus multilocularis has recently been developed as a model for basic and applied studies on trematodes and cestodes. Deep understanding of molecular biology of this parasite is urgently required for efficient and extensive applications such as functional gene probing and anti-helminth drug screening. In our project, we aimed to unveil E. multilocularis miRNA repertoires and the identification of differentially expressed molecules from the mRNA and miRNA transcriptomes using next-generation sequencing technology. Furthermore, one family of E. multilocularis molecules identified, fatty acid-binding proteins that are involved in lipid metabolisms, was phylogenetically and functionally characterized. Our data presented genome-widely developmental expression profiles of E. multilocularis miRNAs. Sixty-five potential miRNAs were predicted with nineteen of them being novel. The majority of differentially expressed miRNAs such as emu-miR-1, 16 and 71 were found to be significantly down-regulated in the primary neoblast-rich cells derived from the vesicles. Comparative analysis revealed that the miR-71/2 cluster was conserved in the platyhelminths. In addition, seven homologues to miRNAs likely linked to planarian neoblasts were present in E. multilocularis with the intact seed regions. Although their expression in E. multilocularis neoblasts remains to be fully certified, these miRNAs are plausible biomarkers for the germline stem cells. For mRNA profiling, we firstly originated an enzymatic approach for the elimination of mitochondrial transcripts to increase the depth of mRNA transcriptomes. In comparison with 2.7% in the control, the frequency of the sequences mapped to the mitochondria was dramatically reduced to 0.04~0.1% in the treated samples without significantly adverse effects on the targets. A number of differentially expressed genes were inferred from E. multilocularis mRNA transcriptomes and FABP-encoding genes were further phylogenetically and functionally characterised. E. multilocularis encoded at least four distinct FABPs and comparative analysis revealed that the current gene set of FABPs may have emerged before speciation of E. multilocularis and E. granulosus. The apparent divergence between Echinococcus and vertebrate FABPs in genomic organizations prompted us to further explore phylogenetically FABP genes across thirty-five invertebrates. The results demonstrated that FABP genes were organized in diverse ways, with a predominant pattern that is commonly present in vertebrates. Moreover, both gene duplication and alternative splicing might be most likely responsible for variety of invertebrate FABP functions. Four E. multilocularis FABPs were developmentally regulated and FABP3 was highly expressed in the vesicles and secreted or released into the hydatid fluid. These proteins appeared to be able to weakly interact with cis-parinaric acid but not a fluorescent fatty acid DAUDA and retinol. In vitro analysis indicated that FABP3 was capable of the induction of cytokine secretion by bone marrow-derived macrophages and dendritic cells probably via Toll-like receptor 2. Additionally, FABP3 was also demonstrated to activate macrophage small reactive molecule production together with a ligand for TLR 2.