S.A. Reid

Trypanosoma evansi control and containment in Australasia

Animal trypanosomosis caused by Trypanosoma evansi is endemic throughout Southeast Asia, where it is an important constraint on the productivity of smallholder livestock. In the past decade, T. evansi has emerged as a serious threat to the viability of smallholder livestock industries in the Philippines and causes severe disease outbreaks with high mortality. Trypanosoma evansi also poses a threat to livestock and native fauna in Australia and Papua New Guinea (PNG) where it is absent, but the risk of it spreading from Indonesia is high. Surveillance for T. evansi in PNG and Australia, and its control in the Philippines is restricted by the poor sensitivity and inadequate validation of existing diagnostic tests and lack of information on the determinants of infection.

Transmission cycles of Giardia duodenalis in dogs and humans in Temple communities in Bangkok—A critical evaluation of its prevalence using three diagnostic tests in the field in the absence of a gold standard

The prevalence and associated risk factors for Giardia duodenalis in canine and human populations in Temple communities of Bangkok, Thailand were determined by evaluating three common diagnostic methods utilised to detect Giardia, namely zinc sulphate flotation and microscopy, an immunofluoresence antibody test and nested polymerase chain reaction (PCR) based on the SSU-rDNA gene. The diagnostic sensitivity and specificity together with the negative and positive predictive values of each test were evaluated in the absence of a gold standard using a Bayesian approach. The median estimates of the prevalence of infection with G. duodenalis in dogs and humans in Thailand were 56.8% (95% PCI, 30.4%, 77.7%) and 20.3% (95% PCI, 7.3%, 46.3%) respectively. PCR and immunofluorescence antibody tests (IFAT) were the most accurate tests overall with diagnostic sensitivity and specificity of 97.4% (95% PCI, 88.5%, 99.9%) and 56.2% (95% PCI, 40.4%, 82.9%) for the PCR and 61.8% (95% PCI, 40.8%, 99.1%) and 94.7% (95% PCI, 87.4%, 99.1%) for IFAT respectively Three cycles, anthroponotic, zoonotic and dog-specific cycles of G. duodenalis were shown to be operating among the human and canine populations in these Temple communities in Bangkok, supporting the role of the dog as a potential reservoir for Giardia infections in humans.

Immunization with recombinant beta-tubulin from Trypanosoma evansi induced protection against T. evansi, T. equiperdum and T. b. brucei infection in mice

The beta‐tubulin gene of Trypanosoma evansi (STIB 806) was cloned and expressed in Escherichia coli. The predicted amino acid sequence of T. evansi beta‐tubulin shows 100%, 99·8%, 99·1%, and 98·6% homology with T. equiperdum, T. b. brucei, T. cruzi and T. danilewskyi, respectively, but is diverse from that of T. cyclops, showing only 51·6% of homology. Recombinant beta‐tubulin was expressed as inclusion bodies in E. coli. It was purified and renatured for immunological studies. Mice immunized with the renatured recombinant beta‐tubulin were protected from lethal challenge with T. evansi STIB 806, T. equiperdum STIB 818 and T. b. brucei STIB 940, showing 83·3%, 70% and 76·7% protection, respectively. Serum collected from the rabbit immunized with recombinant beta‐tubulin inhibited the growth of T. evansi, T. equiperdum and T. b. brucei in vitro. Serum from mice and rabbits immunized with recombinant beta‐tubulin recognized only T. evansi beta‐tubulin and not mouse beta‐tubulin. The results of this study demonstrated that the recombinant T. evansi beta‐tubulin is a potential candidate for the development of a vaccine to prevent animal trypanosomiasis caused by these three trypanosome species.

Trypanosoma irwini n. sp (Sarcomastigophora: Trypanosomatidae) from the koala (Phascolarctos cinereus)

The morphology and genetic characterization of a new species of trypanosome infecting koalas (Phascolarctos cinereus) are described. Morphological analysis of bloodstream forms and phylogenetic analysis at the 18S rDNA and gGAPDH loci demonstrated this trypanosome species to be genetically distinct and most similar to Trypanosoma bennetti, an avian trypanosome with a genetic distance of 0.9% at the 18S rDNA and 10.7% at the gGAPDH locus. The trypanosome was detected by 18S rDNA PCR in the blood samples of 26 out of 68 (38.2%) koalas studied. The aetiological role of trypanosomes in koala disease is currently poorly defined, although infection with these parasites has been associated with severe clinical signs in a number of koalas. Based on biological and genetic characterization data, this trypanosome species infecting koalas is proposed to be a new species Trypanosome irwini n. sp.

Morphological and molecular characterization of Trypanosoma copemani n. sp. (Trypanosomatidae) isolated from Gilbert's potoroo (Potorous gilbertii) and quokka (Setonix brachyurus).

Little is known of the prevalence and life-cycle of trypanosomes in mammals native to Australia. Native Australian trypanosomes have previously been identified in marsupials in the eastern states of Australia, with one recent report in brush-tailed bettongs (Bettongia penicillata), or woylie in Western Australia in 2008. This study reports a novel Trypanosoma sp. identified in blood smears, from 7 critically endangered Gilberts potoroos (Potorous gilbertii) and 3 quokkas (Setonix brachyurus) in Western Australia. Trypanosomes were successfully cultured in vitro and showed morphological characteristics similar to members of the subgenus Herpetosoma. Phylogenetic analysis of 18S rRNA gene sequences identified 2 different novel genotypes A and B that are closely related to trypanosomes previously isolated from a common wombat (Vombatus ursinus) in Victoria, Australia. The new species is proposed to be named Trypanosoma copemani n. sp.

Novel trypanosome Trypanosoma gilletti sp. (Euglenozoa: Trypanosomatidae) and the extension of the host range of Trypanosoma copemani to include the koala (Phascolarctos cinereus)

Trypanosoma irwini was previously described from koalas and we now report the finding of a second novel species, T. gilletti, as well as the extension of the host range of Trypanosoma copemani to include koalas. Phylogenetic analysis at the 18S rDNA and gGAPDH loci demonstrated that T. gilletti was genetically distinct with a genetic distance (± s.e.) at the 18S rDNA locus of 2.7 ± 0.5% from T. copemani (wombat). At the gGAPDH locus, the genetic distance (± s.e.) of T. gilletti was 8.7 ± 1.1% from T. copemani (wombat). Trypanosoma gilletti was detected using a nested trypanosome 18S rDNA PCR in 3/139 (∼2%) blood samples and in 2/29 (∼7%) spleen tissue samples from koalas whilst T. irwini was detected in 72/139 (∼52%) blood samples and T. copemani in 4/139 (∼3%) blood samples from koalas. In addition, naturally occurring mixed infections were noted in 2/139 (∼1.5%) of the koalas tested.

Models for Trypanosoma evansi (surra), its control and economic impact on small-hold livestock owners in the Philippines

Simple demographic and infectious disease models of buffaloes and other domestic hosts for animal trypanosomosis (surra) caused by Trypanosoma evansi were developed. The animal models contained deterministic and stochastic elements and were linked to simulate the benefit of control regimes for surra in village domestic animal populations in Mindanao, Philippines. The impact of the disease on host fertility and mortality were key factors in determining the economic losses and net-benefit from the control regimes. If using a high (99%) efficacy drug in surra-moderate to high risk areas, then treating all animals twice each year yielded low prevalence in 2 years; targeted treatment of clinically sick animals, constantly monitored (monthly), required 75% fewer treatments but took longer to reach a low prevalence than treating all animals twice each year. At high drug efficacy both of these treatment strategies increased the benefit over untreated animals by 81%. If drug efficacy declined then the benefit obtained from twice yearly treatment of all animals declined rapidly compared with regular monitoring and targeting treatment to clinically sick animals. The current control regimen applied in the Philippines of annual sero-testing for surra and only treating sero-positive animals provided the lowest net-benefit of all the control options simulated and would not be regarded as effective control. The total net-benefit from effective surra control for a typical village in a moderate/high risk area was 7.9 million pesos per annum (US

Giardia genotypes in pigs in Western Australia: prevalence and association with diarrhea.

158,000). The value added to buffaloes, cattle, horses, goats/sheep and pigs as a result of this control was US

Identification of zoonotic Giardia genotypes in marsupials in Australia

88,

A Possible Role for Rusa Deer (Cervus timorensis russa) and Wild Pigs in Spread of Trypanosoma evansi from Indonesia to Papua New Guinea

84,