Trichostrongylus tenuis eggs were counted in faeces from individually marked wild red grouse for 8 years. Egg counts varied seasonally and annually. In some years, a sudden increase in mid-April was consistent with delayed maturation of larvae which had overwintered in the birds in a hypobiotic state. A more gradual increase in summer was probably due to uninterrupted maturation of larvae ingested then. Despite 30-fold year-to-year variation in mean egg counts, relative differences in egg counts among known individuals within years tended to persist across years. Rainfall in previous summers explained much of the year-to-year variation in egg counts, probably because parasite recruitment was greatest during wet summers. Grouse density was only weakly related to worm egg counts. The data were not consistent with the hypothesis that the cyclic-type population fluctuation in red grouse numbers observed at the time of this study was caused by the parasites.

The strongest evidence for host specificity of mammalian trypanosomes comes from parasites of the subgenus Trypanosoma (Herpetosoma). Laboratory studies have shown that T. (Herpetosoma) species will not infect an alternative host. However, this has not been demonstrated in wild populations. We screened 560 bank voles (Clethrionomys glareolus) and 148 wood mice (Apodemus sylvaticus) for trypanosomes by PCR amplification of the 18S rRNA gene. In total, 109 (19%) bank voles and 12 (8%) wood mice were infected. A HaeIII restriction site was discovered that could be used to discriminate between T. (H.) evotomys of the bank vole and T. (H.) grosi of the wood mouse. All the parasites in the bank voles were identified as T. (Herpetosoma) evotomys by RFLP-PCR. Out of the 12 wood mouse infections 10 were due to T. grosi. Two of the wood mice were infected with parasites with a novel genotype that was most similar to those of T. evotomys and T. microti of voles. Fifty-six fleas collected from the rodents were also screened for trypanosomes; 9 were infected with T. evotomys and 1 with T. grosi. One of the fleas infected with T. evotomys was collected from a wood mouse.

In the present study, we constructed gender-enriched cDNA libraries for the adult stage of the parasitic nematode Oesophagostomum dentatum (order Strongylida) using suppressive-subtractive hybridization (SSH), sequenced clones from the female-library and male-library (480 from each) and conducted bioinformatic and microarray analyses of the expressed sequence tags (ESTs). In total, 873 ESTs (440 male and 433 female) were obtained, achieving a sequencing success of 91%The nucleotide sequences reported in this article (Tables 1–5) have been deposited in the EMBL, GenBank and DDJB databases under the Accession nos. AM157797–AM158083. Microarray analyses of 516 unique ESTs representing both gender-enriched libraries revealed differential hybridization for 391 of them (75·8%). Of these, 220 (56·3%) had significantly greater signal intensities in the female than in the male, and 154 (70%) of these were predicted to have homologues in C. elegans. These homologues were predicted to be involved in key biological processes, including embryonic nutrition, gametogenesis, molecular binding/transport or metabolism, nucleic acid synthesis and function, and signal transduction. Of the 171 ESTs with statistically higher signal intensities in male O. dentatum, 43·8% had homologues in C. elegans. These homologues included major sperm proteins (MSPs) or MSP-like molecules, keratin-like molecules, molecules involved in metabolism, PDZ domain-containing proteins, sugar binding proteins, protein kinases, serine proteases or protease inhibitors, molecules involved in proteolysis and other proteins, such as enzymes and various putative proteins. Of the 287 ESTs (from both gender-enriched cDNA libraries) with no known homologues in C. elegans, 50 (17·4%) had homologues in other nematodes, 8 had homologues in various other organisms and 104 (36·2%) had no homology to any sequence in current gene databases. The present study lays a foundation for the isolation and molecular, biochemical and functional characterization of selected genes from the gender-enriched cDNA archives established for O. dentatum.

The establishment of Leishmania in mammals depends on the transformation of metacyclic promastigotes into amastigotes within macrophages. The kinetics of this process was examined using mouse macrophages infected with metacyclic promastigotes of L. amazonensis. The appearance of amastigote characteristics, including large lysosome-like organelles called megasomes, stage-specific antigens, high cysteine protease activity and sensitivity to L-leucine methyl ester, was followed over a 5-day period. Megasomes were observed at 48h but probable precursors of these organelles were detected at 12h p.i. The promastigote-specific molecules examined were down-regulated within 5 to 12h after phagocytosis whereas the amastigote-specific antigens studied were detectable from 2 to 12–24h. An increase in the cysteine protease activity and in sensitivity to L-leucine methyl ester of the parasites was detected from 24h. The data indicate that at 48h p.i., parasites exhibit several amastigote features but that complete differentiation requires at least 5 days. The appearance of megasomes or of megasome precursors and the rise in cysteine protease activity correlate quite well with the capacity of parasites to internalize and very likely degrade host MHC molecules. The fact that internalization by the parasites of host cell molecules occurs very early during the differentiation process argues for a role of this mechanism in parasite survival.

Rabbits and mice, but not sheep, guinea-pigs or rats, were suitable hosts for secondary echinococcosis using protoscoleces obtained from Australian sheep. The host specificity is tentatively related to the phylogenetic relationship between the donor host and receptor host.

The development rate of hydatid cysts in mice is described in terms of increase in fresh weight, dry weight, and maximum cyst diameter. Fertile cysts were found after 8 months when the water content of the cysts had increased to more than 95 %. The diameter of fertile cysts up to 14 months ranged from 6 to 30 mm. By 14 months, the average fresh weight of cysts was greater than the body weight of the mice.

This work was carried out during the tenure of an Australian Wool Board Senior Postgraduate Scholarship, and forms part of a thesis to be presented for the degree of Doctor of Philosophy of the Australian National University.

I wish to thank my supervisor, Professor J. D. Smyth, for guidance and criticism.

A technique is described for the efficient feeding of Rhipicephalus appendiculatus nymphae on cattle blood through an artificial membrane bearing tactile and olfactory stimuli. The effect of four anticoagulation methods on the feeding of nymphae was compared and heparinized blood was found to be the most efficacious, followed by defibrinated blood. Blood treated with acid citrate dextrose (ACD) or ethylenediamine tetraacetate (EDTA) inhibited nymphal feeding. Nymphae fed on heparinized and defibrinated blood obtained engorgement weights within the range of ticks fed on mammalian hosts and they subsequently moulted and fed normally as adults and produced viable eggs. Nymphae fed on membranes using either defibrinated or heparinized blood infected with Theileria parva piroplasms developed salivary gland infections as adult ticks and transmitted East Coast fever (ECF) to susceptible cattle. There were indications that T. parva-infected defibrinated blood was not as infective to the feeding nymphae as the infected heparinized blood. When T. parva-infected heparinized blood was used to feed nymphae through membranes in two experiments, it was found that the infections in the resultant adult ticks could be comparable to those of nymphae fed on donor cattle, but were usually lower. The membrane feeding technique will enable the study of factors affecting the tick and T. parva transmission without the complication of host/T. parva interactions and could be useful for both tick maintenance and Theileria parasite isolation and maintenance.

This study examines the ability of an assay which measures the amount of a schistosome specific antigen (CAA) in the host circulation to reliably reflect relative worm burden. Mice were infected with 5 species of schistosome with a range of infection dose. The levels of serum CAA increased during schistosome maturation. In all species tested CAA levels correlated well with adult worm burden once the parasites achieved sexual maturity and remained relatively stable during the establishment of egg production. The amount of CAA produced varied between species but within each species CAA levels were proportional to worm numbers: no density-dependent effects on CAA levels were observed even when mice carried worm burdens that were very large relative to host size. T-cell deprivation of the host had no effect on the CAA/worm burden relationship in either Schistosoma mansoni or S. haematobium infections and the CAA equilibrium was unaltered in intact mice when reduction of worm fecundity occurred. These data support the use of the CAA as an accurate and robust estimate of relative schistosome burden in man.

The life-cycle of Isospora ohioensis in dogs is described. Eight dogs were killed 6, 16, 24, 48, 72, 96, 114 and 120 h after feeding 106I. ohioensis sporocysts to each. First divisional stages occurred in the jejunum at 48 h post-inoculation (p.i.). At 48 h, zoites occurred in pairs in parasitophorous vacuoles of surface epithelial cells of the jejunum. The parasitophorous vacuoles were 7–9 × 6 μm and the zoites were 7–9 × 2·5 μm in sections. At 72 h, uninucleated zoites, multinucleated zoites and meronts containing fully formed merozoites occurred in surface epithelial cells of the jejunum. At 96, 114 and 120 h asexual multiplication occurred throughout the small and large intestine, mostly in the ileum. The number of asexual generations was not determined. At least 2 structurally distinct meronts were identified at 96–120 h p.i. Type I meronts contained larger merozoites (11 × 3 μm) than those (7·5 × 1·5 μm) in Type II meronts. Meronts were merozoite-shaped and contained up to 8 nuclei. Uninucleated, binucleated and multinucleated zoites occurred within the same parasitophorous vacuole. Gamonts occurred in surface epithelial cells of the small intestine, caecum and colon, but predominantly in the ileum, 96–120 h p.i. Macrogamonts were 13–17 × 11–12 (14·5 × 12·8) μm in sections and 21–26 × 17–25 (21·7 × 17·6) μm in smears. Microgamonts were 13–17 × 8–15 (15·3 × 11·4) μm in sections and 24–30 × 15–24 (27 × 19) μm in smears and contained up to 50 microgametes. The cycle was also studied in 6 dogs killed 6, 12, 24, 48, 96 and 120 h after feeding tissues of laboratory mice that had been infected with I. ohioensis oocysts 17 days earlier. Stages were not seen in the dogs killed at 6 and 12 h p.i. Asexual and sexual stages were structurally similar in size and location to the oocyst-induced cycle but their development was faster by 24 h in the mouse-induced cycle. Extra-intestinal stages were not found in sections of mesenteric lymph nodes, spleen, lung, liver, heart, skeletal muscle and brain. However, biological evidence indicated that I. ohioensis invaded the spleens and mesenteric lymph nodes of dogs fed oocysts.

Recent studies of Echinococcus granulosus molecular strain typing have enabled a better understanding of the transmission cycle of cystic echinococcosis. There have been many publications in this area but there is a need for the evaluation of these tools. We have attempted to respond to this need in our study, which assessed 8 DNA fragments of 40 E. granulosus cysts from North Africa. Parasitological material was collected from 5 types of intermediate hosts, in 5 different countries. The primers chosen to amplify DNA targets were defined either in nuclear DNA, or in mitochondrial DNA. After amplification, PCR products were sequenced. The sequences obtained were aligned and comparisons were made within the group and with GenBank sequences. Whether the target was nuclear or mitochondrial, the same 2 main groups of genotypes were found. The first one, the ‘sheep’ strain, was found in the human, sheep and cattle samples collected in North Africa. The second one, the ‘camel’ strain, was found in the camel cysts and cattle and human cysts from Mauritania. These findings further confirm the congruence of the data given by the nuclear and the mitochondrial genome.

A Strain ofHaemonchus contortusshowing moderate levels of resistance to thiabendazole (TBZ) (egg hatch assay LC50 2.1 μM) was subjected, in alternating sequence, to 5 in vitro and 4 in vivo selections with TBZ. This resulted in a significant increase in resistance (egg hatch assay LC50 2.8 μM). The isolates obtained after each combined in vitro and in vivo selection were compared in terms of egg hatch assay, establishment, net egg output, per capita egg output and degree of pathology caused in infected hosts. The increase in resistance was accompanied by a significant increase in larval establishment sheep (22 %) and was associated with more severe pathology, higher egg count and worm recovery 3 months post- infection.

A total of 41 ticks were collected from 15 quokkas on Bald Island and 2 ticks from a Gilbert's potoroo from Two Peoples Bay. Three species of Ixodid ticks Ixodes australiensis, Ixodes hirsti and Ixodes myrmecobii were identified on the quokkas known to have a high prevalence of Trypanosoma copemani. Tick faeces from ticks isolated from 8 individual quokkas and a Gilbert's potoroo were examined with one identified as positive for trypanosomes. Faecal examination revealed trypanosomes similar to in vitro life-cycle stages of T. copemani. In total 12 ticks were dissected and trypanosomes found in sections of their midgut and haemolymph, 49 and 117 days after collection. Tick faeces, salivary glands and midguts from I. australiensis were screened using an 18S rRNA PCR with amplification seen only from the midguts. Sequencing showed 100% homology to T. copemani (genotype A) and 99·9% homology to the wombat (AII) isolate of T. copemani. Trypanosomes were only detected in I. australiensis as neither I. hirsti nor I. myrmecobii survived the initial 30-day storage conditions. We therefore identify a vector for T. copemani as I. australiensis and, given the detection of trypanosomes in the faeces, suggest that transmission is via the faecal-oral route.

The protozoan parasite Toxoplasma gondii is prevalent worldwide and can infect a remarkably wide range of hosts despite felids being the only definitive host. As cats play a major role in transmission to secondary mammalian hosts, the interaction between cats and these hosts should be a major factor determining final prevalence in the secondary host. This study investigates the prevalence of T. gondii in a natural population of Apodemus sylvaticus collected from an area with low cat density (<2·5 cats/km2). A surprisingly high prevalence of 40·78% (95% CI: 34·07%–47·79%) was observed despite this. A comparable level of prevalence was observed in a previously published study using the same approaches where a prevalence of 59% (95% CI: 50·13%–67·87%) was observed in a natural population of Mus domesticus from an area with high cat density (>500 cats/km2). Detection of infected foetuses from pregnant dams in both populations suggests that congenital transmission may enable persistence of infection in the absence of cats. The prevalences of the related parasite, Neospora caninum were found to be low in both populations (A. sylvaticus: 3·39% (95% CI: 0·12%–6·66%); M. domesticus: 3·08% (95% CI: 0·11%–6·05%)). These results suggest that cat density may have a lower than expected effect on final prevalence in these ecosystems.

Physical habitat structure can influence the distribution and abundance of organisms. In rivers, stream drift, a common process originating from the unidirectional water flow, favours the displacement and downstream dispersion of invertebrates. This process could also generate a gradient in infection levels, leading to decreasing numbers of parasites per host as one moves upstream from the river mouth. We tested this hypothesis using 4 trematode species infecting the fish Gobiomorphus breviceps in the Manuherikia River (New Zealand). We analysed the abundance of each trematode infrapopulation as a function of distance from the river junction and fish size by generalized linear models. Our results supported the existence of a longitudinal gradient in trematode abundance along the river with a decreasing downstream-to-upstream continuum. This applied to 3 out of the 4 trematode species studied, suggesting that this might be a common pattern in river populations. Thus, the unidirectional river flow and a major process like drift in lotic systems, that influences the dynamics and distribution of invertebrate hosts, can also affect trematodes. Host properties like habitat preference, and parasite traits, particularly those related to transmission mode can influence the strength of the observed gradient, as may other environmental and biotic factors.

Human African trypanosomiasis (HAT) is caused by Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense and caused devastating epidemics during the 20th century. Due to effective control programs implemented in the last two decades, the number of reported cases has fallen to a historically low level. Although fewer than 977 cases were reported in 2018 in endemic countries, HAT is still a public health problem in endemic regions until it is completely eliminated. In addition, almost 150 confirmed HAT cases were reported in non-endemic countries in the last three decades. The majority of non-endemic HAT cases were reported in Europe, USA and South Africa, due to historical alliances, economic links or geographic proximity to disease-endemic countries. Furthermore, with the implementation of the ‘Belt and Road’ project, sporadic imported HAT cases have been reported in China as a warning sign of tropical diseases prevention. In this paper, we explore and interpret the data on HAT incidence and find no positive correlation between the number of HAT cases from endemic and non-endemic countries. This data will provide useful information for better understanding the imported cases of HAT globally in the post-elimination phase.

Engorged Ixodes ricinus nymphs collected from sheep resident in an upland UK field site were significantly lighter than nymphs that engorged on previously tick-naïve sheep, indicating that site-resident sheep continually exposed to ticks acquired anti-tick resistance. The weights of engorged nymphs that fed on naturally tick-resistant sheep increased significantly, however, when increasingly high numbers of adult female ticks fed on the sheep during seasonal peaks of tick activity. This relationship was unaffected by variations in nymph weight amongst individual sheep, between seasons and years, and potential effects of sheep infection with Ehrlichia phagocytophila; this suggests that high adult tick infestations may directly inhibit the expression of acquired anti-tick resistance by sheep. The length, width and weight of adult ticks and the scutum length of adult females were linearly related to their weight as an engorged nymph. The mean scutum length of adult female ticks feeding on sheep in the field site was greater than that of adult females obtained from engorged nymphs collected from sheep of the same site. This suggests that larger ticks have a survival advantage and that I. ricinus ticks exhibit density-dependent intraspecific facilitation at high infestation levels with potential consequences for the transmission of tick-borne diseases.

Sequences of the first internal transcribed spacer (ITS-1) and the D1-D3 domains of the large subunit (LSU) of the ribosomal DNA (rDNA) were determined for multiple specimens of 4 operational taxonomic units (OTUs) of the monogenean, Pseudorhabdosynochus lantauensis. OTUs were defined based on their collecting localities, host and/or morphological characteristics. All P. lantauensis specimens of one group (OTUs 1 and 3) differed in their sequences of the ITS-1 and partial LSU rDNA when compared with specimens of a second group (OTUs 2 and 4) by 12% and 2%, respectively. Results of the phylogenetic analyses of the LSU rDNA sequence data showed total (100%) bootstrap support for the separation of P. lantauensis into 2 distinct clades. At least 11 of the 18 nucleotide differences in the LSU sequence between the two P. lantauensis clades were derived (i.e. autapomorphic) characters when the morphologically distinct species, P. epinepheli and P. coioidesis, were used as outgroups. Furthermore, there were several autapomorphic character states for each P. lantauensis clade. This provides sufficient evidence to reject the null hypothesis that P. lantauensis represents a single species. Morphological and morphometric differences between these two clades provided additional strong support for the separation of P. lantauensis into two species. These two parasite species were found to co-exist on one of the two species of serranid fish (i.e. Epinephelus coioides) examined in the South China Sea (Guangdong Province, China).

Some Trypanosoma and Leishmania species are multi-host parasites whose distribution overlaps in several parts of the Brazilian Amazon basin. Despite being a common trait among wild mammals, mixed infections and their consequences for the host's health and parasite transmission are still a poorly known phenomenon. Here we describe a triple mixed infection – Trypanosoma cruzi, T. rangeli and Leishmania infantum – in a bone marrow sample from an anteater Tamandua tetradactyla captured in a house backyard from the endemic Abaetetuba municipality in the Amazon basin. T. cruzi was also isolated from blood samples. The mini-exon multiplex PCR characterization detected the infection by T. rangeli and T. cruzi (TcI genotype), while L. infantum infection was confirmed by an ITS-PCR followed by amplicon sequencing. This is the first description of T. rangeli isolation from bone marrow and the first report of L. infantum infection in xenarthrans. The implications of this finding are discussed considering the influence of mixed infections in the role of this mammal species as a putative reservoir host of these 3 trypanosomatid species.

Partial sequences of the DNA polymerase delta (pold) gene from Taenia saginata-like adult worms were sequenced. Phylogenetic analysis revealed that pold gene sequences were clearly divided into two clades, differing from each other in five to seven nucleotides. There is little doubt that T. saginata and Taenia asiatica were once separated into two distinct taxa as has been concluded in previous studies. On the other hand, most of the adult worms, which were identified as T. asiatica using mitochondrial DNA, were homozygous for an allele that originated from the allele of T. saginata via single nucleotide substitution. These results indicate that most of the adult worms, which had been called T. asiatica, are not actually ‘pure T. asiatica’ but instead originated from the hybridization of ‘pure T. saginata’ and ‘pure T. asiatica’.

The immunological differences known to exist between laboratory strains of Eimeria maxima was confirmed. Protection against challenge with different strains or field isolates of the species could be achieved by including small numbers (25 oocysts) of each in the immunizing inoculum. Similar protection was obtained when 4 distinct populations which were allowed to interbreed were used in the immunizing inoculum. This hybrid mixture of E. maxima was used to immunize chickens against challenge with 7 new isolates of E. maxima from poultry houses in different parts of England. The results show that although immunological differences exist within E. maxima good protection against many strains of this species may be achieved by initial infection with the hybrid mixture of E. maxima.

1. An account is given of the development of strains of Trypanosom gambiense in Glossina tachinoides.

Complete development with infection of salivary glands and hypopharynx occupies from 12 to 40 days and upwards, according to the strain of trypanosome used and the temperature conditions during development.

2. It is shown that the first or intestinal phase of development is passed within the extra-peritrophic space and that the lumen or intra-peritrophic space of the gut is not involved.

3. The importance of the proventricular infection as an index of maturing infections is stressed, and it is shown that filamentar crithidia are always present in this organ in the later stages of infection when infection of salivary glands is taking place. It has been proved that these filamentar crithidia may constitute up to 20 per cent. of flagellates in the proventriculus and are partly responsible for the further migration to the salivary glands.

4. An account is given of the infectivity to G. tachinoides of twenty-six strains of T. gambiense from epidemic and endemic sleeping sickness areas in Northern Nigeria. None of the twenty-six was proved to be non-transmissible and twenty-five of the strains gave Total Infectivity Rates in the tsetse of from 0·9–10·5, the Cyclical Developmental Rates ranging from 0·6–9·8.

5. No evidence has been obtained that strains of T. gambiense from chronic cases of human trypanosomiasis in endemic areas are less transmissible by tsetse than strains originating in epidemic areas.

6. An account is given of experiments in which G. tachinoides infected with T. gambiense were exposed to 37° C. for varying lengths of time. The effect of high temperature during the infecting period is seen in a marked and constant increase in the proportion of tsetse infected. In the case of one strain a total infection rate of 77·8 per cent. was obtained with true cyclical development in 73·5 per cent. of the tsetse used.

7. It is suggested that the temperatures at which experimental tsetse are infected in the laboratory with T. gambiense are excessively low, and an investigation is to be made into the optimum climatic conditions for development of the trypanosome in the fly with a parallel attempt to correlate this with the climatic conditions under which wild tsetse become infected in nature.