Experimental infections of canaries with an avian trypanosome, Trypanosoma bouffardi, isolated from West Africa demonstrated that these infections can cause pathological changes in tissues. Enlargement of the spleen coincided with peak parasitaemia but no other gross changes were observed. Histopathological examination of infected and control birds revealed focal myocarditis and lymphoid hyperplasia, in infected birds, which are consistent with trypanosome infection. Avian trypanosome infections are not usually regarded as pathogenic but our results suggest that pathological changes occur which are similar to those observed in mammals infected with salivarian trypanosomes

The life-history of I. pacificus when fed on guinea-pigs under laboratory conditions is described. Feeding of the female proceeds for 10–11 days. Between 120 and 191 mg of concentrated food is found in the female's gut on detaching from this host, and this is directly related to the unfed body weight of the tick.

The egg-laying pattern, in general, shows an initial peak, after which the number of eggs tails off, the whole process lasting approximately 33–40 days. The number of eggs laid by females varies between 790 and 1300. The preoviposition period is about 11–16 days.

Larvae hatch in about 53–55 days, and their feeding time ranges from 4 to 9 days, although in the present experiments larvae emerging from early laid eggs took about 10 days to complete engorgements.

Moulting of larvae to nymphs at 90% R.H. took 37–38 days, and nymphs fed for 7–11 days, with most becoming replete on the ninth.

Very few females emerged from these nymphs, and those that did emerged in just over 30 days. The reasons for this are not clear. No diapause appeared to be necessary to complete the life-cycle, and with due allowance for hardening of the mouthparts the life-history of one generation was about 7 months. Females died within 3 or 4 months if hosts were not available.

From the literature and from hitherto unpublished records some 55 vertebrates have been implicated as hosts to I. pacificus; these hosts include a wide range of mammals, six species of birds and possibly nine species of lizards.

The species has been listed from British Columbia and from five western states of U.S.A. The need for more intensive ecological work on this species is emphasized.

We are indebted to Glen M. Kohls, J. D. Gregson and D. E. Johnstone for permission to use unpublished records from their respective institutions. One of us (D.R.A) is also indebted to the Wellcome Trustees for financial assistance to visit Canada during the preliminary stages of the work.

The life-cycle of canine Isospora bigemina was studied in dogs, cattle, cats and mice. Under experimental conditions dogs served as both definitive and intermediate hosts. Unsporulated oocysts (11 × 12 μm) were shed in the faeces and sporulated outside the host within 12 h at 30–37 °C and 36 to 48 h at 23 °C. Sporulated oocysts measured 12 × 13 μm and contained 2 sporocysts which in turn contained 4 sporozoites. Sporocysts averaged 6 × 9 μm and sporozoites averaged 2 × 6 μm. Although no stages were found microscopically in tissues or in faeces of 23 dogs orally inoculated with sporulated oocysts, the oocysts were infectious because 5 of 14 dogs that ingested tissues of dogs fed oocysts, shed oocysts. Prepatent periods were 7 to 15 days; patency was 1–3 days. No dogs became ill. In the naturally infected dog and one experimentally infected dog, schizonts were found in epithelial cells, distal to the host cell nucleus at the tips of villi throughout the small intestine. Schizonts were 5–7 μm and contained 3–12 merozoites. In the naturally infected dog, gametes were found in the same location. Male gametocytes were 6–8 μm and contained 6–12 microgametes, and female gametes were 7–8 μm.

Sporulated I. bigemina oocysts from a naturally infected dog were not infectious to cattle, cats or mice. Structurally identical oocysts were shed by 2 dogs after ingesting hearts and diaphragm from naturally infected cattle; these oocysts were also not infectious to cattle. Although dogs acted as both intermediate and definitive hosts under experimentation, this is not likely to occur in nature. A canine-bovine-canine cycle would appear to be the natural mode of infection but I. bigemina was not infectious to cattle under experimental conditions.

Unsporulated oocysts were recovered from the allantoic cavity of 5 chicken embryos inoculated with a single sporozoite of Eimeria tenella; some of these oocysts sporulated. Both macrogametes and microgametes are required for the formation of viable oocysts. The results conclusively demonstrate the bisexual nature of the sporozoite, suggest that sexual differentiation is influenced by environmental factors, and show that true clones of Eimeria can be established from individual sporozoites.

A contingency table approach was used to explore the influence of location, host species and time on the genetic composition of a Trypanosoma brucei population in Lambwe Valley, Kenya. Significant differences in zymodeme frequencies were noticed over comparatively short geographical distances suggesting that transmission of T. brucei is somewhat localized. A significant association was observed between zymodeme and the mammalian host from which T. brucei was derived. The association was consistent in different localities in Lambwe valley and remained stable for at least 32 months. These observations indicate that zymodemes are adapted to different host species and that genetic exchange has not disrupted host associations over this time-scale. A major change in the composition of the T. brucei population during a sleeping sickness outbreak in 1980 was confirmed. But while new zymodemes emerged, a decline in overall diversity was noted during times of high sleeping sickness incidence. The results can be explained by selection of T. brucei zymodemes for particular transmission cycles. Although it is not necessary to invoke genetic exchange, sex may help T. brucei to adapt to changes in selection pressures. Such a hypothesis helps to explain why T. brucei appears largely clonal in the short term, even though population studies indicate that sex is responsible for much genetic diversity in the long term. It also explains why neighbouring populations of T. brucei are composed of a different range of zymodemes formed from the same alleles. Such a view implies that genetic exchange has an important role in the microevolution of T. brucei populations.

Malaria remains a major cause of mortality in African children, with no adjunctive treatments currently available to ameliorate the severe clinical forms of the disease. Rosetting, the adhesion of infected erythrocytes (IEs) to uninfected erythrocytes, is a parasite phenotype strongly associated with severe malaria, and hence is a potential therapeutic target. However, the molecular mechanisms of rosetting are complex and involve multiple distinct receptor–ligand interactions, with some similarities to the diverse pathways involved in P. falciparum erythrocyte invasion. This review summarizes the current understanding of the molecular interactions that lead to rosette formation, with a particular focus on host uninfected erythrocyte receptors including the A and B blood group trisaccharides, complement receptor one, heparan sulphate, glycophorin A and glycophorin C. There is strong evidence supporting blood group A trisaccharides as rosetting receptors, but evidence for other molecules is incomplete and requires further study. It is likely that additional host erythrocyte rosetting receptors remain to be discovered. A rosette-disrupting low anti-coagulant heparin derivative is being investigated as an adjunctive therapy for severe malaria, and further research into the receptor–ligand interactions underlying rosetting may reveal additional therapeutic approaches to reduce the unacceptably high mortality rate of severe malaria.

Here we review recent studies on the mode of action of the cholinergic anthelmintics (levamisole, pyrantel etc.). We also include material from studies on the free living nematode Caenorhabditis elegans. The initial notion that these drugs act on a single receptor population, while attractive, has proven to be an oversimplification. In both free living and parasitic nematodes there are multiple types of nicotinic acetylcholine receptor (nAChR) on the somatic musculature. Each type has different (sometimes subtly so) pharmacological properties. The implications of these findings are: (1) combinations of anthelmintic that preferentially activate a broad range of nAChR types would be predicted to be more effective; (2) in resistant isolates of parasite where a subtype has been lost, other cholinergic anthelmintics may remain effective. Not only are there multiple types of nAChR, but relatively recent research has shown these receptors can be modulated; it is possible to increase the response of a parasite to a fixed concentration of drug by altering the receptor properties (e.g. phosphorylation state). These findings offer a potential means of increasing efficacy of existing compounds as an alternative to the costly and time consuming development of new anthelmintic agents.

This cross-sectional study determined the prevalence and distribution of schistosome and soil-transmitted helminth (STH) infections among 1,308 children aged 10–18 years in 34 primary schools in 8 informal urban settlements in Kisumu City, western Kenya. Stool samples were collected and examined for eggs of Schistosoma mansoni and STH (Hookworms, Ascaris lumbricoides and Trichuris trichiura) using the Kato-Katz technique. Haematuria was used as a proxy indicator of urinary schistosomiasis. Schools and water bodies were mapped using a geographical information system. Overall, 34% of children were infected with one or more helminth species whereas 16·2% of children were infected with one or more STH species. Schools in closest proximity to Lake Victoria and River Nyamasaria had the highest S. mansoni prevalence while schools with STH were more homogenously distributed. Mean school prevalence of S. mansoni infection was 21% (range=0–69·7%), S. haematobium 3·6% (range=0–12%), hookworms 6·1% (range=0–20%), A. lumbricoides 4·9% (range=0–18·4%), and T. trichiura 7·7% (range=0–18·6%). Helminth-related morbidities were not associated with infection. Our study demonstrates that schistosomiasis and STH are important health priorities among schools in informal settlements of Kisumu City, and highlights the need for routine deworming in similar settings.

Lambs were infected at daily intervals from 3 weeks of age with increasing doses of infective larvae of Trichostrongylus colubriformis, according to an exponential infection schedule designed to simulate the pattern of infection experienced by grazing lambs during the spring and early summer in Britain. The faecal egg counts and the worm burdens in the lambs followed a similar exponential trend for 12 weeks after initial infection.

The T. colubriformis burdens in these lambs appeared to be cumulative during the first 12 weeks, after which the animals developed a resistance to further reinfection, which was very strong by 16 weeks after initial infection. However, even lambs which were refractory to new infection continued to harbour large numbers of adult worms, which caused clinical disease in most of the lambs.

There was no evidence to suggest that either a turnover of the worm burden, inhibition of larval development or the sudden loss of established parasites played any major role in the regulation of the burdens of T. colubriformis in these young lambs.

The removal of the established burden 16 weeks after initial infection in one lamb appeared to result in a partial loss of resistance to a challenge infection given 10 days later.

Susceptibility to oxidative stress is a well-established feature of the malarial parasite. Pharmacologists have taken advantage of this property to design highly effective pro-oxidant antimalarial drugs. Less well appreciated is the fact that nutritional manipulation of host oxidative stress status by dietary means can have a profound effect on the growth of the parasite. In particular, rapid induction of vitamin E deficiency in mice by feeding highly unsaturated fatty acids (fish oil) strongly suppresses plasmodial growth. Likewise, the status of other antioxidant nutrients (e.g., riboflavin or vitamin C) may also influence the course of malarial infection under certain conditions. A combined nutritional pharmacology approach may offer some promise in controlling malaria.

Nitazoxanide (NTZ) and its deacetylated metabolite tizoxanide (TIZ) exhibit considerable in vitro activity against Besnoitia besnoiti tachyzoites grown in Vero cells. Real-time-PCR was used to assess B. besnoiti tachyzoite adhesion, invasion, and intracellular proliferation in vitro. A number of NTZ-derivatives, including Rm4822 and Rm4803, were generated, in which the thiazole-ring-associated nitro-group was replaced by a bromo-moiety. We here show that replacement of the nitro-group on the thiazole ring with a bromo (as it occurs in Rm4822) does not impair the efficacy of the drug, but methylation of the salicylate ring at the ortho-position in a bromo-derivative (Rm4803) results in complete abrogation of the antiparasitic activity. Treatment of extracellular B. besnoiti tachyzoites with NTZ has an inhibitory effect on host cell invasion, while treatments with TIZ, Rm4822 do not. TEM demonstrates that the effects of Rm4822 treatment upon the parasites are similar to the damage induced by NTZ. This includes increased vacuolization of the parasite cytoplasm, and loss of the structural integrity of the parasitophorous vacuole and its membrane. Thus, Rm4822, due to the absence of a potentially mutagenic nitro-group, may represent an important potential addition to the anti-parasitic arsenal for food animal production, especially in cattle.

The usage of alternative synonymous codons in the completely sequenced, extremely A+T-rich parasite Plasmodium falciparum was studied. Confirming previous studies obtained with less than 3% of the total genes recently described, we found that A- and U-ending triplets predominate but translational selection increases the frequency of a subset of codons in highly expressed genes. However, some new results come from the analysis of the complete sequence. First, there is more variation in GC3 than previously described; second, the effect of natural selection acting at the level of translation has been analysed with real expression data at 4 different stages and third, we found that highly expressed proteins increment the frequency of energetically less expensive amino acids. The implications of these results are discussed.

The mechanisms involved in the pathogenesis of chagasic megacolon are not completely characterized. Although autoimmunity may play a role in the pathogenesis of Chagas' disease, recent studies suggest a positive association of tissue parasitism, inflammation, and severity of lesions. The aim of this study was to evaluate the role of inflammatory cells and the occurrence of fibrosis in the colon of chagasic patients with and without megacolon. Samples from 26 patients were randomly selected and paraffin-embedded tissue blocks were sectioned and evaluated by histology and immunohistochemistry to analyse the occurrence and relation among eosinophils, mast cells, macrophages and fibrosis. Section analyses showed that the presence of eosinophils and mast cells in the analysed inflammatory cells has a direct correlation with fibrosis density in the chagasic megacolon. These data suggest that the megacolon's pathogenesis is based on a continuous process of cell damage. Our data propose that eosinophils, mast cells and macrophages may have a direct connection with the occurrence of fibrosis in the colon of chagasic patients. We believe that potential therapeutic agents against these cells could avoid the fibrosis process and contribute to prevent the development of chagasic megacolon.

Acetylcholine is the major excitatory neurotransmitter controlling motor activities in nematodes, and the enzyme which hydrolyses and inactivates acetylcholine, acetylcholinesterase, is thus essential for regulation of cholinergic transmission. Different forms of acetylcholinesterase are encoded by multiple genes in nematodes, and analysis of the pattern of expression of these genes in Caenorhabditis elegans suggests that they perform non-redundant functions. In addition, many parasitic species which colonise host mucosal surfaces secrete hydrophilic variants of acetylcholinesterase, although the function of these enzymes is still unclear. Acetylcholinesterases have a history as targets for therapeutic agents against helminth parasites, but anti-cholinesterases have been used much more extensively as pesticides, for example to control crop damage and ectoparasitic infestation of livestock. The toxicity associated with these compounds (generally organophosphates and carbamates) has led to legislation to withdraw them from the market or restrict their use in many countries. Nevertheless, acetylcholinesterases provide a good example of a neuromuscular target enzyme in helminth parasites, and it may yet be possible to develop more selective inhibitors. In this article, we describe what is known about the structure and function of vertebrate cholinesterases, illustrate the molecular diversity and tissue distribution of these enzymes in C. elegans, and discuss to what extent this may represent a paradigm for nematodes in general.

Tabanids are haematophagous flies feeding on livestock and wildlife. In the absence of information on the relationship of tabanid flies and protozoan parasites in South Africa and Zambia, the current study was aimed at characterizing tabanid flies collected in these two countries as well as detecting protozoan parasites they are harbouring. A total of 527 tabanid flies were collected whereby 70·2% were from South Africa and 29·8% were from Zambia. Morphological analysis revealed a total of five different genera collected from the sampled areas namely: Ancala, Atylotus, Haematopota, Philoliche and Tabanus. DNA extracted from South African Tabanus par and Tabanus taeniola tested positive for the presence of Trypanosoma congolense (Savannah) and Trypanosoma theileri whilst one member from T. par was positive for Trypanosoma brucei species. DNA extracted from Zambian tabanid flies tested positive for the presence of Besnoitia species at 1·27% (2/157), Babesia bigemina 5·73% (9/157), Theileria parva 30·11% (30/157) and 9·82% (14/157) for Trypanosoma evansi. This study is the first to report on relationship of Babesia and Theileria parasites with tabanid flies. Further investigations are required to determine the role of tabanids in transmission of the detected protozoan parasites in livestock and wildlife in South Africa and Zambia.

We studied patterns of variation in parasite communities of 2 closely related species of Mastomys rodents. These 2 species live in sympatry in South-eastern Senegal, but differ drastically in their habitat choice. We asked (a) whether the host species have the same parasites; (b) whether there is any observable pattern relative to the host species/habitat type in the structure of parasite communities; (c) whether the variability in parasite community for each host species is related to habitat characteristics. We analysed 220 and 264 individuals of each host species, sampled respectively in 10 and 11 trap sites. Twenty parasite taxa were recorded, and the majority were nematodes. Between-host species comparisons showed that helminth communities were slightly more diversified in M. natalensis. Many parasite species were found in both Mastomys. However, various helminth taxa varied in frequency and abundance between host species. Within each host species, helminth diversity, prevalence and/or abundance of some parasites were correlated with habitat or host population factors that may influence parasite life-cycles, such as village structure, or the presence/absence of a pool. Our results suggest that habitat characteristics have a strong impact on helminth community structure.

In the present study we detected Schellackia haemoparasites infecting the blood cells of Lacerta schreiberi and Podarcis hispanica, two species of lacertid lizards from central Spain. The parasite morphometry, the presence of a refractile body, the type of infected blood cells, the kind of host species, and the lack of oocysts in the fecal samples clearly indicated these blood parasites belong to the genus Schellackia. Until now, the species of this genus have never been genetically characterized and its taxonomic position under the Lankesterellidae family is based on the lack of the exogenous oocyst stage. However, the phylogenetic analysis performed on the basis of the 18S rRNA gene sequence revealed that species of the genus Schellackia are clustered with Eimeria species isolated from a snake and an amphibian species but not with Lankesterella species. The phylogenetic analysis rejects that both genera share a recent common ancestor. Based on these results we suggest a revision of the taxonomic status of the family Lankesterellidae.

1. The spores of Eimeria (not the oocysts) are the real infective agents and, as in other Sporozoa, they readily set free their sporozoites in the duodenum or in vitro under suitable conditions.

2. The protective oocyst has double walls: the ectocyst is thick and impervious to fluids but easily fractures by mechanical means, such as drying. Even though cracked, it may continue to protect the thin endocyst and spores until they reach the specially muscular regions of the alimentary canal. Free spores are probably also sometimes ingested.

3. The ectocyst is a continuous envelope without a persistent micropyle. It resembles keratin in many ways.

Vaccination with whole male worm extract (AMA), stichoseome extract (SA) and a short-term inculbation fluid (EXA) of adult Trichuris muris induced a high degree of protective immunity in mice as assessed by reduction in larval worm burden.

Immunodiffusion of AMA against rabbit anti-AMA serum revealed five precipitin lines, one of which was common to both EXA and SA with which only single lines were obtained.

Physico-chemical tests suggested that the antigen is a protein and that carbohydrate, lipid and DNA do not contribute to its antigenicity. Partial fractionation was obtained with ammonium sulphate. All preparations with immunogenic activity showed the common line of EXA on immunodiffusion.

It is concluded that one of the protective immunogens is a protein which can be associated with the precipitin line and originates in the stichosome.

Nine clones were derived from a drug-resistant Trypanosoma congolense stock (IL 2856) and characterized in mice for their sensitivity to isometamidium chloride and diminazene aceturate. All clones were derived from the stock without drug selection and expressed high levels of resistance to isometamidium chloride (50% curative dose [CD50] values ranging from 1·5 to 5·1 mg/kg) and intermediate to high levels of resistance to diminazene aceturate (CD50 values ranging from 5·1 to 21·0 mg/kg). By contrast, the isometamidium chloride and diminazene aceturate CD50 values for a drug-sensitive clone, T. congolense IL 1180, were 0·018 mg/kg and 2·3 mg/kg, respectively. For both drugs, there appeared to be significantly different levels in expression of drug resistance amongst the 9 clones derived from IL 2856. Isoenzyme analysis of 7 enzymes showed that all 9 clones expressed the same electrophoretic variants. Thus, all 9 clones were identical for these phenotypic markers. The clone which expressed the highest level of resistance to isometamidium in mice (IL 3270) was transmitted to Boran cattle via the bite of infected Glossina morsitans centralis. IL 3270 produced an infection rate in tsetse of 5·0%. The resulting infections in cattle were shown to be resistant to intramuscular treatment with 2·0 mg/kg isometamidium chloride and 14·0 mg/kg diminazene aceturate. This contrasts with doses of 0·25 mg/kg isometamidium chloride or 3·5 mg/kg diminazene aceturate which are deemed sufficient to cure fully sensitive infections. Finally, 9 clones (subclones) were derived from IL 3270 and characterized in mice for their sensitivity to isometamidium chloride. Seven of the subclones expressed a significantly lower level of resistance to isometamidium than the parental clone and amongst the subclones there was significant variation in resistance. Thus, expression of a high level of resistance to isometamidium appears to be unstable in the rodent host and at least a component of the genetic determinant(s) for this drug-resistant phenotype is (are) likely to be unstable.