We review the theoretical framework for exploring the impact of individual and spatial heterogeneities in patterns of exposure and contamination and on the basic reproduction number, R0, for human schistosomes. Analysis of water contact data for 5 communities in Zimbabwe and Mali suggests that the impact is substantial, increasing R0 by factors of up to 6·5, mostly due to highly overdispersed distributions of contact rates among individuals. Several practical conclusions emerge: concentration of contacts at a single site should be avoided; the impact of control targeted at certain sites cannot be predicted without knowledge of how individuals' contacts are distributed among sites; control programmes targeted at individuals or sites contributing most to transmission can be very efficient but, conversely, will be ineffective if any of these individuals or sites are missed.

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.

The burden and duration of asymptomatic malaria infections were measured in residents of the malaria endemic village of Gonoa, Madang Province, Papua New Guinea. Plasmodium falciparum, P. vivax and P. malariae infections in people aged 4 years to adulthood were compared. Frequent sampling at 3-day intervals for up to 61 days allowed assessment of individual episodes of infection. Statistical assessment of P. falciparum detection revealed a periodicity consistent with synchronous replication of this species over periods up to 27 days. The duration of P. falciparum episodes was longer across all age groups than that of P. vivax and P. malariae. A trend for decreasing duration with age was also noted in data from each species. This was most prominent in P. falciparum infections: median duration in 4-year-olds was > 48 days compared with a median between 9 and 15 days in older children and adults. The results are consistent with the slow acquisition of immunity to antigenically diverse Plasmodium populations and suggest a faster rate of acquisition to P. vivax and P. malariae than to P. falciparum.

Many pollinators are currently suffering from declines, diminishing their gene pool and increasing their vulnerability to parasites. Recently, an increasing diversity of parasites has been recorded in bumblebees, yet for many, knowledge of their virulence and hence the risk their presence poses, is lacking. The deformed wing virus (DWV), known to be ubiquitous in honey bees, has now been detected in bumblebees. In addition, the neogregarine Apicystis bombi has been discovered to be more prevalent than previously thought. Here, we assess for the first time the lethal and sublethal effects of these parasites during single and mixed infections of worker bumblebees (Bombus terrestris). Fifteen days after experimental exposure, 22% of bees exposed to A. bombi, 50% of bees exposed to DWV and 86% of bees exposed to both parasites had died. Bumblebees that had ingested A. bombi had increased sucrose sensitivity (SS) and a lower lipid:body size ratio than control bees. While dual infected bumblebees showed no increase in SS. Overall, we find that A. bombi exhibits both lethal and sublethal effects. DWV causes lethal effect and may reduce the sub lethal effects imposed by A. bombi. The results show that both parasites have significant, negative effects on bumblebee health, making them potentially of conservation concern.

Following the collapse of the Soviet Union in 1991, there was an increase in the number of cases of human echinococcosis recorded throughout central Asia. Between 1991 and 2001 incidence rates of cystic echinococcosis (CE) increased by 4 fold or more. There also appeared to be increases in prevalence of CE in livestock and prevalences of Echinococcus granulosus reported in dogs. The increase in human echinococcosis was associated with changes in livestock husbandry, decline in veterinary public health services, increases in dog populations and increased poverty, all of which served to promote transmission of E. granulosus. A few years after reports of increased transmission of E. granulosus, the first reports of E. multilocularis infection in dogs were recorded. Further studies indicated that in both Kazakhstan and Kyrgyzstan prevalences of up to 18% were present. Recently there has been a dramatic increase in the number of cases of human alveolar echinococcosis recorded in Kyrgyzstan with over 60 cases reported in 2011.

It has been shown by references to the writings of other workers, and some personal observations in the early part of this paper, that destruction of certain species of bacteria takes place in the alimentary canal of flies—especially in the early developmental stages of the insect—and ticks. In addition, emphasis has been laid on the remarkable sterility of the gut-contents and faeces of some parasitic and semi-parasitic blood-feeders.

That these conditions result from the operation of several factors is possible, and the bactericidal principle described in this paper may reasonably be accepted as one of these factors.

The investigation has been limited to the adult stage of the insects Stomoxys calcitrans, Musca domestica, Anopheles bifurcatus, Aëdes cinereus, Cimex lectularius and Rhodnius prolixus, and the arachnids Argas persicus and Ornithodorus moubata. In all of these a bactericidal principle has been demonstrated in the gut-contents, and, in the case of the insects, also in the faeces.

The bacteria most affected were the common sporing aerobes and the staphylococci, but B. anthracis was destroyed by gut-contents of Stomoxys, and M. catarrhalis and S. haemolyticus by those of Argas.

It may be inferred, from the results of the bactericidal and absorption tests, that the active principle in any one species of arthropod is single, but that different groups of bacteria exhibit differences in susceptibility to its action; some, such as the subtilis group, being very susceptible, are destroyed by concentrations too weak to affect other species, while others, as the staphylococci, require relatively high concentrations for their destruction, and many bacteria appear to be wholly resistant.

The bactericidal principle occurring in different arthropod species showed some differences in the groups of bacteria susceptible to its action, and in the degree of susceptibility of particular groups. Thus, the widest range of action was shown by gut-contents of Argas and Stomoxys, and the least by those of the bugs. The sporing bacilli seemed to be affected more by material from Stomoxys than by that from Argas, but, on the other hand, the staphylococci were apparently more susceptible to the action of Argas material than to that of Stomoxys. The gut-contents of the ticks had a feebly destructive action on B. pestis, while those of some insects actually enhanced its growth. This last observation is of some interest in connection with the case of the plague flea.

With regard to the properties of the active principle, bactericidal action is greater and more rapid at 37° C. than at room temperature. This action is not accompanied by any visible bacteriolysis. The bactericidal principle retains its activity unimpaired for at least six months when kept in the dry state. It is very thermostable, resisting temperatures as high as 120° C. It is not destroyed by tryptic digestion. It is precipitated from solution with proteins by alcohol or acetone, but is not itself affected by these reagents. It is not soluble in the common fat-solvents, ether, chloroform, alcohol or acetone. By allowing it to act upon repeated small doses of bacteria, it rapidly becomes exhausted, and it can be inactivated, possibly through adsorption, by large doses of killed bacteria; even those species which are not destroyed by it. It may also be adsorbed in small amount by bibulous paper.

It exhibits none of the properties of the bacteriophage, and it differs from lysozyme.

In the bactericidal action it is clear that no vital process is involved, in fact it very closely resembles the action of an antiseptic.

Regarding the source of the active principle, there is no doubt that it is formed in the stomach, but whether as a secretion of the gastric cells or as a result of the processes of digestion is not clear. (Nuttall (1908) showed that the destruction of Spirochaeta duttoni in the gut of the bed bug was definitely related to digestion.) An examination of material collected from ticks, kept at 25° C. at various stages of digestion seemed to point to an increase in bactericidal activity during the first two weeks after feeding, and a decline after the sixth or eighth week, but the activity of material from individual ticks, taken at similar intervals after feeding, often varied greatly. Control tests with the blood of the hosts, on which the arthropods were fed, showed that this was not the source of the active principle.

Mice can be partially protected against Schistosoma japonicum by prior exposure to ultraviolet (UV)-attenuated infections which fail to survive to the adult stage and produce no overt pathology in the host. Optimum resistance was induced by parasites exposed to 40 seconds of UV, significantly lower levels of resistance being stimulated by both shorter and longer exposures. No consistent relationship between the degree of resistance induced and the number of irradiated cercariae given could be demonstrated and equivocal results were obtained when comparing the efficacy of single and multiple vaccinations. Vaccinations with UV-attenuated cercariae given intraperitoneally (i.p.) were as efficacious as those given percutaneously but mice were as or more resistant to challenges given by the i.p. route, the possible reasons are discussed. There was no observed delay in the migration of the challenge, vaccinated mice being as resistant when perfused 6 or 3·5 weeks after challenge. Vaccination was species specific since mice exposed to either UV-attenuated S. japonicum cercariae or gamma-attenuated S. mansoni cercariae were resistant to homologous but not heterologous challenge.

The results of experimental infections of Trichuris muris in wild field mice (Apodemus sylvaticus) and laboratory-bred wild house mice (Mus musculus) showed that the parasite elicited an immune response similar to that previously described in strains of laboratory mice. Experiments in laboratory mice showed that the parasite was able to become sexually mature only when small single infections or repeated low-level infections were given. A survey of a population of 43 wild house mice naturally infected with T. muris showed that the pattern of small worm burdens in the majority of mice was consistent with a situation of repeated low-level infection, except in the case of six female mice which harboured larger mature worm burdens. It is suggested that in these mice pregnancy and/or lactation may have suppressed the immune response, allowing the accumulation of a worm burden in excess of the threshold for worm expulsion.

Two groups of sheep born and raised worm-free were dosed with 3000 infective Haemonchus contortus larvae. Group 1 were each given a single dose of infective larvae on day 0 while the group 3 animals were dosed with 100 infective larvae per day for 30 consecutive days. The results of faecal worm-egg counts performed on alternative days, and of differential worm counts carried out on animals slaughtered at crucial times during the course of infection, showed that retardation of development at the 4th larval stage occurred in the group 3 infections and was the major effect of control of the parasitic burden. There was no evidence that egg production per female was affected.

Three animals of the nineteen infected in group 1 died during the course of the experiment owing to the pathogenic effects of the worm burden. Deaths did not occur among the animals of group 3. This finding was correlated with the observation that haematocrit levels were consistently lower among the animals of group 1 than among those of group 3.

The results are interpreted in terms of the theory of threshold behaviour of the immunological response to parasitic infection.

We are greatly indebted to Dr D. F. Stewart for his interest and constructive criticism during the course of these studies, and to Mr E. Teleki and Misses Helen Giller and Lindy Stothart for their valuable technical assistance.

Neospora caninum is a protozoan parasite that causes bovine abortion. The epidemiology of N.caninum is poorly understood and little is known about the genetic diversity of the parasite, or whether individual isolates differ in virulence. Such diversity may, among other factors, underlie the range of pathologies seen in cattle. In this study we analysed biological and genetic variation in 6 isolates of N. caninum originating from canine and bovine hosts by measurement of growth rate in vitro, Western blotting and random amplification of polymorphic DNA (RAPD). This comparative analysis of intra-species diversity demonstrated that heterogeneity exists within the species. The relative growth rate in vitro, as assessed by 3[H]uracil uptake, showed significant variation between isolates. However, no significant differences were detected between the antigenic profiles of each isolate by Western blotting. RAPD-PCR was performed on DNA from the 6 Neospora isolates; 3 strains of Toxoplasma gondii, Sarcocystis sp. and Cryptosporidium parvum were also analysed. Twenty-six RAPD primers gave rise to 434 markers of which 222 were conserved between all the Neospora isolates and distinguished them from the other Apicomplexa. An additional 54 markers were unique for Neospora but were polymorphic within the species and able to differentiate between the individual isolates. The RAPD data were subjected to pair-wise similarity and cluster analysis and showed that the Neospora isolates clustered together as a group, with T. gondii as their nearest neighbour. N. caninum isolates showed no clustering with respect either to host or geographical origin. The genetic similarity between Neospora isolates from cattle and dogs suggests that these hosts may be epidemiologically related, although further analysis of bovine and canine field samples are required. The genetic and biological diversity observed in this study may have important implications for our understanding of the pathology and epidemiology of neosporosis.

The transforming growth factor-β (TGF-β) gene family regulates critical processes in animal development, and plays a crucial role in regulating the mammalian immune response. We aimed to identify TGF-β homologues from 2 laboratory model nematodes (Heligmosomoides polygyrus and Nippostrongylus brasiliensis) and 2 major parasites of ruminant livestock (Haemonchus contortus and Teladorsagia circumcincta). Parasite cDNA was used as a template for gene-specific PCR and RACE. Homologues of the TGH-2 subfamily were isolated, and found to differ in length (301, 152, 349 and 305 amino acids respectively), with variably truncated N-terminal pre-proteins. All contained conserved C-terminal active domains (>85% identical over 115 amino acids) containing 9 cysteine residues, as in C. elegans DAF-7, Brugia malayi TGH-2 and mammalian TGF-β. Surprisingly, only the H. contortus homologue retained a conventional signal sequence, absent from shorter proteins of other species. RT-PCR assays of transcription showed that in H. contortus and N. brasiliensis expression was maximal in the infective larval stage, and very low in adult worms. In contrast, in H. polygyrus and T. circumcincta, tgh-2 transcription is higher in adults than infective larvae. The molecular evolution of this gene family in parasitic nematodes has diversified the pre-protein and life-cycle expression patterns of TGF-β homologues while conserving the structure of the active domain.

Trypanosoma rangeli can infect humans and the same domestic and wild animals and triatomine vectors infected by T. cruzi in Central and South America. This overlapping distribution complicates the epidemiology of Chagas disease because of the cross-reactivity between T. rangeli and T. cruzi antigens. We have studied T. rangeli strains isolated from different geographical regions using the mini-exon gene as a genetic marker. Two pairs of oligonucleotides directed to this gene were designed in order to detect specifically T. rangeli DNA by PCR assays. This assay was highly sensitive, able to amplify the target sequence using the equivalent DNA content of a single parasite as template, and demonstrated no cross-reactivity with T. cruzi DNA. T. rangeli SC-58 strain, isolated in southern Brazil, showed a distinct electrophoretic pattern from the other T. rangeli strains tested. Low stringency single specific primer-PCR (LSSP) assays were able to detect sequence polymorphisms at the mini-exon gene among T. rangeli strains. Sequence comparisons of this gene revealed that the SC-58 strain was genetically distinct from strains isolated in Central America and northern South America. In addition to insertion/deletion events, the presence of microsatellite repeats in the non-transcribed region of the gene contribute to the intra-species variability.

There have been described in detail adaptations of dilution egg counting and direct centrifugal flotation for use on sheep faeces for the ova of Haemonchus contortus, together with certain data on their comparative efficiency. It is suggested that extension of the methods as given, or with appropriate modifications, would permit studies from a variety of angles on the helminthic infestations of the gastro-intestinal tract and its diverticula among domestic animals.

Total lipid from adult male and adult paired Schistosoma mansoni has been fractionated by thin-layer chromatography. Phospholipid, free sterol, and triglyceride were major components of both mixtures. A remarkably small amount of sterol ester was found in adult flukes considering the prevalence of this compound in mouse blood. Free fatty acids were found to be a minor component. Cholesterol was the major free sterol present.

The technical assistance of Mrs Katye Ross Summerlin is gratefully acknowledged.

Thin sections of Trypanosoma cruzi in tissue cultures and from blood agar medium have been examined with the electron microscope.

In the leishmania and crithidia forms of the parasite a neat separation of kinetonucleus and blepharoplast has been obtained and these structures are described. The blepharoplast is the row of basal corpuscles which give origin to the axial fibres of the flagellum.

Division of the parasite seems to begin in the basal corpuscles; it is followed by binary division of the kinetonucleus and only later by fission of nucleus and cytoplasm.

During the development of the leishmania into the crithidia form, the kinetonucleus and basal bodies are dislocated towards the posterior half of the trypanosome. The migration of these two structures and the consequent lengthening of the flagellum causes the surface membrane of the flagellum to be pushed inwards to form a deep invagination, so that the flagellum comes to the surface of the body and is separated from it along its whole length by its own sheath.

In the adult form of the parasite a great vacuole is formed near the posterior tip of the trypanosome. It has no definite structure in the electron microscope, is transparent and seems to be filled with a liquid content. The basal corpuscles are situated in this region near the wall of the vacuole. The typical structure of the kinetonucleus could not be identified with certainty in these forms; it seems to be altered by the formation of the vacuole.

The vacuole is crossed by a fibre system which comes from the body of the trypanosome and is often dislodged by the size of the vacuole. This fibre system ends in a sharply pointed process of varying length.

The cost of reproduction of figures was defrayed by the Instituto de Biofísica da Universidade do Brasil.

Models have predicted that directly transmitted macroparasites may influence the abundance of forage plants in herbivore grazing systems by reducing the food intake of their host. Evidence of parasite-induced alterations in host food intake is, however, limited mainly to sheep, cattle and laboratory rodents. We estimated the effect of naturally acquired parasite infections on the appetite of reindeer. Food intake was significantly lower in infected reindeer compared to animals in which the parasites had been experimentally removed. Among the infected animals there was a significant negative relationship between intensity of the directly transmitted macroparasites (i. e. gastrointestinal nematodes) and mean food intake, indicating that the lower food intake was caused by these parasites. The time-specific onset of depression in food intake is also consistent with seasonality in the pathogenic effect from gastrointestinal nematodes. This shows that parasite-induced changes in herbivore food intake is not restricted to agricultural systems, and implies that parasites may have impact on the dynamics of a wide range of herbivore plant communities.

Haemosporidian parasites are common in birds in which they act as an important selective pressure. While most studies so far have focused on the effect of their prevalence on host life-history traits, no study has measured the effect of parasitaemia. We developed molecular methods to detect, identify and quantify haemosporidia in 2 natural populations of the Blackbird Turdus merula. Three different parasite genotypes were found – 1 Haemoproteus and 2 Plasmodium. A PCR-RFLP screening revealed that only approximately 3% of blackbirds were free of parasites, compared to the 34% of uninfected birds estimated by blood smear screening. A quantitative PCR (q-PCR) assay revealed a weaker parasitaemia in microscopically undetected parasites compared to microscopically detected ones. Large parasitaemia differences were found between parasite species, suggesting either differing parasite life-histories or host resistance. Parasitaemias were also weaker in male hosts, and in urban habitats, suggesting that both host factors (e.g. immunity) and habitat characteristics (e.g. vector availability) may modulate parasite density. Interestingly, these differences in parasitaemia were comparable to differences in parasite prevalence estimated by smear screening. This suggests that previous results obtained by smear screening should be reinterpreted in terms of parasitaemia instead of parasite prevalence.

We characterized 14 trypanosome isolates from sylvatic mammals (9 from primates, 1 from sloth, 2 from anteaters and 2 from opossum) plus 2 human isolates of Brazilian Amazon. These isolates were proven to be Trypanosoma rangeli by detection of metacyclic trypomastigotes in the salivary glands of triatomines and by a specific PCR assay. Polymorphism determined by randomly amplified polymorphic DNA (RAPD) revealed that most (12) of the Brazilian T. rangeli isolates from the Amazon differed from those of other geographical regions, thus constituting a new group of T. rangeli. Four Brazilian isolates clustered together with a previously described group (A) that was described as being composed of being isolates from Colombia and Venezuela. Isolates from Panama and El Salvador form another group. The isolate from Southern Brazil did not cluster to any of the above-mentioned groups. This is the first study that assesses the genetic relationship of a large number of isolates from wild mammals, especially from non-human primates. A randomly-amplified DNA fragment (Tra625) exclusive to T. rangeli was used to develop a PCR assay able to detect all T. rangeli groups.

Muscle tissue from 37 red deer from Norway was examined for sarcocysts. Sarcocysts from 2 reindeer were obtained for comparative studies. Cysts were excised and morphologically classified by light microscopy, scanning electron microscopy, and DNA sequence analysis. Five Sarcocystis species, Sarcocystis hjorti n. sp., Sarcocystis hardangeri, Sarcocystis ovalis, Sarcocystis rangiferi, and Sarcocystis tarandi, were found. All 5 species have previously been identified from either reindeer or moose by their sarcocyst morphology and/or ssu rRNA gene sequence. S. hjorti was the most prevalent species. Multiple variants of the ssu rRNA gene and the first internal transcribed spacer were found in S. rangiferi and S. tarandi from both red deer and reindeer. Phylogenetic analyses indicated that S. tarandi occurs in both red deer and reindeer, but it could not be clearly demonstrated whether the sequence variation within S. rangiferi between hosts was due to different paralogues or/and different species. DNA sequencing was necessary for definitive species identification, since the hair-like protrusions on the cysts of S. hjorti were not always recognizable by light microscopy and since different cervids harbour Sarcocystis species with highly similar cyst morphology of which at least some are not intermediate host specific.

Earlier schemes of classification of the family Heterophyidae have been based in large part on such features as shape of body, presence of oral spines, number and position of testes, and distribution of vitellaria (Witenberg, 1929; Ciurea, 1933; Mueller & Van Cleave, 1932). Price (1940a) was the first to make extensive use of features of the ventrogenital complex (ventral sucker, gonotyl, genital pore, terminal male duct) and excretory bladder, and produced the first reasonable classification of both the family Heterophyidae and the superfamily Opisthorchioidea. In despite of the obvious significance of the rationale of Price's approach, later authors (Morozov, 1952, 1955; Yamaguti, 1958) have largely ignored the ventrogenital complex and recently discovered life-history data, and have used much the same sorts of features as earlier authors.