Isoenzyme-based studies have identified 3 taxa/species/‘phylogenetic complexes’ as agents of visceral leishmaniasis in Sudan: L. donovani, L. infantum and “L. archibaldi”. However, these observations remain controversial. A new chitinase gene phylogeny was constructed in which stocks of all 3 putative species isolated in Sudan formed a monophyletic clade. In order to construct a more robust classification of the L. donovani complex, a panel of 16 microsatellite markers was used to describe 39 stocks of these 3 species. All “L. donovani complex” stocks from Sudan were again found to form a single monophyletic clade. L. donovani ss stocks from India and Kenya were found to form 2 region-specific clades. The partial sequence of the glutamate oxaloacetate transaminase (GOT) gene of 17 L. donovani complex stocks was obtained. A single nucleotide polymorphism in the GOT gene appeared to underlie the isoenzyme classification. It was concluded that isoenzyme-based identification is unsafe for stocks isolated in L. donovani endemic areas and identified as L. infantum. It was also concluded that the name L. archibaldi is invalid and that only a single visceralizing species, Leishmania donovani, is found in East Africa.

Human gastrointestinal bacteria often share their environment with parasitic worms, allowing physical and physiological interaction between the two groups. Such associations have the potential to affect host health as well as the bacterial and helminth populations. Although still in its early stages, research on the interaction between the microbiome and parasitic helminths in humans offers the potential to improve health by manipulating the microbiome. Previously, supplementation with various nutritional compounds has been found to increase the abundance of potentially beneficial gut commensal bacteria. Thus, nutritional microbiome manipulation to produce an environment which may decrease malnutrition associated with helminth infection and/or aid host recovery from disease is conceivable. This review discusses the influence of the gut microbiota and helminths on host nutrition and immunity and the subsequent effects on the human host's overall health. It also discusses changes occurring in the microbiota upon helminth infections and the underlying mechanisms leading to these changes. There are still significant knowledge gaps which need to be filled before meaningful progress can be made in translating knowledge from studying the human gut microbiome into therapeutic strategies. Ultimately this review aims to discuss our current knowledge as well as highlight areas requiring further investigation.

Sexual size dimorphism (SSD) in mammals reveals the extent of sexual selection, which may in turn explain why males are often more infected by parasites than females and that parasites may contribute to the association between SSD and male-biased mortality. Here, we investigated the relationship between SSD in small mammals of Central Europe and the differences in sex infection by fleas. A comparative analysis was conducted for 10 species of rodents and insectivores. We found that males harbour higher flea species richness than females and that the abundance of fleas is higher in males than in females. This difference is not related to male-biased density. However, contrary to our hypothesis, we found that an increase in SSD is not related to an increase in male infection by fleas compared with female infection. We discuss our results in term of sex-differences in immunocompetence and/or sex-differences in behaviour.

Gastrointestinal parasite infections are widespread among wild ungulates. Because many of these parasites infect multiple host species, inter-specific interactions among hosts potentially play an important role in parasite transmission dynamics in ungulate communities. In this study, the effects of inter-specific contact on parasitism rates in 11 sympatric African bovids was examined using habitat overlap among species as a measure of cross-species contact rates. Across individual hosts, strongyle nematode abundance increased with increasing numbers of bovid species occupying a habitat. Furthermore, comparative analyses show a positive association between strongyle prevalence and level of habitat overlap across taxa. These findings suggest that among sympatric bovids, contact between species contributes significantly to the transmission of generalist nematode parasites. For a more host-specific parasite group, coccidia, parasite abundance and individual probability of infection declined in hosts living in bovid rich habitats. This pattern may reflect enhanced inter-specific competition among parasites in these areas. Finally, similar to strongyle abundance, individual parasite richness also increased among hosts occupying habitats with higher numbers of bovid species. No association between habitat overlap and parasite richness was detected at higher taxonomic scales, however, which suggests that contact between host species may not contribute to parasite colonization of new host taxa.

Much evolutionary theory assumes that parasite virulence (i.e. parasite-induced host mortality) is determined by within-host parasite reproduction and by the specific parasite genotypes causing infection. However, many other factors could influence the level of virulence experienced by hosts. We studied the protozoan parasite Ophryocystis elektroscirrha in its host, the monarch butterfly, Danaus plexippus. We exposed monarch larvae to wild-isolated parasites and assessed the effects of within-host replication and parasite genotype on host fitness measures, including pre-adult development time and adult weight and longevity. Per capita replication rates of parasites were high, and infection resulted in high parasite loads. Of all host fitness traits, adult longevity showed the clearest relationship with infection status, and decreased continuously with increasing parasite loads. Parasite genotypes differed in their virulence, and these differences were maintained across ecologically relevant variables, including inoculation dose, host sex and host age at infection. Thus, virulence appears to be a robust genetic parasite trait in this system. Although parasite loads and genotypes had strong effects on virulence, inoculation dose, host sex and age at infection were also important. These results have implications for virulence evolution and emphasize the need for a detailed understanding of specific host-parasite systems for addressing theory.

Inherited bacteria that parasitically distort the pattern of sex allocation of their host, biasing allocation towards female progeny, are found in many arthropods. One such manipulation is male-killing, where male progeny of infected females die during embryogenesis. We here provide evidence for a male-killing bacterium in the coccinellid beetle, Adonia variegata. We then address 3 questions. First, is this male-killing bacterium one that is found in other hosts, or does it represent a new transition to male-killing within the eubacteria? Using the sequence of the 16S rDNA of the bacterium, we found that the male-killing bacterium is a member of the Flavobacteria–Bacteroides group, most closely related to the male-killing bacterium in another ladybird beetle, Coleomegilla maculata. Secondly, is there any evidence that this bacterium affects female host physiology? In a paired test under nutritional stress, we found no evidence for a physiological benefit to infection, and weak evidence of a physiological cost, in terms of reduced fecundity. Thirdly, is there any evidence of host involvement in the transmission of the bacterium to the germ line? We found no evidence of host involvement. Rather, bacteria migrated to the ovariole independently of host cells. We conclude that the bacterium is a parasite, and discuss how 2 different species of ladybird come to be infected with 1 lineage of bacterium, and why case studies of male-killing bacteria have generally found little evidence of any symbiont contribution to host physiological functioning.

In this chapter we summarize the available data on a novel class of ligand-gated anion channels that are gated by the neurotransmitter glutamate. Glutamate is classically thought to be a stimulatory neurotransmitter, however, studies in invertebrates have proven that glutamate also functions as an inhibitory ligand. The bulk of studies conducted in vivo have been on insects and crustaceans, where glutamate was first postulated to act on H-receptors resulting in a hyperpolarizing response to glutamate. Recently, glutamate-gated chloride channels have been cloned from several nematodes and Drosophila. The pharmacology and electrophysiological properties of these channels have been studied by expression in Xenopus oocytes. Studies on the cloned channels demonstrate that the invertebrate glutamate-gated chloride channels are the H-receptors and represent important targets for the antiparasitic avermectins.

Monophyly of all 11 valid Eimeria species from rabbits (Oryctolagus cuniculus Linnaeus, 1758) was revealed based on nuclear 18S rDNA sequence data. This finding implies that these species, which vary considerably in terms of their morphology and biology, diversified on a single host or several closely related species. Phylogenetic analysis divided rabbit Eimeria species into 2 sister lineages, corresponding to the presence/absence of the oocyst residuum. Other morphological or biological traits (oocyst shape and size, presence/absence of oocyst inner structures, pathogenicity, infection site, pre-patent and patent periods, sporulation time, and number of asexual generations) do not explicitly correlate with the phylogeny of rabbit coccidia.

Multiple species infections with parasitic helminths, including nematodes, are common in wild rodent populations. In this paper we first define different types of associations and review experimental evidence for different categories of interactions. We conclude that whilst laboratory experiments have demonstrated unequivocally that both synergistic and antagonistic interactions involving nematodes exist, field work utilizing wild rodents has generally led to the conclusion that interactions between nematode species play no, or at most a minor, role in shaping helminth component communities. Nevertheless, we emphasize that analysis of interactions between parasites in laboratory systems has been fruitful, has made a fundamental contribution to our understanding of the mechanisms underlying host-protective intestinal immune responses, and has provided a rationale for studies on polyparasitism in human beings and domestic animals. Finally, we consider the practical implications for transmission of zoonotic diseases to human communities and to their domestic animals, and we identify the questions that merit research priority.

A monoclonal antibody (MAb G11) species-specific to the Em2 antigen of Echinococcus multilocularis was generated for (i) further biological characterization of the Em2 antigen, (ii) easy affinity-purification of Em2 antigen for immunodiagnostic and immunological investigations and (iii) development of a sandwich-ELISA for the detection of Em2 antigen in diagnostic samples and thus species-specific identification of E. multilocularis metacestode material. The MAb G11 was used in an antibody sandwich-ELISA to detect soluble Em2 antigen with a methodical sensitivity of 80 ng E. multilocularis antigen/ml of solution. MAb G11 specifically detected Em2 antigen in all of 15 E. multilocularis-isolates originating from various geographical areas and in none of other helminth isolates (e.g. Echinococcus granulosus, E. vogeli, and others). Further biological analysis by FITC-labelled MAb G11 demonstrated unique binding activity to the laminated layer of the metacestode. Also, oncospheres were binding FITC-labelled MAb G11 on an outer layer synthesized during cultivation in vitro for 13 days after hatching. Application of the MAb G11 antibody sandwich-ELISA for investigation of solubilized oncospheres confirmed the in vitro synthesis of Em2 antigen by oncospheres on day 13 p.i. Adult stages (somatic antigens) and freshly hatched oncospheres were always MAb G11 negative. Solid-phase MAb G11 was used for purification of the corresponding Em2 antigen by affinity chromatography. A preliminary serological evaluation of the Em2(G11) antigen by ELISA revealed identical immunodiagnostic characteristics, compared to Em2 obtained by classical means, thus suggesting the presented method for future isolation of large-scale Em2 antigen.

1. Studies were made on the morphology, with special emphasis on the taxonomic characters, of 220 mature male and female Ascaridia lineata (Schneider) from chickens at Manhattan, Kansas, U.S.A., and Cambridge, England.

2. Eggs arising in the anterior ovary pass into the posterior uterus; similarly, eggs from the posterior ovary pass into the anterior uterus.

3. In young females eggs may be fertilised at any place in the uteri; in adults, fertilisation occurs near the distal ends of the uteri.

4. With the aid of a micromanipulator, a small structure in one pole of the mature egg, previously decribed as an opening, an opercular plug in the shell, or as an internal thickening of the shell, was found to be a solid, conical appendage of the vitelline membrane, free from the shell.

5. Fertilised eggs develop to the coiled embryo (infective) stage in 16 days when incubated in water at 30° C.; in 1 mm. of water at 33° C., they become infective in 10 days.

6. Water cultures of fertile eggs do not develop when kept at constant temperatures of 0° C., or of 10° C.; at 15° C., development proceeds.

7. Constant refrigeration of fertile eggs in water cultures for 1 month at 0° C. so lower the vitality that, on being incubated subsequently at 30° C., they divide slowly and soon die; keeping the eggs at 10° C. for 1 month has no deleterious effect on them when incubated subsequently at 30° C.

8. In hatching, the embryo may escape from any part of the egg shell, either in the duodenum of the host, or in water cultures; newly hatched larvae swallowed by the chicken seldom become established; infestation normally result from the ingestion of embryonated eggs.

9. The habitat of A. lineata is the duodenum, especially the portion which is a few centimetres posterior to the entrance of the bile ducts; determination of hydrogen-ion concentrations showed that the nematodes live in nearly neutral media (av. 6·7).

10. Penetration of the duodenal mucosa by larvae 10–17 days old is frequent in young chickens; occasionally, a larva goes on through the intestinal wall to the liver and lungs, but ordinarily, after the 17th day the young worms withdraw from the mucosa into the lumen of the intestine.

11. Young A. lineata grew to maturity in 50 days in chickens parasitised when about a month old; in six 8-day periods following hatching of the larvae, the average daily growth in length per period was about 0·12 mm. in the first period, 0·75 mm. in the second, 1·5 mm. in the third, fourth, and fifth periods, and 3mm. in the sixth period.

12. At least three moults occur before the adult from of the nematode is developed. Important morphological changes associated with the moults include: after first moult, presence of pre-anal swelling (males), and of anal prominence in both sexes; after second moult, lips with oral papillae and dentigerous ridges, projecting Iateral folds (alae) present in newly hatched larvae replaced by non-projecting lateral lines, females with vulva and shorter tail proportionately, and males with pre-anal sucker and three pairs of caudal papillae; after third moult, external characters similar to those of the adult A. lineata.

Fifteen children aged from 6 to 12 years who were infected with Ascaris lumbricoides were selected for the study. The number of eggs/g (e.p.g.) of stool was determined using Beaver's direct smear technique, Stoll's dilution egg count technique and Katz's modified thick smear technique. Beaver's direct smear method consistently gave a higher e.p.g. than the other two methods. Results of this study show that Katz's modified thick smear technique was the best of the three methods used. The e.p.g. depends on several factors: the amount of stool passed daily, the concentration of eggs within a stool sample, the daily egg output, the worm load, the age of the worm and the technique used for determining the egg load. The daily average egg output/female A. lumbricoides was estimated to be 238722 (range 134462–358 750) with an average e.p.g. of 3540. The estimation of worm load based on egg count is useful in determining the intensity of infection in an individual. The male/female worm ratio was estimated to be 1: 1·4.

The protozoan parasite Neospora caninum is an important cause of reproductive disease in cattle worldwide. The dog is a definitive host for the parasite and the oocyst stage, shed in the faeces, is a source of infection for cattle through consumption of contaminated feed or water. In addition, transplacental transmission of N. caninum is a very efficient means of the parasite infecting a new host and this can occur in successive pregnancies and over several generations. Neospora parasites may cause disease during pregnancy resulting in death of the foetus or birth of live congenitally infected calves that may show some neurological clinical signs at birth. The stage of pregnancy at which infection/parasitaemia occurs is an important factor in determining disease severity. Neospora infection in the first trimester of pregnancy may have more severe consequences for the foetus compared with infection occurring in the final trimester. The host-parasite relationship during pregnancy is a fascinating interaction and research in this area will improve understanding of disease pathogenesis and the various consequences of the host immune response, being host-protective, parasite protective and contributing to disease pathology. Pregnancy poses an interesting problem for the immune system of the dam as she is essentially carrying a semi-allogeneic tissue graft (the foetus) without immunological rejection taking place. To facilitate the pregnancy the cytokine environment in the placenta favours the regulatory Th-2-type cytokines, whose role is to counteract the pro-inflammatory Th1-type immune responses. Protective immunity to N. caninum, similar to many other intracellular parasites, involves Th1-type immune responses, which may pose problems for the dam trying to control a Neospora infection during pregnancy.

This paper will discuss the host-parasite relationship at different stages of gestation in pregnant cattle and review the implications of this research for our understanding of disease pathogenesis, parasite transmission and host protection.

Several aspects of the coevolutionary dynamics in host-parasite systems may be better quantified based on analyses of population structure using neutral genetic markers. This includes, for example, the migration rates of hosts and parasites. In this respect, the current situation, especially in fluke-snail systems is unsatisfactory, since basic population genetics data are lacking and the appropriate methodology has rarely been used. After reviewing the forces acting on population structure (e.g. genetic drift or the mating system) and how they can be analysed in models of structured populations, we propose a simplified, indicative framework for conducting analyses of population structure in hosts and parasites. This includes consideration of markers, sampling, data analysis, comparison of structure in hosts and parasites and use of external data (e.g. from population dynamics). We then focus on flukes and snails, highlighting important biological traits with regard to population structure. The few available studies indicate that asexual amplification of flukes within snails strongly influences adult flukes populations. They also show that the genetic structure among populations in strongly affected by traits in other than snails (e.g. definitive host dispersal behaviour), as snails populations have limited migration. Finally more studies would allow us to deepen our current understanding of selective interference between flukes and snails (e.g. manipulation of host mating system by parasites), and evaluate how this affect population structure at neutral markers.

The anthelmintic ivermectin has a number of effects on nematodes which result in changes in behaviour, particularly locomotion, including paralysis and an inhibition of feeding. This paper describes the application of an in vitro pharmacological approach to further delineate the action of ivermectin on feeding behaviour. Contraction of Ascaris suum pharyngeal muscle was monitored using a modified pressure transducer system which detects changes in intrapharyngeal pressure and therefore contraction of the radial muscle of the pharynx. The pharynx did not contract spontaneously. However, serotonin (5-HT, 100 μm) stimulated rhythmic contractions and relaxations (pumping) at a frequency of 0·5 Hz. γ-Aminobutyric acid (GABA) and glutamic acid inhibited the pumping elicited by 5-HT. The duration of inhibition was concentration dependent (1–1000 μm) with a threshold of 1 μm and 10 μm respectively (n=8). Ivermectin also inhibited pharyngeal pumping (1–1000 nM). At lower concentrations, ivermectin (1–10 pM) potentiated the GABA and glutamate inhibition, so that inhibition occurred at concentrations which were below threshold in the absence of ivermectin. These data provide evidence that the pharynx is a site for the action of ivermectin. Thus interruption of pharyngeal processes such as, feeding, regulation of hydrostatic pressure and secretion may provide a new site of anthelmintic action.

We have investigated the effect of qinghaosu (QHS, artemisinin) and its derivatives on Leishmania major replication in vitro and on the disease development in mice infected with L. major. Artemisinin is effective against promastigotes in vitro, with an ED 50 (50% effective dose) at 7.5 × 10−7 m. Both artemisinin and artemether are leishmanicidal for amastigotes in infected murine macrophages in vitro, with ED50 at 3 × 10−5 m and 3 × 10−6 m respectively. These compounds have no effect on the viability of macrophages or on the phytohaemaglutinin-induced proliferation of normal spleen cells, even at 10−4 m. BALB/c mice infected in the footpad with L. major developed significantly smaller lesions and parasite loads when treated with the compounds. Intra-lesion injection of the compounds was the most effective route. The intramuscular and oral routes were also effective; however, intravenous injection with artesunate was not effective.

The study of the life-histories of digenetic Trematodes is as yet in its infancy. Many new adult forms are described every year, but very little is known of the life-history of any of these. It is even impossible in the majority of cases to say what sort of larval form is characteristic of any group. Having worked for some years at the study of Trematode larvae I thought it would be of some use to bring together all the work so far done on British marine cercariae in order to form a nucleus round which future work may be centred.

Quantitative and qualitative analysis of lipids has been performed on a rhoptry fraction purified from Toxoplasma gondii tachyzoites. The lipid to protein ratio was estimated to be 0·26. The cholesterol to phospholipid ratio was unusually high at 1·48. Phosphatidylcholine was the major phospholipid; phosphatidylserine, phosphatidylinositol and sphingomyelin were absent whereas significant amounts of phosphatidic acid and lysophospholipids were found. This pelicular composition could be related to functional involvement of the organelle in host-cell invasion.

A Plasmodium falciparum single copy gene predicting a 122 kDa protein belonging to the M1 family of zinc-metallopeptidases was previously reported and related to erythrocytic schizont proteins of 96 (p96) and 68 (p68) kDa. By using protease inhibitors during parasite harvest and enzyme preparations, and polyclonal antibodies specific for 2 peptidic domains deduced from the gene, we identified the 120 kDa precursor and demonstrated its processing into p96 and p68. The N-terminal ends of p96 and p68 were mapped between glycine-123 and lysine-163, both proteins thus containing the catalytic domain. The purified enzyme, here named PfA-M1 (p96/p68), displayed strict aminopeptidase activity, optimal at pH 7·4, with broad substrate spectrum. Its inhibition and reactivation profiles were typical of zinc-metalloaminopeptidases. By Western blotting, PfA-M1 was detected in trophozoites, in addition to schizonts, but not in early rings. PfA-M1 was localized by indirect immunofluorescence confocal microscopy. In trophozoites, the labelling was diffuse in the parasite cytoplasm, with accumulations around the food vacuole. In schizonts, it turned progressively to a vesicle-like pattern, ending as a clear spot in released merozoites. The involvement of PfA-M1 in haemoglobin breakdown and erythrocyte reinvasion is discussed in light of the dual functions recently reported for several P. falciparum proteases.

This paper describes the quantitative aspects of establishment and expulsion of Nippostrongylus brasiliensis in the normal and immune rat. It is proposed that this helminth infection may be described in terms of four phases. These are: (1) loss phase 1 (LP1). A proportion of the infecting dose of larvae is immobilized or destroyed before or during the course of migration via the lungs to the intestine. (2) The plateau phase. The worm number remains static in the predilection site. (3) Loss phase 2 (LP2). This is analogous to the self-cure reaction and during this worms are expelled from the host by an immunological mechanism. The onset of the expulsion is sudden and the process is exponential. It continues at a steady rate until a threshold is reached. (4) Threshold phase. A small residual population of worms is not expelled and survives for a fairly prolonged period.

The pattern is maintained in immune animals undergoing second or third infections but there are quantitative differences. With increasing number of infections LP1 becomes greater, the plateau phase is shortened and the expulsion rate of LP 2 increases. It is suggested that the rate coefficient of this phase is an accurate measure of the immune status of the animal. The threshold phase is of a similar order in first, second and third infections. These results are discussed with regard to their significance as a measure of the immune status of the host, their implication regarding the mechanism of helminth expulsion and their relevance to other helminth infections.

The authors wish to thank the Agricultural Research Council and Allen and Hanburys Ltd. for the financial support which made this work possible. During this period Ellen E. Jarrett held a Horserace Betting Levy Board Scholarship.