The aim of this study was to assess the seroprevalence of the Toxoplasma gondii parasite in pork produced in France, and to determine infection risk factors. An innovative survey was designed based on annual numbers of slaughtered pigs from intensive and outdoor farms in France. A total of 1549 samples of cardiac fluids were collected from pig hearts to determine seroprevalence using a Modified Agglutination Test. Of those, 160 hearts were bio-assayed in mice to isolate live parasites. The overall seroprevalence among fattening pigs was 2·9%. The adjusted seroprevalence in pigs from intensive farms was 3·0%; the highest in sows (13·4%); 2·9% in fattening pigs and 2·6% in piglets. Adjusted seroprevalence in fattening animals from outdoor farms was 6·3%. Strains were isolated from 41 animals and all were genotyped by Restriction Fragment Length Polymorphism as type II. Risk-factor analysis showed that the risk of infection was more than three times higher for outdoor pigs, and that sows’ risk was almost five times higher than that of fattening animals. This study provides further evidence of extensive pork infection with T. gondii regardless of breeding systems, indicating that farm conditions are still insufficient to guarantee ‘Toxoplasma-free pork’.

Salivarian trypanosomes pose a substantial threat to livestock, but their full diversity is not known. To survey trypanosomes carried by tsetse in Tanzania, DNA samples from infected proboscides of Glossina pallidipes and G. swynnertoni were identified using fluorescent fragment length barcoding (FFLB), which discriminates species by size polymorphisms in multiple regions of the ribosomal RNA locus. FFLB identified the trypanosomes in 65 of 105 (61·9%) infected proboscides, revealing 9 mixed infections. Of 7 different FFLB profiles, 2 were similar but not identical to reference West African Trypanosoma vivax; 5 other profiles belonged to known species also identified in fly midguts. Phylogenetic analysis of the glycosomal glyceraldehyde phosphate dehydrogenase gene revealed that the Tanzanian T. vivax samples fell into 2 distinct groups, both outside the main clade of African and South American T. vivax. These new T. vivax genotypes were common and widespread in tsetse in Tanzania. The T. brucei-like trypanosome previously described from tsetse midguts was also found in 2 proboscides, demonstrating a salivarian transmission route. Investigation of mammalian host range and pathogenicity will reveal the importance of these new trypanosomes for the epidemiology and control of animal trypanosomiasis in East Africa.

Plasmodium falciparum resistance to chloroquine has been described in many parts of the world particularly in Africa where malaria is endemic. High levels of chloroquine resistance in our study area, Lambarene–Gabon, has led to the use of an alternative regimen for treatment and prevention of P. falciparum infection. In this study, we examined the in vitro chloroquine sensitivity of 15 isolates from this area and assessed the prevalence of a putative chloroquine resistance associated Pfmdr1 polymorphism (Asn86Tyr) using a novel allele-specific polymerase chain reaction (PCR). Only 4 of the isolates examined were chloroquine sensitive. The allele-specific PCR shows that all 15 isolates carried the variant (86Tyr) codon. Eleven of these were resistant to chloroquine suggesting a 73% agreement between chloroquine resistance phenotype and the point mutation. This molecular marker was examined in a further 73 Gabonese isolates, where 58 (79·5%) showed 86Tyr and 15 (20·5%) showed 86Asn. In all, 4 (4·5%) of the 88 isolates assessed carry both mutant and wild-type codons, suggesting mixed parasite populations. The incomplete agreement found between chloroquine resistance phenotype and Pfmdr1 (86Tyr) polymorphism would support the view that other genetic factors as well as Pfmdr1 may be involved in chloroquine resistance. While our results suggest a high prevalence of 86Tyr polymorphism in Lambarene, the Asp1246Tyr polymorphism (a point mutation which to date has only been associated with South American P. falciparum) seems to be absent in our study area.

The high level of protection which can be induced by vaccination of a range of hosts, from rodents to primates, with live radiation-attenuated schistosome larvae offers great promise for development of a human schistosome vaccine. Studies of the irradiated vaccine models benefitted from significant funding during the 1970–90s and much was learned concerning the inducers, targets and mechanisms of immunity. Less progress was made in definition of the protective antigens involved. The application of new techniques for identifying membrane and secreted antigens has recently provided new vaccine candidates and a new impetus for schistosome vaccine development. This article is intended as an overview of some of the main lessons learned from the studies of the irradiated vaccines as a backdrop to renewed interest in schistosome vaccine development.

Using a combination of morphological, life-history and molecular data, we redescribe Sphaerospora ranae (Morelle, 1929) n. comb. (previously Leptotheca ranae) and emend its taxonomic status. Renal infection was recorded in 2 spp. of frogs (out of 5 amphibian spp. examined), Rana dalmatina (proposed type host) and Rana temporaria, suggesting restricted host specifity of S. ranae. We provide a description of sporogonic stages of S. ranae for the first time and suggest possible modes of its developmental cycle. Phylogenetic analysis inferred from the small-subunit ribosomal DNA revealed a close relationship of S. ranae with piscine Sphaerospora elegans (type species of the genus) and Sphaerospora truttae, forming together with distantly related Leptotheca fugu a ‘Sphaerosporid clade’, the basal branch to all myxosporean species. The close relationship of the 3 Sphaerospora spp. is further supported by the presence of 2 areas with extensive nucleotide insertions in the V4 region of the SSU rDNA (absent in L. fugu), morphology and life-history features. We conclude, that the spore morphology of Sphaerospora s.l., is very simple and probably represents a ‘primitive’, basal morphotype retained in most myxosporean lineages. Based on presented data, we propose emendation of the genus Sphaerospora using morpological, life-history and molecular features.

Cystic echinococcosis, caused by different species of the Echinococcus granulosus complex, is an important zoonotic disease with a particular impact on pastoralist societies. In addition to the widespread taxa with synanthropic transmission, a number of Echinococcus species were described from African wild carnivores early in the 20th century. For lack of study material, most of these were later tentatively synonymized with E. granulosus. Early infection experiments with wildlife isolates gave ambiguous results due to the use of unspecified parasite material, and only recently molecular methods provided the opportunity to shed light on the confusing scenery e.g. by characterizing E. felidis from the African lion. Here we will summarize the convoluted history of Echinococcus research in sub-Saharan Africa and highlight the necessity of molecular surveys to establish the life cycles and estimate the zoonotic potential of these parasites.

The invasion and establishment of Heterodera glycines has been studied for 0–120 h after addition to host roots using a synchronized infection system. A high degree of synchrony in development was noted, with juvenile nematodes reaching their feeding sites adjacent to the root endodermis and discharging their subventral pharyngeal glands by 24 h post-invasion (p.i.). The moult to the 3rd-stage juvenile occurred approximately 89 h after entry to the root. Using an enzyme-linked immunosorbent assay with monoclonal antibodies (MAbs) whose tissue specificities had already been defined by immuno-cytochemistry, it was found that MAbs recognizing intestinal lipid droplets and granules showed a major increase in reactivity 18–24 h p.i. before declining by 39 h p.i. In contrast, the reactivity of two MAbs recognizing adult cuticle increased significantly only after 89 h p.i., following the second moult. MAbs to the two subventral pharyngeal gland cells showed a variety of patterns of changing reactivity during the experiment. One increased within 24 h of invasion before declining whereas a second fell without any initial rise. Two further MAbs specific for the subventral glands initially declined but showed a secondary increase in reactivity after 72 h and could be detected in adult females. This pattern was also seen in a MAb specific for the dorsal pharyngeal gland cell. These changes are discussed in the context of events taking place following invasion, with particular reference to the initiation and maintenance of a feeding syncytium by the developing nematode.

We report the results of experimental studies of the generation of age-prevalence curves for Schistosoma mansoni infections in laboratory populations of Biomphalaria glabrata. Within snail populations of varying sizes and age structures, the net force of infection is shown to be linearly dependent on the rate at which miracidia are introduced into the aquatic habitat of the host. For individual snails, the per capita force of infection is shown to be related to snail age and size, and the death rate of shedding snails is demonstrated to be dependent on the period of time during which a snail has been releasing cercariae. Both factors are important determinants of the proportion of infected snails within populations of hosts and may generate convex age-prevalence curves. Comparisons of snail abundance in populations either exposed, or not exposed, to infection suggest that S. mansoni can act to significantly depress the population growth of its intermediate host. Mathematical models are developed, encorporating the age structure of snail populations, to aid in the interpretation of experimental results.

Adult parasites of Schistosoma mansoni reside within vertebrate mesenteric veins where they consume immense quantities of host glucose after transporting the sugar through their surface syncytium or tegument. Previously we obtained cDNA clones encoding two functional facilitated diffusion glucose transporter proteins expressed by S. mansoni adult worms (Skelly et al. 1994). Antibodies specific for one transporter (SGTP1) have been generated against an extrafacial and an internal domain of the protein and used to localize the protein by light and electron microscopy. By light microscopy both antibodies stain a linear structure approximately 1–5 μm from the surface of the tegument of adult male and female schistosomes. Electron microscopic examination of frozen thin sections show binding of the antibodies to membranes in the base of the tegument and not to the membranes covering the outer surface or their invaginations. Analysis of the gold distribution suggests that the extrafacial domain is disposed toward the interstitial space beneath the tegument and the internal domain faces the syncytial plasm. The localization suggests that SGTP1 may function to transport free glucose from within the tegument and into the interstitial fluids that bathe the internal organs of these parasites.

The antigenic composition of sequential erythrocytic stages of Plasmodium knowlesi has been compared by crossed immuno-electrophoresis using a pool of immune rhesus monkey antiserum. Eleven major parasite antigens have been identified; 9 are stage-independent, and 2 stage-dependent. Differences in the relative amount of the stage-independent antigens have been demonstrated and quantified. The distribution of antigens between parasites and schizont and infected red cell membranes has been examined. Only 6 of the 11 parasite antigens were exhibited by a schizont membrane preparation, all these antigens were also expressed by the intracellular parasite. Antigens exclusive to the schizont membrane were not demonstrated.

The life-cycle of Theileria parva in the gut of the tick Rhipicephalus appendiculatus was investigated in Giemsa-stained smears and in wet preparations under the phase-contrast microscope. The different developmental stages of T. parva were seen in a large proportion of specially-selected R. appendiculatus tickes. After the intra-erythorcytic merozoites were engorged by the ticks during a blood meal, the following development was observed. (1) In the lumen of the gut of infected nymphs, spindle-shaped microgamonts developed out of the ring-forms. These broke up into several thread-like microgameters after nuclear division and development of thread-like cytoplasmic projections. The ring-froms developed into round-forms of 3–4 μm in diameter which are considered to be macrogametes. (2) From the 6th day after repletion, zygotes with a clear zone in the centre appeared in the epithelial cells of the tick's gut. A steady increase in size and a progressively denser cytoplasm were observed up to the 3rd day after moulting to adult ticks. (3) Subsequently a slightly angular, retort-like stage developed by invagination inside the rounded zygotes, and from the 5th day after moulting this stage developed further into a club-shaped kinete. The kinetes propelled themselves in the gut by active gliding movements.

This study quantifies the rate and intensity of re-infection with human hookworm and Schistosoma mansoni infection 12 months following successful treatment, and investigates the influence of socio-economic, geographical and environmental factors. A longitudinal study of 642 individuals aged over 5 years was conducted in Minas Gerais State, Brazil from June 2004 to March 2006. Risk factors were assessed using interval censored regression for the rate and negative binomial regression for intensity. The crude rate and intensity of hookworm re-infection was 0·21 per year (95% confidence interval (CI) 0·15–0·29) and 70·9 epg (95% CI 47·2–106·6). For S. mansoni the rate was 0·06 per year (95% CI 0·03–0·10) and intensity 6·51 epg (95% CI 3·82–11·11). Rate and intensity of re-infection with hookworm were highest among males and positively associated with previous infection status, absence of a toilet and house structure. Rate and intensity of S. mansoni re-infection were associated with previous infection status as well as geographical, environmental and socio-economic factors. The implications of findings for the design of anti-helminth vaccine trials are discussed.

Three species of poeciliids (Gambusia holbrooki, Xiphophorus helleri and X. maculatus) and 15 species of ecologically similar native freshwater fishes (mainly eleotrids, ambassids, melanotaeniids and retropinnids) were examined for parasite richness to investigate parasite flux, qualitative differences, quantitative differences and the structuring factors in parasite communities in the 2 fish types in Queensland, Australia. Theory suggests that poeciliids would harbour depauperate parasite communities. Results supported this hypothesis; poeciliids harboured more species-poor parasite infracommunities and regional faunas than natives (P < 0·0001), despite greater sampling effort for the former. Cluster analysis of presence/absence data for poeciliids and the 6 most-sampled native fishes revealed that parasite communities of the 2 fish groups are qualitatively distinct; the proportion of parasite species with complex life-cycles was lower in poeciliids than in native species, and Myxosporea, Microspora, Coccidia and parasitic Crustacea were all absent from poeciliids. Limited exchange of parasite species has occurred between natives and poeciliids. Logistic ordinal regression analysis revealed that fish origin (exotic or native), environmental disturbance and host sex were all significant determinants of parasite community richness (P < 0·05). Theoretical modelling suggests that poeciliids are at a competitive advantage over native fishes because of their lack of parasites.

In order to better comprehend the putative association between genotype Trypanosoma cruzi II and primates, an evaluation of the infection in free ranging primates and specimens born in captivity from different geographical areas, the Amazon and the Atlantic forest, was carried out. Seroprevalences of the T. cruzi infection among the primates was similar in both biomes (45·5% and 46%). The parasites were isolated from 8 and 4 different species of primates, respectively from the Amazon and Atlantic forest. Multi-locus enzyme electrophoresis (MLEE) typed the isolates from Amazon as zymodeme 1. Mini-exon gene analysis characterized all these isolates as T. cruzi I, the main genotype circulating in the region. In the Atlantic forest, primates infected with TCI and TCII, as well as a mixed infection (TCI and TCII), were detected. These findings prove that primates may maintain stable infections by both genotypes. Moreover, data show that T. cruzi can occur in a wide range of primate genera, independent of their social behaviour, niches or habitats. Considering the high seroprevalence and stability of T. cruzi infection among the primates, these animals play an important role in the maintenance of the parasite in nature.

Ascariasis is one of the most common parasitic diseases in both humans and pigs. It has been shown to cause growth deficits in both species and to impair cognitive development in children. Notwithstanding its substantial impact on pig economy and public health, diagnosis of ascariasis has mostly relied on the detection of eggs in stool and further development of novel, more sensitive methods has been limited or non-existent. Here, we discuss the currently available techniques for the diagnosis of ascariasis in pigs, their caveats, and the implications of a new serological detection technique for the evaluation of both pig and human ascariasis.

Myxozoans are a group of diverse, spore-forming metazoan microparasites bound to aquatic environments. Sphaerospora dykovae (previously S. renicola) causes renal sphaerosporosis and acute swim bladder inflammation (SBI) in juvenile Cyprinus carpio carpio, in central Europe. A morphologically similar species with comparably low pathogenicity, S. angulata has been described from C. c. carpio, Carassius auratus auratus and Carassius gibelio. To clarify uncertainties and ambiguities in taxon identification in these hosts we decided to re-investigate differences in spore morphology using a statistical approach, in combination with SSU and LSU rDNA sequence analyses. We found that developing spores of S. angulata and S. dykovae cannot be distinguished morphologically and designed a duplex PCR assay for the cryptic species that demonstrated S. dykovae is specific to C. c. carpio, whereas S. angulata infects C. a. auratus and C. gibelio. The molecular identification of myxozoan blood stages in common carp and goldfish, which had previously been ascribed to Sphaerospora spp. showed that approximately 75% of blood stages were from non-sphaerosporid coelozoic species infecting these cyprinids and more than 10% were from an alien species, Myxobilatus gasterostei, developing in sticklebacks. We hereby report non-selective myxozoan host invasion and multi-species infections, whose role in SBI still requires clarification.

Here we report on the action of the novel cyclo-depsipeptide anthelmintic, emodepside, on the body wall muscle of the parasitic nematode, Ascaris suum. Emodepside caused (i) muscle relaxation, (ii) inhibition of muscle contraction elicited by either acetylcholine (ACh), or the neuropeptide, AF2 (KHEYLRFamide) and (iii) a rapid relaxation of muscle tonically contracted by ACh. The inhibitory action of emodepside on the response to ACh was not observed in a denervated muscle strip, indicating that it may exert this action through the nerve cord, and not directly on the muscle. Electrophysiological recordings showed emodepside elicited a Ca++-dependent hyperpolarization of muscle cells. Furthermore, the response to emodepside was dependent on extracellular K+, similar to the action of the inhibitory neuropeptides PF1 and PF2 (SDPNFLRFamide and SADPNFLRFamide). Thus emodepside may act at the neuromuscular junction to stimulate release of an inhibitory neurotransmitter or neuromodulator, with a similar action to the PF1/PF2 neuropeptides.

1. Nearly a quarter of a million eggs of Sialis lutaria were collected at Cambridge in 1936. About 0·6 per cent of them were attacked by a parasite.

2. The egg-parasite of Sialis is distinct from Trichogramma evanescens, and is to be called T. semblidis (Aurivillius).

3. The male of Trichogramma semblidis occurs in two forms. Neither consists merely of imperfect or degenerate individuals of the other, for the two forms are equally large and differ constantly and fundamentally in several characters. The species, therefore, exhibits true dimorphism.

4. Rearing experiments involving isolated pure lines show that it is principally the host that determines which form of the parasite shall emerge. Males reared on Sialis are of the apterous form; those reared on three species of Lepidoptera are of the winged form.

5. The dimorphism of T. semblidis is discussed in relation to other examples of dimorphism in the Hymenoptera. It is shown to have several features of special interest.

Two new, nine little known species and nine unnamed ‘species’ of Echeneibothrium Beneden, 1850, have been investigated and observations have been made on the ‘related’ genera, Discobothrium Beneden, 1870, and Pseudanthobothrium Baer, 1956. Brief descriptions of the species are given, but a discussion of the history of each differs greatly from most previous accounts in that few or no synonyms have been listed for the species and in that nine forms are left unnamed; this approach differs greatly from that in a large number of taxonomic papers in helminthology. A detailed discussion of the reasons for adopting this procedure is, therefore, given. A provisional key to nineteen ‘species’ of Echeneibothrium is included. E. minutum sp.nov. closely resembles E. variabile var. exiguum of Euzet (1959), described in this paper as ‘Echeneibothrium sp. of R. clavata’, but occurs only in R. batis and differs in its very small size, in having only about ten proglottids, long peduncles to the bothridia and a myzorhynchus which remains evaginated in fixed specimens. It differs from E. demeusiae Euzet, 1959, mainly in the number of proglottids, testes and loculi. E. elongatum sp.nov. resembles E. dubium from which it differs in the length and form of the strobila, in having only six clearly visible loculi in each bothridium, about twenty testes and in that it occurs in R. circularis. It has, however, more characters in common with a species referred to in this paper as ‘Echeneibothrium sp. from Raja naevus’ and which possesses eight loculi. A list is given of twenty-seven species wrongly allocated by various authors to the genus Echeneibothrium. The comparative morphology of some species of the genus, their ecology and speculations on their life-histories, are described.

It is shown for the first time that Echeneibothrium from species of Raja caught off the British Isles, and presumably from rays of other localities, can be separated into two distinct biological groups. One group includes species which possess shallow open bothridia covering comparatively large areas of the host's mucosa, a variable myzorhynchus which may be either rudimentary or very large and spherical eggs laid singly or in capsules; these occur mainly in rays from shallow waters and in which the mucosa of the intestine lacks prominent villi. The other group includes species in which each bothridium possesses a small opening adapted for attachment to a single villus, the myzorhynchus is consistently cylindrical, the eggs have long polar filaments and the species occur in deep water rays with well-defined villi on the intestinal mucosa. Studies on variation in the form of the eggs in different species of Echeneibothrium in relation to the behaviour and feeding habits of the various host-species suggest that the eggs are eaten by arthropods which are most likely to form part of the diet of the final host. A general trend towards an increase in the complexity of the bothridium and a decrease in the size of the myzorhynchus in Echeneibothrium is thought to be of considerable advantage to the species and an indication that Rhinebothrium, which is restricted to the Myliobatoidea, has evolved from a form like Echeneibothrium which is restricted to the Rajoidea. This supports the view that the Myliobatoidea have evolved from the Rajoidea. The results from this study of Echeneibothrium illustrate the following general rule which can be applied to the Tetraphyllidea, namely, those species which are abundant in any given host are very well adapted for attachment to the host's gut and cause little, if any, damage while rare species are often highly pathogenic.

This work would not have been possible without generous financial aid from the Department of Scientific and Industrial Research and various kinds of other aid and encouragement from a number of persons, in particular Dr Gwendolen Rees.

It was begun in 1958 at the University College of Wales, Aberystwyth, the British Museum (Natural History), University of Montpellier, France, University of Neuchatel, Switzerland, the Plymouth Marine Laboratory and the Aberdeen Marine Laboratory during my tenure of a D.S.I.R. Fellowship. I wish to record my gratitude to Dr Rees who has also suggested improvements to the manuscript and to the following persons: the late Professor T. A. Stephenson, F.R.S., Dr E. E. Watkin, Mr S. Prudhoe, Dr L. Euzet, Professor Jean G. Baer, The Director, Dr F. S. Russell and Staff, in particular Mr J. Green and Mr A. Mattacola, of the Plymouth Marine Laboratory, and the Director and staff, in particular Dr B. B. Rae and Dr Z. Kabata, of the Aberdeen Marine Laboratory.

The work was later continued at the University College, Cardiff, and I am most grateful to Professor James Brough for providing excellent research facilities and to Mr W. O'Grady for technical assistance.

At the Bureau of Helminthology, St Albans, much encouragement from Professor R. T. Leiper, C.M.G., F.R.S., enabled me to put the work together while having free access to his invaluable collection of books and reprints on helminths; it became unnecessary to search elsewhere for the literature. Mr G. Dimmock gave technical assistance for which I am most grateful.

1. Spores of Nosema apis nearly all germinate within 30 min. after entering the ventriculus of the honeybee.

2. The cause of germination in vivo remains unknown, but is probably not a change of osmotic pressure or pH and is probably not due to anaerobic conditions or an enzyme. The agent causing germination is probably unstable and is destroyed when removed from the bee.

3. The anterior end of the ventriculus receives an initial heavy infection and the circumstantial evidence which this provides supports the theory that a free living ‘planont’ stage does not exist but that the parasite is injected directly into the host cell from the spore via the hollow polar filament.

4. The central region of the ventriculus which contains many granules of calcium phosphate receives the lowest initial infection. It is considered possible that the granules inhibit the initial rapid development of the parasite by maintaining an unfavourably high level of pH until they are eventually dissolved by the slowly developing organism.