The fine structure of two kinds of compound presumed sense organs from the heads of three skin parasitic monogeneans Gyrodactylus sp. Entobdella soleae (larva only) and Acanthocotyle elegans is described. One kind of compound receptor consists of a number of associated sensilla, each ending in a single cilium (the spike sensilla of Gyrodactylus and the cone sensilla of E. soleae oncomiracidium).

The other kind of compound organ is made up of one or a few neurones only, each of which bears many cilia (pit organs of E. soleae oncomiracidium and feeding organ sensilla of Acanthocotyle elegans). The spike sensilla of Gyrodactylus have also been studied using a Cambridge Instrument Co. Stereoscan electron microscope and by phase-contrast microscopy. The ciliary endings of all these sense organs are highly modified and have lost the 9 + 2 structure, being packed with many fibres. The fibre arrangement in the cilia of the cone sensillae of E. soleae oncomiracidium and the feeding organ sensilla of A. elegans has been compared with that in the ciliary endings of other invertebrate mechano- and chemoreceptors. The possibility that the spike sensilla of Gyrodactylus may be chemoreceptors has been discussed but it is considered premature to attempt to assign functions to the other sense organs studied. Electron dense membrane-bound inclusions occurring specifically in the nerves supplying the spike sensilla of Gyrodactylus may be neurosecretory.

In spite of its wide distribution among dogs and the evidence of its implication as a zoonotic agent, scant information is available on the biology of Acanthocheilonema reconditum (Spirurida, Onchocercidae). In this study, blood samples from 152 Sicilian dogs were examined for A. reconditum microfilariae at the beginning of the study and 1 year later. The periodicity of microfilaraemia was investigated by bleeding 2 highly microfilaraemic dogs twice a day for 10 days and, later on, every 2 weeks for 1 year and a third animal every 3 h for 96 h. Fleas and ticks infesting dogs were collected and dissected for the detection of A. reconditum larvae. The prevalence of infestation was 11·2% (17/152) and 13·3% (16/120) at the beginning and at the end of the study, with a 1 year cumulative incidence of 5·9%. Although dogs bled twice a day showed a higher number of microfilariae in most of the morning samples, the absence of any circadian rhythm was suggested by data of the third experiment conducted by bleeding a dog every 3 h for 4 days. A. reconditum developing forms were detected in 5·1% (4/78) of dissected fleas, but not in any of the 272 ticks. The study provides new insights into the biology and ecology of this dog filarioid in its definitive and intermediate hosts.

We have used two monoclonal antibodies, 128C3/3 and 504B1, to immunolocalize their carbohydrate epitopes in different developmental stages of Schistosoma mansoni. Both epitopes contain fucose: mAb 128C3/3, as we have shown previously, recognizes fucose in a novel, possibly internal linkage (Levery et al. 1992) while mAb 504B1, as we show here, bound to the Lex epitope, which contains fucose α1 → 3 linked to N-acetyl-glucosamine. The tissue expression of these epitopes was strikingly different and both elicit an immune response in infected hosts. The mAb 128C3/3-defined epitope was exposed on the surface of all larval stages but not on adult worms; however, it was found in the excretory system of adult worms of both sexes. In contrast, surface expression of the Lex epitope was initiated after the transformation of cercariae to schistosomula and was maintained throughout the adult life in both sexes.

The characteristics of intra-lymphocytic Theileria isolated from African buffalo and from cattle that were infected with buffalo-derived parasites were evaluated using anti-schizont monoclonal antibodies (mAbs) and DNA probes. Antigenic differences were revealed by the reactivities of 27 mAbs with the buffalo-derived parasites isolated from different animals. Antigenic diversity was also seen with Theileria-infected lymphoblastoid cell isolates taken from the lymph nodes and blood of the same animals. Two DNA probes, selected from a genomic library of T. parva piroplasm DNA cloned in λgt11, showed specific hybridization to parasite DNA in Southern blots of restriction enzyme-digested, lymphoblastoid cells infected with buffalo-derived theilerial parasites. Genotypic differences between the buffalo-derived parasites were revealed by the restriction fragment length polymorphisms seen with hybridization of those probes to DNA from cloned and uncloned Theileria-infected cell lines. The evaluation of theilerial parasites derived from buffalo and from cattle which underwent typical T. p. lawrencei reactions, after being infected with buffalo-derived theilerial parasites, did not show any specific phenotypic or genotypic characteristics of these parasites that would distinguish them from T. p. parva and T. p. bovis parasites. The validity of these subspecies distinctions is discussed.

The relationship between the ciliated epidermal cells and the subepidermal layer of the miracidium of Fasciola hepatica has been described. Non-ciliated ridge-like extensions of the subepidermal layer separate the ciliated epidermal cells from each other. The sunken portions of the subepidermal layer, each containing a nucleus, lie below the outer body wall muscles of the miracidium and open into the ridge by narrow neck-like connexions. Elongate vesicles, which may be a source of stored plasma membrane similar to that which occurs in the transitional epithelium of other animals, fill most of the ridge. In addition, characteristic round electron dense granules are found in the ridge but the majority are found in the sunken portions of the subepidermal layer.

The development and origins of the tegument of the sporocyst of F. hepatica have been described at the ultrastructural level. When the miracidium is in the process of penetrating the snail host, large vacuoles appear between the ciliated epidermal cells and the basal lamina which overlies the muscles of the body wall. These vacuoles have the effect of loosening the epidermal cells from the basal lamina of the body wall of the miracidium. Possible mechanisms involved in the formation of such vacuoles are suggested and discussed.

During penetration of the snail the ciliated epidermal cells of the miracidium are lost; the ridge, a syncytial layer between the epidermal cells which is connected with the subepidermal layer, spreads over the basal lamina and exposed body wall muscles of the metamorphosing sporocyst to form the new outer covering of the sporocyst.

Cytoplasm passes from the subtegumentary layer into the tegument during this stage of the development of the body wall of the sporocyst. Muscular contraction and microtubules may be involved in the outward movements of this cytoplasm. The nuclei of the subtegumentary layer remain below the muscles of the body wall.

Twenty-four hours after penetration of the snail the outer plasma membrane of the tegument forms folds, which greatly increase the surface area.

Sixty hours after penetration involutions between the folds, which may indicate pinocytosis, are present, and it is suggested that pinocytosis may play a role in food absorption.

The fully formed tegument is a syncytial layer containing numerous electron dense granules, vacuoles, mitochondria and lipid droplets.

The results on the formation of the tegument of the sporocyst have been discussed with reference to the controversy about the origins and terminology of the outer covering of the Platyhelminths.

The asymmetric distribution of phospholipids in the erythrocyte membrane during the intracellular development of the human malaria parasite Plasmodium falciparum was studied. Infected cells of high parasitaemia were treated with phospholipase A2 or sphingomyelinase C, followed by isolation of the host red cell membrane using the Affigel (731) bead method. Additionally, phosphatidylserine on the surface of infected cells was probed using a phosphatidylserine-sensitive prothrombinase assay. Trophozoite-infected cells showed an increase in phosphatidylethanolamine and phosphatidylserine and a decrease in phosphatidylcholine in the outer leaflet. In addition to the changes already present in trophozoite-infected cells, schizont-infected cells showed a decrease in sphingomyelin as well as a further increase in phosphatidylserine in the outer leaflet. The results are discussed with respect to possible mechanisms and consequences of these changes.

1. The life history of N. battus is described, and a comparative description of the life history of N. filicollis is given.

2. The life histories of these two species are compared with those of N. spathiger and N. helvetianus, two closely related species, and are shown to follow the same basic pattern, with minor variations in timing which appear to be specific in nature, and not related to differences in culture methods or host species.

3. The pathogenesis of Nematodirus species is discussed and related to the migration of larvae into the intestinal mucosa during development.

Trypanosoma brucei is exposed to the adaptive immune system and complement in the blood of its mammalian hosts. The aim of this review is to analyse the role and regulation of the proteins present on the external face of the plasma membrane in the long-term persistence of an infection and transmission. In particular, the following are addressed: (1) antigenic variation of the variant surface glycoprotein (VSG), (2) the formation of an effective VSG barrier shielding invariant surface proteins, and (3) the rapid uptake of VSG antibody complexes combined with degradation of the immunoglobulin and recycling of the VSG.

The paper describes a mathematical model for canine leishmaniasis and presents formulae which can be used to estimate the basic reproduction number, R0. The primary concern has been to devise methods of estimation which make best use of those data most easily obtained by fieldwork, e.g. surveys of prevalence in dog (by age) and sandfly populations. A range of formulae are offered which are more or less demanding of data, and which consequently give more or less precise estimates of R0. They include methods for assessing the influence on R0 of heterogeneous biting rates of sandflies on dogs, in which the essence of heterogeneous transmission can be captured merely by measuring relative rather than absolute contact rates.

The proteinases of Trichomonas vaginalis have been analysed by electrophoresis in polyacrylamide gels containing denatured haemoglobin. Seven bands of activity were detected indicating multiple proteinases. All of the enzymes were stimulated by 1 mM dithiothreitol and had inhibitor sensitivities characteristic of cysteine proteinases. The enzymes differed significantly, however, with respect to pH optima and relative sensitivities to inhibitors.

Plasmodium vivax is the most geographically widespread of the malaria parasites causing human disease, yet it is comparatively understudied compared with Plasmodium falciparum. In this article we review what is known about naturally acquired immunity to P. vivax, and importantly, how this differs to that acquired against P. falciparum. Immunity to clinical P. vivax infection is acquired more quickly than to P. falciparum, and evidence suggests humans in endemic areas also have a greater capacity to mount a successful immunological memory response to this pathogen. Both of these factors give promise to the idea of a successful P. vivax vaccine. We review what is known about both the cellular and humoral immune response, including the role of cytokines, antibodies, immunoregulation, immune memory and immune dysfunction. Furthermore, we discuss where the future lies in terms of advancing our understanding of naturally acquired immunity to P. vivax, through the use of well-designed longitudinal epidemiological studies and modern tools available to immunologists.

Praziquantel, the drug of choice against schistosomiasis, disrupts calcium (Ca2+) homeostasis in schistosomes via an unknown mechanism. Voltage-gated Ca2+ channels are heteromultimeric transmembrane protein complexes that contribute to impulse propagation and also regulate intracellular Ca2+ levels. β subunits modulate the properties of the pore-forming α1 subunit of high voltage-activated Ca2+ channels. Unlike other Ca2+ channel β subunits, which have current stimulatory effects, a β subunit subtype found in S. mansoni (SmβA) and S. japonicum (Sjβ) dramatically reduces current levels when co-expressed with Ca2+ channel α1 subunits in Xenopus oocytes. It also confers praziquantel sensitivity to the mammalian Cav2.3 α1 subunit. The Beta Interaction Domains (BIDs) of SmβA and Sjβ lack 2 conserved serines that each constitute a consensus site for protein kinase C (PKC) phosphorylation. Here, we use site-directed mutagenesis of schistosome β subunits to show that these unique functional properties are correlated with the absence of these consensus PKC sites in the BID. Furthermore, a second schistosome β subunit subtype contains both serines in the BID, enhances currents through α1 subunits, and does not confer praziquantel sensitivity. Thus, phosphorylation sites in the BID may play important roles in defining the modulatory properties and pharmacological sensitivities of schistosome Ca2+ channel β subunits.

PF1 (SDPNFLRFamide) is a FMRFamide-like peptide extracted from the free-living nematode Panagrellus redivivus. Here we show that this peptide causes a hyperpolarization of somatic muscle cells of the parasitic nematode Ascaris suum and a relaxation of the somatic muscle strip preparation. We have assessed whether or not the relaxation of Ascaris dorsal muscle strip by PF1 is due to (i) inhibition of the release of the excitatory neuromuscular junction transmitter acetylcholine (ACh), (ii) potentiation of the release of the inhibitory neuromuscular junction transmitter γ-aminobutyric acid (GABA) or (iii) a direct inhibitory action of the peptide on the muscle cells. Under the experimental conditions described here, tonic ACh release does not seem to be involved in determining the resting membrane potential or resting tone of the Ascaris dorsal muscle strip and thus inhibition of tonic ACh release is unlikely to explain the relaxation elicited by the peptide. Furthermore, PF1 (100 nM–1 μM) inhibited the contraction of the muscle strip elicited by bath application of ACh, suggesting either a direct inhibitory action of the peptide on the muscle cells or a potentiation of GABA release. In electrophysiological experiments, the reversal potential for the PF1 hyperpolarization was not the same as that for GABA. Thus, PF1 hyperpolarizes Ascaris muscle by a mechanism that does not involve stimulation of GABA release from inhibitory pre-synaptic terminals.

1. Onchocerca is recorded for the first time from English-bred cattle.

2. A specific diagnosis of O. gutturosa is made, O. bovis being regarded as a synonym. It is suggested that Stiles's description of Filaria lienalis may have referred to this species.

3. The microfilariae occur in the skin of infected cows, and blood-sucking flies of the genus Simulium were found to be the vectors.

4. Complete development was observed in S. ornatum and partial development in S. erythrocephalum. S. ornatum appears to be a very efficient intermediate host, as over 40 per cent became infected.

5. Culicoides nubeculosus, the vector of Onchocerca cervicalis of horses, does not appear to carry the bovine Onchocerca.

Larvae of Boophilus microplus were labelled with [32P] and used to study the rejection of larvae from British breed cattle with different levels of resistance. On animals of high resistance the loss of larvae due to grooming ranged from 9–54% during the first 24 h of infestation, and more time was spent grooming by these animals. Most of the grooming activity could be attributed to the presence of tick larvae and the presence of older tick stages did not increase the percentage of larvae lost. Grooming was directed to attached larvae and these could be removed. Animals of low resistance did not lose a significant number of larvae as a result of grooming, but all previously infested hosts lost a proportion of the larvae (18–39%) which could not be accounted for by grooming. This loss was greater than the total loss of larvae (11%) on animals not previously exposed to B. microplus.

Red fox (Vulpes vulpes) is the most abundant wild canid species in Austria, and it is a well-known carrier of many pathogens of medical and veterinary concern. The main aim of the present study was to investigate the occurrence and diversity of protozoan, bacterial and filarial parasites transmitted by blood-feeding arthropods in a red fox population in western Austria. Blood (n = 351) and spleen (n = 506) samples from foxes were examined by PCR and sequencing and the following pathogens were identified: Babesia canis, Babesia cf. microti (syn. Theileria annae), Hepatozoon canis, Anaplasma phagocytophilum, Candidatus Neoehrlichia sp. and Bartonella rochalimae. Blood was shown to be more suitable for detection of Babesia cf. microti, whilst the spleen tissue was better for detection of H. canis than blood. Moreover, extremely low genetic variability of H. canis and its relatively low prevalence rate observed in this study may suggest that the parasite has only recently been introduced in the sampled area. Furthermore, the data presented here demonstrates, for the first time, the possible vertical transmission of H. canis from an infected vixen to the offspring, and this could explain the very high prevalence in areas considered free of its main tick vector(s).

Modelling wildlife disease poses some unique challenges. Wildlife disease systems are data poor in comparison with human or livestock disease systems, and the impact of disease on population size is often the key question of interest. This review concentrates specifically on the application of dynamic models to evaluate and guide management strategies. Models have proved useful particularly in two areas. They have been widely used to evaluate vaccination strategies, both for protecting endangered species and for preventing spillover from wildlife to humans or livestock. They have also been extensively used to evaluate culling strategies, again both for diseases in species of conservation interest and to prevent spillover. In addition, models are important to evaluate the potential of parasites and pathogens as biological control agents. The review concludes by identifying some key research gaps, which are further development of models of macroparasites, deciding on appropriate levels of complexity, modelling genetic management and connecting models to data.

Partial (DNA) sequences are presented for 2 nuclear (18S and 28S rRNA genes) and 2 mitochondrial (12S rRNA and ND1 genes) loci for 5 species belonging to the Schistosoma japonicum, S. sinensium and S. indicum groups of Asian Schistosoma. Fresh field isolates were collected and cultured for the following taxa: S. incognitum (S. indicum group, central Thailand), S. mekongi (S. japonicum group, southern Laos), S. ovuncatum (S. sinensium group, northern Thailand), S. spindale (S. indicum group, northeast Thailand and central Thailand isolates) and S. sinensium (S. sinensium group, Sichuan Province, China). This represents the first published DNA sequence data for S. ovuncatum and for S. sinensiums.s. from the type locality in China. The paper also presents the first sequence data at the above loci for S. incognitum (except for the 28S sequences) and S. sinensium. Congruence was observed between the phylogenies estimated for each locus, although the relationships of S. incognitum were not so well resolved. Fitch–Margoliash, maximum likelihood (ML) and maximum parsimony methods were used to estimate the phylogenies and the agreement between them was similar to that observed between loci. The ML tree was considered to best represent the data and additional 28S sequences (taken from the GenBank), for S. haematobium, S. japonicum, S. mansoni and Orientobilharzia turkestanicum, were used to construct an overall phylogeny. The S. indicum group taxa showed considerable divergence from the other Asian species and closest affinity with the African group. S. ovuncatum and S. sinensium appeared as sister taxa but their status as sibling species remained supported. The findings are discussed in the context of phylogeographical hypotheses for the origin of Schistosoma. An Asian origin for Schistosoma is also considered.

We compared parasite communities in fish taken from a polluted lake (L. Vatia) and two control lakes before (1986) and after (1995) nine years of markedly reduced chemical and nutrient loading from a pulpmill in central Finland. Discriminant analyses of the 1995 data, using a function based on the 1986 data, showed that the parasite communities in the fish from the two control lakes had changed relatively little, whereas those from L. Vatia had converged on those from the mesotrophic control lake, indicating substantial recovery from the effects of pollution. Only a few species of parasites provided evidence for recovery. These were anodontid glochidia, which had increased markedly in perch, Rhipidocotyle fennica in roach and R. campanula in both fish species. This suggests that the recovery of the polluted lake involved increased populations of anadontid clams in shallow waters. On the other side decrease of Dermocystidium percae on perch fins and Ichthyophthirius multifiliis on roach indicate increased immune responses in the fish, reflecting better water quality. Other parts of the system have apparently not yet recovered.

The initial process in the hatching mechanism of Ascaris eggs is the sudden onset of permeability in the previously impermeable ascaroside membrane. During this change the ascaroside membrane remains intact and no chemical changes can be detected. Using the molecular probe 1-anilino-8-naphthalene sulphonic acid no conformational changes were detected in the ascaroside membrane during the induction of permeability. It is suggested that either the permeability change is due to a very localized chemical or conformational change, not detectable by conventional analytical techniques, or the change is due to mechanical damage of the ascaroside membrane, brought about by the activity of the infective larva.