Monoclonal antibodies reacting with pellicular antigens of Toxoplasma gondii tachyzoites have been selected among hybridomas produced against this organism by immunofluorescence assay. These antigens have been further characterized by immunofluorescence on living zoites, Western immunoblotting and immunoprecipitation of lactoperoxidase surface radio-iodinated tachyzoite lysates. The simultaneous characterization of 5 different surface antigens (P43, P35, P30, P23, P22) some of which have already been studied individually allowed a better definition of these antigens and the characterization of a yet undescribed surface molecule (P23).

The fate of 75Se-labelled parasites and their released pre-synthesized macromolecules has been followed in three murine infection models. Parasite numbers in specific tissues were determined by autoradiography, and released material was estimated by gamma-counting of tissues, with adjustment for the presence of parasite-associated radiolabel. Marked differences were found between the three models. The pattern of migration of normal schistosomula was similar to that previously reported. In addition we have described the transit of parasites through the lymph nodes draining the infection site. Significant quantities of released material were detected in the skin, draining lymph nodes, bloodstream and liver. The circulating material was of parasite origin, macromolecular, and hence potentially antigenic. In comparison to the normal infection, radiation-attenuated parasites (inducing a high level of resistance to challenge) persisted in the skin, draining lymph nodes and lungs, releasing a proportionally greater amount of material in the nodes. In mice exposed to attenuated parasites and treated with the compound ROl1–3128 at 24 h (inducing a low level of resistance) there was an early death and rapid clearance of the parasites whilst still in the skin. This situation resulted in the highest levels of released material in the skin, bloodstream and liver, but negligible levels in the draining lymph nodes. We suggest that the persistence of radiation-attenuated parasites in the skin and draining lymph nodes, together with the prolonged release of antigen in the latter site, compared to the normal situation, are major factors in the induction of resistance.

The development of granulomatous reactions against moulting nymphal pentastomids (Porocephalus crotali) in the tissues of rat and mouse intermediate hosts is described. Adipose tissue and lungs are favoured sites for encystment accounting for 70% of larvae. Six moults separate the primary larva from the final infective stage which first appears about 80 days post-infection (p.i.) and is fully infective by day 120. Larvae, and particularly their cast cuticles, are the foci of granulomatous reactions characterized by an intense eosinophilia. During ecdysis, large numbers of eosinophils permeate the entire lesion but, significantly, degranulation is limited to the underside of cast cuticles where the resultant debris is endocytosed by macrophage/epithelioid cells. A pronounced asymmetry in the granulomatous lesion, evident even in the earliest cysts, results from the accumulation of individual epithelioid granulomas associated with cuticle fragments close to the ventral side of the developing parasite; each is circumscribed by fibrosis. External to this region are extensive tracts of tissue composed of mature plasma cells. Particularly in rats, large numbers of partially degranulated mast cells ( = globule leucocytes) also surround cuticle granulomas, and mast cell granules can accumulate within macrophages and fibroblasts. Inflammation slowly subsides once the infective stage is attained. This 1 cm-long larva resides in a thin, fibrotic, C-shaped cyst and can remain viable for years: uniquely this instar retains its last moulted cuticle as a protective sheath. Nymphal instars II-VI feed predominantly upon eosinophils but we do not yet know whether this requirement is obligate.

Two species of Schizotrypanum, T. (S.) dionisii and T. (S.) vespertilionis, were identified from British bats. Laboratory studies on stocks of isolated trypanosomes from 5 species of bat (Pipistrellus pipistrellus, Nyctalus leisleri, N. noctula, Eptesicus serotinus and Myotis brandti) indicated that the predominant species was T. d. dionisii. Collections and dissection of the bat bug Cimex pipistrelli from bat roosts revealed flagellate infection in a total of 12 out of 20 bugs; 7 of these bugs had metacyclic trypanosomes present. C. pipistrelli and the human bed bug, C. lectularius were reared in the laboratory and allowed to feed on wild-caught bats known to be infected with T. d. dionisii. Development occurred in both species of Cimex. Cimex spp. could be used to detect subpatent Schizotrypanum infections by xenodiagnosis. This technique was used to test the parasitological status of bats collected in the wild or reared in captivity. On a single occasion an apparent transmission of T. d. dionisii to an uninfected (by xenodiagnosis) laboratory reared bat was achieved. A stock of Schizotrypanum isolated from a wild-caught C. pipistrelli collected in a N. leisteri roost was identified by DNA buoyant density centrifugation as T. (S.) vespertilionis. A P. pipistrellus known to be infected with T. d. dionisii was found to have cyst-like structures in thoracic skeletal muscle containing amastigotes. The study provided the strongest evidence yet that C. pipistrelli is the vector of Schizotrypanum in British bats.

Relapse of parasitaemia after drug treatment of trypanosome infections is normally attributed to drug-resistance on the part of the parasite, under-dosage of the drug or reinfection of the host. In addition, inaccessibility of parasites to drug through sequestration in privileged extravascular sites has been shown in the past to occur with Trypanosoma brucei, and we have obtained evidence that extravascular foci of T. vivax can also serve as a source of relapsing infections. Infection of goats with a West African stock of T. vivax resulted in severe illness, which was fatal if untreated. During the terminal stage of an acute infection, clinical signs of central nervous system involvement were apparent. Histologically, the choroid plexus was swollen and oedematous, and in some cases meningitis or meningoencephalitis was seen. Trypanosomes could be detected in the cerebrospinal fluid, and also extravascularly in the choroid plexus and meninges. In three cases they were present in the aqueous humor, associated with corneal cloudiness or opacity. Treatment of 2 goats with the trypanocidal drug diminazene aceturate eliminated parasitaemia, but infections in both relapsed about 6 weeks later, despite trypanosomes being undetectable in the bloodstream during the intervening period. We conclude that the relapse infections were caused by re-emergence of trypanosomes from the CNS and/or the eye, where sequestered parasites may have been inaccessible to the trypanocide.

We have obtained 5 specific DNA probes for African trypanosomes of the subgenera Trypanozoon and Nannomonas. Each probe consists of one repeat unit of the major repetitive DNA (satellite DNA) of each species or intra-specific group. One probe hybridized with all members of subgenus Trypanozoon (except T. equiperdum which was not tested). In subgenus Nannomonas, one probe recognized T. simiae, but 3 probes were needed to identify all stocks of T. congolense available. Each of the 3 latter probes recognized trypanosomes from one of the 3 major groups of T. congolense previously defined by isoenzyme characterization, i.e. savannah, forest and Kenya coast types. As few as 100 trypanosomes could be unequivocally identified by dot blot hybridization and individual trypanosomes could be identified by in situ hybridization. We show how this simple methodology can be used in the field for the identification of immature and mature trypanosome infections in tsetse.

We have determined the molar content of guanine + cytosine (GC content) of DNA of the filarial nematode (Brugia malayi, Brugia pahangi and Dirofilaria imitis) and of the free-living soil nematodes Caenorhabditis elegans and have analysed the DNA for the presence of methylcytosine. Two independent methods, thermal denaturation and direct analysis of base content by HPLC following enzymatic hydrolysis, reveal that the GC content of filarial nematodes is 26–28%. We have been unable to find methylcytosine in the DNA of B. malayi.

A FMRFamide-like peptide has been detected in the nematode Ascaris suum, using the peroxidase-anti-peroxidase (PAP) immunocytochemical technique. Positive reactions were obtained in both the central nervous system and the peripheral nervous system of the worm, the strongest reactions being in the anterior nerve ring, the cephalic papillary ganglia, the lateral ganglia and the dorso-rectal ganglion. Immunoreactivity was observed along the length of the main nerve cords of the worm and, to a lesser extent, in the pharyngeal nerve cords. The possible role of this neuropeptide in the physiology of the nematode is discussed.

The course of primary and secondary infections with Trichostrongylus tenuis in the domestic chicken was investigated. Primary infections were established after the administration of single and trickle doses of infective-stage larvae. The worm burden in the caeca was highest after a single dose of 500 infective-stage larvae; this gave a mean of 87 nematodes per bird on days 8–9 of infection, 20 nematodes on day 14 and 0 on day 28 of infection. Following trickle doses of 60, 100, 200, 300, 400 or 500 infective-stage larvae, there was a rise and then a fall in nematode egg output in all groups. In chickens given a primary dose of 500 infective-stage larvae followed 30 days later by a single secondary dose of 500 infective-stage larvae, the mean worm burden during the secondary infection rose to 57 nematodes on day 9 of infection and then fell rapidly to 18 nematodes on day 15 and to 2 on day 30. Scanning electron microscopy showed changes in the caeca of infected birds, with the caecal surface being covered in a layer of mucus from 12 days after infection. Balls of blood-stained mucus containing nematodes were observed in the caecal droppings from day 9 of infection onwards. It is concluded that chickens rapidly expel an established infection of T. tenuis, unlike the normal host, the red grouse.

The purpose of this study was to determine whether predisposition to Heligmosomoides polygyrus and Aspiculuris tetraptera (Nematoda) exists within a naturally infected population of mice. A breeding mouse population was housed in a spacious arena in which endemic infections of H. polygyrus and A. tetraptera were present. H. polygyrus were over-dispersed in the mouse population. Prevalence reached 100% by the age of 3 weeks; intensity of infection increased to a peak in the 10 to 15-week-old mice, and remained high throughout life. A group of 73 mice was treated with pyrantel pamoate, and the expelled worms were counted. Mice were returned to the arena. Daily egg production was monitored 4, 8, 12 and 14 weeks after treatment. Mice were then killed and numbers of H. polygyrus and A. tetraptera were counted. Significant positive correlations were detected between numbers of H. polygyrus at first treatment and at necropsy, indicating the existence of predisposition. Similar results were obtained for A. tetraptera. Correlations improved when data were analysed by age class of mice. Analyses based on egg-count data during reinfection did not support the hypothesis of predisposition, however. A. tetraptera and H. polygyrus burdens were significantly correlated only in 3 to 4-week-old mice at the time of the treatment.

A continuous succession of egg capsules is formed in the ootype of Protopolystoma xenopodis and released into the urinary bladder of the host, Xenopus laevis. Since egg capsules are passed out with the aquatic toad's urine several times a day, the rate of parasite egg production can be recorded accurately by direct counts. Temperature has a marked influence on the rate of egg output which doubles in response to a rise from 17 to 25 °C. Under constant temperature conditions regular egg collections from isolated toads infested with single Protopolystoma revealed that rates of production remain stable for periods of several weeks. The overall mean rate of egg production was 8·9 eggs/worm/day (e.w.d.) at 20·0±0·5 °C (based on 5 consecutive egg collections from 58 individual parasites). Significant differences were observed, however, between the mean rates of output from different hosts, ranging from 2·1 e.w.d. (s.d. = 0·6) to 17·7 e.w.d. (S.D. = 3·2). Neither differences in worm age nor host-factors could explain the observed range in output. Worms transferred to new hosts resumed egg production at pre-transfer rates, suggesting inherent differences in the capacity for egg production by individual parasites. Total production increased with increasing worm burden, but output per parasite was significantly reduced in groups of 2, 3 or 4. Whilst competition for some finite resource provides one interpretation, this observation could equally reflect a statistical phenomenon associated with the reduced chance of encountering groups of individuals which are heterogeneous for egg production as worm burden increases and sample size decreases.

The effects of duration of infection and of temperature and nourishment in cultures on development of free-living generations of Strongyloides stercoralis were studied quantitatively. Rhabditiform larvae, 228–353 μm long, were collected from infected dogs with or without prednisolone treatment using the Baermann apparatus. Cultures were carried out by the filter paper test-tube method under the following conditions: incubation temperature 15–40°C and faecal dilution 1:0–1:16. Rhabditiform larvae developed predominantly to free-living females at incubation temperatures of 15–30°C and low faecal dilutions, but filariform larvae appeared mainly under extreme conditions such as high temperature. Recovery rates of filariform larvae were inversely related to those of females. It was remarkable that high temperature, but not low faecal dilution, affected development of filariform larvae. Although the appearance of free-living males was constant in various environmental conditions, the present study indicated an increase in free-living males with the duration of infection. Thus, it seems that free-living males are already fixed as male in the egg stage, and potential female rhabditiform larvae differentiate into free-living females or filariform larvae depending on environmental factors. There is no marked difference in the development of rhabditiform larvae into filariform larvae in either the immunosuppressed dog or the intact dog.

Using biochemical genetic methods, we have distinguished 2 sibling species in the complex Bothrimonus nylandicus (Schneider, 1902), which infest 2 congeneric species of sole (Solea lascaris and Solea impar) on European coasts (Atlantic and Mediterranean). Neither of the parasite species is specific for either of the sole species, but one of them is present all year round, whereas the other is absent in the autumn and winter and only appears in the spring, subsequently disappearing at the end of the summer. Only S. impar lives in the Mediterranean, and is equally infested by both cestodes, whereas both species occur in the Atlantic and each of them is preferentially infested by 1 species of cestode. The shortness of the adult stage of the parasite in the definitive host and the presence of 2 life-cycles associated with competition between the 2 hosts in the Atlantic could be responsible for the biological differences observed and for maintaining the sibling species in sympatry.

The major pathways for cysteine catabolism in Hymenolepis diminuta have been investigated. The parasite has an active cystathionine-β-synthase and, as in other tissues, this enzyme has a wide substrate specificity. However, the enzyme from H. diminuta differs significantly from the mammalian enzyme in showing a high serine sulphydrase activity and a high serine lyase activity. There was only low γ-cystathionase activity in H. diminuta and again the enzyme showed a range of substrate specificities. Cysteine aminotransferase activity was readily demonstrated in the tapeworm, but there was no evidence for 3-mercaptopyruvate sulphotransferase activity. An oxidative pathway for cysteine catabolism in H. diminuta was shown by the presence of cysteine dioxygenase and cysteine sulphinate transaminase. The properties of the helminth cysteine dioxygenase were very similar to those of rat liver. H. diminuta was able to reduce cystine to cysteine via a glutathione-cysteine transhydrogenase system.

Total DNAs, isolated from a range of taeniid cestodes (Taenia solium, T. saginata, T. pisiformis, T. crassiceps, T. hydatigena, T. ovis, T. multiceps and T. taeniaeformis), have been subjected to restriction enzyme digestion, Southern transfer and hybridization analysis using cloned fragments of the ribosomal RNA gene of Schistosoma mansoni. Substantial inter-specific genetic differences have been revealed on the basis of characteristic hybridization patterns for each of the taeniid cestode species. Furthermore, a random genomic DNA library has been constructed in the vector plasmid pAT153 using DNA extracted from a pig isolate (Indian origin) of T. solium. A panel of taeniid cestode DNAs including DNA from Echinococcus granulosus, has been used in conjunction with hybridization and restriction enzyme analysis to identify in the library a single recombinant plasmid with a T. solium-specific insert (coded pTS10) and two recombinant plasmids with T. solium inserts having selective specificities for T. solium and T. ovis (coded pTS17) and T. solium, T. saginata, T. ovis and T. multiceps (coded pTS28). These recombinant plasmids and the cloned fragments of the ribosomal RNA gene of S. mansoni have been used in restriction endonuclease, Southern transfer and hybridization analysis to detect intra-specific genetic variation in cysticerci of T. solium from India, Mexico and Zimbabwe. In addition, pTS10 and pTS17 have been used in a simple dot-blot assay to distinguish T. solium from T. saginata.

An official control programme against Echinococcus granulosus and Taenia hydatigena has been in operation in New Zealand for more than 28 years and against Taenia ovis for more than 18 years. This unique effort to control three metazoan parasites at the same time has led to a change from endemic to extinction status for E. granulosus but only a change from hyperendemic to endemic status for T. hydatigena and T. ovis. This has presented problems in determining the most cost-effective future control strategies. To facilitate this, a benefit/cost analysis of 20 options for the combined control of E. granulosus, T. hydatigena and T. ovis in New Zealand was undertaken. This showed that for E. granulosus a future change from the current non-targeted to a targeted approach is strongly indicated. For T. ovis 6 options were cost-effective using a discount rate of 10%. These were (1) a targeted control package using a vaccine in the non-targeted attack phase; (2) a targeted control package using a larvicide in the attack phase; (3) the transfer of all losses due to and responsibility for the control of T. ovis to the producer who administers a larvicide to sheep to be killed for dog food; (4) the transfer of all losses due to and responsibility for the control of T. ovis to the producer who administers praziquantel every 6 weeks to dogs; (5) and (6) two options involving the discontinuation of control. Control of T. hydatigena was assumed to be an incidental outcome of the policies for the other two parasites.

A general mathematical model of a vector-borne disease involving two vertebrate host species and one insect vector species is described. The model is easily extended to other situations involving more than two hosts and one vector species. The model, which was developed from the single-host model for malaria described by Aron & May (1982), is applied to the African trypanosomiases and allows for incubation and immune periods in the two host species and for variable efficiency of transmission of different trypanosome species from the vertebrates to the vectors and vice versa. Equations are derived for equilibrium disease prevalence in each of the species involved. Model predictions are examined by 3-dimensional phase-plane analysis, which is presented as a simple extension of the 2-dimensional phase-plane analysis of the malaria model. Parameter values appropriate for the African trypanosomiases are derived from the literature, and a typical West African village situation is considered, with 300 humans, 50 domestic animals and an average population of 5000 tsetse flies. The model predicts equilibrium prevalences of Trypanosoma vivax, T. congolense and T. brucei of 47·0, 45·8 and 28·7% respectively in the animal hosts, 24·2, 3·4 and 0·15% in the tsetse vectors, and a 7·0% infection of humans with human-infective T. brucei. The contribution to the basic rate of reproduction of the human-infective T. brucei is only 0·11 from the human hosts and 2·54 from the animal hosts, indicating that in the situation modelled human sleeping sickness cannot be maintained in the human hosts alone. The animal reservoir is therefore crucial in determining not only the continued occurrence of the disease in humans, but its prevalence in these hosts as well. The effect of changing average fly density on equilibrium disease prevalences is examined, together with the effect of seasonal changes in fly numbers on disease incidence. In a seasonal situation changes in fly mortality rates affect both future population size and infection rate. Peak disease incidence lags behind peak fly numbers, and that in the less favoured host lags behind that in the more favoured host. Near the threshold fly density for disease transmission disease incidence is more changeable than at higher fly densities and may even exceed equilibrium prevalence at the same average fly density (because most hosts are susceptible at the time that fly numbers begin their annual increase). The implications of the model for disease control are discussed. Identifying the precise role of the animal reservoir may suggest that treating such animals will achieve a greater reduction of human sleeping sickness than direct treatment of the humans alone. Statistically significant results of control campaigns may also be more easily shown by monitoring the non-human reservoirs. The model provides a means by which a correct perspective view can be obtained of the complex epidemiology and epizootiology of the African trypanosomiases.