The biosynthesis and fate of 4 different dense granule proteins of Toxoplasma gondii were studied with 3 monoclonal antibodies raised against tachyzoites and 1 polyclonal antibody raised against a recombinant protein. These proteins have the following molecular weights: 27 kDa (GRA 1), 28 kDa (GRA 2), 30 kDa (GRA 3) and 40 kDa (GRA 4). All four proteins were found in dense granules by immunoelectron microscopy; in T. gondii-infected cells, they were found in the vacuolar network but, in addition, GRA 3 was also detected on the parasitophorous vacuole membrane. Therefore, dense granule contents undergo differential targeting when exocytosed in the parasitophorous vacuole. Metabolic labelling and immunoprecipitation showed that GRA 2 and GRA 3 were processed from lower molecular weight precursors, and that GRA 2 and GRA 4 incorporated [3H] glucosamine and are thus likely to be glycosylated.

A specific enzyme-linked immunosorbent assay, based on a 48 kDa metallo-proteinase isolated from the frontal glands of a porocephalid pentastomid (Porocephalus crotali) in rat intermediate hosts, is described. SDS–PAGE analysis of whole nymphal homogenate, followed by Western blotting and probing with immune rat serum, revealed that only 2 protein bands label consistently; these are the 48 kDa metallo-proteinase and a 150 kDa metallo-proteinase from the body fluid, but only the former labels strongly. Furthermore this response is detected earlier and persists much longer. These characteristics uniquely qualify the 48 kDa metallo-proteinase as a diagnostic antigen. We review the literature concerning the related porocephalid Linguatula serrata from the nasal cavities of domestic dogs, which can zoonotically infect man principally through the ingestion of eggs. We conclude that the high prevalences in dogs in certain areas, most notably the Middle East, must inevitably signal significant human infections. Our ELISA can be readily adapted for the serodiagnosis of linguatulosis and this should establish whether our prediction is true.

A comparison was made of the long-term impact of different methods of administration of chemotherapy (oxamniquine, 30 mg/kg in divided doses; or praziquantel, 40 mg/kg) on prevalence and intensity of Schistosoma mansoni infection in four areas in Kangundo Location, Machakos District, Kenya. In Area A, treatment was offered in October 1983 and again in April 1985 to all infected individuals. In Area H, treatment was offered in April 1985 to individuals excreting ≥ 100 eggs per gram (epg) of faeces. In Area H, treatment was offered in April 1985 to all infected school children, within the framework of the primary schools. In the witness area, Area W, treatment was given in April 1985, for ethical reasons, to a small number of individuals excreting ≥ 800 epg. Prevalence and intensities of infection were subsequently monitored at yearly intervals for three complete post-treatment years. In the Area S schools, clinical examination was also carried out at yearly intervals. Treatment of all infected individuals on two occasions (Area A) was the most effective and long-lasting way of reducing prevalence and intensity of infection. In this area, however, some earlier interventions had been carried out and pre-treatment intensities were lower than in the other areas. Treatment only of infected schoolchildren (Area S) also had a marked and prolonged effect, comparable to or better than treatment of individuals with heavy infections (Area H). Treatment of infected schoolchildren also caused a persistent reduction in the prevalence of hepatomegaly, and there was suggestive evidence from intensities of infection in community stool surveys (but not from incidence rates) of an effect on transmission. In all study areas, reinfection was most rapid and most intense among children. These findings are discussed in the light of theoretical considerations and of results from other studies, both on schistosomiasis and on intestinal helminths. We conclude that, in areas of low morbidity such as Kangundo, chemotherapy of schoolchildren only, at intervals of up to 3 years, is a satisfactory way of producing a long-term reduction in both intensity of infection and morbidity.

The thermal preferences of two strains of the snail Biomphalaria glabrata, one resistant to, and one susceptible to, the parasite Schistosoma mansoni were determined in an aquatic thermal gradient. Snails were tested without exposure to the parasite, and 2 h and 5 weeks after exposure to trematode miracidia. The mean temperature selected by susceptible strain snails 2 h post-exposure tended to be lower than that of unexposed controls, although this was not statistically significant. In this strain, at 5 weeks post-exposure, the preferred temperature dropped by 1.9±0.5°C. The resistant strain displayed a significant drop of 1.8±0.6°C 2 h post-exposure. These results are consistent with the hypothesis that a drop in mean temperatures selected by snails is due to altered levels of endogenous cytokines such as IL-1 or TNF in association with parasite activation of the snail internal defense system.

Variations in the amount of water contact made by individuals and in the amount of water contact made at different sites may have significant impacts on patterns of human schistosome infection. Previous studies have reported variations in the rate of water contact and differences in the sites used between age/sex classes, but there is limited information on variations in individual water contact behaviour. In this paper we report and analyse observations of essentially all water contacts made over a two week period by all individuals in a rural community in eastern Zimbabwe. The mean rate of water contact was 0.43 contacts/person/day. These data were over-dispersed, ranging from zero to 3.3 contacts/person/day; 90% of contacts were made by only 37% of the population. Contact rates were related to age (highest in 8 to 10-year-olds) but not sex, with substantial variation unaccounted for by these variables. Age and sex classes differed in types of water-related activities and the time of day of contact. A greater diversity of sites was used by children than by adults and by males than by females. Individual contact rates were correlated with intensities of infection, although the risk of infection per contact was estimated to be highest in 2 to 4-year-old children and higher for males than females. Five contact sites were used during the study period, with more than 50% of contacts occurring at just 2 sites. Different age and sex classes used different sites and there were additional site-related differences in types of activity and the time of day of use. The implications of these water contact patterns for schistosome epidemiology are discussed. In particular the results provide strong quantitative support for control programmes aimed at heavily used sites (e.g. focal mollusciciding) or at the minority of individuals making most water contacts (e.g. targeted chemotherapy).

In haemolymph of Lymnaea stagnalis, parasitized with the digenetic trematode parasite Trichobilharzia ocellata, a neuropeptide (schistosomin) occurs which antagonizes female gonadotropic hormones, e.g. calfluxin (CaFl). By means of an ultracytochemical hormone-assay, the CaFl assay, it was demonstrated that the occurrence of schistosomin is a general phenomenon in schistosome-infected freshwater snails. Haemolymph of the schistosomiasis-transmitting snail species Biomphalaria glabrata and B. pfeifferi, parasitized with Schistosoma mansoni, also appeared to contain an antagonizing factor, i.e. schistosomin. In contrast, in haemolymph of L. stagnalis parasitized with Diplostomum spathaceum (Diplostomatidae) no schistosomin could be found. This suggests that schistosomin may only occur in snails infected with parasites belonging to the Schistosomatidae. The effect of schistosomin is rather specific. Haemolymph of B. glabrata parasitized with S. mansoni had not the capacity to inhibit the response to CaFl in the target organs for CaFl, the albumen glands of L. stagnalis and Bulinus truncatus. The same holds true for haemolymph of infected L. stagnalis: it did not inhibit the CaFl response in glands of B. glabrata and B. truncatus and even not in those of a related species (L. ovata). Schistosomins in haemolymph of infected B. glabrata and B. pfeifferi, on the other hand, seem more related. Both appeared to inhibit the hormone response in glands of the two Biomphalaria species studied. The results indicate that schistosomin in haemolymph of schistosome-infected pulmonate snails, although functionally related, may differ structurally.

The inheritance of variant surface antigens in Trypanosoma brucei has been determined by identifying variable antigen types (VATs) in each of two cloned parental stocks and then examining the presence and abundance of these VATs in hybrid progeny produced when these stocks undergo genetic exchange during co-transmission through tsetse flies. Nine VATs have been identified from the repertoire of the parental stock STIB 247L and 5 VATs have been identified from the parental stock STIB 386AA; the identified VATs were exclusive to each stock. Their inheritance was elucidated using two assays. In the first, repertoire antisera (RAS) containing antibody specificities to many different VATs were raised in rabbits to the 2 parental stocks and 6 progeny clones. The presence of VAT-specific antibodies in these RAS was then determined by antibody-dependent complement-mediated lysis. In the second assay, the 2 parental stocks and 4 hybrid progeny clones were each independently transmitted through tsetse flies and VATs observed using VAT-specific antisera in indirect immunofluorescence of metacyclic trypanosomes and in bloodstream forms of fly-bitten mice. The results from both assays showed that (1) both metacyclic- and bloodstream-VATs were inherited into the progeny, (2) each hybrid progeny clone contained some VATs from both parents, (3) hybrids did not express all the VATs from either parent, (4) there was little apparent pattern as to which VATs had been inherited and which had not and (5) the VAT repertoires of the hybrid progeny appeared to be larger than those of the parents. In addition, two results indicated that control of VAT expression remains unaltered after genetic exchange. Firstly, the immunofluorescence results showed that VATs present in hybrid trypanosomes were expressed at the same stage during an infection and at approximately the same prevalence as in the parent. Secondly, a double-labelling experiment using direct immunofluorescence indicated that individual hybrid trypanosomes did not generally simultaneously express more than one VAT. Taken together, these results demonstrate that recombinant VAT repertoires are created when trypanosomes undergo genetic exchange and that genetic exchange is a mechanism whereby the generation of new serodemes can occur.

The major glycoprotein of the sheath of Litomosoides carinii microfilariae (gp22) was analysed for its amino acid and amino sugar composition. It is rich in proline, glutamine/glutamic acid and glycine and contains (N-acetyl)galactosamine. The N-terminal amino acid sequence was determined up to position 37. It consists of a group of 6 repeats of the pentapeptide sequence methionine-glycine-proline-glutamine-proline with two minor modifications in repeats 3–6, while the first two repeats follow the general pattern more loosely. Identical N-terminal amino acid sequences were found in at least two other sheath polypeptides (33 kDa, 39 kDa). Antisera prepared against 3 overlapping synthetic peptides corresponding to the amino terminus of gp22 recognized different epitopes. They all reacted with identical patterns of sheath polypeptides. The antisera failed to recognize antigens of 4th-stage larvae of L. carinii. In contrast, cross-reacting epitopes were detected in other parasite stages. Antisera reacted with material surrounding embryos and microfilariae in the uterus of females, and caused patchy fluorescence on the sheath of blood-derived and in vitro-released microfilariae.

Although the relationship between intestinal parasitism, the ingestion and use of energy, and host survival is expected, little work has been done to outline the effect of such organisms upon their host's nutritional requirements in an ecological context. This study is the first to demonstrate that an intestinal helminth previously reported to be of little or no histopathological consequence, Plagiorhynchus cylindraceus, has a significant detrimental impact upon the flow of food energy through a definitive host, the European starling, Sturnus vulgaris. Within both male and female adult European starlings reductions in standard metabolic rates occurred as the result of initial infection, indicating that the host's basal metabolism/thermal regulatory abilities were altered. Moreover, initially infected male starlings, but not females, had an increased consumption and excretion of energy and maintained lower average daily body weights versus controls when temperature stressed. These results appear to be due to either a parasite-mediated alteration in host activity and/or to the disruption of host-digestive abilities. Additionally, these data indicate that, overall, male and female S. vulgaris respond differently to infection and that intestinal helminths normally thought to be of little or no pathological consequence to the host are factors that should be addressed in future studies regarding animal energetics, ecology, and behaviour.

The relationship between host dietary protein and intestinal helminth population biology during primary infection was studied using a mouse–T. muris experimental model. Uninfected CBA/Ca mice fed either a 4% (by mass) or 16% protein diet grew at a similar rate, but both suffered significant growth retardation when infected with T. muris. T. muris initial establishment and fecundity were unaffected by dietary protein content, but were significantly influenced by infection dose, declining at the higher level of infection. Dietary protein deficiency increased parasite survival at high infection levels. Heavily infected hosts fed the 16% protein diet were found to expel almost the whole of their infections around day 21 p.i. By contrast, this response appeared defective in hosts fed the lower 4% protein diet, resulting in the establishment and survival of large, chronic and patent infections. Mean faecal egg output per mouse increased with infection dose in animals fed a low protein diet, but was low and unrelated to infection exposure in hosts fed the 16% protein diet. It is concluded that low protein levels in diet, via alterations in host resistance, may have an important influence on the transmission of T. muris.

Ultrastructural changes associated with exsheathment of the infective juveniles of Haemonchus contortus are described. Hyaline ring formation occurs associated with annulations 10 and 20 and consists of the dissolution of the basal zone and the inner electron-dense layer resulting in the rupture of the sheath. These changes are consistent with the action of a protease-exsheathing enzyme. There is a significant reduction associated with exsheathment in the size of the excretory cells and the number of excretory granules contained within them. No changes in the oesophagus were observed associated with exsheathment. These, and the observations of earlier workers, suggest that it is the excretory cells and not the oesophagus which are the source of exsheathing fluid during exsheathment.

Clavibacter sp. (syn. Corynebacterium rathayi) adhered to both Anguina funesta (syn. Anguina agrostis) and Anguina tritici, but differences in the nature of adhesion were noted. Similar patterns of binding of the bacteria and of anti-wheat germ agglutinin antibody initially led us to believe that the mechanism of bacterial adhesion was related to the presence of wheat-germ agglutinin (WGA) on the outer cuticle of both species of nematodes and its complementary carbohydrate on the bacterial capsule. However, treatment of either species of nematode with sodium metaperiodate inhibited bacterial adhesion but not the binding of anti-WGA antibody. Bacterial adhesion, therefore, is not mediated by WGA on the nematodes' surface. Moreover, differences in patterns of bacterial adhesion to Anguina species, both before and after treatments with NaCl and detergents, suggest basic interspecific differences in the nature of adhesion. Electron microscopy confirmed the contribution of the nematodes' cuticular surface coat (SC) to the process of adhesion, but it is still not clear how the SC interacts with the bacterial capsule or which of its components are involved. While complete removal of the SC with periodate prevented bacterial adhesion, juveniles that naturally resisted bacterial adhesion did not lack a SC. One explanation could be that the SC of individuals, to which bacteria do not adhere naturally, lacks crucial components that cannot be defined by conventional EM.

BALB/c mice were characterized as more resistant to infection with Heligmosomoides polygyrus (Nematoda) than C57BL/6 mice based on lower establishment and survival during a primary infection and stronger protection induced by an immunizing regime. It was hypothesized, therefore, that C57BL/6 mice would be more heavily infected than BALB/c mice when they lived together as a single population in a large indoor arena where transmission occurred through contact between the mice and damp peat trays where parasite eggs developed into larvae. Fifty female mice (including 5 infected mice) of each strain were placed in a 3.2 x 0.8 m arena. Net egg production and numbers of larvae acquired by sentinel mice of each strain were monitored every two weeks. The experiment was replicated twice. The results did not support the hypothesis. No difference was detected between strains of mice in the number of larvae acquired by sentinel mice during 24 h exposure periods, or in the numbers of worms present after 12 or 23 weeks. Net egg production was also comparable between strains. A hypothesis that the unexpected similarity of infection in the two strains was related to differences in rates of contact with the peat trays was not supported by preliminary data on mouse behaviour that revealed equal frequency of contact with peat trays between strains. A second hypothesis that continuous exposure to larvae led to similar infection levels in the two strains (in contrast to the controlled characterization experiments) was also unsupported. Mice were infected weekly with 10, 50 or 100 larvae for 5 or 10 weeks. Net egg production and numbers of worms were consistently higher in C57BL/6 than BALB/c mice. At this time it is not clear why infection in the two strains was virtually identical in the large arenas but clearly distinct in all controlled infection experiments.

The hypothesis that a liver-inhabiting nematode, Capillaria hepatica, can regulate abundance of the house mouse (Mus domesticus) was tested in enclosures, in southeastern Australia. Changes in mouse abundance, and the relationship between mortality and host abundance were compared in three treatment and three control populations. Any effect of C. hepatica on mouse abundance was masked by an unknown regulating factor(s). This factor(s) caused density-dependent mortality in the control and treatment populations and, together with strong seasonal trends in parasite transmission, confounded our test of the ability of C. hepatica to regulate the abundance of house mice. The seasonal trends in transmission have important implications for the potential of this parasite as a biological control agent and for models of the interaction of C. hepatica and mouse populations. Transmission of C. hepatica apparently occurred throughout the 18 month study, further supporting its potential as a biological agent in the control of mouse plagues in this region.

Infective-stage larvae of three different isolates of Toxocara canis were intrinsically ([35S]methionine) labelled in culture, to determine the presence of similarities or differences in the somatic and ES antigens expressed between larvae derived from different hosts and different geographical regions. Two other closely related ascaridids, Toxascaris leonina which infects cats and dogs, and Toxocara vitulorum (Neoascaris vitulorum) which infects cattle, were also compared to T. canis larvae by this method. Overall comparisons were made by 1- and 2-dimensional electrophoresis, while immunological cross-reactivities between the different species were analysed by radio-immunoprecipitation. Our results show that extensive physicochemical characteristics are shared between T. canis isolates, both from different hosts and different geographical locations. A substantial overlap was revealed when T. canis and T. vitulorum antigens were compared, whereas Toxascaris was found to produce a distinct antigen profile: this result was independent of whether methionine- or Iodogen-labelled products were being considered. Antigen recognition by polyclonal antibodies raised to all three species and to the cat ascaridid Toxocara cati, revealed considerable cross-reactivities. The cross-reactions were especially prominent between the Toxocara species, a fact further substantiated when reactivity of T. canis ES-specific monoclonal antibodies were tested against T. leonina and T. vitulorum antigens. The ES antigens of T. leonina were not recognized by the T. canis monoclonals, whereas the majority of these antibodies precipitated antigens of T. vitulorum. One which did not react with T. vitulorum was monoclonal antibody Tcn 2, indicating its species-specific reactivity and therefore its potential for the specific diagnosis of human toxocariasis.

Infectivity of Trichinella pseudospiralis in Chinese hamsters was greater than that of Trichinella spiralis. Concurrent infection with these two species of parasite was accompanied by an increase in infectivity of T. spiralis. The infectivity of T. pseudospiralis, but not that of T. spiralis, was significantly influenced by host age. Regardless of host age, T. pseudospiralis was more infective for the Chinese hamster than was T. spiralis. Early during infection, enteritis (myeloperoxidase activity) in hamsters infected with T. pseudospiralis alone was significantly less than that seen in animals infected with T. spiralis alone or with both species concurrently. Myositis in hamsters infected with T. pseudospiralis alone was significantly less on days 14, 17 and 25 post-infection (p.i.) than that observed in concurrently infected or T. spiralis-infected animals. On days 11 and 14 p.i. inflammation in diaphragm muscle from concurrently infected hamsters was significantly less than that seen in animals infected with T. spiralis alone. Muscle histology supported these chemical data on myositis. Suppression of the host inflammatory response against early muscle-stage larvae of T. pseudospiralis underlies the greater infectivity of this parasite for the Chinese hamster.

The cercariae of Cryptocotyle lingua have a brief but active life during which they do not feed. In order to maximize the probability of infection they must respond to a variety of host-related stimuli yet conserve their finite energy reserves by minimizing unnecessary muscular activity. In laboratory experiments, simulated shadows representing the passage of a fish host were found to increase population activity levels. Evidence was also found for a relationship between shadow duration and the duration of subsequent bursts of swimming activity. Adaptation to continuous shadowing was recorded, its onset and magnitude being dependent on both the frequency and duration of shadows delivered. As cercariae aged they became less responsive to shadow stimuli, especially those of short duration.

This investigation reports for the first time the establishment of a trematode life-cycle in the laboratory using in ovo cultivation of the adult stage. Microphallus primas metacercarial cysts were removed from Carcinus maenas crabs and chemically excysted. Using a modification of the technique developed by Fried (1962) the released metacercariae were grown in fertile hen's eggs to ovigerous adult flukes. These were removed from the under surface of the chorio-allantoic membrane of chick embryos 5 days post-infection and the fluke eggs recovered were used to infect laboratory-reared Hydrobia ulvae snails. When challenged against parasite-free crabs the cercariae released from the experimentally infected snails gave rise to metacercarial cysts that were again identified as M. primas.