The protozoan parasite Toxoplasma gondii produces a family of microneme proteins that are thought to play diverse roles in aiding the parasite's intracellular existence. Among these, TgMIC2 has a putative function in parasite adhesion to the host cell to initiate the invasion process. The invasion process may be localized and inhibited by monoclonal antibodies against the protein(s) involved. Here we report on the construction of a phage-displayed single-chain variable fragment (scFv) library from mice immunized with whole T. gondii parasites. The library was subsequently panned against recombinant TgMIC2 (rpTgMIC2) and 2 different groups of antibody clones were obtained, based on fingerprinting and sequencing data. The expressed recombinant scFv antibody was able to recognize rpTgMIC2 in a Western blot detection experiment. These results show that the phage display technology allows quick and effective production of monoclonal antibodies against parasite antigens. By panning the scFv-displayed library, we should be able to obtain a plethora of multi-functional scFv antibodies towards T. gondii proteins.

In the present study we have used antibodies to Leishmania major promastigote antigens which were eluted from a glutathione-agarose column (LmGbp) and could identify several parasite components among different Leishmania species by using immunoprecipitation and Western blot techniques. The results also showed that some of LmGbp are present among the molecules released into the culture medium. Moreover, immunofluorescence assays clearly demonstrated that LmGbp are expressed by intracellular amastigotes. The electron micrographs of thawed cryosections of L. major-infected cells revealed that the antigens were associated with the membrane of the phagocytic vacuole. Moreover, the Western blot technique allowed us to identify, using other Leishmania species extracts and anti-LmGbp antibodies, a major polypeptide of an apparent molecular mass of 66 kDa. Immunofluorescence studies suggested that the 66 kDa polypeptide is associated with intracytoplasmic vesicles. Cryosections of Leishmania promastigotes improved the fine structure preservation of the organelles and enabled a number of features to be seen, particularly the structures considered as vesicles, which appeared as a complex tubulo-vesicular structure resembling mammalian cell endosomes and Leishmania organelles previously named ‘megasomes’. Further studies using antibodies against the native 66 kDa protein will be needed to investigate the localization of the protein at the ultrastructural level and to follow its intracellular vesicular traffic.

Amblyomma dissimile Koch is well known to science and previous accounts of its biology have already appeared. Newstead (1909) made some observations on its habits in Jamaica. Hooker, Bishopp, and Wood (1912) give an exact account of its life cycle worked out in confinement in the Southern United States of America. These authors do not refer to the occurrence of parthenogenesis in this species.

An attempt was made to evaluate the immunoprophylactic efficacy of Brugia malayi transglutaminase (BmTGA) as a DNA vaccine, for human lymphatic filariasis. BmTGA was cloned and characterized in the DNA vaccine vector pVAX1. Further, the tissue distribution study of the DNA construct, pVAX-TGA was carried out in mice and the DNA vaccine was shown to be efficiently distributed to all the organs, was accessible to the immune system, and at the same time was metabolized quickly and did not pose problems of toxicity. Intramuscular immunization in mice showed significant antibody production and splenocyte proliferation upon antigenic stimulation. The immune responses were biased towards the Th1 arm, as evaluated in terms of isotype antibody distribution and cytokine profile. Thus, analysis of the humoral and cellular immune responses indicated that BmTGA is a potent immunogen. However, protection studies as determined by the micropore chamber method using live microfilarial larvae, showed that the DNA vaccine could confer only partial protection in the mouse model. We conclude that despite the induction of sufficient humoral and cellular immune responses, BmTGA as a DNA vaccine could not confer much protection against subsequent challenge and other aspects of the immune responses need to be further investigated.

1. Observations have been made on the life cycles of Cercaria paracauda, Iles, G. tetraglandis, Iles, and C. Apatemon gracilis minor, Yamaguti. The development of the tetracotyle metacercaria of C. tetraglandis has been followed and also that of the metacercaria and adult of A. g. minor. The pathology and distribution of the metacercariae of the last two species within the leech, Erpobdella testacea (Sav.) has been studied.

2. It was noted that development of the metacercariae within the same host and resulting from the same infection was not simultaneous and that it was improbable that a 100 % encystment of the metacercariae would occur. Temperature was found to have a marked effect on the length of time required for encystment.

3. Data are given on the infectivity and excystation of the metacercariae of A. g. minor and on the biology and morphology of the adult of this subspecies.

4. Stress is placed on the importance, in the systematics of the digenetic trematodes, of considering both the morphological and biological characters of the adult and those of the related larval stages.

To test the homogeneity of 18S sequences within Trypanosoma cruzi, riboprint profiles were separated by temperature gradient gel electrophoresis (TGGE). Upon interpretation of melting curves of fragments within a riboprint profile, there appeared to be two 18S sequence types within each stock examined. Two similar types were also observed within outgroup taxa Trypanosoma conorhini, Trypanosoma rangeli and Leishmania braziliensis. From DNA hybridization studies, these fragments were shown to have homology to the 18S V1 region. There are therefore two 18S V1 regions, differing in sequence, present in all taxa examined. When only a single 18S sequence is used to represent each taxa for phylogenetic inference, comparisons may be between paralogous and not orthologous copies of this region, such that, inferred relationships may merely reflect a gene history. This seriously questions the current molecular phylogeny of these protozoa using 18S data.

The study was designed to determine comparatively the prognostic value of immunoblotting and ELISA in the serological follow-up of young cystic echinococcosis (CE) patients exhibiting either a cured or a progredient (non-cured) course of disease after treatment. A total of 54 patients (mean age 9 years, range from 3 to 15 years) with surgically, radiologically and/or histologically proven CE were studied for a period up to 60 months after surgery. Additionally, some of the patients underwent chemotherapy. Based on the clinical course and outcome, as well as on imaging findings, patients were clustered into 2 groups of either cured (CCE), or non-cured (NCCE) CE patients. ELISA showed a high rate of seropositivity 4 to 5 years post-surgery for both CCE (57·1%) and NCCE (100%) patients, the difference found between the two groups was statistically not significant. Immunoblotting based upon recognition of AgB subcomponents (8 and 16 kDa bands) showed a decrease of respective antibody reactivities after 4 years post-surgery. Only sera from 14·3% of CCE patients recognized the subcomponents of AgB after 4 years, while none (0%) of these sera was still reactive at 5 years post-surgery. At variance, immunoblotting remained positive for AgB subcomponents in 100% of the NCCE cases as tested between 4 and 5 years after surgical treatment. Immunoblotting therefore proved to be a useful approach for monitoring post-surgical follow-ups of human CCE and NCCE in young patients when based upon the recognition of AgB subcomponents.

Regional differences in the tegumentary tissue of Moniezia expansa, as revealed at the light-microscope level by histological and histochemical techniques, are described and evidence for secretory activity by the interproglottidal glands is presented.

In very immature proglottides the interproglottidal glands are at the ‘precryptic’ stage. Gland cells may be differentiated from other tegumentary cells by their high RNA content and in certain gland cells the presence of an alcian blue staining material.

In mature proglottides the glands consist of rosette-like clusters of cells around crypt-like intuckings of the tegument. Two types of cells are found in the gland, small alcian blue-staining cells which are most numerous in the neck region of the crypt, and larger cells, the predominant gland cells, which do not stain with alcian blue but possess non-specific esterase activity. No other tegumentary cells in Moniezia exhibit this activity. Esterase and phosphatase activity is found in the tegument and crypt of the glands and in the interproglottidal folds.

The non-enzyme histochemistry confirms and extends the observations of previous workers.

Cytochrome oxidase and succinic dehydrogenase were detected in the tegumentary cells and tegument. Very strong reactions were given in the neck and scolex, with a progressive diminution of activity posteriorly along the strobila. Very low activities were recorded in the tegument of the glands.

The blood and broncho-alveolar leucocyte (BAL) responses, to a primary Necator americanus infection, were studied in male BALB/c and NIH mice. Following percutaneous infection, a significant blood leucocytosis occurred in both the BALB/c and NIH strains. The peak response occurred, in both strains, on day 10 p.i. and this was reflected in the eosinophil response which peaked at the same time-point. The change in blood eosinophil numbers, as a result of infection, was found to be the greatest recorded for any cell type. In BALB/c mice, however, infection elicited a significantly more intense total leucocyte, lymphocyte and eosinophil response than in NIH mice. In contrast, the BAL response was more intense in the NIH strain. Peak BAL responses were observed between days 12 and 16 p.i., in both strains, and the change in broncho-alveolar eosinophil numbers, as a result of infection, was found to be the greatest recorded for any cell type. The relationship between the observed leucocyte responses and resistance to the migrating larvae of a primary infection is discussed.

Bidirectional signalling between Trypanosoma brucei brucei and CD8+ T cells involves reciprocal action of the parasite-derived trypanosome derived lymphocyte triggering factor (TLTF) and lymphocyte-derived IFN-γ. Herein we further characterize this relationship and report quantitation of TLTF secretion into culture supernatants using a newly developed ELISA. Secretion is induced by IFN-γ in a dose–response manner, with 100 U/ml giving maximal yields of bioactive TLTF as assessed by ELISA and ELISPOT. This is a constitutive and active secretion. Furthermore, IFN-γ-induced secretion was dependent on tyrosine protein kinase activity. Specific blockers of this signalling system resulted in lower yields of TLTF in the culture supernatants.

The stages of spermatogenesis and the structure of the primary oocyte of Sphaerostoma bramae were studied in material fixed in Bouin and Flemming's fluid and in preparations treated according to the Kolatchev and the Feulgen techniques.

Chromosome counts of primary and secondary spermatocytes indicate that the diploid number is twenty-four.

The stages of spermatogenesis, in general, conform to the usual pattern of this process in digenetic trematodes. The spermatogonia, spermatocytes and early spermatids are not connected together by central strands, nor by a central disk, as is claimed for some other species. A study of sections stained in haematoxylin and of Feulgen preparations showed that the spermatozoon is composed of an elongate nucleus and a tail. It was not possible, with the methods employed during the present investigation, to determine the fine structure of the tail.

The primary ovarian oocytes are in the interphase or in early prophase of the first maturation division. There is evidence that material is extruded from the nucleus to the cytoplasm.

The Golgi elements of the male germ-cells are revealed in Kolatchev preparations as short rods and filaments. The Golgi elements of the spermatid are eliminated in the residual cytoplasm. Mitochondria, in the form of granular bodies and short rods, were visible in spermatogonia, spermatocytes and spermatids. Those of the spermatid remain in the residual cytoplasm.

Short, rod-like Golgi bodies are present in the primary oocytes. In the young cells they form a compact mass situated close to or in contact with the nuclear membrane. Later, the elements spread out through the cell. Granular and rod-shaped mitochondria are concentrated in a single mass at one pole of the nucleus or in two masses at opposite sides of the nucleus. A few mitochondria are scattered through the cytoplasm.

The differential permeability of cell membranes to various electrolytes and non-electrolytes represents one of the most fundamental characteristics of life. In the context of parasitology, differences in transport properties when parasite and host cells are compared represent an important avenue for chemotherapy. For example, a recent success in this area exploits the difference in nucleoside transport systems between host and parasite to produce combination therapy of Schistosoma japonicum by tubercidin and nitrobenzylthioinosine-5′-monophosphate (Elkouni, Diop & Cha, 1983). The morning session of this symposium was devoted to various aspects of membrane structure and function, particularly in the context of characterizing parasite transport systems. It is therefore probably worth while to present a simple classification of membrane transport systems, based on their complexity, and energy requirements. Table 1 summarizes the principal types of transport to be considered, starting with simple (Stokesian) diffusion. This will depend on permeant size, charge and lipid solubility. Classically, transmembrane transport of such molecules as aliphatic alcohols, ureas and small sugars has been studied, in simple systems such as the red cell (see Stein (1986) for references). In fact this route is important for intracellular delivery of a variety of hydrophobic molecules, for example phenylalanine benzyl ester transport as an antisickling agent (Acquaye, Young, Ellory, Grecki & Wilchek, 1983), or quin-2 and fura-2 for intracellular calcium measurements (Tsien, 1981).

To determine if, as is the case with Trypanosoma brucei and T. congolense, serodemes of T. vivax could be distinguished on the basis of immunity to the metacyclic stages of the parasite, attempts were made to immunize goats by infection with infected tsetse, followed by chemotherapy or eventual ‘self-cure’. Thirty goats were infected by tsetse with either clones or stocks of T. vivax from East or West Africa. Twenty-four goats were treated with diminazene aceturate (Berenil, Hoechst A.G.) 2–6 weeks after infection and 6 goats were allowed to self-cure. Infection, followed by treatment, induced immunity to a first homologous challenge by infected tsetse in only 2 of 24 goats (one immune to the East African stock, and the other to a clone of the West African stock). Immunity to a clone of the East African stock was induced in 3 or 4 animals after a second infection and treatment and in the fourth animal of the group following a third infection and treatment. One of 2 goats infected with the clone of the East African stock was immune to challenge at 16 weeks, following self-cure without treatment, and 1 of 4 goats infected with the parent stock was similarly immune when challenged at 40 weeks post-infection. Goats susceptible to infection with East African T. vivax showed evidence of partial immunity by delayed pre-patent periods and depressed parasitaemias after challenge. Goats infected with the relatively more virulent West African T. vivax were, however, completely susceptible to infection after homologous challenge, and showed only a slight delay in pre-patent period. A similar result was obtained in a further 8 goats primed and challenged by large numbers of tsetse (20 or 100 infected tsetse/goat) with the West African T. vivax. In further experiments using a very short treatment interval, infections following challenge were clearly shown to be the result of a lack of immunity rather than relapse following treatment. Lytic antibody activity to cultured metacyclic trypanosomes could not be detected during infection but such activity against bloodstream forms was detected after 2 weeks of infection. It is suggested that the primary reason for the erratic induction of immunity to T. vivax employing this methodology is the low number of metacyclics transmitted by infected tsetse, and thus poor antigenic stimulus encountered by goats upon tsetse challenge.

The development of in vitro culture systems that allow the maintenance, and support the development of Echinococcus, Giardia and Cryptosporidium in the laboratory have had a significant impact on their biology and taxonomy and the epidemiology of infections they cause. This short retrospective review demonstrates how radical shifts in our understanding have occurred as a result of being able to grow these organisms in culture, and how molecular tools have helped in the interpretation of such research that often reflects the observations of earlier workers.

The present report is confined to the species found in Chinese amphibians by Faust in his field studies during the period from 1920 to 1925. Two new species of the genus Acanthocephalus are described in this paper and notes on their geographical distribution and host relationships are included along with observations on the occurrence of undetermined species of encysted larval Acanthocephala in amphibian hosts.

The copepod described in this paper has been found in Strangford Lough, Co. Down, an area which has already proved rich in ascidicolous copepods of the suborder Notodelphyoida (Gotto, 1952). The present species, however, belongs to the section Poecilostoma of the suborder Cyclopoida, many members of which are associated with marine animals of several different groups. The extremes of retrogressive specialization which characterize many of the notodelphyoids are not, on the whole, so apparent in these cyclopoids, their associative habits often being inferred from general structure rather than proved occurrence with any particular host. Thus a fair number of the types described have been found unattached in dredged material, the possible host being any one of several species taken in the same haul. Associations of an apparently loose nature are therefore sufficiently frequent to render the present form of particular interest since it has so far been found exclusively in the compound ascidian Trididemnum tenerum (Verrill), in which it is localized in the extensive cloacal cavities. Further search may of course reveal alternative hosts, since the family Lichomolgidae, to which it undoubtedly belongs, has a wide range of acceptable hosts. Indeed the genus Lichomolgus is known to occur in association with sponges, polychaets, nudibranchs, gasteropods, cephalopods, holothurians and ascidians—and one species, L. agilis (Leydig) has been recorded from a score of different nudibranchs.

Surface antigens of Schistosoma haematobium were identified by 125I-surface labelling of schistosomula followed by immunoprecipitation of the solubilized, labelled surfaces. The major antigens, after electrophoresis, formed a continuous smear corresponding to a molecular weight in the range 35–24000; in addition, a 17000 antigen was also identified. These surface antigens, in contrast to somatic antigens, were species-specific, as judged by immunoprecipitation with human anti-S. mansoni serum and serum from mice vaccinated with highly irradiated S. mansoni cercariae. S. haematobium surface antigens, however, were recognized to some extent by serum from mice chronically infected with S. mansoni. It is suggested that this cross-reactivity may reflect the heterologous immunity demonstrated experimentally between these two species, whilst the species-specificity of vaccine sera to surface antigens may mirror the highly specific immunity induced by vaccination.

A survey was carried out to determine values for the prevalence and intensity of infection of intestinal helminths in children living in Cocle Province, Republic of Panama. The data set consisted of the results of microscopic examinations (modified Kato Katz technique) of stool samples collected in November 1987 from 661 children attending primary schools in 4 communities in the province. The overall prevalences of Ascaris lumbricoides, hookworm and Trichuris trichiura infections were found to be 18·2, 12·0 and 27·5% respectively. There were significant differences between the infection prevalence values for children attending the different schools, but not with respect to age or sex. Positive associations were detected between particular pairs of infections and these were most evident when Trichuris trichiura was involved. The data revealed evidence of a positive association between siblings at one school only for each helminth infection.

Patterns of genetic variation among populations can reveal the evolutionary history of species. Pinworm parasites are highly host specific and form strong co-evolutionary associations with their primate hosts. Here, we describe the genetic variation observed in four Trypanoxyuris species infecting different howler and spider monkey subspecies in Central America to determine if historical dispersal processes and speciation in the host could explain the genetic patterns observed in the parasites. Mitochondrial (cox1) and ribosomal (28S) DNA were analysed to assess genetic divergence and phylogenetic history of these parasites. Sequences of the 28S gene were identical within pinworms species regardless of host subspecies. However, phylogenetic analyses, haplotype relationships and genetic divergence with cox1 showed differentiation between pinworm populations according to host subspecies in three of the four Trypanoxyuris species analysed. Haplotype separation between host subspecies was not observed in Trypanoxyuris minutus, nor in Trypanoxyuris atelis from Ateles geoffoyi vellerosus and Ateles geoffoyi yucatanensis. Levels of genetic diversity and divergence in these parasites relate with such estimates reported for their hosts. This study shows how genetic patterns uncovered in parasitic organisms can reflect the host phylogenetic and biogeographic histories.

Schistosomiasis and malnutrition are often overlapped in poor communities, resulting in disproportionately high mortality rates. Currently, fragmented data make it difficult to define the relationship between diet and schistosomiasis. Thus, we systematically review the preclinical evidence on the impact of diet in Schistosoma mansoni infection. From a structured search, we recovered 27 original articles. All studies used mice and most of them investigated hypoproteic (70.37%), hyperlipidic (22.22%) or vitamin-deficient (7.41%) diets. Diets based on carbohydrate, zinc or milk supplementation were investigated at a reduced frequency (3.70% each). Hypoproteic diets attenuated parasitic load and granulomatous inflammation, but also reduced host resistance to S. mansoni infection, determining higher mortality rates. By stimulating steatohepatitis, parasitic load and granulomatous inflammation, hyperlipidic diets increase organ damage and mortality in infected animals. Although a high-sugar diet and vitamin restriction potentiate and zinc supplementation attenuates S. mansoni infection, the current evidence for these diets remains inconclusive. Analysis of methodological quality indicated that the current evidence is at high risk of bias due to incomplete characterization of the experimental design, diet composition and treatment protocols. From the bias analysis, we report methodological limitations that should be considered to avoid systematic reproduction of inconsistent and poorly reproducible experimental designs.