The causal prophylactic activity of the novel hydroxynaphthoquinone, S66C80, was assessed against the exo-erythrocytic (EE) stages of Plasmodium berghei cultured in the human hepatoma cell line, HepG2. 566C80 was found to be highly active as an inhibitor of EE development and was more active than the established causal prophylactic pyrimethamine. A 566C80 concentration of 1·85 × 10−9 M, added 3 h after sporozoite invasion, reduced the numbers of EE forms visible at 48 h by 50%, while the equivalent concentration of pyrimethamine was 1·95 × 10−8 M.

We have investigated the effect of qinghaosu (QHS, artemisinin) and its derivatives on Leishmania major replication in vitro and on the disease development in mice infected with L. major. Artemisinin is effective against promastigotes in vitro, with an ED 50 (50% effective dose) at 7.5 × 10−7 m. Both artemisinin and artemether are leishmanicidal for amastigotes in infected murine macrophages in vitro, with ED50 at 3 × 10−5 m and 3 × 10−6 m respectively. These compounds have no effect on the viability of macrophages or on the phytohaemaglutinin-induced proliferation of normal spleen cells, even at 10−4 m. BALB/c mice infected in the footpad with L. major developed significantly smaller lesions and parasite loads when treated with the compounds. Intra-lesion injection of the compounds was the most effective route. The intramuscular and oral routes were also effective; however, intravenous injection with artesunate was not effective.

Protocols for in vitro excystation of oocysts of Cryptosporidium parvum, including different chemical pre-incubation steps, were compared to examine some of the biochemical triggers involved in excystation and to define an in vitro excystation protocol of a reproducibly high efficiency. Pre-incubation steps which increased the permeability of the oocysts were found to enhance excystation dynamics and pre-treatment of oocysts with saliva was found to decrease the permeability and reduce excystation. Although excystation was maximal after incubation for 4 h, sporozoites tended to lyse over this period, and maximum sporozoite recovery occurred after 30 min. The results obtained are discussed in relation to excystation protocols adopted by different research groups and a number of recommendations are given for in vitro excystation of C. parvum oocysts.

The characterization of four Trypanosoma vivax isolates from Colombia in South America showed that although minor phenotypic differences existed between them, these parasites are antigenically related and belong to a single serodeme. Characterization by isoenzyme assay, karyotyping and DNA probe analysis, showed the Colombian isolates to be more similar to the West African than to Kenyan T. vivax. There was, however, little serological cross-reactivity between South American and African groups of T. vivax. Although the T. vivax isolates from Colombia were pathogenic for dairy calves which showed the typical sign of progressive emaciation, these parasites failed to infect mice or tsetseand could not be cultivated as bloodstream forms in vitro.This study represents initial attempts to establish the phenotypic and serological diversity amongst T. vivax isolates from South America.

Aerobic resistance of Trichomonas vaginalis to metronidazole was induced in vitro by anaerobic cultivation of drug-susceptible trichomonads with low concentrations of the drug (2–3 μg/ml) for 50 days. Minimal lethal concentrations (MLC) for metronidazole of the resistant derivatives were high in aerobic susceptibility assays (MLC = 216–261.5 μg/ml) but low in anaerobic assays (MLC = 4.2–6.3 μg/ml), surpassing MLC values of their parent strain approximately 50-fold and 3-fold under aerobiosis and anaerobiosis, respectively. Sensitivity to metronidazole under anaerobic conditions and activity of the hydrogenosomal enzyme pyruvate: ferredoxin oxidoreductase indicated that the resistance was of the aerobic type. Dependence of the resistance manifestation on O2 was further confirmed by susceptibility assays in vitro performed in defined gas mixtures of different oxygen content (1–20%). Five percent concentration of O2 proved to be the threshold required for resistance demonstration and the MLC values further increased with increasing O2 concentrations. The in vitro-induced resistance was also demonstrated in vivo by subcutaneous mouse assay. The dose of metronidazole needed to cure 50% of infected mice (DC50) was 223 mg/kg × 3 for resistant derivative MR-3a but 6.6 mg/kg × 3 only for its drug-susceptible parent strain. The metronidazole – resistant strains developed in this study correspond by their properties to drug-resistant T. vaginalis strains isolated from patients refractory to treatment, and promise to be a useful tool in the study of 5-nitroimidazole aerobic resistance.

The population dynamics of Sphaerospora dicentrarchi and S. testicularis, myxosporean parasites of sea bass (Dicentrarchus labrax), was studied in different groups of cultured and wild fish. S. dicentrarchi prevalence was higher for wild fish (100%) than for cultured ones (70.5%). Infection levels increased with host age and there were no significant sex differences in cultured fish. Among parasitized organs, the gall bladder and intestine registered the highest infection levels in wild and cultured fish. A progressive increase in infection levels of S. testicularis was observed in cultured males during the spawning season in four consecutive periods. The evolution of the percentage of the different parasitic stages is pointed out. The infection seems to be related to water temperature and gonadal maturation.

Adults of the porocephalid pentastomid Porocephalus crotali infect the lung of rattlesnake definitive hosts and larvae develop in rat intermediate hosts. In the latter, nymphs encyst within a variety of tissue sites (commonly abdominal fat bodies and lungs) and each becomes the focus of an eosinophilic granuloma. From an early stage in infections, granulomas become increasingly infiltrated by mast cells which, using conventional histology and paired immunofluorescence against mast cell proteases, appear to be exclusively of the mucosal phenotype. Mucosal mast cells are concentrated along the dorsal region of the parasite and in a plug of tissue containing degenerating cuticles within independent granulomas, which is located between its head and tail. ELISAs against the rat mast cell proteases I and II (RMCP I and II), extracted from abdominal fat, lung, spleen, liver and kidney granulomas at various intervals post-infection, reveal a substantially elevated concentration of RMCP II in all lesions. In fat, concentrations increase up to about 100 days post-infection, at which time moulting ceases and inflammatory responses subside. RMCP II was scarcely detectable in matched control tissues. Unlike infections with certain nematode parasites, where enteric mucosal mast cells secrete RMCP II systemically, concentrations of RMCP II in the serum of infected rats were significantly reduced when compared with age-matched uninfected controls. These results confirm that P. crotali can selectively recruit mucosal mast cells to a variety of tissue sites, most of which are non-mucosal. Possible mechanisms are discussed.

The presence of phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31), an enzyme at the branchpoint of glycolysis and the Krebs cycle was detected in the Filaria Molinema dessetae. This enzyme has not previously been identified in Helminths, which have so far been found to only possess a phosphoenolpyruvate carboxykinase (EC 4.1.1.32). This enzyme had a level of activity comparable to that of pyruvate kinase, and was relatively less active than enzymes such as malate dehydrogenase or lactate dehydrogenase. We propose here a method of purification of M. dessetae PEP-carboxylase. When purified to electrophoretic homogeneity, the enzyme had a molecular weight of 64 kDa. Kinetic studies indicated that the carboxylation reaction had an optimal pH of 5·8. The enzyme was inhibited by cations such as Fe2+, Zn2+, Cd2+, Cu2+ but required the presence of Mg2+ or Mn2+. The enzyme was thermostable. The apparent Km value of 2·38 mmol for phosphoenolpyruvate for the carboxylation reaction was higher than previously reported values. The Km value for KHCO3 was found to be 1·6 mmol. PEP-carboxylase did not catalyse the reverse reaction.

Groups of South African, farm-reared merino lambs about 4 months old were vaccinated with extracts of adult Haemonchus contortus enriched with H11, an integral membrane protein from the parasite's intestinal microvilli, or with proteins soluble in Tween 20. The lambs were challenged with 5000 infective 3rd-stage larvae. Compared to the adjuvant-injected controls, lambs vaccinated with 350 μg/kg liveweight of the H11-enriched extract showed an 89% reduction in parasite egg production and an 88% reduction in total worm burden at post-mortem 35 days post-challenge. The H11-enriched extract contained some protein also present in the Tween 20 extract. Lambs vaccinated with 600 μg/kg of protein soluble in Tween 20 showed a 40% reduction in faecal egg counts and 51% reduction in total worm numbers 35 days post-challenge. Animals injected with 20 μg/kg of a sub-fraction containing H11 obtained from the H11 extract, showed a 71% reduction in egg output and total worm numbers.

5-HT is a candidate for the excitatory transmitter at the neuromuscular junction in trematodes including Fasciola hepatica. This study has determined the response of a muscle strip preparation from Fasciola hepatica to 5-HT and a range of agonists that distinguish between the vertebrate receptor 5-HT subtypes. 5-HT increased the resting tone and the rhythmic activity of the muscle strip. Of the 19 compounds tested, only 10 had an effect similar to 5-HT and all but 2 of these were tryptamine compounds. 5-HT was more potent than tryptamine whilst 4-OH-tryptamine had no effect, suggesting that the response is mediated by a 5-HT rather than a tryptamine receptor. 5-Fluorotryptamine and 5-carboxyamidotryptamine were the most potent agonists. 8-OH-DPAT also mimicked the effect of 5-HT, though less potently. Assuming that these agents elicit their response through a common receptor, this suggests the presence of a 5-HT receptor with similar properties in terms of agonist recognition as the vertebrate 5-HT1 class of receptor involved in controlling Fasciola muscle motility.

Four monoclonal antibodies (MAb) to Echinococcus granulosus Antigen 5 (Ag5) and Antigen B (AgB) were assessed in an enzyme-linked immunosorbent assay (ELISA) for detection of circulating antigens (CAg) in sera of human patients with E. granulosus infection. Around 5·5–8% of 200 sera from 42 surgically proven hydatid patients contained detectable CAg by individual MAb. The combined detection rate for CAg, using four MAb, was 19% (38/200). Although hydatid CAg was detected by MAb in at least one serum sample from 21 of 42 patients, some patients remained negative in the assay regardless of the time when serum samples were taken (pre- or post-operatively), or of the continuing presence of hydatid cysts, their location or fertility. In addition, it was observed that the binding capacity of MAb for sheep hydatid cyst fluid antigen (SHCF) was somewhat reduced in the presence of normal human serum. The CAg detection assay would only be useful for assessment of hydatid infection status in patients with detectable CAg in serum samples.

Substance P immunoreactivity (SP-IR) was detected in the nervous system of the gull-tapeworm Diphyllobothrium dendriticum. The distribution of the SP-IR neurons in the plerocercoid differs from that of other peptidergic and aminergic neurons in the worm. As well as occurring in the ganglionic commissure and along the two main nerve cords, SP-IR neurons are located laterally to the main nerve cords but not dorsally or ventrally. The SP-IR neurons have projections extending to the surface. Bipolar SP-IR neurons with processes to the surface also occur in the tip of the scolex. A sensory function for the SP-IR neurons is suggested.

The investigation of Price & Clancy (1983), which demonstrated a significant positive correlation between total helminth species number per host species and geographical range of freshwater fish host species in Britain, was re-examined using a different measure of parasite species richness. Re-calculation of the correlations between the two parameters after controlling for the effect of the composition of the list of fish by excluding, on biological and distributional grounds, 2 species of agnathans and 7 species of introduced teleosts, and for the effect of sampling effort by using helminth richness in the richest component community of each fish species rather than check-list data, reveals no significant relationship between helminth species richness and host range. Habitat and an omnivorous host diet now appear more significant determinants of helminth richness than the accumulation of parasites by predation. The findings provide little support for the interpretation of the relationship between helminth species richness and host range in terms of island biogeographic theory, but do support an alternative explanation in terms of the colonization time hypothesis, i.e. that helminth species richness is related to the time since the fish host arrived in Britain.

Brown trout (Salmo trutta) from four lakes within an area of approximately 40 km2 were found to harbour nine helminth species. Crepidostomum metoecus could be classified as core species, while Crepidostomum farionis had an intermediate position. All the others could be classified as satellite species. Salmon (Salmo salar) fingerlings were grown to smolts in an earthpond over the summer. The fingerlings became infected with Crepidostomum spp. which may indicate a superior colonization ability in the core species. The results support the ‘rescue effect hypothesis’ rather than the ‘carrying capacity hypothesis’.