The fine structure of egg shells of four different genera belonging to the order Tylenchida has been examined. The species examined were Meloidogyne javanica, Rotylenchulus reniformis, Tylenchulus semipenetrans and Pratylenchus minyus. They are all similar in their basic structure, being composed of vitelline membrane, chitin and lipid layers, but there is considerable variability in the thickness of these layers.

We have retained the conventional nomenclature because of its convenience, but it is clear that these layers have a variety of chemical components. However, they do appear to contain the compounds from which they take their name. Thus chitin occurs in the chitin layer, and lipid in the lipid layer. The latter is removed by the technique used in isolating the shell from the egg. Chemical analysis of the hydrolysis products of these shells has revealed a high (35 %) proline content which appears to be a characteristic of those nematode egg shells which have been examined so far. These analyses and treatment with enzymes indicate that the chitin layer is a chitin–protein complex.

Experiments on the permeability of eggs of M. javanica at different temperatures indicate that changes in permeability are not due to the melting of a single lipid with a distinct melting point as had been thought in the past. We have found that Arrhenius activation energies calculated from the two slopes of an Arrhenius plot were 17·8 kcal/mol and 43·0 kcal/mol respectively, the transition from one to the other taking place at 62°C. We think that these changes are due to changes in the properties of lipoprotein membranes in the lipid layer. These membranes appear to be of paramount importance in controlling the permeability of the nematode egg shell.

The entire ribosomal DNA repeat unit of a steinernematid species (Nashes isolate) was cloned as three separate EcoR I fragments in the plasmid pUC18. An equimolar cocktail of these three clones was used to identify Steinernema species on Southern blots as each species displays its own unique restriction fragment length polymorphisms. The clones also identified two new species isolated in a soil survey of coastal regions of Britain. One of the clones (pSn4.0) can detect length heterogeneities in the rDNA repeat unit of various isolates of some of the species, particularly the most common in the United Kingdom, S. feltiae. These differences in the rDNA repeat unit length remained constant over several years for one isolate of S. feltiae, but were different for each of the geographical isolates studied to date.

Acanthocephaloides geneticus sp.nov. is described from Arnoglossus laterna (Bothidae) from the Mediterranean littoral (Sète-France). The new species most closely resembles Acanthocephaloides propinquus (Dujardin, 1845) Meyer, 1932, whose type host is Gobius niger (Gobiidae). A genetic comparison of the parasites collected from these two Teleostean fishes has confirmed the value of morphological differences which seemed too slight to raise the populations to species status.

1. Experiments on the liberation of cercariae from their snail hosts have shown that under normal conditions C. limbifera (Seifert) and Cercaria “Z” n.sp. emerge during the day-time only, while in C. cambrensis I (Wright) the emergence takes place during the whole twenty-four hours, but considerably more so during the night-time.

Continual darkness causes a decrease in the numbers emerging, and a lowering of the temperature partially or completely checks the escape of the larvae.

In Cercaria “Z” there is a double maximum between 11 a.m. and 4 p.m. in the numbers liberated.

The invasion of L. truncatula by the larval stages of Fasciola hepatica leads to the disintegration of the host's tissues, the appearance of black pigment and the reduction of the liver cells to a thin layer of protoplasm containing the nuclei.

Developmental rates for Wuchereria bancrofti larvae maturing in the vector Aedes polynesiensis were estimated by analysing stage-frequency data consisting of counts of larval stages in mosquitoes reared at 20, 22·5, 25, 27·5, 30 and 32 °C. Base temperatures (i.e. low temperature thresholds) for W. bancrofti development were estimated by the x-intercept method and the model of Lactin et al. (1995). Resulting values were similar with both methods and were ≈12·5 °C for microfilariae (mf) in thorax, ≈17 °C for L1, 15·5 °C for L2 and 16·5 °C for L3. Upper thresholds estimated by the Lactin et al. model were 29·3 °C for mf, 29·1 °C for L1, 32·2 °C for L2 and 31·5 °C for L3. In addition, an original method was devised for computing the L3 upper threshold, by modelling L3 length shrinkage with temperature. It gave a value of 31·4 °C. At 32 °C, L2 and L3 stages exhibited altered morphology, larvae being shorter and wider than expected. The model of Lactin et al. described adequately the non-linear relationship between developmental rate and temperature, but a linear degree-day approach may be applied for temperatures below 27–28 °C.

The present study was designated to ascertain the anthelmintic activity of the rhizomes of Paris polyphylla and to isolate and characterize the active constituents. The methanol extract from rhizomes of P. polyphylla showed significant anthelmintic activity against Dactylogyrus intermedius with the median effective concentration (EC50) 22·5 mg L−1. Based on this finding, the methanol extract was fractionated by silica gel column chromatography in a bioassay-guided fractionation yielding 2 bioactive compounds, the structures of these compounds were elucidated as formosanin C and polyphyllin VII. The in vivo tests revealed that formosanin C and polyphyllin VII were significantly effective against D. intermedius with EC50 values of 0·6 and 1·2 mg L−1, respectively. The acute toxicities (LC50) of formosanin C and polyphyllin VII for grass carp were 2·8 and 2·9 mg L−1, respectively. The overall results provide important information for the potential application of formosanin C and polyphyllin VII in the therapy of serious infection caused by D. intermedius.

Few papers have been published on tick lectins so far, and therefore more data are needed to complete the mosaic of knowledge of their structural and functional properties. Tissue-specific lectin/haemagglutinin activities of both soft and hard ticks have been investigated. Some tick lectins are proteins with binding affinity for sialic acid, various derivatives of hexosamines and different glycoconjugates. Most tick lectin/haemagglutinin activities are blood meal enhanced, and could serve as molecular factors of self/non-self recognition in defence reactions against bacteria or fungi, as well as in pathogen/parasite transmission. Dorin M, the plasma lectin of Ornithodoros moubata, is the first tick lectin purified so far from tick haemolymph, and the first that has been fully characterized. Partial characterization of other tick lectins/haemagglutinins has been performed mainly with respect to their carbohydrate binding specificities and immunochemical features.

The physiology of the neuromuscular systems in Hymenolepis diminuta and H. microstoma was studied in vitro using intact, adult worm and strips of worm body wall. Intact worms were insensitive to ionic changes in the in vitro buffering system. However, strips of body wall containing longitudinal muscles were extremely sensitive to ionic manipulation. In intact worms tension generated in the strobila had two components; small brief tension peaks up to 500 mg amplitude are superimposed on larger, longer peaks of up to 1200 mg amplitude. Removal of the scolex and neck region either failed to show significant changes in tension, or showed a reduction in amplitude but not of frequency. Muscle contraction of both H. diminuta and H. microsoma were qualitatively similar. In split-worm preparations the concentration of Ca2+ in the bathing solution significantly affected both spontaneous and evoked contractions in H. diminuta and H. microstoma; the addition of CaCl2 greatly reduced the amplitude and frequency of the contractions. The chloride salts of cobalt, barium, cadmium and manganese elicited prolonged contractions of the longitudinal musculature of both H. diminuta and H. microstoma. While CoCl2 was the most effective in stimulating muscle contraction, the magnitude of the response varied with the concentration of Ca2+ in the bath. The results indicate that peripheral inhibition is extremely important in cestode motor control and that extracellular calcium ions may regulate the peripheral inhibitory mechanisms.

A successful attempt was made to induce immunological tolerance to N. brasiliensis in laboratory rats by exposing them to larval infections before the age of immunological maturity.

The adult capacity of rats to eliminate a primary infection in 3 weeks and to acquire resistance to reinfection was developed by 5 weeks of age. On the other hand, primary infections of 250–660 larvae, originating before 4 weeks of age, survived in 61 of 83 rats up to 10 weeks. These rats were not resistant to challenge at 5 or 7 weeks of age, nor did the challenge infection undergo immune elimination. Such tolerance was evident even in rats whose primary worm burden was chemically abbreviated prior to challenge.

It was not always possible to distinguish between tolerant and immune animals on the basis of egg output patterns. Suppression of egg output occurred even in tolerant rats harbouring relatively stable female populations.

For immunological experiments on N. brasiliensis infection in rats, the determination of a ‘take ratio’ is advocated: (20 day take/10 day take) × 100. This ratio eliminates the error inherent in variability of takes and renders comparable the results of different experiments.

The work was carried out in 1965 during tenure by one of us (T.K.) of an Exchange Scholarship under the British–Hungarian Cultural Agreement. Supervision by Professor W. Mulligan and the co-operation of colleagues in the Wellcome Laboratories are gratefully acknowledged.

Cystosporogenes operophterae, a microsporidian parasite of the winter moth, Operophera brumata, was cultured in a Spodoptera frugiperda cell line. The infected cultures were achieved by addition of haemolymph from larvae infected with C. operophterae. The infection rates for the cell line ranged from 80 to 95% after incubation for 10 days at 25 or 20 °C. There was no development in cultures incubated at 30 °C. Light and electron microscopy were used to observe the development of C. operophterae in vitro.

Sera from donors exposed to malaria were tested for their ability to block the transmission of isolates from Cameroonian Plasmodium falciparum gametocyte carriers. Sera were selected from amongst Cameroonian and Gambian donors who had positive antibody reactivity against the surface of activated gametes and against epitopes of Pfs 48/45 (a potential transmission-blocking vaccine candidate antigen). Aliquots of washed blood from gametocyte carriers were resuspended in test and control sera and fed to An. gambiae mosquitoes via a membrane feeder. Comparisons of the prevalence and intensity of infections in dissected mosquitoes showed variations in the ability of sera to block the transmission of the different isolates. Sera were identified that had little or no blocking effect on the transmission of isolates unless the isolate was poorly infectious. Some sera completely blocked the transmission of some isolates whilst having little or no effect on others. The observed variation in transmission-modulating activity may have implications for the development of a transmission-blocking vaccine.

Early in the history of schistosomiasis research, children under 5 years of age were known to be infected. Although this problem was recognized over 100 years ago, insufficient action has been taken to address this issue. Under current policy, such infected children only receive their first antiparasitic treatment (praziquantel – PZQ) upon entry into primary school as current mass drug administration programmes typically target school-aged children. For many infected children, they will wait up to 6 years before receiving their first medication and significant schistosomiasis-related morbidity may have already established. This inequity would not be accepted for other diseases. To unveil some of the reasons behind this neglect, it is paramount to understand the intricate historical relationship between schistosomiasis and British Imperial medicine, to underline its lasting influence on today's public health priorities. This review presents a perspective on the historical neglect of paediatric schistosomiasis, focusing on important gaps that persist from the early days after discovery of this parasite. Looking to end this inequity, we address several issues that need to be overcome to move forward towards the lasting success of schistosomiasis control and elimination efforts.

In view of the mandate from the World Health Organization (WHO) for developing novel drug candidates against human lymphatic filariasis, dihydrofolate reductase (DHFR) inhibitors are explored as potential antifilarial agents. The in vitro biological evaluation of an in-house library of 12 diverse antifolate compounds with 2,4-diaminopyrimidine and 2,4-diamino-s-triazine structural features against Brugia malayi is reported. To confirm the DHFR inhibitory potential of these compounds, reversal studies using folic acid and folinic acid were undertaken. Inhibition of DHFR can induce apoptosis; in this light, preliminary evidence of apoptosis by test compounds was detected using ethidium bromide–acridine orange staining and the poly(adenosine diphosphate-ribose) polymerase (PARP) inhibition assay. Among the evaluated compounds, 3 showed significant activity against both microfilariae and adult worms. The effects of 2 of these compounds were mostly reversed by folic acid, validating DHFR inhibitory activity. Partial reversal of the effect of 2 compounds by folinic acid and non-reversal of the effect of the third compound both by folic and folinic acids are discussed. This study opens new avenues for the discovery of lead molecules by exploiting the folate pathway against one of the major neglected tropical diseases, filariasis.

The biology and life-cycle of Holostephanus lühei is briefly described. The similarities in morphology between the adhesive organ gland cells of H. lühei and Cyathocotyle bushiensis are stressed. Alkaline and acid phosphatases were demonstrated in the adhesive organ gland cells and cuticle. Acid phosphatase also occurs in the caecal cells. Non-specific esterase, sensitive to E600 10−5M and Mipafox 10−3M, is present in the gland cells. The caeca contain a non-specific esterase sensitive to AgNO3 10−2M and PCMB 10−4M. Leucine aminopeptidase is present in the gland cells. The in vitro studies confirmed that the secretory products pass to the exterior of the parasite and have a histolytic action.

The four strigeids studied, Cyathocotyle bushiensis, Holostephanus lühei, Diplostomum spathaceum and Apatemon gracilis minor, are compared and the biological role of the secreted enzymes are discussed.

My sincere thanks are due to Dr D. A. Erasmus for suggesting this line of investigation and for his encouragement during the study. I am also grateful for his performing, on my behalf, the infection experiments and criticizing the manuscript. I wish to thank Professor J. Brough for his interest in the progress of the work, and the University College, Cardiff, for a grant from William E. Morgan's Bequest.

The nature of antigens present on the surface coat of Trypanosoma brucei brucei (serotype AnTat–1) trypanosomes has been investigated by light and electron microscopy using indirect immunofluorescence and immunoenzymatic labelling techniques. The use of a monospecific anticomponent V serum has shown that the variant-specific antigen, component V, is the major constituent of the surface coat. Antigens of heterologous serotype, of culture form trypanosomes, of host serum proteins or host red blood cells were not demonstrated on the surface coat.

Interferon-gamma (IFN-γ) has a crucial role for host defence against parasite infection. It is not clear, however, how IFN-γ affects the parasite-infected host cells. The effect of IFN-γ on Neospora caninum-infected cells was investigated in murine fibroblasts and canine kidney cells in vitro. In the presence of IFN-γ, the viability of the infected host cell was decreased and apoptotic cell death occurred, as analysed by DNA stainings with propidium iodide and a terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labelling (TUNEL) and DNA fragmentation. The percentage of apoptotic cells depended on the dose of IFN-γ. Flow cytometric analysis indicated a significant increase of FasL expression on the IFN-γ-treated cells following N. caninum infection. Moreover, IFN-γ treatment down-regulated Bcl-2 expression in the cells cultured with N. caninum while parasite infection up-regulated Bcl-2 expression. The present study suggests that the IFN-γ induced increases of FasL expression and down-regulated Bcl-2 expression in N. caninum-infected cells are associated with apoptosis in vitro.

The genus Acanthobothrium van Beneden, 1850, seems to be in a state of slight confusion due, possibly, to some inadequate descriptions which have led to subsequent misidentifications of some of the species. The type species A. coronatum was first recorded and very briefly described by Rudolphi (1819) as Bothriocephalus coronatus from ‘Squali stellaris, Squali squatinus and Torpedinis marmoratae“. Since then specimens have been assigned to this species which probably belong elsewhere; the list of synonyms and the host records are extensive. Baer (1948) divided the species of the genus into two groups, namely, those parasitic in sharks and those in rays. This seems to be a reasonable distinction as specificity is very marked in cestodes and especially in those of elasmobranchs. It was thought that a detailed study of the type species, presented here, would be a useful preliminary to a consideration of the genus as a whole.

Specimens of A. coronatum were obtained from the spiral valve of Scyliorhinus stellaris (L.) caught in Cardigan Bay in July 1956 and at Plymouth in August 1958 and August and September 1959. The number of worms, complete with scoleces varied from 6 to 10 per fish and in one instance 14. Many fragments of two to three proglottids and many individual proglottids were also present. The scoleces were attached in the first turn of the spiral valve and the strobila adhered feebly along the whole length, sometimes as far back as the rectal gland, leaving on removal a distinct groove in the mucosa.

(i) Amoebae closely resembling Entamoeba coli (of Man) have been studied in macaques (M. rhesus, M. sinicus, M. nemestrinus) and in a marmoset (Hapale jacchus).

(ii) Pure strains of these amoebae have been isolated and cultivated, and carefully compared with similar pure strains of E. coli from Man (E. coli hominis).

(iii) The complete life-cycles of these various strains—amoebae, cysts, and all stages of precystic and metacystic development—have been studied in vitro.

(iv) The “E. coli” of macaques (E. c. macacorum) was experimentally transmitted to a man, who ingested pure culture-cysts of a strain derived from M. rhesus.

(v) The “E. coli” of the marmoset (E. c. jacchi) was likewise transferred to M. sinicus, and from this host again to M. rhesus and Man.

(vi) The natural infections observed in M. sinicus, M. rhesus, and H. jacchus all died out spontaneously after persisting for various periods (4½ years, 2 years, and 1 year respectively). The experimental infection with E. coli macacorum induced in Man also disappeared spontaneously (after about 6 weeks).

(vii) Attempts to infect kittens experimentally with E. coli jacchi were uniformly unsuccessful.

(viii) All infections with “E. coli” from every source—whether natural or experimental—have proved to be completely innocuous.

(ix) Culture-cysts of various strains of “E. coli” have been found able to live at low temperatures for all times up to about 4½ months (maximum 135 days at 1°–2° C., for a strain from H. jacchus).

(x) The “E. coli” of H. jacchus—and all its experimental derivatives—ingests human red blood-corpuscles in vitro as readily as E. histolytica.

(xi) Several strains of E. coli hominis, isolated directly from naturally infected human beings, have been found to possess the same property.

(xii) Since strains of E. coli thus exist—both in men and in monkeys—which eat human red corpuscles avidly, this faculty can no longer be considered as peculiarly characteristic of E. histolytica; and the importance of this observation, for the diagnosis of amoebic dysentery, is therefore emphasized. The statement that E. histolytica ingests red corpuscles, while E. coli does not, is untrue. It is not possible to distinguish these species by this character alone.

(xiii) As it has been found impossible to distinguish various strains of “E. coli” living in macaques, a marmoset, and men, by any recognizable specific feature—all of them being so closely alike morphologically, physiologically, culturally, and in their ability to inhabit these different hosts—it is concluded that no specific difference actually exists between any of them. The “E. coli” of the Macaques and of the Common Marmoset is probably Entamoeba coli itself—the species which naturally inhabits Man.

The population dynamics of Ascaridia galli was studied in 70 ISA Brown layer pullets, 42 of them were each experimentally infected with 500 embryonated A. galli eggs and 28 chickens were kept as uninfected controls. Six chickens from the infected group and 4 from the control group were necropsied at 3, 7, 10, 14, 21, 28 and 42 days post-infection (d.p.i.). The mean worm recovery varied from 11–20% of the infection dose with the highest recovery at 3 d.p.i. and the lowest at 21 and 42 d.p.i. (P < 0·05). More larvae were recovered from the intestinal wall than from the content (P < 0·0001) and intestinal content larvae were longer than those from the wall (mean length 1·6 and 1 mm, respectively, P < 0·0001). Although larvae were growing over time, a population of small-sized larvae (length  < 1 mm) was recovered at all d.p.i. During the first week of infection most of the larvae were located in the anterior half of the jejunoileum but they moved posteriorly with the age of infection. Thus, a subpopulation of larvae mainly in the lumen grew with time while another subpopulation remained small and associated with the mucosa. During the infection both subpopulations moved to a more posterior localization in the gastrointestinal (GI) tract.

Despite the great interest to quantify the structure of host–parasite interaction networks, the real influence of some factors such as taxonomy, host body size and ecological opportunity remains poorly understood. In this paper, we investigate the general patterns of organization and structure of interactions in two anuran–parasite networks in the Brazilian Pantanal (seasonally flooded environment) and Atlantic Forest (non-flooded forest). We present theoretical models to test whether the structures of these host–parasite interaction networks are influenced by neutrality, host taxonomy and host body size. Subsequently, we calculated metrics of connectance, nestedness and modularity to characterize the network structure. We demonstrated the structure networks were influenced mainly by body size and taxonomy of the host. Moreover, our results showed that the seasonally flooded environment present networks with higher connectance/nestedness and lower modularity compared to the other environment. The results also suggest that seasonal floods may promote ecological opportunities for new species associations.