Young rabbits were infected by mouth with sporulated Eimeria stiedae oocysts and killed at 1, 3, 5, 8, 12, 16, 24, 30, 48, 54, 72, 84, and 96 h after infection. Sporozoites were found in the lamina propria of the duodenum between 5 and 8 h after infection. Sporozoites were present in the mesenteric lymph nodes, both free and within lymphatic monocytes at between 12 and 84 h after infection. No sporozoites were observed in duodenal portal blood. These observations provide morphological evidence compatible with a lymphatic route of migration between duodenum and mesenteric lymph nodes, probably within lymphatic monocytes. The route to the bile duct epithelium is discussed and a subsequent migration in the portal blood suggested.

I wish to express my gratitude to Dr D. F. Kelly of the School of Veterinary Medicine, Cambridge, for his guidance and constructive criticism, and to Dr Elaine Rose of the Houghton Poultry Research Station, St Ives, for her invaluable advice on technical problems and for donating the oocysts for the study. I should also like to thank Dr E. A. Pierce of the Department of Animal Physiology, Babraham, and Mr P. Long of Houghton for their advice.

This study was carried out as a Research Project during the Natural Sciences Tripos, Part II Pathology course 1965, in the Department of Pathology, Cambridge.

Praziquantel is widely used for treatment of schistosomiasis. Elimination of intravascular worms involves a combination of drug-induced changes at the worm surface and a host immune response against exposed parasite components. Loss of spines is a typical feature of schistosomes obtained from praziquantel-treated infected animals. The mechanism for this is unknown. We have considered the possibility that praziquantel treatment involves exposure of surface spines consisting of ‘paracrystalline’ actin and a subsequent host response against actin through mechanisms established in the host independently from schistosomal infection. Drug-induced exposure of actin was demonstrated by fluorescence microscopy using anti-actin antibodies and phallacidin, an actin-binding mushroom toxin. Actin spines remained intact at the schistosome surface after in vitro exposure, but spine morphology was lost after in vivo exposure to praziquantel. Disintegration of spines in vivo was associated with binding of host antibodies. In vitro spine destruction could be seen in the presence of normal human serum. The effect was linked to calcium-dependent binding of actin depolymerizing factor, gelsolin.

We analysed the patterns of tick distribution on 2274 adult toads from Venezuela and Brazil, to explore whether these ectoparasites have any impact on the survival of Bufo marinus. A maximum-likelihood analysis showed that aggregation levels of ticks decreased significantly with the mean intensity of infection. This decline could be attributed to a density-dependent reduction of ticks within toads, density-dependent tick-induced toad mortality and/or density-dependent tick-induced changes in toad susceptibility. However, the relationship between the rate of change in tick loads and tick burdens from recaptured toads indicated that neither the loss of ticks within toads nor the toad susceptibility to further infection were dependent upon tick burdens. Therefore, we can indirectly infer that density-dependent tick-induced toad mortality is responsible for the observed decline in aggregation levels with tick age and burdens. On the other hand, a significant negative relationship between tick burdens and the size-specific weight of toads suggested that ticks may also have a significant impact on the patterns of weight deposition of adult toads. This evidence suggests that these ectoparasites may play an important role in regulating the densities of B. marinus in native habitats.

The variable antigen repertoire expressed by metacyclic Trypanosoma brucei rhodesiense is not influenced by the anamnestic expression whereby the variable antigen type (VAT) ingested by a tsetse fly is present at high levels in early bloodstream populations of fly-infected mice. This has been demonstrated by feeding to Glossina morsitans a trypanosome line expressing a VAT which is normally a component of the metacyclic repertoire. The VAT did not constitute a significantly increased proportion of the resultant metacyclic population which would have occurred had anamnestic expression and metacyclic expression been linked. Five other metacyclic VATs were also present at control levels. We conclude that the mechanisms of expression of VATs in the fly and in the mammal are independently controlled.

Three satellite DNAs previously isolated from the entomopathogenic nematodes Steinernema carpocapsae, Heterorhabditis bacteriophora and Heterorhabditis indicus give hybridization signals only with the S. carpocapsae, H. bacteriophora and H. indicus populations tested, indicating that these satellite sequences are species-specific. Because of their reiteration and their variabilities, we have shown that these sequences are able to discriminate at the interspecific level between the Steinernema and Heterorhabditis species, but also at the intraspecific level between S. carpocapsae strains. Furthermore, in simple squashed nematode experiments, we are able to unambiguously identify S. carpocapsae, H. bacteriophora and H. indicus populations. This last procedure is effective even on a single infective juvenile, with the main advantage that it avoids time-consuming DNA extractions.

The mechanism whereby praziquantel produces a contraction and subsequent flaccid paralysis (a loss of sensitivity to subsequent stimuli) of Schistosoma mansoni in a medium containing an elevated Mg2+:Ca2+ ratio was investigated. In RPMI, praziquantel produced a concentration-dependent tonic contraction of the parasite with an EC50 of 200 nM. Magnesium inhibited the contraction in such a manner as to convert the tonic contraction to a phasic one without altering the peak force generated. The Mg2+-dependent block was non-competitive with praziquantel but was competitive with extracellular Ca2+, ratios of 7·5:1; Mg2+: Ca2+ being needed to inhibit the tonic contraction and to induce flaccid paralysis. Flaccid paralysis was associated with a reduced ability of the parasite to take up 45Ca2+ from the bath compared to parasites that had not entered into flaccid paralysis and flaccid paralysis was reversible. Recovery from flaccid paralysis was accelerated by treatments that are expected to increase Ca2+ uptake by the parasite. At a concentration of 500 nM, praziquantel produced 2 distinct phasic contractions in intact parasites incubated in an elevated [Mg2+] medium but only 1 phasic contraction in parasites lacking their surface tegumental membranes. In zero Ca2+ I-RPMI, 10 μM praziquantel produced a phasic contraction of intact parasites but did not stimulate contraction of detegumented parasites until Ca2+ was reintroduced into the bath. These results indicate that praziquantel interacts with specific Ca2+-permeable sites in the tegumental and sarcoplasmic membranes of the parasite and that under these conditions of elevated Mg2+:Ca2+ ratios, these sites become blocked by Mg2+, leading to flaccid paralysis of the parasite.

Helminths are extremely numerous and cosmopolitan parasites of man and domesticated animals and they are responsible for considerable clinical and sub-clinical disease and tremendous economic loss. Thus, immunization against the parasitic helminths of man and animals would be of major health importance and to considerable economic advantage.

Five normal rhesus monkeys were infected with Plasmodium knowlesi sporozoites (A-strain); two developed rapidly fatal malaria and three chrinic relapsing infections. Vaccination with P. knowlesi (W-strain) merozoites (unmodified or formol-treated and freeze-dried) in Freund's complete adjuvant (FCA) did not inhibit pre-erythrocytic parasite development after challenge with A-strain sporozoites. However, the subsequent blood-stage infection was terminated in nine out of ten vaccinated monkeys even though the challenge strain was different form that used for vaccination. The degree of parasitaemia (0·01–0·70 %) and brevity of infection (1–12 days) in six animals vaccinated with untreated merzoites was similar to that observed after direct challenge with blood-stage parasites. Monkeys were equally resistant to sporozoite challenge given as the post-vaccination infection or administered 6 months after blood challenge. These results are discussed in relation to the development of a human malaria vaccine.

The transmission-blocking monoclonal antibody 13.1, which recognizes the ookinete surface antigen Pbs21 of Plasmodium berghei, and an IgG2a isotype control antibody 26.37 were purified by caprylic acid and ammonium sulphate precipitation. Fab fragments were prepared by papain digestion. IgG but not Fab from antibody 13.1 reduced ookinete formation by P. berghei in culture by as much as 94% at a concentration of 100 μg/ml. There was little difference in antibody efficacy in the range 6·25–400 μg/ml in this assay. The parasite was most sensitive to antibody activity in the first 6–9 h of culture, i.e. the gamete/zygote and early retort stages. Peripheral blood leucocytes (PBL) were essential to achieve maximal inhibition by mAb 13.1 (activity was abrogated totally if PBL were removed). Together the data suggest that one of the mechanisms of action of this antibody is antibody-mediated PBL killing. Phagocytosis of parasites was noted in these experiments in all cultures. We have not attempted in this study to distinguish between Fc-mediated opsonization, as opposed to antibody-dependent cellular cytotoxicity.

The development of the free-living infective larvae of a homogonic strain Strongyloides ratti is described.

The larvae develop only between 15 and 34 °C. Transfer experiments show the temperature block to be in the preparation for the second moult.

Within the temperature range 15–34 °C, increasing the temperature speeds up the rate of development of all the larval stages equally, the Q10 for development being 2·5.

The maximum percentage development occurs at 20 °C. The percentage development is highest in faeces–peat culture (95% development at 20 °C), whilst the percentage development in charcoal and vermiculite cultures is about the same (75% development at 20 °C.).

Larvae grown on charcoal cultures are larger than those grown on vermiculite, which are larger than those grown on peat. No significant difference was found in the length:oesophagus and length:width ratios or in the variability of larvae grown at different temperatures or on different culture media.

Different worm densities in the cultures of from 2000 to 10000 larvae per g of culture did not affect either the size of the infective larve or the percentage development.

The optimum temperature for survival is 15 °C. Worms grown at 20 °C lived longer than worms grown at any other temperature. There was no evidence of temperature adaptation by the larvae.

The infective larvae are positively thermotactic, and show maximum activity at 37 °C.

I should like to thank my supervisor, Dr Tate, for his advice and encouragement. The work was carried out during the tenure of a Medical Research Council Scholarship.

The great importance of lice as parasites and vectors of disease in man led one of us (Nuttall), soon after the outbreak of the Great War, to undertake a detailed study of these insects. It was known, prior to the war (1909–14), that lice transmit typhus and relapsing fever and during the war (1917–18) it was established that they also serve as vectors of trench fever. Beginning with February 1917, a number of papers dealing with various aspects of the louse problem were published in Parasitology (see vols. X–XIII), the study of their structure, systematics and biology being actively pursued whilst the best methods of combating them were sought for with a view to aiding the armies in the field. Of these papers, that dealing with the structure and function of the genitalia in Pediculus (II, 1917) bears directly on the subjects to which this Iconography relates.

Trichomonas vaginalis is responsible for the most common non-viral, sexually transmitted infection, human trichomoniasis, and is associated with an increased susceptibility to HIV. An escalation in resistance (2.5–10%) to the clinical drug, metronidazole (MTZ), has been detected and this compound also has adverse side-effects. Therefore, new treatment options are urgently required. Herein, we investigate the possible anti-T. vaginalis activity of 1,10-phenanthroline-5,6-dione (phendione) and its metal complexes, [Ag(phendione)2]ClO4 and [Cu(phendione)3](ClO4)2·4H2O. Minimum inhibitory concentration (MIC) against T. vaginalis ATCC 30236 and three fresh clinical isolates and mammalian cells were performed using serial dilution generating IC50 and CC50 values. Drugs combinations with MTZ were evaluated by chequerboard assay. A strong anti-T. vaginalis activity was found for all test compounds. IC50 values obtained for [Cu(phendione)3](ClO4)2·4H2O were similar or lower than those obtained for MTZ. In vitro assays with normal cells showed low cytotoxicity and [Cu(phendione)3](ClO4)2·4H2O presented a high selectivity index (SI) for fibroblasts (SI = 11.39) and erythrocytes (SI > 57.47). Chequerboard assay demonstrated that the combination of [Cu(phendione)3](ClO4)2·4H2O with MTZ leads to synergistic interaction, which suggests distinct mechanisms of action of the copper–phendione complex and avoiding the MTZ resistance pathways. Our results highlight the importance of phendione-based drugs as potential molecules of pharmaceutical interest.

Faecal egg counts were examined in 2 flocks of naturally infected Scottish Blackface sheep in southern and central Scotland. The distribution of mean counts was right skewed and similar to a gamma distribution. The counts varied with month, with mean counts rising from May to July, then falling but rising again in October, although data within each year did not always show such a clear pattern. There was no significant difference in mean egg count between the 2 farms examined. The distribution of egg count variances was also right skewed and conformed to a gamma distribution. There was a strong relationship between the mean and the variance for each population, implying that variation among populations in variances largely mirrored variation in mean egg counts. Populations with high mean egg counts and variances did not necessarily have more adult nematodes but had a greater number of adult nematodes from species other than Teladorsagia circumcincta, particularly Cooperia spp., Trichostrongylus axei and Trichostrongylus vitrinus. The contribution of different parasite species to the egg count explains the relatively poor and inconsistent fit of the negative binomial distribution to faecal egg counts in lambs.

The entomopathogenic nematode Steinernema carpocapsae is mutualistically associated with the bacterium Xenorhabdus nematophila. Infective Juveniles (IJs) transport X. nematophila cells that provide them with good conditions to reproduce within the insect. In the laboratory, long term stationary-phase culture conditions sometimes lead X. nematophila's variant 1 cells, which were previously isolated from the worms, to spontaneously and irreversibly change into a new phenotypic variant (variant 2). In this paper, we tested the ability of each phenotypic variant to (i) be transmitted by IJs, (ii) to optimize the worm's fitness within the insect, and (iii) to counteract the effect of closely related antagonistic bacteria previously shown as being able to totally prevent S. carpocapsae's reproduction within the insect. We found that IJs did associate with cells of both phenotypes but that the variant 2 cells were preferentially retained by the nematodes when both variants were present in the insect. Both phenotypic variants led to the same fitness of S. carpocapsae in insects not infected by antagonistic bacteria. In insects infected by antagonistic bacteria, both variants were able to provide protection to S. carpocapsae. Nevertheless, this protection depended on the phenotypic variant and the antagonistic bacteria that were co-injected into the insect. Further analysis conduced in vitro showed that this variability could be partly linked to the sensitivity of each antagonistic bacterium to xenorhabdicin, produced by X. nematophila.

We compared the clinical outcome and vertical transmission of six canine Neospora caninum isolates using a pregnant BALB/c model. Four of the isolates were obtained from oocysts of naturally infected dogs (Nc-Ger2, Nc-Ger3, Nc-Ger6 and Nc-6Arg) and two were from diseased dogs with neurological signs (Nc-Bahia and Nc-Liv). The dams were inoculated with 2×106 tachyzoites of each isolate at day 7 of pregnancy. Morbidity, mortality and the antibody responses were evaluated in both the dams and the offspring, as was parasite transmission to the progeny. The mortality rates varied from 100% in Nc-Bahia and Nc-Liv-infected pups to 19% or less for those infected with the isolates from oocysts. The vertical transmission rates varied from 9 to 53% for N. caninum from oocysts, compared with 100% for the Nc-Liv and Nc-Bahia isolates. All dams showed specific IgG responses against tachyzoite and rNc-GRA7 antigens, confirming Neospora infection. The highest IgG levels were detected in mice inoculated with the Nc-Liv and Nc-Bahia isolates. These results demonstrate marked differences in virulence between the N. caninum isolates obtained from oocysts and neurologically affected dogs. This variability could help us to explain the differences in the outcome of the infection in definitive and intermediate hosts.

Due to side-effects and inefficiency of the drugs used in malaria treatment, finding alternative medicine with less side-effects has attracted much attention. In this regard, in the present study, nanocomposite synthesized and its effects on the metabolites of P. falciparum were investigated. Subsequent to synthesis of nanocomposites, characterization was carried out using nuclear magnetic resonance (NMR), liquid chromatography-mass spectrometry (LC-MS), scanning electron microscopy, dynamic light scattering and Fourier-transform infrared tests. Solubility and drug release were measured and its toxicity on Vero cell was assessed using the MTT assay. The antiparasitic effect of the nanocomposite on the metabolites of P. falciparum was investigated by 1H NMR spectroscopy. Among synthesized nanocomposites, the average size of 239 nm showed suitable solubility in water as well as slow drug release. The MTT assay showed no toxicity for Vero cell lines. Concentrations of 2.5 μg mL−1 of nanocomposite eliminated 82.6% of the total parasites. The most effected metabolic cycles were glyoxylate and dicarboxylate metabolism. In this study, 1H NMR spectroscopy was used with untargeted metabolomics to study the effect of the nanocomposite on P. falciparum. Playing an essential role in understanding drug-target interactions and characterization of mechanism of action or resistance exhibited by novel antiprotozoal drugs, can be achieved by targeting metabolic using LC-MS.

We compared the distribution of the digenean Pholeter gastrophilus in the stomach of 27 harbour porpoises, Phocoena phocoena, 27 striped dolphins, Stenella coeruleoalba, 18 bottlenose dolphins, Tursiops truncatus, and 100 long-finned pilot whales, Globicephala melas. The stomach of these species is composed of 4 chambers of different size, structure and function. In all species, P. gastrophilus was largely restricted to the glandular region of the stomach, but the parasite tended to favour the fundic chamber in bottlenose dolphins and harbour porpoises, the pyloric chamber in pilot whales, and none in striped dolphins. However, predictability at infrapopulation level was generally low, suggesting a weak preference of P. gastrophilus for any of the chambers. Three hypotheses were tested to investigate a common cause for the distribution of P. gastrophilus in all host species, namely, colonization of chambers was (1) sequential, (2) dependent on chamber size, or (3) dependent on the passage time of food through the whole stomach. The latter hypothesis was indirectly tested by assuming, based on previous evidence from other vertebrates, that the greater the size of the stomach and/or the energy content of prey, the greater the delay of food passage. We found no compelling evidence that chamber colonization was sequential, or related to chamber size in any species. However, the distribution of P. gastrophilus was significantly more anteriad when the host species had larger stomachs and, particularly, when hosts fed on prey with higher caloric content. Accordingly, the stomach distribution of P. gastrophilus at this scale seems to be passively driven by features of the diet and digestive physiology of each host species. This study provides a general framework to formulate null hypotheses in future studies on microhabitat choice by parasites.

The transformation of cercariae of Schistosoma mansoni into schistosomula has previously been achieved by passage through membranes of biological origin. This transformation has now been effected in the absence of a membrane. Chemical stimulus for transformation results from contact with human or rat skin lipid and fractionation of the latter led to localization of the active component in a fraction containing phospholipids. The series of processes involved in transformation was subsequently found to be set in motion by the action of egg lecithin-water mixtures on an aqueous suspension of cercariae and yields of 90% of live schistosomula may be obtained reproducibly with crude egg lecithin-water in the ratio 1:50. The juvenile worms were found, by accepted criteria, to be identical to those produced by the normal penetration procedure. Purified lecithin does not produce the same stimulus and the effect of the physical state and of possible impurities present in crude egg lecithin is discussed.

This work was supported by the Conselho Nacional de Pesquisas (Brazil), the Banco Nacional de Desenvolvimento Econâmico FUNTEC contracts nos. 99 and 110 (Brazil), U.S. Army Grants nos. DAHC-19-70-G-0017 (to B.G.) and DAHC-19-70-G-0070 (to J.P.) and by the FORGE Foundation. Contribution no. 18 from the Schistosomiasis Research Unit, Institute of Biological Sciences, Federal University of Minas Gerais, Brazil.

In a very important revision of the Streblidae, Speiser (1900) classified them into two subfamilies: (1) the Nycteriboscinae and (2) the Streblinae. To the first subfamily he referred all the genera of the world, with the exception of Strebla and Euctenodes. Upon the presence of the ctenidium and some other characters these two American genera were included in the second subfamily. This classification of the subfamilies was also adopted by Kessel (1924, 1925).

The effect of temperature (19–36 °C) and snail host density (0.014–10 snails/1) on the snail-finding capacity of Echinostoma caproni cercariae is described. The initial swimming speed increased whereas the length of the infective period decreased with increasing temperature. The combined effect resulted in the E. caproni cercarial snail-finding capacity being temperature independent in the range 19 to 36 °C at a snail density of 0·014 snails/1. A moderate temperature dependency was, however, seen at a lower snail density. The cercarial snail-finding capacity was snail-host density dependent in the density range 0.014 to 1 snail/1. The findings from this study show that a relatively low and biologically realistic snail host density must be used in experimental studies if realistic estimates of the dynamics of cercarial transmission are to be obtained.