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.

Community data on the prevalence of helminth infections is important for guiding health policy, but expensive to collect. As a result most surveys focus on school-aged children, frequently using schools as a sentinel population. Since there already exists a vast amount of data on infection levels in school-aged children, but limited community-based data, we undertook a literature search on age-stratified infection data for intestinal nematode infections and schistosomiasis in Africa, to investigate whether estimates of the prevalence of infection in school-aged children could provide an index for determining community prevalence. The observed data on prevalence of infection in infants, school-aged children and adults were fitted using linear and logistic regression models which take into account variation in sample prevalences. Despite the wide variation in study sites, the observed relationship between community prevalence and school-aged prevalence was remarkably consistent for each parasite species. The prevalence of infection in school-aged children alone was shown to be higher than the predicted prevalence in the community, but the degree of overestimation was dependent on the parasite species and the level of infection. The results suggest that the prevalence of infection in school-aged children could provide a cost-effective predictive tool which can be used at a district/national level to identify target areas for control and to evaluate the numbers at risk of infection.

The work reported here describes the comparative ability of albendazole (ABZ), fenbendazole (FBZ) and triclabendazole (TCBZ) to penetrate through the tegument of mature Fasciola hepatica, and the influence of the physicochemical composition of the incubation medium on the drug diffusion process. The data obtained from the trans-tegumental diffusion kinetic studies were complemented with the determination of lipid-to-water partition coefficients (octanol-water) for the benzimidazole (BZD) anthelmintic drugs assayed. Sixteen-week-old F. hepatica were obtained from untreated artificially infected sheep. The flukes were incubated (37 °C) over 60 and 90 min in incubation media (pH 7·4) prepared with different proportions of ovine bile and Krebs' Ringer Tris (KRT) buffer (100, 75, 50, 25 and 0% of bile) containing either ABZ, FBZ or TCBZ at a final concentration of 5 nmol/ml. After the incubation time expired, the liver fluke material was chemically processed and analysed by high performance liquid chromatography (HPLC) to measure drug concentrations within the parasite. Additionally, the octanol-water partition coefficients (PC) for each molecule were calculated (as an indicator of drug lipophilicity) using reversed phase HPLC. The 3 BZD molecules were recovered from F. hepatica at both incubation times in all incubation media assayed. The trans-tegumental diffusion of the most lipophilic molecules ABZ and FBZ (higher PC values) tended to be greater than that observed for TCBZ. Interestingly, the uptake of ABZ by the liver flukes was significantly greater than that measured for TCBZ, the most widely used flukicidal BZD compound. This differential uptake pattern may be a relevant issue to be considered to deal with TCBZ-resistant flukes. Drug concentrations measured within the parasite were lower in the incubations containing the highest bile proportions. The highest total availabilities of the 3 compounds were obtained in liver flukes incubated in the absence of bile. Altogether, these findings demonstrated that the entry of the drug into a target parasite may not only depend on a concentration gradient, the lipophilicity of the molecule and absorption surface, but also on the physicochemical composition of the parasite's surrounding environment.

Both verbal and mathematical models of parasite virulence predict that genetic diversity of microparasite infections will influence the level of costs suffered by the host. We tested this idea by manipulating the number of co-existing clones of Plasmodium mexicanum in its natural vertebrate host, the fence lizard Sceloporus occidentalis. We established replicate infections of P. mexicanum made up of 1, 2, 3, or >3 clones (scored using 3 microsatellite loci) to observe the influence of clone number on several measures of parasite virulence. Clonal diversity did not affect body growth or production of immature erythrocytes. Blood haemoglobin concentration was highest for the most genetically complex infections (equal to that of non-infected lizards), and blood glucose levels and rate of blood clotting was highest for the most diverse infections (with greater glucose and more rapid clotting than non-infected animals). Neither specific clones nor parasitaemia were associated with virulence. In this first experiment that manipulated the clonal diversity of a natural Plasmodium-host system, the cost of infection with 1 or 2 clones of P. mexicanum was similar to that previously reported for infected lizards, but the most complex infections had either no cost or could be beneficial for the host.

Research was undertaken to clarify the true taxonomic position of the terrestrial tortoise apicomplexan, Haemogregarina fitzsimonsi (Dias, 1953). Thin blood films were screened from 275 wild and captive South African tortoises of 6 genera and 10 species between 2009–2011. Apicomplexan parasites within films were identified, with a focus on H. fitzsimonsi. Ticks from wild tortoises, especially Amblyomma sylvaticum and Amblyomma marmoreum were also screened, and sporogonic stages were identified on dissection of adult ticks of both species taken from H. fitzsimonsi infected and apparently non-infected tortoises. Parasite DNA was extracted from fixed, Giemsa-stained tortoise blood films and from both fresh and fixed ticks, and PCR was undertaken with two primer sets, HEMO1/HEMO2, and HepF300/HepR900, to amplify parasite 18S rDNA. Results indicated that apicomplexan DNA extracted from tortoise blood films and both species of tick had been amplified by one or both primer sets. Haemogregarina  fitzsimonsi 18S rDNA sequences from tortoise blood aligned with those of species of Hepatozoon, rather than those of species of Haemogregarina or Hemolivia. It is recommended therefore that this haemogregarine be re-assigned to the genus Hepatozoon, making Hepatozoon fitzsimonsi (Dias, 1953) the only Hepatozoon known currently from any terrestrial chelonian. Ticks are its likely vectors.

Encephalitozoon cuniculi is an obligate intracellular pathogen that has a wide host distribution, but primarily affects rabbits. The aim of this study was to characterize both the cell-mediated and the antibody response in rabbits after experimental infection using 2 different infection routes: oral and ocular. SPF rabbits were infected with low (103 spores) and high (107 spores) infection doses. Monitored parameters included clinical signs, detection of spores in urine, antibody response detected with ELISA, and cell-mediated immunity detected by antigen-driven lymphocyte proliferation. At week 13 post-infection, half of the rabbits in each group were suppressed by intramuscular administration of dexamethasone. At week 18 post-infection, animals were euthanized. Clinical signs were mild with exacerbation after immunosuppression. Spores in urine and antigen-specific cell-mediated immunity were detected from weeks 5 and 4 post-infection, respectively. Specific IgM was detected 1 week after infection, and IgG antibodies followed 1 week later in rabbits infected with the high dose. Immunological responses were dose dependent. The authors can conclude that both oral and ocular experimental infection with E. cuniculi resulted in an immune response of the infected animals. Rabbits could be used as an experimental model for the study of ocular microsporidiosis.

This study applies flexible statistical methods to morphometric measurements obtained via light and scanning electron microscopy (SEM) to discriminate closely related species of Gyrodactylus parasitic on salmonids. For the first analysis, morphometric measurements taken from the opisthaptoral hooks and bars of 5 species of gyrodactylid were derived from images obtained by SEM and used to assess the prediction performance of 4 statistical methods (nearest neighbours; feed-forward neural network; projection pursuit regression and linear discriminant analysis). The performance of 2 methods, nearest neighbours and a feed-forward neural network provided perfect discrimination of G. salaris from 4 other species of Gyrodactylus when using measurements taken from only a single structure, the marginal hook. Data derived from images using light microscopy taken from the full complement of opisthaptoral hooks and bars were also tested and nearest neighbours and linear discriminant analysis gave perfect discrimination of G. salaris from G. derjavini Mikailov, 1975 and G. truttae Gläser, 1974. The nearest neighbours method had the least misclassifications and was therefore assessed further for the analysis of individual hooks. Five morphometric parameters from the marginal hook subset (total length, shaft length, sickle length, sickle proximal width and sickle distal width) gave near perfect discrimination of G. salaris. For perfect discrimination therefore, larger numbers of parameters are required at the light level than at the SEM level.

Myxosporeans are among the most common parasites of marine fish. Their economic importance is mainly as pathogens of both wild and farmed fish, but they have also been used as biological tags in population studies of their fish hosts. Here we review the literature and show the distribution of different families of Myxosporea infecting marine fishes in the world's oceans – the North Atlantic, South Atlantic, North Pacific, South Pacific and Indian. We also analyse their distribution in different orders of marine fishes. New families, genera and species of marine Myxosporea are continually being described and many more await description. Some regions, in particular the North Atlantic, have been more thoroughly investigated than others, so the analyses we present may not reflect the true distributions and we acknowledge that these may change considerably as other regions are investigated more fully. The distribution of myxosporean families in different taxonomic groups of marine fishes can indicate phylogenetic relationships between parasite and host and suggest the origins of different myxosporean taxa. We present some examples, while recognizing that new molecular information on phylogenetic relationships within the Myxozoa will lead to major changes in classification.

Temporal changes in the prevalence of antigenic variants in Plasmodium falciparum populations have been interpreted as evidence of immune-mediated frequency-dependent selection, but evolutively neutral processes may generate similar patterns of serotype replacement. Over 4 years, we investigated the population dynamics of P. falciparum polymorphisms at the community level by using 11 putatively neutral microsatellite markers. Plasmodium falciparum populations were less diverse than sympatric P. vivax isolates, with less multiple-clone infections, lower number of alleles per locus and lower virtual heterozygosity, but both species showed significant multilocus linkage disequilibrium. Evolutively neutral P. falciparum polymorphisms showed a high turnover rate, with few lineages persisting for several months in the population. Similar results had previously been obtained, in the same community, for sympatric P. vivax isolates. In contrast, the prevalence of the 2 dimorphic types of a major antigen, MSP-2, remained remarkably stable throughout the study period. We suggest that the relatively fast turnover of parasite lineages represents the typical population dynamics of neutral polymorphisms in small populations, with clear implications for the detection of frequency-dependent selection of polymorphisms.

Research on the interactions between the field voles (Microtus agrestis) of Kielder Forest and their natural parasites dates back to the 1930s. These early studies were primarily concerned with understanding how parasites shape the characteristic cyclic population dynamics of their hosts. However, since the early 2000s, research on the Kielder field voles has expanded considerably and the system has now been utilized for the study of host–parasite biology across many levels, including genetics, evolutionary ecology, immunology and epidemiology. The Kielder field voles therefore represent one of the most intensely and broadly studied natural host–parasite systems, bridging theoretical and empirical approaches to better understand the biology of infectious disease in the real world. This article synthesizes the body of work published on this system and summarizes some important insights and general messages provided by the integrated and multidisciplinary study of host–parasite interactions in the natural environment.

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.

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.

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.

The total number and distribution of schistosomula of Schistosoma mansoni and S. haematobium in all tissues and organs of mice from infection to 14–27 days was determined by compressed tissue autoradiography. Attrition of schistosomula, manifested as a decrease in the number of autoradiographic foci, was observed in organs other than the liver. Attrition commenced about 2 days after cercariae entered the skin, and conformed to a single exponential function with a rate constant (± standard error) of 7·0±0·5%/day for S. mansoni and 3·2±0·7%/day for S. haematobium. The temporal distribution of schistosomula of S. mansoni and S. haematobium differed quantitatively. In the case of S. mansoni, concomittant with a decrease in skin counts, the lung curve rose rapidly to a peak centred on day 6 and thereafter decreased more or less parallel to the total body curve. Significant accumulation in the liver was not observed until day 7, whereupon liver counts rose steadily to a plateau that extended from about day 14 to the end of the experiment and approximated the number of adult worms recovered from the hepatic portal vessels on day 42. A maximum of 26% and mean of 12% of all foci in the body were counted on autoradiograms of tissues other than the skin, lung and liver. The pelt averaged 14% of the body weight yet schistosomula were detected only in the area initially exposed to cercariae. The eviscerated carcass averaged 54% of the body weight yet contained only 0·8% −3·4 % of the schistosomula during the period of accumulation in the liver. Between day 6 and day 14, the ratio of schistosomula in the pulmonary circulation to schistosomula in the systemic circulation did not remain constant, as would be the case if schistosomula circulated passively and randomly, but instead displayed a statistically significant decrease from 0·92 and 0·85. For these reasons, it was considered unlikely that schistosomula had circulated randomly and repeatedly through the pulmonary and systemic circulations and entered the hepatic portal system by chance, as hypothesized by Miller & Wilson (1980). Instead it was considered more probable that schistosomula migrating from lungs to liver had followed a directed path through intervening vessels (Kruger, Heitman, van Wyk & McCully, 1969) or tissues (Wilks, 1967). Schistosoma haematobium distribution patterns differed from those of S. mansoni in slower movement of schistosomula from skin, their peak accumulation in lungs at about day 8 followed by a low rate of decrease, and a very low liver plateau which approximated the number of adult worms recovered from the hepatic portal vessels on day 42.

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.

The Index of Association (IA) has been proposed by Maynard Smith et al. (1993) as a general method for characterizing the population structures of microorganisms as either: clonal, epidemic, cryptic species or panmictic. With reference to the current debate surrounding the mode of reproduction in parasitic protozoa, this study explores (i) the suitability and limitations of the IA for characterizing populations of Trypanosoma brucei s.l., and (ii) the idea that the significance of genetic differences between populations may be better understood if the evolution, spread and temporal stability of certain parasite genotypes are also considered. Four populations of T. brucei from Côte d'Ivoire, Uganda and Zambia are analysed using the IA and a complementary test for linkage disequilibrium, test f of Tibayrenc, Kjellberg & Ayala (1990). The two populations from Uganda are characterized as epidemic, while the others appear more or less clonal; the merits of the two methods are compared. The implications of the various population classifications are discussed with reference to genotype longevity in each region; the evolutionary and biomedical consequences of the genetic non-homogeneity of T. brucei are reviewed.

An account is given of the spontaneous occurrence in the Anglo-Egyptian Sudan of strains of the camel trypanosome, Trypanosoma evansi, devoid of kinetonuclei.

These cases, of which four are recorded, were observed in widely separated localities within the enzootic area of camel trypanosomiasis.

A critical review is given of the question of “akinetonucleate” trypanosomes in general—in strains produced experimentally and in those occurring spontaneously—with special reference to their mode of origin and their significance in the production of new races.

The prevalence of “akinetonucleate” forms in normal strains of trypanosomes has been estimated in the case of a number of species and races hitherto not examined from this point of view, all the existing data being brought together in tabular form.

While the factors causing the appearance of “akinetonucleate” individuals and strains under natural conditions could not be ascertained, irregular division of the trypanosomes is held to be mainly responsible for the elimination of the kinetonucleus and for the persistence of the aberrant condition.

1. Aphidius matricariae Hal. is an internal parasite of Myzus persicae Sulz.; it was also found parasitizing M. circumflexus Buckt. and Aulacorthum solani Kalt. (Myzus pseudosolani Theob.).

2. The percentage of aphides successfully parasitized increased with an increased number of strikes per host. A single female can lay up to 350 eggs, and one female successfully parasitized 309 M. persicae.

3. The female parasite reaches its maximum activity during the 2nd or 3rd day of oviposition, and this activity has no correlation with light intensity.

4. The disappearance of supernumerary larvae due to cannibalism has been observed.

5. The species is arrhenotokous. Mating takes place from 2 hr. after emergence. In the progeny of early matings the females preponderated; in later matings the sex ratio was approximately 1:1.

6. The parasite appears to fly under the same conditions as those generally favourable for aphides. It is generally disseminated by flight of the adult itself and through localized movements of parasitized apterous aphides.

7. The host is unable to reproduce if ‘struck’ in the 3rd instar or earlier.

This paper describes further characteristics of the immunization of sheep against schistosomes using live, irradiated schistosomula. Sheep immunized with a non-virulent strain of Schistosoma mattheei were protected against a more virulent strain of the same species for over a year. As there was no evidence that the irradiated parasites were able to persist this long, it was concluded that the vaccine had induced a sterile resistance. Heterologous vaccination, using irradiated S. mattheei schistosomula to immunize against S. bovis or irradiated S. mansoni schistosomula to immunize against S. mattheei, failed to induce any protection.

The amount of [14C]methionine in the stomach, anterior and posterior intestine wall and lumen, blood and caecum of a rat, and in the anterior and posterior parts of the tapeworm Hymenolepis diminuta in the intestine, was measured at intervals from 10 min to 10 h after feeding a meal of 2 g glucose and 37.5 mg methionine by stomach tube.

Nearly all the meal left the stomach within 5 h; the amount entering the tapeworm depended on the position of the worm. That part lying in the anterior half of the intestine was subjected to a rapid influx reaching a maximum concentration of at least 8 mM within 1–3 h after the meal was administered. The level in the worm then dropped to about 20 % of the peak level, although the stomach was still emptying and the level of methionine in the intestine wall was still rising. This was a stable state, and equal to the concentration in the posterior part of the worm which was reached 2 h after the rat was fed.

The reason why the anterior part of the worm should show a big influx and efflux is discussed and related to changes in concentration of methionine in the lumen. The lack of such a flux into and out of the worm further down the intestine is related to the host's homeostatic control of amino acid balance in the intestine.

The results prove that dietary imbalance of amino acids affect the amino acid pool in a tapeworm for about 3 h, if it lies in the anterior of the gut. A dietary regime is suggested which should produce a prolonged effect and thereby retard growth of the worm.

The weights of the parts of the worms found in the anterior and posterior half of the small intestine revealed an unexpected phenomenon; the majority of the tapeworm, by weight, lies in the anterior of the intestine for about 4 h following a meal, but in the posterior of the intestine after the stomach has emptied.

It is a great pleasure to thank Miss Gillian Moore and Miss Patricia Grant for technical assistance, and Shell International Petroleum Company, London, for financial support.