Histological studies using horse-radish peroxidase (HRP) as a marker, injected iontophoretically through a recording electrode, have indicated the origins of potentials encountered upon advancement of the electrode into and beneath the dorsal surface of adult male Schistosoma mansoni. The first potential encountered, having a value of – 51 ± 0·6 mV originates across the outer tegumental membrane. The next potential has a value of — 28 ± 0·6 mV and originates in the muscle masses underlying the tegument. Finally, a potential having the value — 10 ± 0·5 mV originates within the basal lamina and the interstitial fibres and extracellular space surrounding the muscle. Altering ion concentrations in the bathing medium (i.e. high K+, low Na+, zero Ca2+, low Cl−, high Li+) depolarizes all three potentials. External applications of ouabain and the anti-schistosomal, praziquantel, also cause depolarization of the potentials. It appears that the muscle potential and the tegumental potential are primarily K+-dependent. The depolarizing effects of ouabain, LiCl and low Na+ suggest that active transport is important in the maintenance of the muscle potential, just as is the case for the tegumental potential. There appears to be a close correlation between changes in the tegumental, muscle and extracellular space potentials. The correlation between tegument and muscle potential changes might be explained by junctional complexes between tegumental cell bodies and muscle cell bodies.

The influence of the late components of complement (C) on percutaneous primary infections with Schistosoma mansoni was studied in inbred mice genetically deficient or intact in C5. Worm recoveries were diminished in C5-deficient male mice as compared to C5-intact animals. Twenty-four-day-old parasites were also shorter following growth in C5-deficient animals. At 7 weeks after infection, female schistosomes, but not male parasites, were shorter and fewer eggs/schistosome pair were deposited in the livers of C5-deficient as compared to C5-intact male mice. In addition to the presence of C5, the mouse sex was found to influence the outcome of an infection. Schistosomes showed a reduced infection rate and were relatively stunted after 24 days of growth in female mice of the two strains as compared to male mice of the corresponding strains. The results suggest that C5 plays no role in defence against a primary infection in mice.

1. Suspensions made from infected caecal material, derived from chickens which had been artificially infected with Histomonas meleagridis by the administration of embryonated Heterakis ova, can be used to transmit the organisms to other chickens when administered per os.

2. Infections with histomonad suspensions can only be obtained in chickens that have been starved, or in chickens that are feeding but have received an alkali mixture immediately before the suspension is administered. The highest incidence of infections is obtained by administering suspensions to starved chickens that have received 1 g. alkali mixture beforehand.

3. Successful infection appears to depend upon the pH of the gizzard and, possibly, of the upper intestine. The incidence of lesions produced by feeding suspension is increased as the contents of the gizzard become alkaline. It is also suggested that the consistency of the food may play a part, as coarse food, along with the infective material, is retained in the gizzard until it is completely ground up and mixed with the gastric juice.

4. In one series of passages the suspension became attenuated after fourteen passages through chickens. In other cases suspensions became attenuated after only six passages through chickens.

5. The courses of infections produced by histomonad suspensions and Heterakis ova are compared.

6. The advantages of the method of using histomonad suspensions are discussed in their relation to the screening of chemotherapeutical agents.

E. praeox has been isolated in Britain for the first time and is similar in regard to its oocyst size, prepatent time, life-cycle and the development of host immunity to the reports of Tyzzer et al. (1932). Oocysts had mean dimensions of 20·4 × 17·45 μm and the first oocysts were discharged 83½ h after infection.

Oocyst production and life-cycle studies suggest that at least three generations of schizogony precede gametogony and that at least one further generation of schizogony is needed to explain the oocyst reproduction resulting from a small dose of oocysts. Young chickens 1½–3 weeks of age were not such suitable hosts as older chickens (6 weeks) and the reproduction of the parasite was seriously reduced when large numbers (e.g. 105, 106) were used to infect chickens. A high degree of resistance to reinfection occurred after a single exposure to infection.

Sporozoite infections of the caeca or cloaca resulted in the infection of the usual small intestine site; there was no development at the site of inoculation and attempts to infect the embryo allantois did not succeed.

I wish to thank Dr M. Elaine Rose for the surgery required for the caecal infections, for her interest throughout and for help with the manuscript; Dr R. F. Gordon for help with the manuscript and permission to publish and Mr B. J. Millard for skilled technical assistance.

Human–wildlife interactions have reached unprecedented levels, and humans are influencing the earth's ecosystems more rapidly and extensively than ever before. This situation is cause for serious concern, especially since disease interactions between wildlife and humans have been recognized as major conservation threats. In this study, long-tailed macaques, Macaca fascicularis, from 2 forest parks located in north-eastern Thailand were investigated to determine the influence of habitat modification by humans on helminth parasite associations in non-human primates. Macaque populations with contact to anthropogenically modified environments were compared with sylvatic groups in nearby natural environments. In order to test for human–non-human primate transmission of parasites, the local human populations were also examined. Humans were infected with a number of potentially pathogenic parasites, including Opisthorchis viverrini and Strongyloides stercoralis. However, eggs of these helminths were not detected in macaque feces. Thus, no direct parasite transfer from humans to non-human primates could be confirmed. However, macaque groups with more frequent contact with human-modified habitats, and a higher portion of human-provided food in their diet, had significantly higher prevalences and intensities of Strongyloides fuelleborni and of an intestinal fluke (probably Haplorchis sp.) than sylvatic groups. Positive correlations were found between the time foraging on the ground and infection with S. fuelleborni, and the amount of human-provided food and intestinal fluke infection. Human alteration of habitat and associated modifications in non-human primate behaviour are likely to play a role in determining the occurrence, prevalence and intensity of zoonotic helminth infection of wild non-human primates.

Malaria remains one of the most significant global public health burdens, with nearly half of the world's population at risk of infection. Malaria is not however a monolithic disease – it can be caused by multiple different parasite species of the Plasmodium genus, each of which can induce different symptoms and pathology, and which pose quite different challenges for control. Furthermore, malaria is in no way restricted to humans. There are Plasmodium species that have adapted to infect most warm-blooded vertebrate species, and the genus as a whole is both highly successful and highly diverse. How, where and when human malaria parasites originated from within this diversity has long been a subject of fascination and sometimes also controversy. The past decade has seen the publication of a number of important discoveries about malaria parasite origins, all based on the application of molecular diagnostic tools to new sources of samples. This review summarizes some of those recent discoveries and discusses their implication for our current understanding of the origin and evolution of the Plasmodium genus. The nature of these discoveries and the manner in which they are made are then used to lay out a series of opportunities and challenges for the next wave of parasite hunters.

Factors such as the particular combination of parasite–mosquito species, their co-evolutionary history and the host's parasite load greatly affect parasite transmission. However, the importance of these factors in the epidemiology of mosquito-borne parasites, such as avian malaria parasites, is largely unknown. Here, we assessed the competence of two mosquito species [Culex pipiens and Aedes (Ochlerotatus) caspius], for the transmission of four avian Plasmodium lineages (Plasmodium relictum SGS1 and GRW11 and Plasmodium cathemerium-related lineages COLL1 and PADOM01) naturally infecting wild house sparrows. We assessed the effects of parasite identity and parasite load on Plasmodium transmission risk through its effects on the transmission rate and mosquito survival. We found that Cx. pipiens was able to transmit the four Plasmodium lineages, while Ae. caspius was unable to transmit any of them. However, Cx. pipiens mosquitoes fed on birds infected by P. relictum showed a lower survival and transmission rate than those fed on birds infected by parasites related to P. cathemerium. Non-significant associations were found with the host–parasite load. Our results confirm the existence of inter- and intra-specific differences in the ability of Plasmodium lineages to develop in mosquito species and their effects on the survival of mosquitoes that result in important differences in the transmission risk of the different avian malaria parasite lineages studied.

The lectin-inhibitory sugars D-glucosamine (GlcN) and N-acetyl D-glucosamine (GlcNAc) are known to enhance susceptibility of the tsetse fly vector to infection with Trypanosoma brucei. GlcNAc also stimulates trypanosome growth in vitro in the absence of any factor derived from the fly. Here, we show that GlcNAc cannot be used as a direct energy source, nor is it internalized by trypanosomes. It does, however, inhibit glucose uptake by binding to the hexose transporter. Deprivation of D-glucose leads to a switch from a metabolism based predominantly on substrate level phosphorylation of D-glucose to a more efficient one based mainly on oxidative phosphorylation using L-proline. Procyclic form trypanosomes grow faster and to higher density in D-glucose-depleted medium than in D-glucose-rich medium. The ability of trypanosomes to use L-proline as an energy source can be regulated depending upon the availability of D-glucose and here we show that this regulation is a graded response to D-glucose availability and determined by the overall metabolic state of the cell. It appears, therefore, that the growth stimulatory effect of GlcNAc in vitro relates to the switch from D-glucose to L-proline metabolism. In tsetse flies, however, it seems probable that the effect of GlcNAc is independent of this switch as pre-adaptation to growth in proline had no effect on tsetse infection rate.

The in vitro activity of artemether against 63 African isolates of Plasmodium falciparum from Libreville, Gabon was evaluated using an isotopic drug susceptibility semi-microtest. The 50% inhibitory concentration (IC50) values for artemether were in a narrow range from 0·8 to 34·8 nm (mean IC50 5·0 nm) and the 95% confidence interval (CI95%) was 3·6–6·3 nm. In vitro decreased susceptibility or resistance were observed with artemether (14%), to chloroquine (90%), to quinine (32%). Isolate susceptibility to amodiaquine and halofantrine was high i.e. 100% and 98%, respectively. There was a significant positive correlation between responses to artemether and amodiaquine (r2=0·45, P<0·001), artemether and chloroquine (r2=0·36, P<0·001), artemether and quinine (r2=0·31, P<0·001), and artemether and halofantrine r2=0·19, P<0·01). Positive correlation between these drugs suggests in vitro cross-resistance or at least common features in drug uptake and/or mode of action or resistance.

Soil-transmitted helminthiasis (STH), among the most common neglected tropical diseases, is a major public health problem in Malaysia with a possible impact on the nutritional status and school participation of rural children. This study was carried out among Aboriginal schoolchildren, living in an endemic area for STH in Malaysia, to determine the possible relationship between intestinal helminthiasis and school absenteeism. We also evaluated whether successful treatment of the infection will affect school attendance among the subjects. Stool analysis revealed that more than 90% of the subjects were infected with at least 1 helminth species, with Ascaris lumbricoides and Trichuris trichiura infections being most prevalent. Infection of moderate-to-heavy worm burdens, low level of fathers' education and anaemia were identified as the significant predictors of high absenteeism among the subjects (P<0·05). Following treatment of the infected children, it was found that school absenteeism was reduced significantly (P<0·01). In conclusion, STH continues to have significant impacts on public health, particularly in rural communities with a negatively significant effect on the school participation of Aboriginal children. A school-based de-worming programme should be introduced and incorporated in the current educational assistance targeted towards the Aboriginal communities, under the auspices of the government.

The aims of the study were (1) to describe the Dactylogyrus communities living on selected South European Barbus species, (2) to analyse morphometric variability of their attachment and reproductive organs, and (3) to perform molecular phylogenetic analyses, in order to investigate the mode of speciation in Dactylogyrus species parasitizing congeneric hosts. In Bulgaria, Dactylogyrus crivellius, D. dyki and D. petenyi were found on B. balcanicus, and D. dyki on B. cyclolepis. In Spain, Dactylogyrus carpathicus and D. dyki were detected on B. meridionalis. Morphometric analyses of D. dyki revealed significant differences in the attachment and reproductive organs when individuals from different Barbus species were compared. Two monophyletic groups were recognized from the molecular phylogenetic analyses: the first included D. carpathicus and D. crivellius which have large body size and anchors, with a weakly supported basal position of D. malleus from B. barbus; the second included D. dyki and D. petenyi which have small body and anchor sizes. The comparison of host and parasite phylogenies did not indicate the intrahost speciation. Intraspecific molecular variability was found between individuals of D. dyki and D. carpathicus from different Barbus species, suggesting the need for a taxonomic revision for these species.

Gall bladders from 273 otter carcasses, collected throughout England and Wales, were screened to assess the status of gall bladder parasites in the Eurasian otter, Lutra lutra. The digenean Pseudamphistomum truncatum had previously been found in UK otters collected between 2000 and 2007. The parasite was established in Somerset and Dorset but its distribution elsewhere in the UK was largely unknown. In the current study, P. truncatum was also found to be abundant in south Wales, with occasional cases elsewhere, but appears to be absent from the north of England. Overall, 11·7% of otters were infected with 1–238 P. truncatum. A second digenean, Metorchis albidus, previously unreported in British otters, was found in the biliary system of 6·6% of otters. M. albidus appears well established in Suffolk, Norfolk and north Essex but was recorded elsewhere rarely. Both parasites are associated with pathological damage to the otter gall bladder. The recent discovery of these two non-native parasites provides a unique opportunity to assess their impact on native British fauna.

Sarcoptes scabiei is an important parasitic mite in both man and animals. Little is known about the molecular interactions between this pathogen and its host. This is in part explained by the paucity of mite-derived material, including antigens. To extend the knowledge of the molecular repertoire in S. scabiei, we have performed a gene survey by an expressed sequence tag (EST) analysis. A total of 1020 ESTs were generated from an S. scabiei cDNA library. The average sequence read was 510 bp after editing and the overall sequencing success was 89%. Clustering of the sequences resulted in 76 clusters, comprising 36% of the ESTs. Sequence similarity searches showed that almost half of the S. scabiei ESTs could be assigned a putative identity. Many of these transcripts shared similarity with genes involved in basic metabolism and cellular organization. In the data set we also identified several proteases and other types of potential allergens implicated in various disease mechanisms. A relatively large fraction of the ESTs coded for different proteins carrying protease inhibitor-like domains. The clones with no similarity to previously identified genes constituted 11% of our transcripts. The EST data generated in this study will be a valuable resource in further studies of the biology of S. scabiei and in the identification of genes that can serve as potential targets in the control of the parasite.

Human strongyloidiasis is a serious disease mostly attributable to Strongyloides stercoralis and to a lesser extent Strongyloides fuelleborni, a parasite mainly of non-human primates. The role of animals as reservoirs of human-infecting Strongyloides is ill-defined, and whether dogs are a source of human infection is debated. Published multi-locus sequence typing (MLST) studies attempt to elucidate relationships between Strongyloides genotypes, hosts, and distributions, but typically examine relatively few worms, making it difficult to identify population-level trends. Combining MLST data from multiple studies is often impractical because they examine different combinations of loci, eliminating phylogeny as a means of examining these data collectively unless hundreds of specimens are excluded. A recently-described machine learning approach that facilitates clustering of MLST data may offer a solution, even for datasets that include specimens sequenced at different combinations of loci. By clustering various MLST datasets as one using this procedure, we sought to uncover associations among genotype, geography, and hosts that remained elusive when examining datasets individually. Multiple datasets comprising hundreds of S. stercoralis and S. fuelleborni individuals were combined and clustered. Our results suggest that the commonly proposed ‘two lineage’ population structure of S. stercoralis (where lineage A infects humans and dogs, lineage B only dogs) is an over-simplification. Instead, S. stercoralis seemingly represents a species complex, including two distinct populations over-represented in dogs, and other populations vastly more common in humans. A distinction between African and Asian S. fuelleborni is also supported here, emphasizing the need for further resolving these taxonomic relationships through modern investigations.

In order to reveal the mechanisms underlying nurse cell formation during Trichinella spiralis infection, the expression of the factors of tumor necrosis factor-alpha (TNF-α)/TNF receptor 1 (TNFR-1) signalling pathway mediating apoptosis was investigated. The analysed factors included TNF-α, TNFR-1, TNF receptor-associated death-domain (TRADD), caspase 3, caspase 8, TNF receptor associated factor-2 (TRAF2) and receptor interactive protein (RIP), all of which are involved in the TNF-α/TNFR-1 signalling pathway-mediated apoptosis. The quantitative RT-PCR indicated that the infected muscle tissues up-regulate the expression of pro-apoptosis genes (TNF-α, TNFR-1 and TRADD, caspase 3 and caspase 8), and anti-apoptosis genes (TRAF2 and RIP) at the beginning of cyst formation. The expression returned to the normal level after cyst formation. The quantitative RT-PCR analysis of mRNA from tissue samples isolated by laser capture micro-dissection confirmed that the up-regulation of these genes was restricted in infected muscle cells, was not in the inflammation cells around infected muscle cells nor in normal muscle cells. The in situ localization study of pro-apoptosis (TRADD, caspase 3) and anti-apoptosis gene products (TRAF2) indicated that these were expressed in the basophilic cytoplasm (infected muscle cell origin) of the nurse cells. Thus the present study suggests that the TNF-α/TNFR-1 signalling pathway is involved in nurse cell formation.

Amblyomma birmitum sp. nov. is formally described as a new record from 99 Ma old Burmese amber from Myanmar. This confirms the presence of the extant hard tick genus Amblyomma C.L. Koch, 1844 (Ixodida: Ixodidae) in the Late Cretaceous. This discovery is placed in its wider context and some reports of fossil hard ticks, such as a Hyalomma C.L. Koch, 1844 in Eocene Baltic amber, are misidentifications. The genus Amblyomma belongs to the clade Metastriata, a group which probably also accommodates two extinct genera, Cornupalpatum Poinar and Brown, 2003 and Compluriscutata Poinar and Buckley, 2008, also found in Burmese amber. All three fossils are thus only a little younger than published molecular divergence time estimates (ca. 124 ± 17 Ma) for the Metastriata lineage. Amblyomma has a largely Gondwanan distribution today. However, in some biogeographical scenarios, e.g. the Samafrica model, its predicted radiation time postdates the dissolution of the original Gondwana supercontinent raising questions about how its current distribution pattern was achieved.

1. Methods are described for the isolation and maintenance of a pure strain of Eimeria mitis.

2. The prepatent period of the infection in chicks was 101 hr. The mean dimensions of the oocysts were 15·38 × 13·83μ, and sporulation occurred after 48 hr. at 60° F. (15·5° C.). The production of oocysts following infection with known numbers of oocysts is recorded.

3. When 500,000 or more oocysts of E. mitis were given to chicks aged 6, 17 or 26 days the growth rate of the birds was markedly reduced. Young chicks were most susceptible and a few deaths occurred when 6-day-old chicks received 2·5 million oocysts. E. mitis is unlikely to be of pathological importance under normal conditions of good husbandry practice.

4. Pathological lesions due to this species of Eimeria were not found in sections of intestinal tissues.

5. Developmental stages of this parasite were found throughout the whole of the small intestine.

6. The growth-retarding effects and oocyst production by E. mitis were controlled when 0·2% sodium sulphadimidine was given in the drinking water.

The response of sheep given six sensitizing infections each of 3000 infective larvae of H. contortus at fortnightly intervals to a challenge infection of 3000 infective larvae has been studied. While a large proportion of the challenge infection (ca. 3000 parasites) persisted to termination of the experiment on day 141 in sheep (group 3) in which the challenge was superimposed upon the sensitizing infections, only a mean of 437 worms, including both 4th-stage larvae and adults, was recorded at this time in another group (group 5) in which the sensitizing infections were terminated by anthelmintic treatment on day 90, i.e. 8 days before challenge. The proportion of the total worm burdens of the challenge infections arrested at the 4th larval stage was increased for both groups (25% for group 3 and 18% for group 5) by comparison with unsensitized control groups (4% for group 6 and 2% for group 7).

As the effect of immunological control was expressed more vigorously in the group (group 5) in which the sensitizing infections were terminated before challenge it seems likely that recovery of immunological competence occurred in these animals following removal of the sensitizing infections. This conclusion suggests that, owing to the persistence of the sensitizing infections and therefore exposure of the immune mechanism to continuous antigenic insult, the animals which received the superimposed challenge infection had entered a phase of immunological exhaustion during the challenge period.

The ectoparasitic copepod Lepeophtheirus pectoralis exhibited a regular annual cycle of abundance on its hosts. Both the incidence and intensity of infection were greatest in August or September and both declined over winter to their minimum values in April. Very similar cycles were observed in both 1972 and 1973. Analysis of the age structure of the parasite population showed that the increase in abundance coincided with the onset of the breeding season. Although females always outnumbered males the sex ratio approached unity during the breeding season, which extended from May to October. Copulating pairs were most commonly observed during this period.

There was a marked bimodal peak of egg production each year, with the first mode occurring around May and the second mode around August or September. This pattern was produced by a system of alternation of generations operating within the parasite population, with a summer type generation (distinguishable by its rapid maturation rate and reduced longevity) alternating with an overwintering type of generation.

Resistance to E. necatrix induced by complete life-cycle infections was transferable from duodenum to caeca and vice versa.

Complete life-cycle infections with E. tenella protected against subsequent attempts to infect with 2nd-generation merozoites. A similar experiment using E. necatrix showed a partial effect.

E. necatrix gametogony-only infections did not protect against subsequent gametogony-only infections and did not protect against schizogony in the duodenum. They did suppress oocyst production resulting from entire life-cycle infections.

Satisfactory oocyst production from the intra-caecal inoculation of second generation E. necatrix merozoites was not obtained.

Repeated oral infections with E. tenella or E. necatrix reduced subsequent cross-infections with E. necatrix or E. tenella respectively by approximately half. Both schizogony and gametogony stages were affected.

A similar result was obtained with E. tenella oocyst inoculation when previous infections were initiated by the intra-caecal injection of E. necatrix sporozoites.

Oocyst production from the oral administration of E. tenella oocysts to birds previously infected with gametogony stages only of E. necatrix was much reduced but the schizogony stages of the challenge infection were not appreciably affected. Previous repeated caecal inoculation of E. necatrix merozoites did not reduce oocyst production from subsequent inoculations of E. tenella merozoites or of E. necatrix merozoites.

Thanks are due to Mr P. L. Long for his interest and advice. The excellent technical assistance of Mr Brian Millard and other members of the Parasitology Department is also gratefully acknowledged.