We analysed 3 independently collected datasets of fully censused helminth burdens in wood mice, Apodemus sylvaticus, testing the a priori hypothesis of Behnke et al. (2005) that the presence of the intestinal nematode Heligmosomoides polygyrus predisposes wood mice to carrying other species of helminths. In Portugal, mice carrying H. polygyrus showed a higher prevalence of other helminths but the magnitude of the effect was seasonal. In Egham, mice with H. polygyrus showed a higher prevalence of other helminth species, not confounded by other factors. In Malham Tarn, mice carrying H. polygyrus were more likely to be infected with other species, but only among older mice. Allowing for other factors, heavy residual H. polygyrus infections carried more species of other helminths in both the Portugal and Egham data; species richness in Malham was too low to conduct a similar analysis, but as H. polygyrus worm burdens increased, so the prevalence of other helminths also increased. Our results support those of Behnke et al. (2005), providing firm evidence that at the level of species richness a highly predictable element of co-infections in wood mice has now been defined: infection with H. polygyrus has detectable consequences for the susceptibility of wood mice to other intestinal helminth species.

The epidemiology of Strongyloides stercoralis was studied in families of clinical (reference) cases and their neighbours at endemic foci in Jamaica. Thirteen foci were studied based on the place of residence of a reference case. For each household of a reference case, the 4 most proximal neighbourhood households (spatial controls) were included in the study. Out of 312 persons contacted 244 were followed up using questionnaires, stool examination and serology. Prevalence of infection based on stool examination was 3·5% and on ELISA 24·2%. Prevalence increased with age but was not related to gender. Reference cases were significantly older than the general study population. The prevalence of infection based on both serology and stool examination was significantly higher in reference than in neighbouring households (the reference cases, themselves, were not included in the analysis). Furthermore, prevalence of infection was highest among persons who shared a bedroom with a reference case and decreased significantly with increasing spatial separation. This is indicative of close contact transmission which has not been previously shown for a geohelminth, but which is common among microparasites.

Hosts strongly influence parasite fitness. However, it is challenging to disentangle host effects on genetic vs plasticity-driven traits of parasites, since parasites can evolve quickly. It remains especially difficult to determine the causes and magnitude of parasite plasticity. In successive generations, parasites may respond plastically to better infect their current type of host, or hosts may produce generally ‘good’ or ‘bad’ quality parasites. Here, we characterized parasite plasticity by taking advantage of a system in which the parasite (the yeast Metschnikowia bicuspidata, which infects Daphnia) has no detectable heritable variation, preventing rapid evolution. In experimental infection assays, we found an effect of rearing host genotype on parasite infectivity, where host genotypes produced overall high or low quality parasite spores. Additionally, these plastically induced differences were gained or lost in just a single host generation. Together, these results demonstrate phenotypic plasticity in infectivity driven by the within-host rearing environment. Such plasticity is rarely investigated in parasites, but could shape epidemiologically important traits.

The following list, compiled during my tenure of a grant given by the Nuffield Foundation to the Wildfowl Trust, Slimbridge, Gloucestershire, records species of Protozoa, helminths and Arthropoda found in anatid birds, either by the authors of the papers listed in the References, or by others to whose work these authors refer. It will be seen that there are, after the names of all the parasites, numbers which indicate, by reference to the corresponding numbers in the References, the authors and publications in which the species concerned are recorded. Thus Cotylurus brevis (142, 145*) means that this trematode was recorded from Aix galericulata, the Mandarin Duck, by Dubois (1953) and that Dubois & Rausch (1950a) reported an experimental infection of this duck with it.

1. The mode of attachment of Echinococcus granulosus to the dog intestine was examined.

2. In an attached worm, the scolex was found to penetrate deeply into the crypts of Lieberkühn, with the rostellum fully extended, the hooks penetrating the epithelium lightly and each sucker holding a ‘plug’ of mucous cells in its cavity.

3. The expansion of the rostellar tip was aided by a muscular disk, the rostellar pad, which probably assists attachment by forcing the hooks outwards.

4. The epithelium in the region of the suckers showed eosinophilic reactions which may indicate a mild host reaction.

5. The existence of a small group of spindle-shaped secretory cells which form a rostellar gland, lying immediately anterior to the rostellar pad, is reported. This gland was found to be the source of secretion droplets seen to be released in vitro from the scolex region.

6. The secretion droplets were found to be exceptionally labile and not preserved in the rostellar gland cells by routine fixation methods. Prolonged fixation (5–12 months) in formol (7% formaldehyde) gave excellent preservation but globules were fairly well preserved after 3 months fixation. Fixation in Baker's formol-calcium for 2 weeks followed by controlled chromation was also found to retain the secretion reasonably well but not as adequately as the prolonged fixation.

7. The secretion was found to be negative to the PAS, bromo-phenol blue (with Hg), and ninhydrin-Schiff reactions. The areas adjacent to the globules gave a protein reaction and the periphery of the globules appeared dark by phase contrast microscopy.

8. The secretion was weakly positive to performic acid-Schiff and gave strong reactions with Sudan Black after controlled chromation, both in frozen and wax sections. These results suggest that the secretion may be a liproprotein or a lipid-protein coacervate but it is recognized that confirmation by biochemical methods is desirable.

9. The possible immunological implications of these results are discussed.

This work was supported by the Australian Wool Research Committee and the U.S. Public Health Service Grant No. E. 4707–02. The technical assistance of Mrs A. L. Warrener and Mr A. B. Howkins is gratefully acknowledged.

(-)-Epigallocatechin-3-gallate (EGCG) is the major tea catechin and accounts for 50–80% of the total catechin in green tea. (-)-Epigallocatechin-3-gallate has antioxidant, anti-inflammatory, anti-microbial, anti-cancer, and anti-trypanocidal activities. This report describes the inhibitory effect of (-)-Epigallocatechin-3-gallate on the in vitro growth of bovine Babesia parasites and the in vivo growth of the mouse-adapted rodent babesia B. microti. The in vitro growth of the Babesia species was significantly (P<0·05) inhibited in the presence of micromolar concentrations of EGCG (IC50 values=18 and 25 μM for B. bovis, and B. bigemina, respectively). The parasites showed no re-growth at 25 μM for B. bovis and B. bigemina in the subsequent viability test. The drug significantly (P<0·05) inhibited the growth of B. microti at doses of 5 and 10 mg/kg body weight, and the parasites completely cleared on day 14 and 16 post-inoculation in the 5 and 10 mg/kg treated groups, respectively. These findings highlight the potentiality of (-)-Epigallocatechin-3-gallate as a chemotherapeutic drug for the treatment of babesiosis.

Standard indirect immunocytochemical techniques have been interfaced with confocal scanning laser microscopy (for whole-mount preparations) and epifluorescence microscopy (for cryosections) to investigate the occurrence and distribution of serotoninergic and peptidergic nerve elements in adult H. diminuta. Serotonin (5-HT)-immunoreactivity (IR) was widespread throughout the worm, occurring in the paired cerebral ganglia, transverse commissure, the 10 longitudinal nerve cords and in a plethora of small nerve fibres of the peripheral nervous system. An abundance of serotoninergic nerve cell bodies was found in association with the lateral nerve cords. The genital atrium and accessory reproductive ducts were richly innervated with serotoninergic nerve fibres. Thirty-five antisera to 20 vertebrate regulatory peptides and 1 invertebrate peptide (FMRFamide) were used to screen the worm for neuropeptide IR. Immunostaining was obtained with antisera raised to pancreatic polypeptide (PP), peptide YY (PYY), neuropeptide Y (NPY), substance P (SP), peptide histidine isoleucine (PHI), xenopsin (XP) and FMRFamide. The most extensive pattern of IR occurred with antisera to PP and PYY, IR being evident in the cerebral ganglia, transverse commissure, longitudinal nerve cords and in small nerve fibres that ramified throughout the parenchyma. A series of bipolar nerve cell bodies between the median nerve cords displayed PP/PYY-IR. The distribution of FMRFamide-IR was reminiscent of the PP/PYY pattern but was less extensive. Comparison of the serotoninergic and peptidergic nervous systems has revealed general similarities and some distinct differences, especially with regard to the distribution of immunoreactive nerve cell bodies. Quantitative data are presented on the levels of PP-, SP-, PH1-, and gastrin-releasing peptide (GRP)-immunoreactivities demonstrable in acid-alcohol extracts of whole worms. The highest level of peptide IR determined was recorded for PP.

The ability of the nematode Anguillicola crassus to infect eel larvae (glass-eel stage) was tested. The results show that glass-eels fed on infected copepods, the natural intermediate host of the nematode, can be infected. Light microscopical examination of the infected developing swimbladder tissue revealed that the infection results in a significant thickening of the connective tissue. The basolateral labyrinth of gas gland cells is very much reduced in infected swimbladders, and the distance of gas gland cells to blood capillaries is enlarged. Critical swimming speed, defined as the speed where the larvae were no longer able to swim against the current, was similar in infected and uninfected animals. At intermediate speeds (about 60–80% of critical swimming speed) infected eels showed a slightly higher swimming activity than control animals. Resting oxygen consumption, measured as an index of metabolic activity, within the first 2 months of infection was higher in control animals, which may be due to a reduced rate of activity in infected glass-eels. By 4–5 months after the infection, however, it was significantly higher in infected animals. This may indicate that at this stage a higher activity of the animals is required to compensate for the increase in body density, but swimming performance of infected and non-infected glass-eels was not significantly different. Oxygen consumption during swimming activity, measured in a swim tunnel at 50% of maximal swimming speed, also was not affected. The results thus show that even glass-eels can be infected with A. crassus, and this probably contributes to the rapid spread of the nematode in Europe. While aerobic metabolism during swimming activity is not affected at this stage of infection, the swimbladder tissue shows severe histological changes, which most likely will impair swimbladder function.

Novel technologies have prompted a new paradigm in disease surveillance. Advances in computation, communications and materials enable new technologies such as mobile phones and microfluidic chips. In this paper we illustrate examples of new technologies that can augment disease detection. We describe technologies harnessing the internet, mobile phones, point of care diagnostic tools and methods that facilitate detection from passively collected unstructured data. We demonstrate how these can all assist in quicker detection, investigation and response to emerging infectious events. Novel technologies enable collection and dissemination of epidemic intelligence data to both public health practitioners and the general public, enabling finer temporal and spatial resolution of disease monitoring than through traditional public health processes.

Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica on the East Coast of the United States. Transmission dynamics of this parasite were investigated in situ for 2 consecutive years (May through October) at 2 lower Chesapeake Bay sites. Compared to previous studies where seasonal infection patterns in oysters were measured, this study also provided parasite water column abundance data measured using real-time PCR. As previously observed, salinity and temperature modulated parasite transmission dynamics. Using regression analysis, parasite prevalence, oyster mortalities and parasite water column abundance were significantly positively related to salinity. Perkinsus marinus weighted prevalence in wild oysters and parasite water column abundance both were significantly related to temperature, but the responses lagged 1 month behind temperature. Parasite water column abundance was the highest during August (up to 1200 cells/l) and was significantly related to P. marinus weighted prevalence in wild oysters, and to wild oyster mortality suggesting that parasites are released in the environment via both moribund and live hosts (i.e. through feces). Incidence was not significantly related to parasite water column abundance, which seems to indicate the absence of a linear relationship or that infection acquisition is controlled by a more complex set of parameters.

Introduction. Evaluation of Leishmania drug susceptibility depends on in vitro SbV susceptibility assays, which are labour-intensive and may give a biased view of the true parasite resistance. Molecular markers are urgently needed to improve and simplify the monitoring of SbV-resistance. We analysed here the gene expression profile of 21 L. braziliensis clinical isolates in vitro defined as SbV-resistant and -sensitive, in order to identify potential resistance markers. Methods. The differential expression of 13 genes involved in SbV metabolism, oxidative stress or housekeeping functions was analysed during in vitro promastigote growth. Results. Expression profiles were up-regulated for 5 genes only, each time affecting a different set of isolates (mosaic picture of gene expression). Two genes, ODC (ornithine decarboxylase) and TRYR (trypanothione reductase), showed a significantly higher expression rate in the group of SbV-resistant compared to the group of SbV-sensitive parasites (P<0·01). However, analysis of individual isolates showed both markers to explain only partially the drug resistance. Discussion. Our results might be explained by (i) the occurrence of a pleiotropic molecular mechanism leading to the in vitro SbV resistance and/or (ii) the existence of different epi-phenotypes not revealed by the in vitro SbV susceptibility assays, but interfering with the gene expression patterns.

The aim of this work was to study the structure of the parasite communities of Digeneans of 2 families of Teleost fishes (Sparidae and Labridae) of the Mediterranean sea. We tried to quantify the importance of both the microhabitat requirements of the parasite species and the effect of host biological factors on the parasite communities. We applied, for the first time in parasite community studies, the Canonical Correspondence Analysis (CCA) to analyse (i) the spatial distribution of parasite species within the digestive tract of the hosts; (ii) the host's biological factors (such as diet, host length, gregariousness and abundance) that may influence this spatial distribution of parasite species. Our results showed that potential microhabitats were vacant in the 2 host families studied revealing a lack of niche saturation because either there was little inter- and/or intraspecific competition or there were enough available space and resources within the host. Our results also indicated that the position of the parasite in the digestive tract is much more important than host biological factors for the structure of parasite community. Finally, we highlight the potential use of the CCA method for controlling for phylogenetic constraints in multi-species analyses.

The experimental studies of Brucei group trypanosomes presented here demonstrate that the balance of host and parasite factors, especially IFN-γ GPI-sVSG respectively, and the timing of cellular exposure to them, dictate the predominant MP and DC activation profiles present at any given time during infection and within specific tissues. The timing of changes in innate immune cell functions following infection consistently support the conclusion that the key events controlling host resistance occur within a short time following initial exposure to the parasite GPI substituents. Once the changes in MP and DC activities are initiated, there appears little that the host can do to reverse these changes and alter the final outcome of these regulatory events. Instead, despite the availability of multiple innate and adaptive immune mechanisms that can control parasites, there is an inability to control trypanosome numbers sufficiently to prevent the emergence and establishment of virulent trypanosomes that eventually kill the host. Overall it appears that trypanosomes have carefully orchestrated the host innate and adaptive immune response so that parasite survival and transmission, and alterations of host immunity, are to its ultimate benefit.

1. The adults of Fasciola hepatica feed mainly, if not exclusively, on blood.

2. The oxyhaemoglobin of the blood is converted to haemoglobin, and then to acid haematin, the pH of the empty gut being about 6·4.

3. The blood is concentrated by the removal of fluid, and haemolysis eventually occurs in the lumen of the gut. Some of the haematin is absorbed by the epithelial cells, and the remainder is concentrated as faeces.

4. The gut epithelium is responsible for extracellular digestion and absorption, but there is little evidence that the major changes in its appearance are associated with particular phases of this process.

5. Attempts at increasing survival times in vitro by using blood as food have failed to date because of bacterial contamination.

During asexual development Plasmodium schizonts undergo a series of complex biochemical and structural changes. Using tightly synchronized cultures of 2 P. falciparum lines (clone C10 and strain ITO4) for light microscopy and fluorescence imaging we monitored the timing and sequence of expression of proteins associated with invasion-related organelles. Antibodies to rhoptry, micronemal and dense granule proteins (Rhoptry Associated Protein 1, Apical Membrane Antigen 1, Erythrocyte Binding Antigen 175, Ring-infected Erythrocyte Surface Antigen) and to pellicle-associated proteins (Merozoite Surface Protein 1, PfMyosin-A) were used. Clone C10 developed faster than ITO4; this difference was also found in the timing of protein expression seen by immunofluorescence. Light microscopic data were combined with transmission electron microscopic analysis using serial sectioning of ITO4 schizonts to determine nuclear number and organellar development. Thus a timetable of schizont structural maturation was established. Generally, the timing of organelle-specific antigen expression correlates well with the ultrastructural data. Rhoptries are formed mainly between second and fourth nuclear divisions, micronemes between the end of the fourth nuclear division and merozoite separation from the residual body, while dense granules are generated mainly after the micronemes. PfAMA-1 appears in micronemes before EBA-175, suggesting micronemal heterogeneity.

1. Procercoids of Schistocephalus solidus reared in copepods were found to infect marine and freshwater varieties of Gasterosteus aculeatus, and Pungitius pungitius, but not Barbus ‘Schuberti’.

2. In primary infections growth of S. solidus plerocercoids in P. pungitius was much slower than in G. aculeatus and ceased after 6–8 days; plerocercoids were dead by day 11 at 19·5 °C, and by day 14 at 10 °C.

3. A challenge infection, following two previous infections given 14 and 28 days before, becomes established in G. aculeatus, but in P. pungitius plerocercoids are rejected within 3–5 days.

4. The possible rejection mechanism involved in P. pungitius to primary and tertiary infections is discussed and electronmicrographic evidence presented of a degenerative change in the tegument visible by 8 days in a primary infection. Prior to 8 days the survival and growth of S. solidus plerocercoids surgically transferred to G. aculeatus, indicated no irreversible damage had occurred.

The evolutionary significance of chromosome size polymorphism was explored in a representative panel of 26 Trypanosoma cruzi stocks. We tested a progressive model (aCSDI) assuming that the larger the size difference between homologous chromosomes, the more divergent the parasites are. This was contrasted with a non-progressive model (Jaccard's distance), in which any chromosome size difference has the same weight. ACSDI-based dendrograms were very similar to those built-up from multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA (RAPD) data: structuring in 2 major lineages (T. cruzi I and T. cruzi II) and 5 small subdivisions within T. cruzi II was identical, and branching was very similar. Furthermore, a significant correlation (P<0·001) was observed between aCSDI and phenetic distances calculated from MLEE and RAPD data. In contrast, analysis of chromosome size polymorphism with Jaccard's distance generated dendrograms with relatively long branches, causing most branching points to cluster close together, which generates statistically uncertain branching points. Our results thus support a model of progressive chromosome size-variation and show that despite an extensive polymorphism, chromosomal sizes constitute valuable characters for evolutionary analyses. Furthermore, our data are consistent with the clonal evolution model previously proposed for T. cruzi.

The prevalence of Taenia saginata (Cestoda) among 146 predominantly Pokot tribesmen in western Kenya was studied in an attempt to assess the reliability of detecting infections by faecal examinations. Firstly, a single faecal sample was microscopically examined after being processed by a standard ether sedimentation technique. Secondly, all subjects were treated with niclosamide and a purgative and the tapeworms were recovered. In this way the efficiency of diagnosing infections by faecal examinations was estimated. T. saginata eggs were found in the faeces of 68% of the men who were eventually found to be infected. Information was collected by interview from all subjects about whether they were or ever had been infected with tapeworms and about the form of treatment previously sought or taken. Eleven out of 25 men were apparently unaware of their tapeworm infections while no evidence of infection was found among 10 of the 24 reportedly infected men. A change in the pattern of treatment for tapeworms was reported which may have resulted in a recent decline in the prevalence of infection.

The majority of Haemogregarina species have been based on the morphology of their erythrocytic stages and supposed strict host specificity. The quantity of species with a limited number of overlapping diagnostic traits has led to a considerable mess in haemogregarine taxonomy and significant synonymy. We analysed host specificity, intra- and interspecific variability, evolutionary relationships, and the distribution of the type species of the genus Haemogregarina – H. stepanowi. The morphology of blood stages and 18S rDNA sequences of this haemogregarine from four western Palaearctic hard-shelled freshwater turtles (Emys orbicularis, Mauremys caspica, Mauremys leprosa and Mauremys rivulata) were compared with Haemogregarina balli. Additional sequences of 18S rDNA of Haemogregarina-like isolates collected from three species of African hinged terrapins (genus Pelusios) were used to enlarge the dataset for phylogenetic analyses. Thirteen sequences (1085 bp) of Haemogregarina representing all four western Palaearctic turtle species were identical, corresponding to H. stepanowi, which is closely related to the Nearctic species H. balli. In our analyses, Haemogregarina spp. constituted a monophyletic clade sister to the genus Hepatozoon. Haemogregarina stepanowi possesses a wide distribution range from the Maghreb, through Europe, Turkey and the Middle East to Iran. We consider that the genus Haemogregarina has a low host specificity crossing the family level of its vertebrate hosts and that its distribution is likely to be linked to the vector and definitive host – the leech.

Random amplified polymorphic DNA markers (RAPD) were used to visualize the genetic diversity within and between infrapopulations of Schistosoma mansoni recovered from the natural vertebrate host, Rattus rattus, trapped at an insular Guadeloupean focus. Phenotypes were characterized by the sex of the parasites and by 8 polymorphic markers generated by 3 primers. Among the 212 parasite individuals recovered from 10 infected rats, 78 genotypes were characterized. All the hosts naturally infected harboured multiple parasite genotypes with a maximum diversity of 28 genotypes/host. Phenotypic and genotypic diversity calculated by Shannon-Wiener's indices and Lynch and Milligan's estimators respectively is, on average, greater within than between hosts. Considering the very low snail infection rates observed in this focus and the rapid turnover of the vertebrate hosts, our results suggest that the high mobility of the vertebrate host and/or plurimiracidial snail infections could be factors responsible for parasite genetic diversity within hosts.