The D1-D2 domains of LSU rDNA were used to reconstruct the phylogenetic relationships within the Ancyrocephalinae (Monogenea: Dactylogyridae) utilizing maximum-parsimony (MP), maximum-likelihood (ML), minimum evolution (ME) and neighbour-joining (NJ) methods. A total of 32 monogenean taxa were examined in the present study, including 9 Haliotrema species and 13 other species representing the Ancyrocephalinae, 4 Thaparocleidus species representing the Ancylodiscoididae, and 6 species representing the Diplectanidae which were used as multiple outgroups. All 4 analyses (i.e. MP, ML, ME and NJ) inferred the same interrelationship pattern: (Diplectanidae, (Ancylodiscoididae, Dactylogyridae)) with high bootstrap support. However, 9 Haliotrema species were dispersed to form 4 clades together with species from other genera, indicating the apparent non-monophyly of Haliotrema. Three major groups were defined based on reconstructed phylogenetic trees to explain the radiation of Haliotrema species. The morphology of the reproductive organ, particularly the male copulatory organ (MCO), was discussed to further understand the formation of each group. (1) Results of the present study indicated an intimate relationship among Metahaliotrema (2 species), Protogyrodactylus (4 species) and Haliotrema (2 of 9 species), and notably, all these species share vagina-absence. (2) Based on the present molecular analyses and the morphological characters of the MCO, we propose to transfer H. spirotubiforum and the undetermined Haliotrema sp. ZHDDb to Euryhaliotrema as new combinations. (3) We propose to erect a new genus to accommodate the Haliotrema species with horn-like shaped MCO. Taxonomic implications of the present molecular phylogenetic analyses are discussed. A wider range of taxa and more DNA markers displaying various evolutionary rates should be used to estimate phylogenetic relationships among species within the Ancyrocephalinae and Ancylodiscoididae in further studies.

We have examined Plasmodium falciparum gametocyte prevalence, density and their genetic complexity among children of 2 sympatric ethnic groups (Mossi and Fulani) in villages in Burkina Faso. The 2 groups are known to have distinct differences in their susceptibility and immune responses to malaria. We used RT-PCR and sequence-specific probes to detect and type RNA of the gametocyte-specific protein Pfs48/45. There were no differences in detection rates of asexual forms and gametocytes among the 2 groups, using PCR and RT-PCR, respectively. However, there were significant differences in densities of asexual forms and gametocytes, which were both higher among Mossi than Fulani. Both asexual forms and gametocyte densities were influenced by age and ethnicity. Multiple-clone infections with more than 1 gametocyte genotype were equally prevalent among Fulani and Mossi. These differences can most probably be attributed to genetic differences in malaria susceptibility in the 2 ethnic groups.

A succession of antimalarial drugs has been deployed to treat human falciparum malaria but each has, in turn, been nullified by the spread of drug resistance. The consensus view has always been that, once present, resistance will inevitably rapidly increase to 100%. However, recent field evidence has shown this is not inevitable, and that drug resistance may initially spread and then stabilize at relatively low frequencies. It is proposed that intense competition between separate malaria clones co-infecting the same human can generate complex dynamics capable of explaining this observation. Standard population genetic analysis confirms this assertion. The dynamics underlying the evolution of antimalarial resistance may therefore be much more complex than previously realized, and can resolve the apparent paradox between field data and the underlying theory of the evolution of resistance. This explanation is novel and the results are equally applicable to other parasitic species where multiple infections of the same host are common.

The impact of Toxocara canis infection of Balb/c mice mothers on the future immune response of their offspring towards reinfection with the same parasite was studied. Two groups of offspring, the first originating from the mothers infected with a single dose of 1000 Toxocara canis eggs and the second from non-infected mothers, were both challenged with 500 T. canis eggs per animal at 6 weeks of age. The proportions of spleen CD4+ and CD8+ T lymphocytes, the level of serum cytokines IFN-γ and IL-5, eosinophilia in peripheral blood, the production of specific antibodies and the number of migrating larvae were monitored. In both groups of offspring, the challenge infection resulted in an increase in CD4+ T-cell subtype in comparison with the non-infected healthy control, although after an initial decline a subsequent increase in CD8+ was observed. The immunoregulation index (CD4+/CD8+) was lower in the group of mice originating from infected mothers throughout the whole experiment compared to the offspring of non-infected mothers as well as in healthy control mice of the same age. In the offspring of infected mothers mainly, a reduced production of IFN-γ and of IL-5, suppressed eosinophilia and a higher level of protective antibodies was detected, compared to the control second group, in which the INF-γ concentration significantly increased after day 42 p.i. In the first group of offspring before challenge, 12·7±2·5 larvae in the brains and 32±2·1 larvae in the muscles transmitted from the infected mothers were detected. There was a significant reduction in larval recovery from brain on days 42 and 49 p.i. (56·7 and 56·8%, respectively), while from muscles in the same time there was a reduction of 46·7 and 39%, respectively, compared to the offspring of non-infected mothers. These results indicate a significant protective memory of immune mechanisms against T. canis induced in offspring of Toxocara-infected mother mice.

Toxocariasis is a common helminthic infection that has a worldwide distribution. However, data from Lebanon about the prevalence of this infection are non-existent. We conducted a Toxocara seroprevalence study with 150 subjects attending the outpatient clinics at the American University of Beirut Medical Center between May and June 2004. Serum specimens were tested for anti-Toxocara antibodies by enzyme-linked immunosorbent assay and confirmed by Western blot. Multivariate analysis was performed to identify risk factors for infection. The seroprevalence rate of toxocariasis was 19%. Male gender and below high school education were significantly associated with a positive serological test (odds ratios=3·1 and 2·8, respectively). Higher numbers of persons in the household, and low family income during childhood, were significant on bivariate analysis only. Toxocariasis is common in Lebanon. A large population-based survey is needed to confirm these results.

PTPases have been reported as a virulence factor in different pathogens. Recent studies suggest that PTPases play a role in the pathogenesis of Leishmania infections through activation of macrophage PTPases by the parasite. We report here the presence of a membrane-bound PTPase in Leishmania major promastigotes. We detected differences in the PTPases present in the procyclic and metacyclic stages of promastigotes. In metacyclic promastigotes, the PTPase activity was totally inhibited by specific PTPase and serine/threonine inhibitors, whereas in procyclic promastigotes the PTPase activity was inhibited only with PTPase inhibitors. Two antibodies against the catalytic domains of the human placental PTPase1B and a PTPase from Trypanosoma brucei cross-reacted with a 55–60 kDa molecule present in the soluble detergent-extracted fraction of a Leishmania homogenate. Metacyclic promastigotes expressed more of this molecule than parasites in the procyclic stage. Yet the specific activity of the enzyme was lower in metacyclic than in procyclic promastigotes. Ultrastructural localization of the enzyme showed that it was more membrane-associated in metacyclic promastigotes, whereas in procyclic promastigotes it was scattered throughout the cytoplasm. This is the first demonstration of a PTPase present in Leishmania major promastigotes that differs in expression, activity and ultrastructural localization between the procyclic and metacyclic stages of the parasite's life-cycle.

We are studying early events in the establishment of Trypanosoma brucei in the tsetse midgut using fluorescent trypanosomes to increase visibility. Feeding flies with the lectin-inhibitory sugars D-glucosamine (GlcN) or N-acetyl-glucosamine (GlcNAc) has previously been shown to enhance fly susceptibility to infection with trypanosomes and, as expected, we found that both sugars increased midgut infection rates of Glossina morsitans morsitans with T. brucei. However, GlcNAc did not show the inhibitory effect on salivary gland infection rate reported previously for GlcN. Both sugars significantly slowed the movement of the bloodmeal along the midgut. GlcN also significantly increased the size of the bloodmeal taken and fly mortality. The most surprising finding was that GlcNAc stimulated trypanosome growth not only in the midgut, but also in vitro in the absence of any factor derived from the fly. Thus our direct comparison of the effects of GlcN and GlcNAc on the trypanosome-tsetse interaction has shown that these sugars impact on trypanosome growth and tsetse physiology in different ways and are not interchangeable as suggested in the literature. The sugars cause multiple effects, not restricted solely to the inhibition of midgut lectins. These findings have implications for current models of tsetse susceptibility to trypanosome infection.

This study investigated why the susceptible or resistance phenotype to the nematode Heligmosomoides polygyrus was lost when susceptible (C57BL/6) and resistant (Balb/c) strains of mice were housed together in indoor arenas with continuous transmission of the parasite larvae present in peat trays (Scott, 1991). First, both strains expressed their normal phenotype when given a controlled challenge while living in arenas, and when experimentally infected with only 5 parasite larvae. To test whether chronic exposure to peat altered the resistance phenotype, mice were given a challenge infection while living on peat. C57BL/6 mice living on peat had higher egg production and higher worm numbers than Balb/c mice, except at 2 months post-challenge. Finally, natural transmission rates were increased in arena experiments through either regular replacement of arena mice with naïve mice or direct introduction of additional larvae. A transient difference in infection levels between strains was detected in response to a modest increase in transmission whereas a 10-fold increase in transmission allowed C57BL/6 mice to exhibit the typical profile of high egg production and elevated worm numbers. These data indicate that C57BL/6 mice are less able to regulate parasite numbers at high transmission rates compared with lower transmission rates.

Extracts of plants, such as papaya, pineapple and fig, are known to be effective at killing intestinal nematodes that inhabit anterior sites in the small intestine, such as Heligmosomoides polygyrus. In this paper, we demonstrate that similar in vitro efficacy also occurs against a rodent nematode of the large intestine, Trichuris muris, and confirm that the cysteine proteinases present in the plant extracts are the active principles. The mechanism of action of these enzymes involved an attack on the structural proteins of the nematode cuticle, which was similar to that observed with H. polygyrus. However, not all plant cysteine proteinases were equally efficacious because actinidain, from the juice of kiwi fruit, had no detrimental effect on either the motility of the worms or the nematode cuticle. Papaya latex was also shown to significantly reduce both worm burden and egg output of mice infected with adult T. muris, demonstrating that enzyme activity survived passage to the caecum and was not completely inactivated by the acidity of the host's stomach or destroyed by the gastric or pancreatic proteinases. Thus, the cysteine proteinases from plants may be a much-needed alternative to currently available anthelmintic drugs due to their efficacy and novel mode of action against different gastrointestinal nematode species.

In the present study, we constructed gender-enriched cDNA libraries for the adult stage of the parasitic nematode Oesophagostomum dentatum (order Strongylida) using suppressive-subtractive hybridization (SSH), sequenced clones from the female-library and male-library (480 from each) and conducted bioinformatic and microarray analyses of the expressed sequence tags (ESTs). In total, 873 ESTs (440 male and 433 female) were obtained, achieving a sequencing success of 91%The nucleotide sequences reported in this article (Tables 1–5) have been deposited in the EMBL, GenBank and DDJB databases under the Accession nos. AM157797–AM158083. Microarray analyses of 516 unique ESTs representing both gender-enriched libraries revealed differential hybridization for 391 of them (75·8%). Of these, 220 (56·3%) had significantly greater signal intensities in the female than in the male, and 154 (70%) of these were predicted to have homologues in C. elegans. These homologues were predicted to be involved in key biological processes, including embryonic nutrition, gametogenesis, molecular binding/transport or metabolism, nucleic acid synthesis and function, and signal transduction. Of the 171 ESTs with statistically higher signal intensities in male O. dentatum, 43·8% had homologues in C. elegans. These homologues included major sperm proteins (MSPs) or MSP-like molecules, keratin-like molecules, molecules involved in metabolism, PDZ domain-containing proteins, sugar binding proteins, protein kinases, serine proteases or protease inhibitors, molecules involved in proteolysis and other proteins, such as enzymes and various putative proteins. Of the 287 ESTs (from both gender-enriched cDNA libraries) with no known homologues in C. elegans, 50 (17·4%) had homologues in other nematodes, 8 had homologues in various other organisms and 104 (36·2%) had no homology to any sequence in current gene databases. The present study lays a foundation for the isolation and molecular, biochemical and functional characterization of selected genes from the gender-enriched cDNA archives established for O. dentatum.

The comparative development of Echinococcus multilocularis was studied in its definitive hosts, the fox, dog, cat and raccoon dog, beyond the pre-patent period to 90 days post-infection. All host species, apart from cats were susceptible to infection and capable of supporting substantial worm burdens. Although worms in cats matured and produced thick-shelled eggs, their overall development was retarded compared to that in other species in which the parasite matured rapidly producing large populations of gravid worms. E. multilocularis matured rapidly in foxes and raccoon dogs and this was sustained in raccoon dogs but not in foxes in which maturation of worms declined during the later stages of infection, in contrast to that in both raccoon dogs and dogs. These populations were sustained for longer in raccoon dogs and dogs compared to foxes. Cats would appear to have only a minor role in the maintenance of E. multilocularis in endemic areas, and infections in cats may be of minimal public health significance. In contrast, foxes, dogs and the recently recognized definitive host the raccoon dog, are all capable of playing significant roles in the epidemiology of alveolar echinococcosis. This study also demonstrated that the developmental processes of growth, segmentation, proglottization and maturation in adult Echinococcus are independent and can be influenced by environmental factors thus confirming earlier in vitro observations.

Niche apportionment models have only been applied once to parasite communities. Only the random assortment model (RA), which indicates that species abundances are independent from each other and that interspecific competition is unimportant, provided a good fit to 3 out of 6 parasite communities investigated. The generality of this result needs to be validated, however. In this study we apply 5 niche apportionment models to the parasite communities of 14 fish species from the Great Barrier Reef. We determined which model fitted the data when using either numerical abundance or biomass as an estimate of parasite abundance, and whether the fit of niche apportionment models depends on how the parasite community is defined (e.g. ecto, endoparasites or all parasites considered together). The RA model provided a good fit for the whole community of parasites in 7 fish species when using biovolume (as a surrogate of biomass) as a measure of species abundance. The RA model also fitted observed data when ecto- and endoparasites were considered separately, using abundance or biovolume, but less frequently. Variation in fish sizes among species was not associated with the probability of a model fitting the data. Total numerical abundance and biovolume of parasites were not related across host species, suggesting that they capture different aspects of abundance. Biovolume is not only a better measurement to use with niche-orientated models, it should also be the preferred descriptor to analyse parasite community structure in other contexts. Most of the biological assumptions behind the RA model, i.e. randomness in apportioning niche space, lack of interspecific competition, independence of abundance among different species, and species with variable niches in changeable environments, are in accordance with some previous findings on parasite communities. Thus, parasite communities may generally be unsaturated with species, with empty niches, and interspecific interactions may generally be unimportant in determining parasite community structure.

Host snail demography and trematode parasitism were followed for one host generation in a shallow brackish lagoon of the western Baltic Sea. In addition, a laboratory experiment was simultaneously conducted to quantify the effects of parasitic infection on host fecundity. Hydrobia ventrosa of the cohort of 1996 had a maximum life-span of up to 2 years and reproduced between May and November of their second calendar year in 1997. Snails died after reproduction. The first trematode infections appeared in May 1997 when the snails started to mature. Total trematode prevalence peaked in summer and declined during winter to the lowest level in early spring 1998. Eight taxa of larval trematodes were found. Egg production of females with trematode infections was significantly reduced. Among females with pre-patent infections, about 20% were still able to produce eggs. Among females with patent infections merely 9% could lay eggs, compared to an average of about 51% in uninfected females. Taking into account a summer prevalence of about 25%, parasitic infections caused an overall reduction in egg production of the snail host population of about 15%. The reduction in host fecundity as a result of larval trematode infection did not measurably affect the population dynamics of H. ventrosa, because other environmental factors, especially winter severity and available food supply, were concluded to be much more relevant.

This study used light and electron microscopy to provide observations and morphometric details of the life-cycle of the gymnophallids (Trematoda, Digenea), Parvatrema margaritense (Ching, 1982) n. comb., the parthenogenetic metacercariae (‘germinal sacs’) of which were previously described by Ching (1982) as Cercaria margaritensis. The research was instigated by the discovery, on the Barents Sea coast, of a high prevalence of gymnophallid sporocysts and cercariae in the bivalve Turtonia minuta and an equivalent presence of distinctive gymnophallid metacercariae in the gastropod Margarites helicinus. Experiments and data obtained from naturally infected M. helicinus demonstrated that cercariae released from the bivalves invaded the gastropods to give rise to the metacercariae. Two generations (M1 and M2) of these parthenogenetic metacercariae were formed in the extrapallial cavities of their bivalve hosts and they, in turn, gave rise to a third generation (M3) which was shown to infect marine ducks such as the eider (Somateria mollissima). As only small numbers of cercariae are released from T. minuta, it was concluded that the inclusion of parthenogenetic metacercariae in the life-cycle is particularly significant. It allows each cercaria that infects M. helicinus to give rise to over 2000 invasive metacercariae. Evidence suggests that the parthenogenetic metacercariae are commensal rather that parasitic in the pallial cavities of their hosts. Implications of this for theories of early digenean evolution are discussed.