To understand possible factors controlling transmission of trematode larvae between first and second intermediate hosts we examined the impact of ambient fauna on parasite transmission in a marine intertidal parasite-host association. Cockle hosts (Cerastoderma edule) kept together with selected co-occurring macrozoobenthic species in mesocosms acquired a lower parasite load compared to cockles kept alone, when targeted by cercariae of the trematode Himasthla elongata. The reduction of parasite load in the cockles differed between the 7 macrozoobenthic species tested and was between 35 and 91%. Three different types of reduction could be distinguished: (1) predators (Carcinus maenas, Crangon crangon) actively preying upon cercariae, (2) non-host filter feeders (Crepidula fornicata, Mya arenaria, Crassostrea gigas) filtering cercariae but not becoming infected and (3) alternative hosts (Mytilus edulis, Macoma balthica) becoming infected by the cercariae and thus distracting cercariae from the target hosts. In addition, interference competition may occur in the form of disturbance of cockles by ambient organisms resulting in lower filtration rates and subsequently lower parasite loads. Our results suggest that the species composition and relative abundance of the ambient fauna of parasite-host systems play an important role in controlling trematode transmission rates in benthic marine systems.

When third-stage trichostrongyle larvae exsheathed, they released an exsheathing fluid which eroded the sheath at the anterior end. This led to the formation of a refractile ring and ultimately to the fracture of the sheath which enabled the larva to escape.

Exsheathment was induced by ovine rumen fluid. The exsheathing activity of rumen fluid was impaired by centrifuging, filtering, boiling and aeration, but activity could be restored by the addition of reducing agent. The activity in vitro of either fresh rumen fluid or freeze-dried rumen fluid, after addition of a reducing agent, was comparable with that obtained in vivo.

After stimulation, the process of exsheathment continued in the absence of rumen fluid. The ability of rumen fluid to stimulate larvae was dependent upon temperature, hydrogen-ion concentration and oxidation-reduction potential.

Exsheathing fluid was present in larvae both before and after exsheathment. Larvae which had been stimulated to exsheath released exsheathing fluid for a brief interval and further release could be obtained only after the stimuli had been temporarily withdrawn.

The mechanisms for reception of the stimulus to exsheath and for storing and releasing exsheathing fluid were located in a region between the base of the oesophagus and the excretory pore. The fluid was probably released from the excretory pore.

Exsheathing fluid from one species was not necessarily as active against sheaths from other species as against its own. The activity of exsheathing fluid was dependent upon the presence of a heat-stable cofactor of low molecular weight. Activity of the dialysed fluid could be restored by addition of MnCl2 or MgCl2. Values of Q10 for the action of the fluid on the sheath lay between 1·3 and 1·9. Activity was inhibited by Hg2+, Fe3+ and iodoacetic acid.

Despite the ubiquitous presence of parasites, parasitism has not been considered among the list of regulatory factors in animal populations until recently. A detailed long-term study on the impact of the direct life-cycle nematode Heligmosomoides polygyrus on a breeding population of laboratory mice provides a clear example of the ability of helminths to regulate host abundance. In the absence of the parasite, the mouse population equilibrated at a density of 320 mice/m2 as a result of density-dependent effects on recruitment. When the parasite was added and transmission was maintained at high levels, infected mouse populations equilibrated at densities of < 18 mice/m2. Reduced rates of parasite transmission and elimination of the parasite from the system both resulted in an increase in mouse density. These results have implications for both ecology and parasitology as they demonstrate a potentially important but often ignored component of host populations that may well influence host abundance and community structure.

The mode of action of bioactive plants on gastrointestinal nematodes remains obscure. Previous in vitro studies showed that exsheathment was significantly disturbed after contact with tannin-rich extracts. However, the role of important factors (extract concentration, parasite species) has not been assessed and no information is available on the occurrence in vivo. These questions represent the objectives of this study. The model incorporated the parasites Haemonchus contortus and Trichostrongylus colubriformis with sainfoin as the bioactive plant. A set of in vitro assays was performed, measuring the changes observed, after 3 h of contact with increasing concentrations of sainfoin, on the rate of artificial exsheathment. The results indicated that sainfoin extracts interfered with exsheathment in a dose-dependent manner and the process overall was similar for both nematodes. The restoration of control values observed after adding PEG to extracts confirms a major role for tannins. A second study was performed in vivo on rumen-cannulated sheep fed with different proportions of sainfoin in the diet to verify these in vitro results. The consumption of a higher proportion of sainfoin was indeed associated with significant delays in Haemonchus exsheathment. Overall, the results confirmed that interference with the early step of nematode infection might be one of the modes of action that contributes to the anthelmintic properties of tanniniferous plants.

The biological membranes of Trypanosoma brucei contain a complex array of phospholipids that are synthesized de novo from precursors obtained either directly from the host, or as catabolised endocytosed lipids. This paper describes the use of nanoflow electrospray tandem mass spectrometry and high resolution mass spectrometry in both positive and negative ion modes, allowing the identification of ~500 individual molecular phospholipids species from total lipid extracts of cultured bloodstream and procyclic form T. brucei. Various molecular species of all of the major subclasses of glycerophospholipids were identified including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol as well as phosphatidic acid, phosphatidylglycerol and cardolipin, and the sphingolipids sphingomyelin, inositol phosphoceramide and ethanolamine phosphoceramide. The lipidomic data obtained in this study will aid future biochemical phenotyping of either genetically or chemically manipulated commonly used bloodstream and procyclic strains of Trypanosoma brucei. Hopefully this will allow a greater understanding of the bizarre world of lipids in this important human pathogen.

1. The egg shells of Fasciola are derived from globules or granules contained in the, vitelline cells. These globules contain an orthodihydroxyphenol and protein.

2. The anatomy and histology of the Mehlis's gland region have been described, and the site of liberation of the above granules has been isolated.

3. Mechanical forces, due to the churning action of the uterus, are believed to be responsible for freeing most of the granules. This liberation can be correlated with the presence of spermatozoa in the uterus. The spermatozoa can penetrate the vitelline cells, but this does not free the globules.

4. Active spermatozoa, in alkaline solutions, tend to form aggregates, and incorporate into these aggregates any solid material in the vicinity. This is apparently responsible for the arrangement of cells within the egg, and for the formation of a shell round it from fused vitelline cell globules.

5. The polyphenol of the newly formed egg shell is oxidized into a quinone in the anterior uterine coils. Polyphenol oxidases have not been conclusively demonstrated. The relative uterine coils are surrounded by an especial concentration of a generally distributed tissue haemoglobin.

6. The final egg shell is composed of a quinone tanned protein, similar to the sclerotin of the cockroach ootheca.

7. Suggestions are advanced as to the possible functions of Mehlis's gland and Laurer's canal.

Anthropogenic influence on ecosystems, such as habitat fragmentation, impacts species diversity and interactions. There is growing evidence that degradation of habitats favours disease and hence affects ecosystem health. The prevalence of haemoparasites in the Common Fruit Bat (Artibeus jamaicensis) in a tropical lowland forest in Panamá was studied. We assessed the relation of haemoparasite to the general condition of the animals and tested for possible association of haemoparasite prevalence to habitat fragmentation, with special focus on trypanosomes. Overall, a total of 250 A. jamaicensis sampled from fragmented sites, here man-made, forested islands in Lake Gatùn, and sites in the adjacent, continuous forest in and around the Barro Colorado Nature Monument were examined. Using microscopy and DNA-sequencing 2 dominant types of haemoparasite infections, trypanosomes and Litomosoides (Nematoda) were identified. Trypanosome prevalence was significantly higher in bats from forest fragments, than in bats captured in continuous forest. We attribute this to the loss of species richness in forest fragments and specific characteristics of the fragments favouring trypanosome transmission, in particular changes in vegetation cover. Interestingly, the effect of habitat fragmentation on the prevalence of trypanosomes as multi-host parasites could not be observed in Litomosoides which probably has a higher host specificity and might be affected less by overall diversity loss.

Trypanoplasma borreli and Trypanosoma carassii are kinetoplastid parasites infecting cyprinid fish. We investigated the role of nitric oxide (NO) in immune modulation during T. borreli and T. carassii infection of carp. Phagocytic cells from different organs produced NO and serum nitrate levels increased, demonstrating that T. borreli activates NO production in vivo. In contrast, T. carassii did not induce NO production in vivo and inhibited LPS-induced NO production in vitro. Production of NO was detrimental to the host as T. borreli-infected carp treated with the inducible NO synthase inhibitor aminoguanidine had a higher survival than infected control carp. This detrimental effect can be explained (in part) by the toxicity of NO to cells in vitro as NO inhibited the proliferative response of blood and spleen leukocytes. Head-kidney phagocytes were resistant to the immunosuppressive effects of NO in vitro. The NO-inducing activity of T. borreli may be an adaptation developed to ensure survival and immune evasion in the fish host. Apparently, T. carassii has adopted another strategy by deactivating specific functions of phagocytes. Both strategies may ensure long-term survival of the parasite.

Wistar-Furth strain male rats and CFW strain male mice were immunized against Trichinella spiralis using an antegenic fraction derived from a cell-free homogenate of mature muscle larvae.

In rats, animlas immunized with 250 μg of antigen harboured singnificantly fewer (135000) muscle larvae 30 days after oral challenge than controls (231000). Furhter 7-day-old adult worms derived from immunized rats shed 48% fewer (P < 0·001) newborn larvae over a 24 h period in vitro than adult worms from non-immunized animals.

Mice were injected with either 10 or 100 μg of antigen. In comparison with non-immunized controls, mice immunized with 100μg of antigen harboured significantly fewer adult worms at days 7 and 9 after oral challenge infection, while female worms recovered from immune mice on days 6–10 after challenge shed fewer newborn larvae in vitro. Finally, mice immunized with 100μg of antigen harboured significantly fewer (10391) muscle larvae at 30 days after challenge than did controls (47750).

Immunization of mice with 10μ of antigen did not induce a statistically significant reduction in adult worms at either days 7 or 9 after challenge (P < 0·5). However, adult female worms from mice receiving 10μg of antigen still shed fewer larvae than did adults from control mice (P < 0·05). Mice immunized with 10μg of antigen harboured significantly fewer (13700) recoverable muscle larvae than did controls at 30 days after challenge (39000).

Vaccinia virus, a member of the Poxviridae, expresses many proteins involved in immune evasion. In this review, we present a brief characterisation of the virus and its effects on host cells and discuss representative secreted and intracellular proteins expressed by vaccinia virus that are involved in modulation of innate immunity. These proteins target different aspects of the innate response by binding cytokines and interferons, inhibiting cytokine synthesis, opposing apoptosis or interfering with different signalling pathways, including those triggered by interferons and toll-like receptors.

The development of nasal avian schistosomes of the genus Trichobilharzia in their final host is poorly known. Therefore, an experimental infection of ducklings (Anas platyrhynchos f. dom.) by T. regenti was performed. The infection resulted in leg paralysis and orientation/balance disorders of birds. The examination of the duck's spinal cord and brain confirmed the presence of developing parasites in pre-patent as well as patent periods. The absence of the worms in other tissues strongly supports our hypothesis that the parasite migrates through the central nervous system (CNS) to its final location in bird nasal mucosa. The injury level is probably dependent on number of parasites as well as yet unknown host factors. The affinity to the CNS seems to be high; also by exposure of experimental animals to low cercarial doses the growing worms in the CNS were found. In addition to the generally accepted view that bird schistosomes may cause cercarial dermatitis of mammals (including man), there is evidence of a partial development of T. regenti in mouse CNS; in certain cases leg paralysis was also recorded. Therefore, the pathogenesis spectrum caused by bird schistosomes in birds/mammals needs to be reconsidered.

Faecal egg counts are often used to measure Schistosoma mansoni infection, but the considerable variation between successive counts complicates their interpretation. The stochastic model described in this paper gives a description of observed egg counts in a population and can be used as a tool to gain an insight into the underlying worm load distribution. The model distinguishes between two sources of variation in egg counts: (1) variation caused by the difference in worm load between individuals, and (2) the variability of egg counts for an individual with a given worm load. Empirical data, single and repeated measurements, from surveys in five villages in Burundi and Zaire have been used to fit and validate the model. We have discussed possible mechanisms that explain the differences in estimated values between the villages. The model indicates that the expected number of eggs in a stool sample per S. mansoni worm pair is lower than suggested by autopsy data and that, possibly as a consequence of immunity, the inter-individual variation in worm loads decreases with age.

The anthelmintic properties of tannin-rich plants are being explored as an alternative to chemical drugs. Most data have been acquired on legume forages, but only few on browse plants. The present study aimed to (i) screen the in vitro effects of extracts from 7 Mediterranean plants on Haemonchus contortus, (ii) verify the role of tannins using an inhibitor, polyvinyl polypyrrolidone (PVPP) and (iii) verify the in vivo effects of extracts from 4 plants. Significant inhibition was shown in vitro using a larval migration inhibition (LMI) assay for all extracts except that from Olea europaea var. koroneiki. After adding PVPP, the LMI values were restored to control levels for all plants except Pistacia lentiscus and Ceratonia siliqua, confirming a role for tannins in the activity. In the in vivo experiment, 48 lambs composed 6 groups, depending on diet. On Day 0, groups G1–G5 received H. contortus and Trichostrongylus colubriformis larvae and G6 remained uninfected. The various diets were distributed from Days 14 to 45: P. lentiscus (G1), Quercus coccifera (G2), C. siliqua (G3), Onobrychis viciifolia (G4), or Medicago sativa for the 2 control groups (G5, G6). Egg excretion, packed cell volumes (PCVs) and inorganic phosphate were measured weekly throughout the entire experimental period. At slaughter, the worms were enumerated and their fecundity assessed. Consumption of the 4 browser plants did not provoke differences in pathophysiological measurements but there were significant decreases in egg excretion, mainly explained by significant decreases in worm fecundity for both species, without any statistical difference in worm numbers.

Cane toads (Bufo marinus) were introduced to Australia in 1935 and have since spread widely over the continent, generating concern regarding ecological impacts on native predators. Most Australian cane toad populations are infected with lung nematodes Rhabdias pseudosphaerocephala, a parasite endemic to New World (native-range) cane toad populations; presumably introduced to Australia with its toad host. Considering the high intensities and prevalence reached by this parasite in Australian toad populations, and public ardour for developing a control plan for the invasive host species, the lack of experimental studies on this host-parasite system is surprising. To investigate the extent to which this lungworm influences cane toad viability, we experimentally infected metamorph toads (the smallest and presumably most vulnerable terrestrial phase of the anuran life cycle) with the helminth. Infected toads exhibited reduced survival and growth rates, impaired locomotor performance (both speed and endurance), and reduced prey intake. In summary, R. pseudosphaerocephala can substantially reduce the viability of metamorph cane toads.

The prevalence of various species and subgroups of trypanosomes in the Sinfra area of C⊚te d'Ivoire was determined using the polymerase chain reaction (PCR). Using this technique to amplify specific satellite DNA targets, it was possible to identify developmental-stage trypanosomes in the midguts and the proboscides of tsetse without expansion of parasite populations. The predominant tsetse species in the area was Glossina palpalis, while G. pallicera and G. nigrofusca were also present. Microscopical examination of 811 non-teneral flies revealed an infection rate of 14% in midguts and/or proboscides. Three subgroups of Trypanosoma congolense (Savannah, Forest & Kilifi), T. simiae, T.godfreyi, West African T. vivax and T. brucei ssp. were identified using PCR. T. congolense Forest was the most abundant of the Nannomonas trypanosomes. Approximately 40% of all infections were mixed, and there was a significantly higher prevalence of apparently mature T. brucei ssp. trypanosomes than has previously been reported. The present study demonstrates that PCR facilitates the easy identification of mature trypanosome infections in tsetse, providing a reliable estimation of trypanosomiasis challenge.

The life-history of Aggregata, eberthi is summarized in the accompanying diagram (Text-fig. 3). The stages shown above the dotted line occur in Sepia officinalis, those below in Portunus.

Merozoites (R), when swallowed by the cuttlefish, pass through the epithelium and enter the submucous tissue of the caecum or intestine, where they develop into young (undifferentiated) parasites (A) which grow into males (B) or females (E). The former, after undergoing repeated nuclear divisions (C), produce broods of microgametes (D)—each of which (D*) has a complicated structure. Females (E), on the other hand, are bodily converted into macrogametes (F), which undergo a complex series of nuclear changes during maturation. No reduction occurs during gametogenesis.

Fertilization (G) is effected by the union of one microgamete with one macrogamete, and is followed by the formation of a fertilization membrane which, in part, gives rise to the oocyst. The zygote (H) becomes a sporont, which, after repeated division of its nucleus (I, J), ultimately segments—leaving no residuum—into a large number of spherical uninucleate sporoblasts (K). Bach of these is then converted into a spore (L) containing three sporozoites and a small sporocystic residue.

The ripe spores pass out of the intestine with the cuttlefish's excrement; and when swallowed by a Portunus they open in its intestine and liberate their contained sporozoites (M). These penetrate the epithelium of the crab's midgut, and enter the subepithelial connective tissue, where they grow into schizonts (N, O). Finally, the schizonts undergo schizogony, producing large broods of merozoites (P, Q). No further development takes place unless the schizogonic cysts, in the crab's coelome, are eaten by a cuttlefish: in which case they liberate their contained merozoites (R) and the development just described then begins anew.

A. eberthi is, at every stage but one in the whole life-cycle, a haploid organism. Every nuclear division is mitotic: and every nucleus (except the zygote nucleus) contains a single set of six differentiated and unpaired chromosomes. In the zygote nucleus alone there are 12 chromosomes—six homologous pairs—derived from the two gamete nuclei: but at the first division of this nucleus (first division in sporont, Text-fig. 3, I) the chromosomes are again halved to six. Reduction (meiosis) thus occurs immediately after fertilization. No halving of the chromosome number occurs during gametogenesis or schizogony; and males, females, and asexual parasites all possess an identical chiomosomic constitution. Every nucleus in Text-fig. 3 is therefore an x nucleus, with the exception of stage H, which is 2x. (Between stages G and I many nuclear changes occur, but only one—the so-called “fertilization spindle”—is figured (H) in the diagram.)

On the basis of known DNA sequences of Dirofilaria repens and D. immitis we designed specific primers for the amplification by Polymerase Chain Reaction (PCR) of the DNA from the two species. The PCR-based identification was found to be unambiguous and allowed specific diagnosis of microfilariae in blood samples, of developing larvae in the mosquito vector and of immature adults in bioptic material, overcoming the serious constraints of the morphological separation of these filarial parasites at the pre-adult stages. The technique was found to be very sensitive and applicable to samples stored either dry or in various preservation media, with the exception of formalin. The reliable identification of D. repens and D. immitis from bioptic material is expected to greatly enhance the chances of detecting human infections and to further clarify the role of the two parasites as pathogens of man. The possibility of routine identification of developing larvae in the vector will substantially improve the perspectives for epidemiological investigations, particularly in Southern European regions, such as Italy, where the two nematode species are largely sympatric.

Parasites often exploit more than one host species at any stage in their life-cycle, but the extent to which these host species are used varies greatly. Parasites typically achieve their highest prevalence, intensity and/or abundance in one host species (the principal host), whereas infection levels in auxiliary hosts range from relatively high to very low. The present study examines what influences the distribution of parasite individuals among their different host species, using metazoan parasites that use freshwater fish as their definitive or only host. Specifically, I test the hypothesis that differences in relative infection levels by a parasite among its auxiliary hosts are proportional to the taxonomic distance between the respective auxiliary hosts and the parasite's principal host. Taxonomic distance among hosts is a surrogate measure of their similarity in terms of ecology, physiology and immunology. Using data on 29 parasite species and 6 fish communities, for a total of 47 parasite-locality combinations, it was found that taxonomic distance between the auxiliary hosts and the principal host had no real influence on infection levels in auxiliary hosts, measured as either prevalence, intensity or abundance. The analysis revealed differences in the degree of specialization among major groups of parasites: in terms of abundance or intensity, auxiliary hosts were less important for cestodes than for nematodes and copepods. The lack of an effect of taxonomic distance may indicate that ecological similarity among host species, arising from convergence and not from relatedness, is more important than host phylogeny or taxonomy. Although the results are based on a limited number of parasite taxa, they suggest that parasites may be opportunistic in their colonization of new hosts, and not severely constrained by evolutionary baggage.

Evidence from many sources indicates that, in several parasite–vector systems, flight ability, fecundity or gonotrophic concordance of vectors, are affected by parasites (including bacteria, rickettsia and viruses) and in some cases parasites cause well-documented pathogenic effects (see reviews, for example, by Lavoipierre (1958a) and Hawking & Worms (1961) on filaria; Molyneux (1977, 1983) for trypanosomatids; Killick-Kendrick (1979) for Leishmania).

The parameters involved in the induction of resistance against Schistosoma mansoni by injection of irradiated, artificially transformed schistosomula were studied in mice. Single intramuscular injections of 500 schistosomula exposed to radiation doses in the range 2·3 to 160 krad. resulted in significant protection (in the range 20 to 50% as assessed by reduced worm burdens) against a challenge infection administered at intervals from 3 to 24 weeks post-vaccination. However, schistosomula irradiated with 20 krad. consistently resulted in better protection than those exposed to either higher or lower radiation doses despite the persistence of stunted adults from the infections irradiated with 2·3 krad. Vaccination with 40 krad. schistosomula resulted in significant protection in terms of reduced worm and tissue egg burdens and increased survival following lethal challenge. Varying the number of irradiated schistosomula, the frequency and route of their administration, the site of challenge and the strain of host all failed to enhance the level of resistance. However, percutaneously applied, irradiated cercariae were found to be more effective in stimulating resistance (60%) than intramuscularly injected, irradiated schistosomula (40%).