The amounts of ammonia excreted by two endoparasitic flatworms have been compared and, weight for weight, the sheep liver fluke Fasciola hepatica produces ten times as much ammonia as the rat tapeworm Hymenolepis diminuta. This is thought to reflect a difference in diet, the fluke feeding mainly on the blood and tissue proteins of its host, and the tapeworm mainly on carbohydrates from the food materials in the gut of the host. However, the addition of glucose to the saline in which the flukes are cultured produces a marked decrease in the ammonia and an increase in the lactic acid excreted, suggesting that flukes are not obligatory protein feeders but that they may utilize carbohydrate when it is available. The relatively low production of lactic acid when proteins are being metabolized suggests that the flukes are producing an alternative end-product of respiratory metabolism. It is possible that this is the neutral lipid found in the excretory system of Fasciola and the possibility of an alternative metabolic pathway for the production of such lipid is discussed. The respiratory metabolism appears to be independent of oxygen, and the possible role of oxygen in other aspects of the metabolism of the fluke is discussed.

I should like to express my sincere gratitude to Dr J. Llewellyn for his constant advice and encouragement during the course of this work and the preparation of the manuscript. I also thank Professor 0. E. Lowenstein, F.R.S., for allowing me the use of the facilities in his department during the tenure of an S.R.C. Research Studentship. Finally, my thanks are due to Mr E. Wilson, the Veterinary Officer at the Birmingham City Meat Market, for his constant co-operation in obtaining supplies of liver flukes.

A comparison is given of the ultrastructure of the vitelline cell in Schistosoma mansoni, S. haematobium, S. japonicum and S. mattheei. Four stages in development of the vitelline cell have been categorized as follows: Stage 1, the undifferentiated cell; Stage 2, the developing cell showing the beginning of synthetic activity; Stage 3, the developing cell showing active protein synthesis; Stage 4, the fully mature vitelline cell. These stages in development have been defined morphologically and Stages 1, 2 and 3 are very similar in all 4 species. Lipid is present in the Stage 4 cells of all species but appears earlier at Stage 3 in S. haematobium and S. mattheei. There are several differences as to the intercellular inclusions of the Stage 4 cells, the most marked of these being the absence of calcareous corpuscles from S. japonicum as compared with the other 3 species.

Although parasites are reported to alter host responses to predators, little is known about the neurochemical mechanisms involved. Using an odour preference test, we examined the effects of an acute, subclinical infection with the naturally occurring, single host, enteric protozoan parasite, Eimeria vermiformis, on the responses of male laboratory mice, Mus musculus, to a predator. Uninfected mice avoided the odour of a predatory cat, spending a minimal amount of time in a Y-maze in the vicinity of the cat odour. In contrast, mice infected with E. vermiformis, spent a significantly greater amount of time in the proximity of the cat odour, showing a reduced avoidance of the cat odour and a reduction in predator-induced fear or anxiety. This was not related to augmented opioid activity and decreased pain sensitivity in the infected mice, as neither treatment with the exogenous opiate, morphine, nor restraint stress-induced augmentation of endogenous opioid activity, had any significant effects on the responses of uninfected mice to cat odour. The altered responses of the infected mice to the cat odour were reduced by peripheral administration of the gamma-aminobutyric A (GABAA) antagonists, bicuculline and picrotoxin, but were not significantly affected by either the benzodiazepine antagonist, Ro 15–1788, the opiate antagonist, naloxone, or the excitatory amino acid, N-methyl-D-aspartate (NMDA) antagonist, MK-801. These results indicate that infection with E. vermiformis in mice reduces the avoidance of predator odour through neurochemical systems associated with anxiety involving, at least in part, GABAA receptor mechanisms.

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT determine the anti-parasitic effects against the main cattle nematodes. We used in vitro tests targeting L1 larvae (feeding inhibition assay) and adults (motility assay) of Ostertagia ostertagi and Cooperia oncophora. In the larval feeding inhibition assay, O. ostertagi L1 were significantly more susceptible to all CT fractions than C. oncophora L1. The mean degree of polymerization of CT (i.e. average size) was the most important structural parameter: large CT reduced larval feeding more than small CT. The flavan-3-ols of prodelphinidin (PD)-type tannins had a stronger negative influence on parasite activity than the stereochemistry, i.e. cis- vs trans-configurations, or the presence of a gallate group. In contrast, for C. oncophora high reductions in the motility of larvae and adult worms were strongly related with a higher percentage of PDs within the CT fractions while there was no effect of size. Overall, the size and the percentage of PDs within CT seemed to be the most important parameters that influence anti-parasitic activity.

1. The egg distributions of five chalcid parasites of the knapweed gall-fly have been studied in a small area.

2. A distinction is made between search for hosts by random movements, and random search in the sense of Nicholson; these terms are defined. Random movements may give rise to a distribution of parasitism different from that expected if search were purely random.

3. The species discussed do not all distribute their eggs at random amongst the hosts. Some species superparasitize the hosts more, and one (Eurytoma curta) much less than would be expected if the egg distribution were purely random, owing to peculiarities in oviposition behaviour.

4. Parasitism by E. curta is unevenly distributed in space, being higher in areas of high host density. This suggests that search is not exactly random, but is also concentrated in space. A tentative explanation is advanced based on the assumption that search is by random movements.

5. Parasitism by E. robusta is very patchy. This non-random distribution is attributed to random movements performed by a very few parasites.

6. It is concluded that these spatial discrepancies do not seriously affect the numerical accuracy of the assumption that search is random in small areas, and that the theory of Nicholson & Bailey may be accurate to a first approximation.

Eight 7-month-old sheep were dosed continuously with Trichostrongylus colubriformis larvae and 4 sheep were killed at 5 and 10 weeks from the commencement of infection. Flattening of the mucosa and villous atrophy were commonly present at slaughter, and parasites were often found in superficial channels parallel with the luminal surface. At 10 weeks the mucosa was thickened and highly cellular. Leucine aminopeptidase, alkaline phosphatase and maltase activity were reduced in the proximal third of the small intestine. Liver vitamin A was reduced in 3 of 4 infected sheep at 10 weeks, but serum vitamin A levels were comparable with those of worm-free control sheep throughout the experiment.

Behavioural, parasitological and immunological data were obtained from 48 children up to 6 years old, resident in a Schistosoma haematobium endemic area in Zimbabwe. The children averaged more than 1 contact with infective water bodies every 3 days and all showed immunological evidence of exposure (an anti-cercarial and/or anti-egg antibody response). IgM was the dominant isotype and appeared in the youngest children, followed by IgA, IgE and IgG3. However, only 38 children showed evidence of infection (an anti-egg response or eggs in urine) and only 14 were excreting eggs. The best estimates from these data are that less than 1 in 100 contacts result in infection and less than 1 in 1000 result in egg output. This suggests that there may be substantial attrition of invading cercaria even in naïve individuals.

In this study we assessed three technologies for silencing gene expression by RNA interference (RNAi) in the sheep parasitic nematode Haemonchus contortus. We chose as targets five genes that are essential in Caenorhabditis elegans (mitr-1, pat-12, vha-19, glf-1 and noah-1), orthologues of which are present and expressed in H. contortus, plus four genes previously tested by RNAi in H. contortus (ubiquitin, tubulin, paramyosin, tropomyosin). To introduce double-stranded RNA (dsRNA) into the nematodes we tested (1) feeding free-living stages of H. contortus with Escherichia coli that express dsRNA targetting the test genes; (2) electroporation of dsRNA into H. contortus eggs or larvae; and (3) soaking adult H. contortus in dsRNA. For each gene tested we observed reduced levels of mRNA in the treated nematodes, except for some electroporation conditions. We did not observe any phenotypic changes in the worms in the electroporation or dsRNA soaking experiments. The feeding method, however, elicited observable changes in the development and viability of larvae for five of the eight genes tested, including the ‘essential’ genes, Hc-pat-12, Hc-vha-19 and Hc-glf-1. We recommend the E. coli feeding method for RNAi in H. contortus and provide recommendations for future research directions for RNAi in this species.

Timely diagnosis of the nematode Angiostrongylus vasorum in dogs is important in view of severe and permanent lung and cardiovascular lesions that may occur. The performance of the classical Baermann coprological method was compared with ELISAs for the serological detection of circulating antigen and specific antibodies and with Polymerase chain reaction (PCR) performed on EDTA blood, feces and tracheal swabs of serial samples from experimentally inoculated dogs over 13 weeks post inoculation (wpi) (n = 16) and following anthelmintic treatment (n = 6). Patency was observed from 6·7 to 7·6 wpi in all dogs, Baermann results were then mostly positive (116/119, 97%) during the patent period, with wide variations in the numbers of first stage larvae numbers. Blood PCR was tested positive on 1–2 occasions in 11/16 dogs in the pre-patent period, while all tested positive by antibody-detection ELISA by 6 wpi. The proportion of dogs testing positive by fecal PCR and antigen-detection ELISA rose early in the patent period. Tracheal swabs were occasionally DNA-positive in 3/16 dogs starting from 10 wpi. Following treatment, larval excretion stopped within 3 weeks and blood PCR results became negative within 1 week (5/6 dogs), while 4/6 dogs were positive for parasite DNA in tracheal swabs. Parasite antigen and specific antibodies both persisted in the blood for 3–9 weeks after treatment, with average optical densities and the proportion of positive dogs falling gradually, while results using other tests were much more variable. Results indicate that the earliest and most consistent results are obtained by the ELISAs, which can also be used for monitoring dogs after anthelmintic treatment.

In the eastern Tibetan plateau both human cystic and alveolar echinococcosis (AE) caused by infection with Echincoccus granulosus or Echinococcus multilocularis, respectively are highly endemic. The domestic dog plays a key role in zoonotic transmission in this region. Our primary objective was to investigate the role of domestic dogs in maintaining transmission of E. multilocularis in Shiqu county, Sichuan. A cohort of 281 dogs was followed up over one year after a single treatment with praziquantel followed by re-infection surveillance at 2, 5 and 12 months post-treatment. Faecal samples were tested by an Echinococcus genus-specific coproantigen ELISA and two species-specific copro-PCR tests. Total Echinococcus coproantigen prevalence in Shiqu at baseline was 21% and 9·6% after 2 months. E. multilocularis copro-PCR was positive in 11·2% of dogs before treatment (vs 3·6% with E. granulosus copro-DNA), 2·9% at 2 months post-treatment, and 0% at 5 month and 12 months. The results suggest that dogs may have the potential to maintain E. multilocularis transmission within local pastoral communities, and thus dog dosing could be an effective strategy to reduce transmission of E. multilocularis as well as E. granulosus in these co-endemic Tibetan communities.

The Houghton (H) strain of Eimeria acervulina was attenuated by serial passage through chickens of the first oocysts produced during infection. This selection pressure resulted in a reduction in the pre-patent period of the parasite, shown to be due to the selection of a line predominantly with only 3 instead of 4 generations of schizonts. The precocious line had a reproductive potential much lower than that of the parent strain and it was significantly less pathogenic. Chickens given oocysts of the precocious line were almost completely immune to challenge with the Houghton strain.

Quinacrine resistance has been induced in 3 Giardia duodenalis laboratory stocks and 4 lines resistant to other drugs. The quinacrine-resistant lines were maintained in normally lethal levels of 5 µM quinacrine and some lines are viable in 20 µM levels of the drug. Fluorescence studies indicated that quinacrine was taken up by sensitive cells but was actively excluded from resistant trophozoites. The nuclei were not a site of drug accumulation and no specific fluorescence in the trophozoite could be attributed to any structure. Blebs of concentrated drug appeared prior to disintegration of the membrane of drug-sensitive trophozoites exposed to drug overnight. Parasite lines already resistant to furazolidone adapted more readily to quinacrine exposure than drug-sensitive stocks. This multiple drug resistant phenotype was not as marked with metronidazole-resistant lines.

The life cycle and pathogenicity of a strain of Eimeria isolated in Great Britain from turkey poults by single cell inoculation are described and, using the criteria laid down by Tyzzer, the species is identified as E. adenoeides.

The life cycle is of the same general pattern as in other Eimeria species, consisting of two asexual and one sexual generations.

The organism is highly pathogenic for young poults, a dose of 200,000 oocysts producing 100 % mortality in 3-week-old birds and smaller doses causing reduced weight gain. Birds 11 weeks old resisted a dose of 3 million oocysts.

The gross and microscopic pathology of the infection is described. No changes were found in the blood picture.

Different parasite species sharing the same intermediate host species may have similar or conflicting interests, depending on whether they are at the same stage in their life-cycle or whether they share the same definitive host. In the New Zealand cockle, Austrovenus stutchburyi, metacercariae of the digenean Meiogymnophallus sp. are positively associated with metacercariae of Curtuteria australis. This relationship is found in different cockle samples, and is independent of cockle shell size, which suggests that it is not merely the product of metacercariae accumulation over time. Both digenean species have the same definitive host, oystercatchers. Metacercariae of C. australis manipulate the phenotype of cockles, impairing the cockle's ability to burrow in the sediments. This makes the host more susceptible to oystercatcher predation. Thus Meiogymnophallus sp. can benefit by associating with C. australis and may hitch a ride with the manipulator parasite. This is supported by the finding that cockles impaired by C. australis and lying at the sediment surface harbour greater numbers of Meiogymnophallus than buried cockles. A third digenean species, whose sporocysts are found in cockles and which is not transmitted by predation, occurred only in surface cockles. Finally, a parasitic copepod with a direct life-cycle was found evenly distributed among buried and surface cockles, independently of their metacercarial loads. These results show that different parasite species do not use cockles in a random fashion, and that not all patterns of host use are consistent with shared or conflicting interests among parasites.

Sex steroids play a significant role in regulating the parasite load in experimental intraperitoneal Taenia crassiceps cysticercosis of male and female Balbc/anN mice. Briefly, oestrogens increase parasite loads and androgens decrease them (1) by acting directly on the parasite, favouring or hindering its reproduction, respectively, and (2) by biasing the hosts' immune response towards a parasite-permissive Th2 or a parasite-restrictive Th1 response. The infected male host also undergoes drastic endocrinological and behavioural changes that may impinge upon the course of infection, and the host's mating behaviour and its exposure to predators. In addition, at different times of infection, significant changes occur in the expression of c-fos in the host's hyppocampus, hypothalamus and preoptic area. Thus, the host's brain seems to sense and/or react to intraperitoneal infection. The physiological domains of the network affected by the infection, which classically included the hypothalamus-pituitary-axis and the immune system, must now incorporate the host's sexual hormones and other areas of the brain. The network's complex circuitry and functions may help understand some basic questions of parasitology (i.e. the hosts' sexual dimorphism in parasite infections, host-parasite specificity, heterogeneity in the course and outcome of infections at different stages of parasite and host development). The plurality of elements and the complexity of the network that regulates the host-parasite relationship also point to additional strategies for the treatment and control of infections.

The gp63 encoding genes were characterized by PCR–RFLP in 35 isolates representative of the Leishmania donovani complex (L. infantum, L. donovani, L. archibaldi and L. chagasi), with special attention to Mediterranean L. infantum from different geographical origins, and in separate groups from Old World Leishmania (L. major, L. tropica and L. aethiopica). The aim was to evaluate how the possible selective pressure by the host on these important surface proteins would influence structuring of our sample. Comparison was carried out with the structure obtained (i) from reported isoenzyme data, characters supposed to vary neutrally, and (ii) from PCR–RFLP analysis of gp63 inter-genic regions, containing non-translated spacers and regulatory genes. Polymorphism within the gp63-encoding region, was much higher than in gp63 inter-genic regions. In the gp63 intra-genic dendrogram, the 4 species of L. donovani complex were discriminated and quite distinct from outgroups. Within L. infantum, geographical structuring was observed and did not overlap with the structure built-up from isoenzymes and inter-genic data. These results support the idea of a strong host-selection on gp63, at vector level but most of all at vertebrate (human or dog) immunological level. Furthermore, they illustrate how the nature of genetic characters may influence the perception of population structuring.

Although clam populations in France are known to be infected with protozoans of the genus Perkinsus, no molecular characterization was previously performed on these parasites. Considering that several members of this genus have been associated with mortalities of molluscs worldwide, a study was undertaken in order to characterize these parasites in France. For that purpose, clams, Ruditapes philippinarum and R. decussatus, collected from different production areas and found to be infected with Perkinsus sp. in thioglycolate culture medium, were selected for PCR-RFLP tests and sequencing. Perkinsus olseni was detected in all the investigated areas and results also suggested the presence of P. chesapeaki in Leucate, a lagoon on the Mediterranean coast and in Bonne Anse in Charente Maritime, on the Atlantic coast. Clonal cultures from both detected species were produced in order to describe and compare in vitro stages. Differences in size between both Perkinsus spp. were noticed especially for schizonts and zoosporangia. Lastly, in situ hybridization tests allowed confirmation of the presence of both species in the same R. decussatus population and even in same clams. This is the first detection of P. chesapeaki in Ruditapes species and outside North America, which questions its introduction into Europe.

The action of intra- and interspecific competition, mediated by density-dependent effects on growth, was investigated among the 3 helminth species found in the alimentary tract of 104 cormorants, Phalacrocorax carbo sinensis. Intraspecific density-dependent effects on worm sizes were observed in the abundant nematode Contracaecum rudolphii, as shown by a negative correlation between mean worm size and intensity of infection. Higher intensities of infection by C. rudolphii were also associated with more variable worm sizes in the nematode Syncuaria squamata, suggesting a one-sided and density-dependent interspecific effect. There was also clear evidence of some form of negative interaction between the nematode S. squamata and the acanthocephalan Southwellina hispida from two fronts. First, there was a strong negative correlation between the intensities of infection of the 2 species across hosts. Second, sizes of worms of 1 species became more variable as the number of worms of the other species per host increased, and vice versa. This interspecific density-dependent effect on growth was thus apparently symmetrical. We also found evidence that worm size is a predictor of egg output in the 3 helminth species, indicating that intra- and interspecific density-dependent effects on growth can affect population dynamics in these worms. These results illustrate the complex nature of density dependence in helminth growth, and how its effects can act both within and among species.

1. Optimum growth of E. invadens occurred in HSre + S in the presence of Bact. coli, at 24–30° C. The amoebae could not be ‘trained’ to grow above 35° C.

2. The amoebae ingested normal rat erythrocytes, but not those of chick or frog.

3. Mass encystation occurred only under conditions which favoured the growth of the amoebae, but was not dependent on an acceleration in growth rate; in certain strains, an increased production of cysts was correlated with the depletion of starch in the medium.

4. The amoebae were always killed by treatment with N/20-HCl for 1 hr. at room temperature, although usually they did not survive after treatment for only 10 min. At room temperature, the cysts survived treatment with N/10-HC1 for at least 6 days, and with 0·002% HgCl2 for at least 4 hr.

5. The amoebae of E. invadens were adversely affected by aerobic conditions.

6. The amoebae could survive for up to 35 days at 8° C. but for not more than 7 days at 37° C. The cysts survived for at least 14 days at 8° C., for 7, but not 11, days at 24° C., and for less than 7 days at 37° C.

7. It is suggested that, in view of the morphological identity of E. aulastomi, E. invadens, E. moshkovskii and E. histolytica, and the fact that the amoebae of these groups form a series growing at increasingly higher temperatures, these four organisms constitute subspecies rather than full species. The taxonomic status of E. ranarum is still open to question.

Leptocotyle minor is a monogenean parasite specialized for living on the denticlecovered skin of the dogfish Scyliorhinus canicula. Each adult parasite attaches itself to the outer surface of a single denticle. Small postlarvae are found on the inner surfaces of the denticles. The denticles have hard enamel on both inner and outer exposed surfaces. Haptoral hooks, such as are present in other monogenean oncomiracidia, would be unable to penetrate this hard surface and are absent in postlarval and adult parasites which attach themselves by cement. The oncomiracidium possesses six haptoral spicules which may be homologues of the marginal hooks of other monogeneans, but these spicules disappear early in postoncomiracidial development.

Adult parasites feed on the host's epidermis which lies between the denticles. The position of small postlarvae on the inner surfaces of the denticles enables them to reach the epidermis for feeding purposes. Growth then permits young parasites to migrate away from the epidermis on to the outer surfaces of the denticles.

Adult parasites move from one denticle to another in a leech-like manner, and copulation occurs between parasites which meet as a result of these movements.

I am grateful to the Director and Staff of the Plymouth Laboratory for their hospitality and particularly to Mr J. E. Green for his interest and co-operation.