Larvae of Taenia pisiformis were cultured in vitro in medium containing 2·5, 5 or 20% (v/v) of normal rabbit serum (NRS). Greatest development occurred in 20% NRS, and the potency of antigens collected in medium from each culture tested by intradermal (i/d) skin tests in infected rabbits paralleled the in vitro growth rate of larvae. ‘Culture’ antigens from 5% NRS stimulated good immunity in rabbits to a challenge infection with T. pisiformis eggs, although they were poorly reactive in skin tests.

T. pisiformis larvae were also cultured in 10% (v/v) of nitrates of serum reduced to one-half of its volume by passage through 300 000 MW cut oif (XM300F) or 100000 MW cut off (XM100F) ultrafiltration membranes. Larvae cultured using XM300F had growth rates comparable with those cultured in 20% NRS, and the antigens released into the culture medium had equal potency in i/d tests and in stimulating protective immunity in rabbits. Larvae did not develop in XM100F orproduce skin-reactive or protective antigens.

Crude ‘culture’ antigen from cultures in 20% NRS was separated into 4 fractions by nitration on Sephadex G200. All of these fractions gave i/d skin reactions in infected rabbits. Fraction 3 (F3) was the most active, but was shown by acrylamide gel electrophoresis and immunoelectrophoresis to be highly contaminated with rabbit serum proteins. F3 was separated into fractions on DEAE-Sephadex A50, and the third fraction from this was as active as the original culture medium in i/d skin tests, but had only 5% of the original protein concentration. Electrophoresis demonstrated few serum contaminants, and 2 indistinct protein bands that were not present in a similar fraction of NRS.

Neither Sephadex G200 F3 nor DEAE-Sephadex F3 stimulated protective immunity in rabbits, suggesting that antigens stimulating immunity against the establishment of T. pisiformis in rabbits and those provoking cell-mediated immune reactions may be different.

It is generally accepted that free-living stages of parasitic helminths are dependent on aerobic degradation of endogenous energy sources for their energy generation. This concept, however, is not the result of extensive experimental evidence, but originated mainly intuitively as oxygen is widely available in their habitat and these stages generally have a small size. Schistosoma mansoni, the sole parasitic helminth whose energy metabolism has been studied throughout its life-cycle indeed has aerobically functioning free-living stages. However, large differences exist in energy metabolism between adult stages of distinct parasitic helminths, and caution should be taken in predicting that all free-living stages of all parasitic helminths have the same, aerobic energy metabolism. Hence, this report studied the energy metabolism of Fasciola hepatica miracidia and demonstrated that F. hepatica miracidia are also dependent on aerobic degradation of their endogenous glycogen stores by glycolysis and on Krebs cycle activity for energy generation. However, in contrast to S. mansoni, F. hepatica miracidia cannot function anaerobically, as inhibition of the respiratory chain blocked motility and carbohydrate degradation, and finally resulted in death of the miracidia. Therefore, this report demonstrated that differences exist between miracidia of distinct species, in pre-adaptation of their energy metabolism to the occasional hypoxic conditions within their next host.

We studied the prevalence and intensity of the parasitic copepod Myicola ostreae in 2 closely related oysters Crassostrea angulata and C. gigas and their F1 hybrids. The effects on host and host reaction were also analysed to better understand host-parasite relationships between copepods and bivalve molluscs. Full reciprocal crosses were carried out between C. angulata and C. gigas and the progenies were reared in the wild in Ria Formosa Lagoon (Portugal), allowing natural infestation by M. ostreae. Prevalence and intensity were significantly higher in C. angulata than in C. gigas. The parasite level of F1 hybrids was similar to C. angulata and significantly higher than in C. gigas. The results of our study support a hypothesis of dominantly inherited susceptibility to M. ostreae infestation. Moreover, copepods were observed on the gill surface of C. gigas engulfed by a capsule-like structure. Histological analyses revealed that the copepods were surrounded by a massive agglomerate of haemocyte-like cells encircled by a thin layer of fibroblast-like cells. This encapsulation response was not observed in C. angulata or in F1 hybrids. These results suggest that the differential susceptibility to M. ostreae between C. angulata and C. gigas may be ascribed to host defence factors.

By means of light and electron microscopical techniques, the organization of the reproductive tract of female Moniliformis dubius is described. The tract consists of an efferent duct system associated with 2 ligament sacs present in the body cavity. The efferent duct system consists of uterine bell, uterus and vagina. The uterine bell contains a complex system of routes and is associated with the ligament sacs in a way that facilitates egg sorting. The dorsal and ventral ligament sacs exist with separate cavities at the posterior end, where they are attached to the efferent duct, but merge into a single sac at the anterior of the trunk.

The in vitro interaction between rat peritoneal macrophages and Dipetalonema viteae microfilariae in the presence of amicrofilaraemic rat immune serum was studied by transmission electron microscopy. The probable sequence of events leading to the killing of D. viteae microfilaria by macrophages is as follows. (a) Rat peritoneal macrophages in the presence of amicrofilaraemic rat immune serum adhere to the parasite surface, (b) the macrophages extend their pseudopodia around the parasite, (c) the ‘lysosome-like’ granules discharge their contents on to the parasite surface, (d) the lytic activity of these products begins at the parasite surface and (e) subsequent breaking of the microfilarial cuticle occurs, exposing the parasite intracellular material.

Electrophysiological and pharmacological experiments suggest the presence of an electrogenic glutamate transporter in the motornervous system of the parasitic nematode Ascaris suum. This putative transporter occurs in hypodermis (a tissue in some respects analogous to glia) and in DE2 motorneurons, a dorsal excitatory motorneuron class which receives excitatory glutamatergic post-synaptic potentials. Glutamate application to hypodermis produced non-conductance mediated depolarizations that were smaller in amplitude and slower in rate of rise than DE2 responses where a glutamate-activated conductance occurs. The hypodermal response is sodium dependent and calcium independent. Excitatory amino acid ionotropic receptor agonists (kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid and N-methyl-D-aspartate) were ineffective in eliciting hypodermal responses. The ionotropic receptor antagonist, 6,7-dinitroquinoline-2,3-dione, had no effect on hypodermal glutamate responses. The L- and D-forms of glutamate, aspartate and homocysteate produced hypodermal and DE2 depolarizations consistent with the pharmacological profile for glutamate transporters in other systems. Glutamate transport inhibitors (L-trans-pyrrolidine-2,4-dicarboxylate and beta-hydroxyaspartate) elicited electrogenic depolarizations in hypodermis and DE2. These results suggest that the hypodermal glutamate response has an electrogenic transporter component, while the DE2 response has 2 components, one conductance-mediated and the other due to an electrogenic transporter.

The larvae of Aspiculuris tetraptera were found in the mid-colon of mice within hours of infection. When the colon was divided into 10 equal sections the larvae were mainly found in sections 5, 6 and 7 during the first 6 days of the infection. The worms entered the crypts of Lieberkühn in this region on day 1 and remained there until day 4 or 5. After this time they left the crypts and returned to the lumen of the colon. On day 7 the worms emigrated anteriorly and thereafter were recovered only from the proximal region of the colon (sections 1–4), although in heavier infections a few larvae remained in the mid-colon.

The initial establishment site was the same in both Rattus norvegicus and in Apodemus sylvaticus, but an infection with A. tetraptera in the abnormal hosts, R. norvegicus and A. sylvaticus, was characterized by less than 7% of the inoculum becoming established, a slower rate of growth and a wider distribution centering around the preferred site. The small number of established larvae was lost from the rat before day 12 and from A. sylvaticus before day 8, whereas larvae persisted in laboratory mice for a longer period.

Six samples of rodent blood infected with malaria parasites were isolated from Cameroon. Of these, 2 contained mixed infections of Plasmodium vinckei and P. yoelii, 3 contained P. vinckei alone and 1 P. chabaudi alone. Each isolate was cloned and the resulting lines examined for morphology of blood and mosquito forms, and for electrophoretic variation in enzymes. The P. chabaudi and P. yoelii lines were morphologically and enzymically identical to isolates of the Central African Republic. Similarly, 1 P. vinckei line was identical to an isolate of the Central African Republic. The remaining 4 P. vinckei lines showed considerable variation, some enzymes being like those in isolates of surrounding regions, while others were unique to Cameroon.

The effect of ambient temperature on the ability of Moniliformis dubius to become established in the intermediate host Periplaneta americana, and the effect of temperature on the rate of development of the parasite, has been investigated. Development is postponed at temperatures below 20 °C and infection does not normally occur at 37 °C, the parasites possibly being unable to avoid the defence mechanisms of the host. Both temperatures are tolerated by the host. A marked temperature dependence of rate of development is observed.

Cod (Gadus morhua), an important fish species in the Baltic Sea, is the paratenic host for many parasite species, including the zoonotic nematodes, Anisakis sp. and Contracaecum osculatum. We aimed to identify which invertebrate species (found in situ in the fish stomach) are responsible for infection of cod with zoonotic nematodes. We found that Crangon crangon and Gammarus sp., both invertebrate prey species of cod, were infected with Anisakis simplex and C. osculatum, respectively. These host–parasite systems are reported here for the first time, implicating C. crangon and Gammarus sp. as sources of infection of Baltic cod with zoonotic nematodes.

Sequences of the first and second internal transcribed spacers (ITS1 + ITS2) of nuclear ribosomal DNA were employed to determine whether the congeneric assemblages of species of the strongyloid nematode genus Cloacina, found in the forestomachs of individual species of kangaroos and wallabies (Marsupialia: Macropodidae), considered to represent species flocks, were monophyletic. Nematode assemblages examined in the black-striped wallaby, Macropus (Notamacropus) dorsalis, the wallaroos, Macropus (Osphranter) antilopinus/robustus, rock wallabies, Petrogale spp., the quokka, Setonix brachyurus, and the swamp wallaby, Wallabia bicolor, were not monophyletic and appeared to have arisen by host colonization. However, a number of instances of within-host speciation were detected, suggesting that a variety of methods of speciation have contributed to the evolution of the complex assemblages of species present in this genus.

We have previously shown that the C-terminal region of the intermediate subunit of Entamoeba histolytica galactose- and N-acetyl-D-galactosamine-inhibitable lectin (C-Igl) is a useful antigen for serodiagnosis of amebiasis. An immunochromatographic kit was developed using fluorescent silica nanoparticles coated with C-Igl prepared in Escherichia coli. Samples for examination were added to the freeze-dried particles and then applied to the immunochromatographic device, in which a test line on the membrane was also coated with C-Igl. Fluorescent intensity was measured using a hand-held reader. In an evaluation of the kit using a human monoclonal antibody, the minimum amount of C-Igl specific antibody showing positive results was 100 pg. In the evaluation of serum samples with different antibody titers in indirect immunofluorescent antibody tests in the kit, 20 µL of serum was sufficient to obtain positive results at 30 min. Serum samples from symptomatic patients with amebic colitis and amebic liver abscess and those from asymptomatic E. histolytica-cyst carriers showed positive results in the kit. Based on evaluation using sera from healthy controls and patients with other infectious diseases, the sensitivity and specificity of the kit were 100 and 97.6%, respectively. Therefore, we conclude that the newly developed kit is useful for rapid serodiagnosis of amebiasis.

Many of the fundamental concepts in studying infectious diseases are rooted in population ecology. We describe the importance of population ecology in exploring central issues in infectious disease research including identifying the drivers and dynamics of host-pathogen interactions and pathogen persistence, and evaluating the success of public health policies. The use of ecological concepts in infectious disease research is demonstrated with simple theoretical examples in addition to an analysis of case notification data of pertussis, a childhood respiratory disease, in Thailand as a case study. We stress that further integration of these fields will have significant impacts in infectious diseases research.

In theory, the age at which maturation occurs in parasitic nematodes is inversely related to pre-maturational mortality rate, and cross-species data on mammalian nematodes are consistent with this prediction. Immunity is a major source of parasite mortality and parasites stand to gain sizeable fitness benefits through short-term adjustments of maturation time in response to variation in immune-mediated mortality. The effects of thymus-dependent immune responses on maturation in the nematode parasites Strongyloides ratti and Nippostrongylus brasiliensis were investigated using congenitally thymus-deficient (nude) rats. As compared with worms in normal rats, reproductive maturity of parasites (presence of eggs in utero) in nude rats occurred later in S. ratti but earlier in N. brasiliensis. Immune-mediated differences in maturation time were not associated with differences in worm length. Thymus-dependent immunity had no effect on pre-maturational mortality. Results are discussed in relation to theoretical expectations and possible explanations for the observed patterns in parasite maturation.

Certain species of parasites have the apparent ability to alter the behaviour of their host in order to facilitate the completion of their own life cycle. While documented in hairworms (phylum Nematomorpha), the ability for mermithid parasites (from the sister phylum Nematoda) to force hosts to enter water remains more enigmatic. Here, we present the first experimental evidence in a laboratory setting that an insect which normally never enters open water (the European earwig Forficula auricularia) will readily enter the water when infected with a mermithid nematode (Mermis nigrescens). Only adult mermithids appear capable of inducing this polarising shift in behaviour, with mermithid length being a very strong predictor of whether their host enters water. However, mermithid length was only weakly associated with how long it took an earwig to enter water following the beginning of a trial. Considering the evidence presented here and its alignment with a proteomic investigation on the same host–parasite system, this study provides strong evidence for adaptive behavioural manipulation and a foundational system for further behavioural and mechanistic exploration.

This longitudinal study explored whether aspects of subsistence agriculture were associated with presence and intensity of Ascaris and hookworm in preschool children in rural Panama. Questionnaires were used to collect data on household socio-demographics, child exposure to agriculture and household agricultural practices. Stool samples were collected from children (6 months–5 years) at 3 time points, with albendazole administered after each to clear infections, resulting in 1 baseline and 2 reinfection measures. A novel Agricultural Activity Index (AAI) was developed using principal components analysis to measure the intensity of household agricultural practices. Zero-inflated negative binomial regression models revealed baseline hookworm egg counts were higher if children went to the agricultural plot and if the plot was smaller. Baseline and reinfection Ascaris egg counts were higher if children went to the plot and households had higher AAI, and higher at baseline if the plot was smaller. Caregiver time in the plot was negatively associated with baseline Ascaris egg counts, but positively associated with baseline hookworm and Ascaris reinfection egg counts. Children who spent more time playing around the home were less likely to be infected with Ascaris at baseline. We conclude that preschool child exposure to subsistence agriculture increased Ascaris and hookworm intensity.

The commonly accepted view regarding the manner of entry of larvae of Wuchereria bancrofti (Cobb.) into the body cavity of its mosquito host is that microfilariae from human blood enter the stomach of the mosquito along with the blood meal and after casting their sheath in the stomach, they penetrate through the wall of the stomach, enter the haemocele and then find their way into the thoracic muscles. The time taken for the penetration of the stomach wall according to Looss (1914) is 6–12 hours. According to Bahr (1912) the worms appear in the thoracic muscles in 24 hours, while Lebredo (1905) recorded that in exceptional cases, worms were seen in the thorax 13 hours after feeding, but ordinarily the time interval was longer.

1. Three new species of eugregarine are described from the mid-gut of the larva of Costelytra zealandica (White).

2. Euspora zealandica occurs in the anterior region of the mid-gut near the gut caeca. Slictospora costelytrae occurs just posteriorly to E. zealandica. Euspora sp. occurs posteriorly to the malpighian tubules, but was found only rarely.

3. A description of the species and the life-cycle of two species are given.

4. The life-cycle of S. costelytrae takes about 8 weeks and that of E. zealandica 6 weeks.

5. Gametocysts will develop only at a humidity of 95–100 %.

6. The bi-associative species, E. zealandica, is more abundant in the 1st and 2nd instars than the large species, S. costelytrae. Both species are equally abundant in the 3rd instars, but the peak for incidence was in May for E. zealandica and June for S. costelytrae. This is related to the longer period of time taken for the life-history of S. costelytrae.

7. The incidence of gregarines builds up in each instar to over 90% then falls off, the time of fall off corresponding with the production of gametocysts which pass out with the faeces of the grub.

8. The gregarines mostly complete their cycle before the instars moult. This is borne out by the observation that cysts were not found in the smallest size group of each instar. There were many cases in the larger size group of all three instars where cysts only occurred.

9. Gametocysts are produced mainly in the autumn.

10. Gregarines were not found in grubs approaching metamorphosis (prepupae) nor in pupae nor adults.

11. It seems unlikely from the results that the gregarines have an adverse effect on the grubs as, in general, it was found that the larger grubs had the greater number of gregarines present, but further work on this is needed.

12. The life-history of the gregarines is closely correlated with the life-history of the beetle.

13. The presence of the gregarines only in the larval stages is related to the different habitat and behaviour of larva and adult.

I am grateful to the University Grants Committee for financial support.