Helminthologically naive 8-week-old lambs were administered 50000 infective-stage larvae of the trichostrongyle nematode Nematodirus battus. Initial mean adult worm burdens were significantly reduced (P < 0·01) at day 21 post-infection (p.i.). Peripheral blood eosinophil counts showed a significant increase (P < 0·001) by day 25 p.i. when compared with uninfected control animals. Sections of small intestine taken at post-mortem from the area of adult worm establishment over the course of infection showed a significant increase in mucosal eosinophil and mast cell numbers during the period of adult worm rejection, when compared with uninfected control animals. These results show that young lambs develop significant increases in the numbers of cells associated with an inflammatory reaction in the intestine during the development of a partially protective immune response to this parasitic nematode.

Excretory–secretory (E–S) products, released by Trichobilharzia ocellata cultured in vitro, were assessed for their modulatory effects on the internal defence system of the freshwater snail, Lymnaea stagnalis. E–S products were concentrated and fractionated using high-pressure liquid chromatography (HPLC) and bacterial clearance activity of haemocytes was taken as a parameter for the internal defence system. Two main fractions of E–S products were produced by T. ocellata, a low molecular fraction (low MW) and a high molecular fraction (high MW), weighing approximately 2 kDa and 40 kDa, respectively. Stage-dependent release of E–S products by transforming miracidia and mother sporocysts was indicated by the difference in proportions of these two fractions, which varied over time. After a culture time of 0–33 h, the smaller fraction was prominent whilst E–S products released between 33 and 72 h consisted of a higher proportion of the high MW fraction. After a culture of 72–96 h, lower and comparable quantities of both fractions were produced by the same batch of cultured parasites. Assaying the 2 fractions released during 0–33 h revealed that the high MW fraction had a suppressing effect on the bacterial clearance activity of haemocytes from juvenile snails and the low MW fraction activated haemocyte activity. Combined fractions obtained during 0–33 h had an activating effect on haemocyte activity but fractions separated from media collected at 33–72 h culture had a suppressing effect on the bacterial clearance activity of the haemocytes. The combined E–S fractions from the 72–96 h incubation had no effect on haemocyte activity. The present study shows that T. ocellata modulates the internal defence system of L. stagnalis by actively interfering with haemocyte function. MALDI-MS analysis of the low MW fraction revealed several components ranging from 450 to 1600 Da. Although it is as yet not clear which factor is responsible for the activating effect it is supposedly a peptide as it is pronase-sensitive and resistant to TFA and acetonitrile.

Based on substrate preferences, cercariae of Schistosoma mansoni were seen to express both cathepsin L and cathepsin B cysteine proteinases, although the former activity was many-fold greater. Two cathepsin L activities identified in cercarial extracts by zymography co-migrated with activities in extracts of 3 h and 24 h schisotosomula and in extracts of adult worms. Since these enzymes have been implicated in haemoglobin digestion by adult worms, they may perform a similar function in schistosomula. Immunolocalization using scanning electron micrographs showed that cathepsin L and cathepsin B proteinases were present in the cercarial post-acetabular glands. In addition, cercarial serine proteinase activities considered to facilitate skin penetration efficiently cleaved the substrates Z-Gly-Pro-Arg-NHMec and Z-Gly-Pro-Lys-NHMec. Cercariae release most of this serine proteinase activity when induced to secrete the contents of their acetabular glands. In contrast, newly transformed 3 h and 24 h schistosomula did not express this activity.

To study the infection dynamics of metacercariae of the digenean Cryptocotyle lingua, wild living Atlantic cod, Gadus morhua, were caged for 18 months close to the shore. Here they were exposed to naturally occurring transmission stages of the parasite. First, both the abundance and the variance to mean ratio of metacercariae increased, but during the second half of the study the abundance levelled out, and the variance to mean ratio showed a significant decrease. Host mortality was negligible throughout the study. Based on the relationship between pigment spots and metacercariae observed by skin digestion, there was no indication of density-dependent parasite mortality. We conclude that the infrapopulations of metacercariae on the caged cod probably were regulated by density-dependent host responses acting against the cercariae.

Infection with developing metacestodes of the rat tapeworm, Hymenolepis diminuta, is known to retard the accumulation of the yolk protein, vitellin, in the terminal ovarian follicles of the intermediate host, Tenebrio molitor. It is probable that this is the result of competitive inhibition of juvenile hormone binding at a microsomal binding site in the beetle follicular epithelium. Experiments were designed to test the hypothesis that inhibitor molecules were circulating in the haemolymph of infected beetles. Whole haemolymph, collected from male or female beetles at various stages post-infection, was injected into non-infected female recipients 2 days post-emergence. Ovaries were removed 3 days later and the vitellin content of the same sized follicles measured using an ELISA. The vitellin content of follicles from recipients of haemolymph from females infected with metacestodes at stage 1 and stage 3–4 was significantly reduced (24 and 27·9%) compared to sham-infected females. However, haemolymph from females infected with mature metacestodes did not affect the vitellin content. Results were thus comparable to those obtained by monitoring ovarian vitellin levels in female T. molitor with bona fide infections. Haemolymph from infected males did not affect ovarian vitellin content. These results indicate that molecules that can modulate vitellogenesis may be present in the haemolymph of females infected with developing metacestodes but that these factors disappear later in infection.

The distribution of Varroa jacobsoni on clustered, overwintering workers of Apis mellifera was investigated. The majority of mites were found between the 3rd and 4th ventro-lateral tergites of the abdomen with a significant preference for the left side of the host. This position would enable the mites to place their mouthparts in close proximity to the central portion of the bees' ventriculus. This may allow the mites access to nutrients at higher concentrations than would occur elsewhere in the haemolymph. At the start of winter, most infested bees carried only a single mite but towards the end increasing numbers of bees carried 2 or more mites. There was also an increase in the mite : bee ratio and more mobile mites (i.e. those moving about on the bees) were recorded. These results suggest that the mites suffer a lower overwinter mortality than the bees and are capable of transferring to a new host either before the original host dies or before it falls to the floor of the hive. This will have consequences for the transfer of diseases by Varroa within the hive.

Schistosoma mansoni egg counts by faecal examination vary considerably and are not very sensitive, so prevalences are underestimated. The distribution of egg counts can adequately be described by a stochastic model which distinguishes variation in counts between persons and variation in repeated counts within a person. Based on this model a pocket chart has been developed which predicts the proportion of individuals harbouring at least 1 S. mansoni worm pair – the ‘true prevalence’ – from a simple single survey prevalence and geometric mean egg count (using common duplicate 25 mg Kato–Katz smears). The current paper describes the validation of this chart by comparing predicted true prevalences with prevalences observed after 5–7 repeated Kato–Katz faecal examinations (Burundi), by examination of a large quantity of stool using the Visser filter (Brazil) or a selective sedimentation–filtration method (Surinam). Because 5–7 repeated examinations do not suffice to measure all infections, predictions have been made of the cumulative proportion positives over 5–7 surveys – the ‘approximate true prevalence’ – as well. After dividing the data into age groups, 12 different subsets were considered for validation. In all 12 cases, predicted true prevalences (or approximate true prevalences for the Burundi data) agree well with those observed. The overall agreement depends only slightly on the assumed relationship between worm numbers and mean egg counts, with a good fit for a productivity between 0·8 and 4·4 eggs per gramme faeces (EPG) per worm pair (WP). This interval includes the most plausible value from the literature, i.e. 1·0 EPG/WP, which has been applied in the initial pocket chart. These findings support the validity of the chart to predict true prevalences for a wide range of productivity assumptions, and reinforces the applicability of its underlying stochastic model to describe egg count variation. However, as predictions appear to vary importantly when using only part of the data, it is also concluded that the pocket chart never compensates for limited validity of initial single survey prevalences and geometric means in consequence of small sample sizes.

Parasite life-history traits should reflect past environmental and host-related selective pressures acting to produce strategies that maximize transmission success. The evolution of adult body size and egg production in 804 species of trematode parasites was investigated using independent contrasts derived from a phylogeny of trematodes. Contrasts in trematode body size were positively correlated with contrasts in egg size, and almost significantly correlated with contrasts in numbers of uterine eggs. After controlling for body size, no relationship existed between egg size and egg numbers, suggesting that there is no trade-off between the two components of egg production. Average host body mass and latitude of the sampling site did not correlate with either trematode body size or egg size. Contrasts between trematode taxa exploiting ectotherm hosts and their sister taxa exploiting endotherms showed no consistent difference in either body size or egg size. The effect of other variables on trematode life-histories, such as the nature of the habitat in which eggs are released, the site of attachment within the host's body, or the number of hosts involved in the life-cycle, could not be evaluated statistically. The similarity in life-history traits among members of given clades suggests that phylogenetic constraints may have acted to limit or mask any adaptive changes expected from changes in host-related or environmental conditions.

Developmental rates for Wuchereria bancrofti larvae maturing in the vector Aedes polynesiensis were estimated by analysing stage-frequency data consisting of counts of larval stages in mosquitoes reared at 20, 22·5, 25, 27·5, 30 and 32 °C. Base temperatures (i.e. low temperature thresholds) for W. bancrofti development were estimated by the x-intercept method and the model of Lactin et al. (1995). Resulting values were similar with both methods and were ≈12·5 °C for microfilariae (mf) in thorax, ≈17 °C for L1, 15·5 °C for L2 and 16·5 °C for L3. Upper thresholds estimated by the Lactin et al. model were 29·3 °C for mf, 29·1 °C for L1, 32·2 °C for L2 and 31·5 °C for L3. In addition, an original method was devised for computing the L3 upper threshold, by modelling L3 length shrinkage with temperature. It gave a value of 31·4 °C. At 32 °C, L2 and L3 stages exhibited altered morphology, larvae being shorter and wider than expected. The model of Lactin et al. described adequately the non-linear relationship between developmental rate and temperature, but a linear degree-day approach may be applied for temperatures below 27–28 °C.

The blood and broncho-alveolar leucocyte (BAL) responses, to a primary Necator americanus infection, were studied in male BALB/c and NIH mice. Following percutaneous infection, a significant blood leucocytosis occurred in both the BALB/c and NIH strains. The peak response occurred, in both strains, on day 10 p.i. and this was reflected in the eosinophil response which peaked at the same time-point. The change in blood eosinophil numbers, as a result of infection, was found to be the greatest recorded for any cell type. In BALB/c mice, however, infection elicited a significantly more intense total leucocyte, lymphocyte and eosinophil response than in NIH mice. In contrast, the BAL response was more intense in the NIH strain. Peak BAL responses were observed between days 12 and 16 p.i., in both strains, and the change in broncho-alveolar eosinophil numbers, as a result of infection, was found to be the greatest recorded for any cell type. The relationship between the observed leucocyte responses and resistance to the migrating larvae of a primary infection is discussed.

The lateral diffusion (DL) properties of the fluorescent lipid probe 5-N (octadecanoyl) aminofluorescein (AF18) inserted into the surface of muscle-stage larvae of Trichinella spiralis were investigated by fluorescence recovery after photobleaching. AF18 was not free to diffuse laterally in dormant larvae, and this remained unchanged after larval activation in vitro with trypsin and bile. However, a significant increase in surface fluidity of the probe was demonstrated (%R = 74·5; DL = 11·5 × 10−9 cm2/sec) when larvae invaded intestinal epithelial tissue following oral infection of mice. Membrane-permeant photoactivatable caged cyclic AMP was used to analyse the putative mechanism responsible for this increase in lateral diffusion in the parasite surface. Although incubation of larvae with 1–50 μM caged cAMP had no effect on surface fluidity, incubation with 100 μM caged cAMP induced a substantial increase in the lateral mobility of AF18 (%R = 64·3; DL = 8·3 × 10−11 cm2/sec) immediately following photo-activation of the caged messenger. This induced fluidity, however, was transient and the larval surface reverted to immobility within 15 min. These observations constitute the first reported measurement of the fluid properties of the surface of intracellular parasites, the first demonstration of the parasite surface fluidity altering as a result of host cell invasion and the first indication of a mechanism underlying changes in surface fluidity in parasitic helminths.

Oocysts of Cryptosporidium parvum showed relatively low levels of SOD activity. The SOD which had a pI of 4.8 and an approximate molecular weight of 35 kDa appeared to be iron dependent. Catalase, glutathione transferase, glutathione reductase and glutathione peroxidase activity could not be detected, nor could trypanothione reductase. No NADH or NADPH oxidase activity could be detected, nor could peroxidase activity be demonstrated using o-dianisidine, guaiacol, NADPH or NADH as co-substrates. However, an NADPH-dependent H2O2 scavenging system was detected in the insoluble fraction.

This paper describes the localization of paramyosin immunoreactivity in Schistosoma japonicum and represents the first comparative immunolocalization study among schistosome adult, cercariae and lung schistosomula by electron microscopy. A polyclonal antibody was utilized to immunolabel paramyosin or paramyosin-like proteins. Paramyosin was localized within the muscle layer of all 3 developmental stages. Furthermore, paramyosin was localized within granules of the post-acetabular glands of cercariae, and within the tegument matrix and surface of lung schistosomules. Adults and cercariae did not display any detectable paramyosin on the surface or within the tegument. The possible functions of paramyosin within S. japonicum and the relevance of these findings in relation to the reported protective properties of paramyosin as an anti-schistosome vaccine target molecule are discussed.

The equine intestinal cestode Anoplocephala perfoliata has been the subject of recent epidemiological and immunological studies because of its suspected association with intestinal disease in the horse. We have previously shown that the IgG(T) subtype antibody response to the 12/13 kDa component of the parasite excretory/secretory (E/S) antigen is positively correlated with parasite intensity. In this study, we utilize that correlation to examine the changes in natural infection intensity with age. Infection intensity based on IgG(T) responses showed a triphasic age-dependency pattern with peak mean worm burden in the 6 months–2 years age group, falling to a lower plateau level from 3 to 15 years, and rising again in older age groups. Anti-E/S total IgG was found to have a convex age-dependency curve, with maximal response in the 6 months–2 years old age group. IgG(a) showed a triphasic response similar to the age-intensity profile of IgG(T); IgG(c) showed steadily increasing levels of antibody with age. The IgG(b) age-dependency profile was intermediate between IgG(a) and IgG(c). Age-specific correlation coefficients between anti-12/13 kDa IgG(T) (as a measure of infection intensity) and IgG(a) and IgG(b) revealed statistically significant values for many age groups. The relative importance of exposure to infection and the development of acquired immunity as determinants of the observed age-intensity pattern is considered.

The intermediate hosts for Taenia saginata and T. solium are cattle and pigs (and humans for the latter), respectively. In vitro-hatched (but not activated) oncospheres of both Asian Taenia (T. saginata asiatica, a new subspecies of T. saginata or T. asiatica, a new species) and T. solium injected subcutaneously into the backs of mice with severe combined immunodeficiency (scid) developed into fully matured cysticerci. Five-month-old metacestodes of Asian Taenia had no hooklets and were bigger in size than those previously reported and similar to those of T. saginata. Their morphology suggested that the cysticerci were more advanced than those in the intermediate host animals. It is suggested that scid mice are valuable experimental animal models for studying human taeniid cestode infections.

We have used nuclear (ribosomal ITS1) and mitochondrial (ND1) sequences to characterize human and pig isolates of Echinococcus granulosus collected by fine needle aspiration biopsy (FNAB) in Poland. The data indicate clearly that the Polish patients were not infected with the common sheep strain (G1 genotype) of E. granulosus, normally associated with human cystic hydatid infection. Instead, the hydatid parasite infecting the Polish patients shares very similar ND1 sequence with the previously characterized pig (G7) genotype but it also exhibits some clear differences. In particular, E. granulosus DNA from the Polish patients amplified a single ITS1 fragment in PCR and distinct ITS1–RFLP patterns were obtained after restriction digestion. The form of hydatid isolated from the Polish patients appears, therefore, to represent a distinct, previously undescribed genotype (designated G9) of E. granulosus.

The microsporidian Encephalitozoon cuniculi can infect a wide variety of mammals including man. In this study, E. cuniculi isolates of animal origin were compared with 6 isolates obtained from HIV-infected patients. Based on results of Western blot analysis, random amplified polymorphic DNA (RAPD) and the sequence of the rDNA internal transcribed spacer (ITS) the isolates were classified into 3 groups with the repeated element 5′GTTT-3′ in the ITS being a reliable genetic marker. Five isolates from Swiss patients were found to be homologous to isolates from Swiss rabbits (strain I). The sixth isolate from a patient from Mexico differed by all methods and could be attributed to E. cuniculi strain III that has been described from 2 dogs from the USA. All of these isolates were distinguished from isolates from blue foxes from Norway (strain II). Intraspecific nucleotide divergence of the SSU rRNA gene of E. cuniculi belonging to the 3 strains was in the same low range (0·00–0·15%) as was found for the corresponding sequence of 2 E. hellem isolates. Groups of 2 rabbits were infected by oral inoculation of 107E. cuniculi spores (2 isolates from strain I of human and rabbit origin, 1 from strain III) as shown by antibody responses and the re-isolation of the parasites from brain material. The results provide further evidence that per oral transmission of the parasite between various hosts is feasible.

A characteristic feature of infections with the nematode parasite of mice Heligmosomoides polygyrus, is a marked IgG1 hypergammaglobulinaemia. A possible source for this immunoglobulin has recently been demonstrated, through evidence that H. polygyrus adult worm homogenate (AWH) can induce the in vitro production of non-specific IgG1 from mouse lymphocytes. To determine the interactions between this immunoglobulin and the parasite, the ability of IgG1 to bind to AWH of H. polygyrus was investigated. Protein (Western) blotting indicated that mouse monoclonal antibodies are able to bind non-specifically to selected parasite antigens. Furthermore, by binding H. polygyrus adult worm homogenate to cyanogen bromide (CNBr)-activated Sepharose CL-4B, an affinity column was prepared which could be used to efficiently purify mouse IgG1 monoclonal antibodies. These antibodies were eluted from the affinity column and still retained their original specificity. These results indicate that H. polygyrus not only induces the production of non-specific IgG1 by the host, it can also bind this immunoglobulin to its own specific proteins. Thus, it is possible that IgG1 produced during a primary infection with H. polygyrus may not entirely benefit the host.

The role of T-cells in development of experimental cerebral malaria was analysed in C57B1/6J and C57B1/10 mice infected with Plasmodium berghei K173 or Plasmodium berghei ANKA by treatment with anti-CD4 or anti-CD8 mAbs. Mice were protected against cerebral malaria (CM) when anti-CD4 or anti-CD8 mAbs were injected before or during infection. Even in mice in end-stage disease, i.e. with a body temperature below 35·5 °C, treatment with anti-CD4 or anti-CD8 antibodies or the combination protected against CM, whereas chloroquine treatment was completely ineffective in inhibiting further development of the cerebral syndrome.

Colchicine, a drug which poisons the polymerization of microtubules, was assayed for effects on the invasion of Plasmodium falciparum merozoites into red cells in order to investigate if merozoite microtubules have a function in invasion. Culture conditions and concentrations of colchicine were established where the maturation and rupture of schizonts was unaffected by the drug. This was judged first by light microscopy, including morphology and counts of nuclear particle numbers, then by ultrastructural studies which excluded deranged organellogenesis as a cause of merozoite failure, and finally by diachronic cultures in which both recruitment and loss of schizonts could be counted. Specific invasion inhibition was seen when 10 μM–1 mM colchicine was present. Red cells pre-incubated in colchicine and then washed showed no reduction in their extent of invasion, and neither red cell lysis, sphering nor blebbing were apparent. We conclude that intact microtubules are necessary for successful merozoite function.