A gene encoding a protein of 646 amino acid residues with a molecular mass of 71·3 kDa showing homology to the cytoplasmic form of the 70 kDa heat shock protein was cloned and sequenced from the nematode parasite Trichinella britovi (Tb). The gene was expressed in vitro as a protein of 71 kDa that was immunoprecipitated by a Trichinella-infected rabbit serum. Monospecific polyclonal antibodies raised against the recombinant Tb Hsp70 expressed in Escherichia coli, recognized a protein of 70 kDa by Western blot analysis of Tb soluble antigen (muscular stage). Tb Hsp70 was located in the nuclei of the muscle larvae as determined by the indirect immunofluorescent pattern on cross-sections of the worm. The expression of this protein was not detected in adult worm nuclei suggesting a differential expression of Hsp70 between the 2 stages of Trichinella.

Localization of the type 2 Alu sequence (B2), a highly repetitive DNA sequence in the mouse genome, was examined by in situ polymerase chain reaction (in situ PCR) in schistosomes. The signals to the B2 sequence were detected in the cytoplasm of the tegumental membrane and in the nuclei of the mesenchymal, testicular, ovarian and vitelline cells of 8- week Schistosoma japonicum. In contrast, it was difficult to detect any signals of this sequence in 8-week S. mansoni, whereas in 24-week male S. mansoni the signals were observed in the cytoplasm of the tegumental tubercles and in the nuclei of the mesenchymal and testicular cells. On the other hand, in 24-week female S. mansoni the signals were found in the nuclei of the mesenchymal, ovarian and vitelline cells but not found in the tegument. On the contrary, no hybridization band of the B2 sequence was detected in the amplified DNA of 3-week schistosomula of either species. These observations proved that the host DNA sequences existed in restricted schistosome cells and were accumulated in the schistosome body during their development.

Ivermectin is a promising drug for the treatment of lymphatic filariasis. A meta-analysis of trials investigating the effects of a single treatment suggested a dose-dependent effect on the production of microfilariae (mf) by adult Wuchereria bancrofti parasites. A mathematical model that describes the parasite dynamics in the human host and the impact of ivermectin treatment is presented and its outputs compared with these trials. The calculated trend in mf density after treatment appears to be particularly sensitive to the assumption about the mean life-span of mf. Adopting 0.5–2 years as a range of plausible values for this mf life-span, the model is used to estimate the impact of treatment on the parasite. It is found that irrespective of dosage, ivermectin eliminates 100% of the blood mf from a patient. Furthermore, at a dosage level of 400 μg/kg a single treatment irreversibly reduces the mf production of the adult parasites by at least 65%. For a dosage of 200 μg/kg this reduction is at least 35%. No such effect can be concluded from the results of trials using lower dosages.

The planorbid snail Biomphalaria glabrata responded to exposure to either the compatible digenetic trematode Echinostoma paraensei or the incompatible species Echinostoma trivolvis by producing increased amounts of several distinctive plasma polypeptides. These polypeptides characteristically precipitated from plasma when mixed with secreted–excreted products (SEP) of sporocysts or rediae from either digenean species. In contrast, control snails, or snails that had been wounded or infected with bacteria (Serratia marcesens or Staphylococcus epidermidis) showed no obvious plasma alterations and no precipitates formed when their plasma was mixed with SEP. Another planorbid species, Helisoma trivolvis, which displays reverse compatibility for the echinostome species used, also responded to exposure to both echinostomes by increased production of plasma polypeptides that precipitated in the presence of SEP. With some individual variation, these 2 snail species synthesized SEP-reactive plasma polypeptides forming diffuse bands centred at 53, 65, 80–120 and 200 kDa (the latter absent in Helisoma trivolvis). The 53 kDa polypeptides had not been observed before, whereas the others have been noted from B. glabrata. The diffuse 65 kDa band was strongly bound by anti-fibrinogen antibodies, supportive of earlier studies indicating it contains fibrinogen-related domains. The other specified polypeptides were also bound by these antibodies raising the possibility that they too contain fibrinogen domains. The results are suggestive of a general ability of these 2 planorbid snails to detect the presence of echinostomes even if the latter are subsequently incapable of development. The complex response they then mount, one not evoked by other challenges such as wounding or bacterial infection, may represent a dedicated response to a frequently encountered group of pathogenic parasites, the digeneans (echinostomes).

The initial infection process of nematode-trapping fungi is based on an interaction between the trapping structure of the fungus and the surface of the nematode cuticle. A bioassay was designed to investigate the predatory response of several isolates of nematode-trapping fungi against 3 mutants of Caenorhabditis elegans (AT6, AT10 and CL261), which have been reported to differ in the reaction of their cuticle to antibodies and lectins. The bioassay was also applied to infective larvae of animal (Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta and Trichostrongylus axei) and plant (Meloidogyne spp.) parasitic nematodes. Differences in trapping ability were most marked in the first 24 h, and were density dependent. Although the isolate of Arthrobotrys responded very rapidly in the first 24 h, Duddingtonia flagrans was generally the most effective isolate and Monacrosporium responded relatively poorly throughout all experiments. All the fungi tested trapped the srf mutants of C. elegans more efficiently than the wild type, and there were differences between the different srf mutants of C. elegans. Differences in trapping ability were also observed between different isolates of D. flagrans; similarly, differences in trapping behaviour were observed not only amongst the different species of plant-parasitic nematodes, but also between the sheathed and exsheathed larvae of the animal-parasitic nematodes.

Detection of Onchocerca volvulus larvae in vector populations is of prime importance in the assessment of the effectiveness of onchocerciasis control programmes. Traditionally, detection of larvae is attained by the dissection of flies, but this time-consuming method cannot easily discriminate between species of Onchocerca. The genome of all Onchocerca species has a unique 150 bp repeat, which can be amplified by PCR, and O. volvulus-specific DNA probes can detect these products by Southern blot (SB). This study optimizes a PCR/SB assay, and compares it with fly dissection to estimate the prevalence (p) and intensity of infection (m) in the local vector population of a Mexican community that has become hypoendemic as a result of 7 years of treatment with ivermectin and nodulectomy. The PCR detected 1 infected fly in a pool of 99 uninfected flies, but the optimal pool size was 50 flies. At the community level, 1 out of 10550 flies was positive (p=0·0095%, 95% confidence intervals CI=0·00024–07·05280%; m=0·00027 larvae/parous fly, CI=−0·00026–0·00081) by PCR, and 4 out of 10772 flies (p=0·0371%, CI=0·01012–0.09505%; m=0·00107 larvae/parous fly, 95% CI=0·00002–0·00212) by dissection (observed m=0·0005). Both methods produce statistically similar estimates of the prevalence and intensity, indicating that pool screening is a viable alternative for entomological surveillance in areas where the intensity of transmission is becoming extremely low as a result of control interventions.

A total of 121 European eels (Anguilla anguilla) from 2 sampling sites on the River Rhine were investigated in respect of their parasite communities. Special attention was given to the swim bladders, intestines, gills and fins of the fish. Twelve different parasite species were found to live in and on the eels. Data from each sampling site were kept separate. Parasites found in descending order of prevalence were: Anguillicola crassus, Trypanosoma granulosum, Myxobolus sp., Paratenuisentis ambiguus, Pseudodactylogyrus sp., Bothriocephalus claviceps, Myxidium giardi, Pomphorhynchus laevis, Trichodina sp., Raphidascaris acus, Acanthocephalus lucii and Acanthocephalus anguillae. Significantly different prevalences were reported for L3 larvae of A. crassus, adult P. ambiguus, B. claviceps and Myxobolus sp. at the 2 sampling sites. The highest number of parasite species was recorded from the intestine, which contained up to 6 different helminths. The coexistence of the acanthocephalans P. laevis and P. ambiguus, which showed clear patterns of distribution within the intestine of the respective hosts, was reported for the first time. Up to 3 different helminth species were found in the intestine of individual fish. Among those, acanthocephalans were the most prevalent worms with the eel-specific parasite P. ambiguus as the dominant species not only of the intestinal but also of the total component communities. Both infra and component communities exhibited low diversity and were dominated by this single species. The evenness reached only approximately 50% or less and it remained unclear why the helminth communities of the eels from the River Rhine with its huge catchment area exhibit such a low parasite diversity and high dominance.

A cDNA library constructed from 3 day post-infective L3 of the filarial nematode Brugia pahangi was screened by differential hybridization with cDNA probes prepared from different life-cycle stages. Five cDNA clones hybridizing selectively to the mosquito-derived L3 probe were isolated and characterized. Northern blot analysis of 4 of the clones confirmed that each was most highly expressed in the mosquito-derived L3. The expression of each mRNAduring parasite development in the mosquito vector was investigated using RT–PCR, and all were shown to be abundant in the immature L3. Four of the 5 cDNAs cloned coded for structural proteins: 2 cuticular collagens, and the muscle proteins tropomyosin and troponin. Further studies on troponin using an antiserum raised to the recombinant protein demonstrated that the protein, unlike the mRNA, was present in all life-cycle stages examined, while immunogold labelling demonstrated that it was localized to the muscle blocks.

This paper reports on the influence of maternal exposure to Ascaris suum on worm burden distributions in experimentally infected piglets. In the first study, sows were inoculated before and during gestation (6 months, long-term exposure) with 10000 A. suum eggs twice weekly. In a second study, sows were inoculated during gestation only (3 months, short-term exposure) with increasing doses of eggs (10000–40000 eggs twice weekly). Helminth-naive sows served as controls in both studies. The third study used the same design as the short-term exposure study, but piglets from exposed and control sows were cross-suckled within 4 h of birth before colostrum uptake. All piglets were inoculated 2 or 3 times with 50 A. suum eggs on days 4 and 7 (and 14) after birth, and left with the sows. At 10 weeks of age all piglets were necropsied, and liver lesions and worm burdens were recorded. Surprisingly, in piglets born to long-term exposed sows, the prevalence of A. suum infection and the mean worm burden were significantly higher than those in piglets from control sows. In contrast, neither worm burdens nor prevalence were significantly different between piglets from short-term exposed sows compared with their controls. In the cross-suckling experiment, 67% of piglets suckling control sows harboured worms at slaughter, compared with 15%of piglets suckling exposed sows. Maximum likelihood analysis of worm burden distribution and the degree of parasite aggregation showed 3 distinctly different types of overdispersed distributions: worm counts in piglets from control sows, in piglets from short-term exposed sows and in piglets from long-term exposed sows. When the worm burden data were analysed including the cross-suckled piglets by biological mother, it appeared that the control and short-term distributions converged and that only the long-term exposure was significantly different. Overall, the degree of parasite aggregation in piglets infected with A. suum decreased with exposure of the sows. A non-linear relationship was observed between prevalence of infection and mean worm burden, which was different for piglets from exposed and control sows, and similar to relationships of this type that previously have been found in human A. lumbricoides infections. It was concluded that in porcine A. suum infections maternal exposure alters the distribution of worms in their offspring, in which the duration of exposure appeared to be an important influence. The results of the cross-suckling further suggest that maternal factors, e.g. antibodies, are transferred via colostrum.

The efficacy of an isolate of the nematophagous fungus Duddingtonia flagrans against gastrointestinal nematodes of cattle was examined at 2 dose levels on 2 permanent pastures, with high and low stocking rates, respectively. Thirty calves, experimentally infected with Ostertagia ostertagi, were divided into 3 comparable groups and allocated to 3 similar paddocks in each of the 2 trials. Two of the 3 groups received fungal material once per day during the initial 2 months, either at high dose (106 fungal spores/kg body weight) or low dose (5×105 or 2·5×105 fungal spores/kg body weight). The third group remained as an untreated control group. Faecal, blood, and herbage samples were collected and animals were weighed every month from May to September. The pasture grazed at a high stocking rate had a large number of overwintering infective larvae, while the pasture grazed at a low stocking rate had a low overwintering herbage larval infectivity. The results showed that, at a high stocking rate, the recovery of infective larvae on pasture was diminished and calves were prevented from clinical ostertagiosis by using the D. flagrans Troll A-isolate. At low stocking rate, the parasite burden seemed not to be very heavy, and a conclusive effect of the fungi at the dose-level used could not be detected.

Cysteine proteinases have been implicated in the protection conferred by vaccination with detergent-soluble extracts of Haemonchus contortus. In the present study, antisera from sheep refractory to Haemonchus challenge following vaccination with a ‘proteinase-enriched’ Haemonchus gut membrane extract, were employed to screen a cDNA expression library of the adult parasite. This resulted in the isolation of 3 cDNAs (designated hmcp1, 4 and 6) encoding cathepsin B-like cysteine proteinases. Immunocytochemical studies specifically localized the products of these genes to the microvillar surface of the parasite's gut and RT–PCR experiments revealed that these were developmentally regulated, being expressed exclusively during the blood-feeding parasitic stages. In addition, a generic PCR approach was adopted in order to identify the predominant cysteine proteinases in a UK strain of Haemonchus. A panel of 5 cDNAs, including hmcp1 and 4, was amplified in this way. Genomic Southern blot analysis indicated that some of these enzymes were encoded by single-copy genes, whereas others were encoded by multi-copy genes. Subsequent sequence analysis revealed that the proteases identified in this study were distinct from those previously reported in USA strains of the parasite.

The survival capacities of Muellerius capillaris (Nematoda: Protostrongylidae) free-living larval stages (L1) of an Israeli desert isolate (Nubian isolate) (92 mm rain per annum) and a French isolate from a temperate habitat (650 mm rain per annum) were compared under humid and extreme dry conditions. Under the dry conditions (7% relative humidity [r.h.] and 33% r.h. at 23 °C), both isolates exhibited the same remarkable survival capacity for periods of 17 and 28 days, respectively (>92% survival, P>0·1). However, during and after recovery from the anhydrobiotic state, different behaviour patterns of L1 were observed in each isolate. Under humid conditions (97% r.h.) for 10 days, both isolates exhibited similarly low survival percentages (13·4±4·6%, S.E., survival for the Nubian isolate, 3±3% for the French isolate, P>0·05). In water, the French isolate was less active, survived better (52±8·4% vs 28·7±3·7, by day 28, P<0·05) and retained a better morphological appearance (shrunk less) than the Nubian isolate. Larval development of the French isolate in the land snail Theba pisana was significantly faster than that of the Nubian isolate, reaching a higher proportion of infective stages (L3) on day 36 post-infection (78·6% vs 56% P>0·05). However, similar infection intensities were recovered in snails infected with either of the isolates (59·8±10·3, S.E., n=14, for the French isolate, 53±6, n=13, for the Nubian isolate, P<0·05).

In Denmark, Finland, Iceland, Norway and Sweden, 413 sow herds were randomly selected for sampling. Faeces from pigs of 7 age groups/categories were examined for helminth eggs (11233 individual samples), and an accompanying questionnaire was completed at each visit. In total, 1138 pigs on 230 farms were found to be positive for Ascaris suum. Considerable differences in the occurrence of A. suum could be observed directly for several of 20 independent variables at the herd or category level. However, given that univariate analyses may be severely affected by confounding of covariates resulting in spurious inference, additional multivariate analyses were undertaken. An ordinary logistic regression on Ascaris positive/negative farms showed that Denmark had the highest frequency of infected herds, while Iceland and Finland had the lowest frequencies and that herds using ‘late weaning’ and ‘Class 2’ drugs (pyrantel, levamisole) were most often infected. Because many herds were found to be totally negative for A. suum, mixed hierarchical logistic-normal regression models (both the penalized quasi-likelihood and the Markov Chain Monte Carlo methods) were developed for both a full (all herds) and a reduced (the 230 infected herds) data set using either a cut-off of >0 eggs per gram (epg) or >200 epg to counter for false-positive egg counts. Estimates for identical models, but where the animal level variance was constrained to the binomial assumption, were also calculated. Significant covariates were robust to model development with ‘Age group’, ‘Country’, ‘Weaning age’, ‘Water system’ and simple interactions between the latter two and ‘Age group’ being significantly associated with the occurrence of A. suum, while all variables concerning anthelmintic drug, anthelmintic strategy, floor type, bedding, dung removal, washing and disinfection were not. These findings are discussed in the light of the complex relationship between A. suum and its pig host.

It has been reported that infection with Nippostrongylus brasiliensis induces villus atrophy with various histological alterations. In N. brasiliensis-infected rats, villus length in the jejunum was reduced significantly at day 10 p.i., when serum levels of rat mast cell protease (RMCP) II had increased significantly. To determine whether the villus atrophy is associated with enhancement of apoptosis, apoptotic nuclei were labelled using the nick end-labelling method. Numbers of labelled cells were markedly increased in the villus epithelium at 7–10 days p.i., while the numbers returned to normal 14 days p.i. when worms were rejected from the intestine and villus length became normal. Examination of the expression of the adhesion molecule E-cadherin showed granular immunoreactivity in the cytoplasm of atrophic villus epithelium with loss of normal localization to epithelial cell borders. In mast cell-deficient Ws/Ws rats, villus length was reduced as significantly as in +/+ counterparts at day 10 p.i. with marked increases in the numbers of apoptotic cells. These results suggested that villus atrophy was closely associated with enhanced apoptosis and loss of adhesion in epithelial cells. Mast cell activation appears not to be involved in these alterations.

The excretory–secretory antigen of Fasciola hepatica (ESA) is involved in the suppressive phenomena of cellular immune responses in rats. The ESA can depress the proliferative response of spleen mononuclear cells and inhibit nitric oxide (NO) production by peritoneal cells. In the present study we identified ESA proteins of ca 24 kDa, which shared significant sequence homology to glutathione-S-transferase (GST) obtained from homogenates of F. hepatica adults, other helminths and different mammals. When the dimeric form of these proteins ca 48 kDa was cultured with rat spleen cells, a significant decrease of proliferative response to Con A was detected, starting from 20 μg/ml of ESA protein (P<0·03). We also observed a significant inhibition of nitrite production by incubation with the dimeric form in normal peritoneal macrophages (P<0·04). These results indicated that the GST secreted by the parasite could be involved in evasion of the parasite from the host immune response.

Pig faeces were deposited on experimental plots in the spring, summer, autumn and winter to study development and survival of Ascaris suum and Trichuris suis eggs under outdoor conditions. Faeces were placed either in short grass or 2 cm below the surface of bare soil, imitating pastures used by nose-ringed, grazing pigs or normally rooting pigs, respectively. The numbers and developmental stages of the eggs were recorded in faeces and soil for up to 50 weeks post-deposition. Embryonation took place only during the summer months and seemingly was independent of the microclimate. The majority of A. suum and T. suis eggs, which are generally considered to be extremely resistant and long-lived, seems to disappear rather fast. The disappearance rate for A. suum eggs was higher than for T. suis eggs, and both egg types disappeared significantly faster in the summer months than in the winter months, and when placed in short grass than when buried in soil (less exposed). We discuss how knowledge on egg development and survival may be used in the planning of pasture strategies for control of helminth infections in outdoor pigs.

Protective immunity against larval Taenia crassiceps has been shown to rely on T cells; however, the roles of the specific subsets of T cells during infection are not known. To investigate a possible role for γδT cells, this study investigated larval infection in δ-chain knock-out C57BL/6 (deltaKO) and wild-type C57BL/6 mice. It was found that deltaKO mice and C57BL/6 mice were equally susceptible to infection suggesting γδT cells do not play a major role in protective immunity. Cytokine production by concanavalin A (ConA)-stimulated spleen cells from infected deltaKO mice and C57BL/6 mice were determined. All infected mice demonstrated an increased IL-10 production suggesting a Th1-inhibitory function. Cells from infected deltaKO mice and C57BL/6 mice did not show increases in IL-4 production. Heavily-infected C57BL/6 mice showed a decrease in IFN-γ production compared to deltaKO mice. These observations suggest that an increase in IL-10 production best correlates with a non-protective immune response. To make comparisons between in vitro cytokine production and systemic immune responses, cytokine levels in serum were determined. C57BL/6 mice and deltaKO mice showed increases in serum levels of IL-4 and IFN-γ at 52 days post-infection. The systemic immune response of these mice, therefore, is a mixed Th1/Th2-type response and γδT cells are apparently not responsible for the systemic increases in these cytokines.

In Strongyloides ratti-infected rats, 2 peaks of egg excretion were observed; a large one with maximum egg production on days 7–8 of infection and a small more inconspicuous one around day 25. The second peak, which had been ignored in most studies, was produced by adults in the caecum and the colon. The adults were larger in length and had more embryonated eggs in the uterus compared with adults in the small intestine at day 25 post-infection. It is suggested that parasitic adults once expelled from the small intestine resettle and recover in the large intestine. Filter paper faecal culture carried out for 9 days at different days post-infection revealed that the total number of infective larvae that developed during the second peak was twice the number that developed during the first peak, despite the fact that total egg output during the second peak was less than one twentieth of the first peak. The results suggest that the small second peak was as important as the first one in the transmission of S. ratti.