This paper reviews the immunomodulatory effects (immunosuppression or immunoactivation) of various anthelmintics including levamisole, fenvalerate, dieldrin, carbofuran, aminocarb, thiabendazole, fenbendazole, oxfendazole and ivermectin. The induced modulation of immune function may occur via direct and/or indirect mechanisms. The immunomodulatory effects of these anti-parasitics have been studied in a variety of bacterial (e.g. brucellosis, salmonellosis, paratuberculosis, mastitis), viral (e.g. infectious bovine rhinotracheitis, Herpes, foot and mouth disease), parasitic (e.g. onchocerciasis, coccidiosis, ascariasis, schistosomiasis) and neoplastic diseases. Some antiparasitics have also been used to boost immunity in a number of human diseases including leprosy, Hodgkin's disease, rheumatoid arthritis, and in adjuvanted therapy of colorectal cancer. The ability to stimulate the immune response of animals offers a new means of disease intervention. Future research on immunomodulatory effects of anti-parasitics, for humans and domestic farm animals, will provide additional methods of treating immunosuppressed subjects. The immunopotentiating or immunosuppressing activity of anti-parasitics will dictate whether co-administration of vaccines and anthelmintics or administration of vaccines during the window of immunoactivation is justified or not.

Toxoplasma gondii is an obligate intracellular protozoan parasite. The objective of the present study was to examine the behaviour of Toxoplasma gondii RH Ankara strain tachyzoites in a cell culture environment. The study represents the first step in determining whether T. gondii RH Ankara strain tachyzoites, grown in cell culture, are of sufficient quality to allow cessation of in vivo tachyzoite production for diagnostic assays. In the present study, T. gondii RH Ankara strain tachyzoites were continuously produced in myeloma X63.Ag8.653, HeLa, Hep-2, and Vero cell cultures for 2 months. The average size of the tachyzoites was 3×5·7 μm prior to the first inoculation but after continuous production, a marked decrease was noted in average tachyzoite size. The smallest tachyzoite size, was 1×2·1 μm after 2 months, in myeloma cell cultures even though the yield of tachyzoites increased. With other cell cultures, tachyzoite yields were not as high as myeloma cell culture although decrease in size was less. The smallest decrease in tachyzoite size, averaging 2×3·8 μm after 2 months, was observed in tachyzoites produced in HeLa cell cultures. A virulence assay in small groups of BALB/c mice, using tachyzoites derived from cell cultures, was also conducted. The preliminary results of the virulence assay suggest that as the size of the tachyzoites decreased, the virulence in mice decreased. Future research will focus on the effect of the size of cell culture-derived T. gondii RH Ankara strain tachyzoites on the virulence, protein expression, and the reliability of diagnostic assays. Ultimately, the behaviour of tachyzoites from various T. gondii strains will be observed in cell culture to determine if size is altered.

Damage to the surface membrane of adult Schistosoma mansoni, and the activity of the excretory system, as shown by resorufin fluorescence, was observed following treatment with praziquantel and incubation with other molecules. Praziquantel treatment induced damage to the surface membrane as measured by the use of a variety of fluorescent compounds. The excretory system of the male worm was inhibited immediately after praziquantel treatment, but fully recovered after culture for 2 h following removal of praziquantel. The excretory system of the female, observed to be minimally active in untreated worm pairs, was often greatly activated in paired females, as shown by intense resorufin labelling, after praziquantel treatment, and this continued during recovery of the male excretory system. In experiments with normal worm pairs, the female could be activated by inhibiting the metabolic rate of the pair by a cooling procedure. The effects on the excretory system of changes in culture conditions (such as changes in pH, concentrations of bacterial lipopolysaccharide, cytokines, reactive oxygen species, compounds which remove cholesterol, such as beta-methyl cyclodextrin, and damaging basic poly-L-lysine) were also assessed. It is concluded that the extensive excretory system of the adult worm is responsive to drug treatment and to certain changes in environmental conditions. Its activity seems to be strongly linked to the integrity of the surface membrane.

Human parasites are often distributed in metapopulations, which makes random sampling for genetic epidemiology difficult. The typical approach to sampling Schistosoma mansoni involves laboratory passage to obtain individual worms with small sample size and selection bias as a consequence. By contrast, the naturally pooled samples from egg output in stool or urine directly represent the genetic composition of current populations. To test whether pooled samples could be used to estimate population allele frequencies, DNA from individual cloned parasites was pooled and amplified by PCR for 7 microsatellites. By polyacrylamide gel analysis, the relative band intensities of the products from the major alleles in the pooled samples differed by 0–6% from the summed intensities of the individual clones (mean=2·1%±2·1% S.D.). The number of PCR cycles (25–40) did not influence the accuracy of the estimate. Varying the frequency of 1 allele in pooled samples from 32 to 69% likewise did not affect accuracy. Allele frequency estimates from aggregate samples such as eggs will be a better foundation for studies of parasite population dynamics as well as the basis for large-scale association studies of host and parasite characteristics.

The clinical spectrum of Trichomonas vaginalis infection varies from asymptomatic to mild, moderate or severe vaginitis. Nitric oxide and other reactive nitrogen radicals produced by immune effector cells are important cytotoxic and cytostatic mediators against several microorganisms including parasites. In the present study, inducible nitric oxide synthase (iNOS) and reactive nitrogen intermediates (RNI) were determined in leucocyte cultures (stimulated with T. vaginalis in vitro) and vaginal washes (VWs) of 22 symptomatic and 20 asymptomatic T. vaginalis-infected and 20 healthy women by immunoblotting and Griess method respectively. The iNOS protein was detected in leucocytes and VWs of all the symptomatic and asymptomatic women, but was not detected in any of the samples from healthy women. Mean iNOS protein band intensity was significantly higher in leucocytes as compared to VWs (P<0·001) of both symptomatic and asymptomatic women and was also higher in leucocytes of asymptomatic as compared to symptomatic women (P<0·05). Mean RNI concentration was also significantly higher in leucocytes (P<0·01) and VWs (P<0·05) of asymptomatic as compared to symptomatic women, and was also higher in samples of infected as compared to healthy women (P<0·001). These results suggest that reactive nitrogen radicals may have a role in limiting T. vaginalis infection in asymptomatic women.

With the aim to study proteinases released to the culture medium during Trypanosoma cruzi metacyclogenesis, the presence of cysteine proteinases (CPs) was analysed in culture supernatants obtained throughout the differentiation induced by stimulation of epimastigotes with Triatoma infestans hindgut homogenate. In SDS-gelatin containing gels, an important endopeptidase activity with apparent molecular weight range between 97 and 116 kDa was encountered at pH 6, which was abolished by the specific cysteine proteinase inhibitor E-64 and TLCK, but not by pepstatin, 1,10 phenantroline or PMSF. This novel CP, named TcCPmet, showed affinity to cystatin-Sepharose, denoting its thiol-proteinase character as well as to ConA-Sepharose, indicating it contains N-linked oligosaccharides. However, it presented a different elution pattern on ConA-Sepharose than cruzipain and, in addition, it was not recognized by anti-cruzipain serum, facts that strongly suggest the different nature of both CPs. Moroever, evidence is presented indicating that TcCPmet was able to hydrolyse the same chromogenic peptides as cruzipain at optimal alkaline pH values, although with a different order of effectiveness. Our results indicate the presence of a novel CP secreted by metacyclic trypomastigotes and reinforces the important role of these enzymes in metacyclogenesis.

This paper investigated the intestinal distribution and fecundity of 2 species of Diplostomum parasites, D. spathaceum and D. pseudospathaceum, in 2 species of definitive hosts, herring gull (Larus argentatus) and common gull (L. canus), using both empirical field data and experimental infections. At the level of individual hosts, the parasite species occupied different parts within the intestine, but the fecundity of the worms, measured as the number of eggs in the uterus, did not differ between the parasite species except in wild common gulls. Interestingly, egg numbers in individual hosts were positively correlated between the parasite species suggesting that some birds provided better resources for the parasite species. At the host population level, fecundity of the worms did not differ between the host species or between adult birds and chicks. Both parasite species were also aggregated to the same host individuals and it is likely that aggregation is transferred to gulls from fish intermediate hosts. Individual differences in suitability and parasite numbers between hosts provide important grounds and implications for epidemiological model-based parasite prevention strategies.

The parasite community of animals is generally influenced by host physiology, ecology, and phylogeny. Therefore, sympatric and phylogenetically related hosts with similar ecologies should have similar parasite communities. To test this hypothesis we surveyed the endoparasites of 5 closely related cheilinine fishes (Labridae) from the Great Barrier Reef. They were Cheilinus chlorourus, C. trilobatus, C. fasciatus, Epibulus insidiator and Oxycheilinus diagramma. We examined the relationship between parasitological variables (richness, abundance and diversity) and host characteristics (body weight, diet and phylogeny). The 5 fishes had 31 parasite species with 9–18 parasite species per fish species. Cestode larvae (mostly Tetraphyllidea) were the most abundant and prevalent parasites followed by nematodes and digeneans. Parasites, body size and diet of hosts differed between fish species. In general, body weight, diet and host phylogeny each explained some of the variation in richness and composition of parasites among the fishes. The 2 most closely related species, Cheilinus chlorourus and C. trilobatus, had broadly similar parasites but the other fish species differed significantly in all variables. However, there was no all-encompassing pattern. This may be because different lineages of parasites may react differently to ecological variables. We also argue that adult parasites may respond principally to host diet. In contrast, larval parasite composition may respond both to host diet and predator-prey interactions because this is the path by which many parasites complete their life-cycles. Finally, variation in parasite phylogeny and parasite life-cycles among hosts likely increase the complexity of the system making it difficult to find all-encompassing patterns between host characteristics and parasites, particularly when all the species in rich parasite communities are considered.

Widespread use of ivermectin (IVM) as part of the Onchocerciasis Control Program (OCP) in West Africa could influence the evolution of the human filarial parasite Onchocerca volvulus. Use of IVM, in some areas for 15 years, may have restricted genetic diversity of O. volvulus, resembling effects attributed to a population bottleneck. Large population-based chemotherapy programmes, such as the OCP, may impose strong selection pressure on parasites and an examination of possible genetic selection by IVM in O. volvulus is warranted. IVM is a substrate for P-glycoprotein; a homologue from O. volvulus (OvPGP) has been linked with IVM sensitivity. Linkage disequilibrium (LD) patterns of 28 genetic markers spanning the OvPGP locus were examined in 4 O. volvulus populations from the Volta Region of Ghana, West Africa. Reduced gene diversity, increased heterozygosity and an increase in the number of markers not in Hardy-Weinberg equilibrium were associated with increasing IVM treatment. The number of regions in LD decreased with treatment and with time. However, between 1999 and 2002, seven regions of OvPGP were always in complete LD, while surrounding areas showed a reduction in genetic variation. The use of IVM for onchocerciasis control has imposed strong selection on O. volvulus populations, reducing genetic variation and disrupting LD.

We describe the epidemiology of the protostrongylid parasites Parelaphostrongylus odocoilei and Protostrongylus stilesi in Dall's sheep (Ovis dalli dalli) from the Mackenzie Mountains, Northwest Territories, Canada (65 °N; 128 °W). Peak numbers of 1st-stage larvae of both parasites were shed by Dall's sheep on their winter range from March until May. In larval development experiments in the Mackenzie Mountains, peak numbers of infective 3rd-stage larvae of P. odocoilei were available in gastropod intermediate hosts in August–September. For both protostrongylids, the majority of transmission likely occurs on the winter range, with infection of gastropods when they emerge from hibernation in spring, and infection of Dall's sheep upon their return in fall. We validated a degree-day model for temperature-dependent development of larval P. odocoilei in gastropods, and applied degree-day models to describe and predict spatial and temporal patterns in development of P. odocoilei and P. stilesi in northern North America. Temperature-dependent larval development may currently limit northward range expansion of P. odocoilei into naïve populations of Dall's sheep in the Arctic, but climate warming may soon eliminate such constraints. In Subarctic regions where both P. odocoilei and P. stilesi are endemic, the length of the parasite ‘growing season’ (when temperatures were above the threshold for larval development) and amount of warming available for parasite development has increased over the last 50 years. Further climate warming and extension of the seasonal window for transmission may lead to amplification of parasite populations and disease outbreaks in host populations.

The effects of cold storage of infective third-stage larvae (L3) of different isolates of the parasitic nematode Haemonchus contortus were studied with respect to infectivity, pre-patent period and propensity for larval arrestment. Two complementary experiments were conducted with 2 groups of lambs, each animal being inoculated with 2000 L3 of either Swedish or Kenyan origin. In a first experiment, L3s were cold treated at 5 °C for 9 months prior to infection, whereas in a second experiment larvae were newly hatched. Individual faecal egg counts (FECs), and worm burdens were determined for each experiment. The results showed that the greatest differences were associated with the pre-treatment of larvae. The pre-patent period and the FECs differed significantly between the experiments but not between the isolates used in each experiment. However, the extent of hypobiosis was significantly different between the two isolates when fresh larvae were used (36% Kenyan isolate and 70% Swedish). The storage of H. contortus at 5 °C had no apparent effect on the infectivity of L3s, as high establishment ranging from 43 to 74% were observed, irrespective of isolates used. This study showed that H. contortus exhibited similar phenotypic traits regardless of geographical origin. Thus, there was limited evidence for adaptations to temperate climatic conditions.

Infective larvae of an intestinal nematode, Strongyloides venezuelensis, enter rodent hosts percutaneously, and migrate through connective tissues and lungs. Then they arrive at the small intestine, where they reach maturity. It is not known how S. venezuelensis larvae develop during tissue migration. Here we demonstrate that tissue invasion ability of S. venezuelensis larvae changes drastically during tissue migration, and that the changes are associated with stage-specific protein expression. Infective larvae, connective tissue larvae, lung larvae, and mucosal larvae were used to infect mice by various infection methods, including percutaneous, subcutaneous, oral, and intraduodenal inoculation. Among different migration stages, only infective larvae penetrated mouse skin. Larvae, once inside the host, quickly lost skin penetration ability, which was associated with the disappearance of an infective larva-specific metalloprotease. Migrating larvae had connective tissue migration ability until in the lungs, where larvae became able to settle down in the intestinal mucosa. Lung larvae and mucosal larvae were capable of producing and secreting adhesion molecules.

The aim of this study was to investigate the effects of grazing different bioactive forages on acquired immunity against Teladorsagia circumcinta infection. The development of immunity was assessed by following the response of trickle-infected lambs grazing chicory (Cichorium intybus; IC), sulla (Hedysarum coronarium; IS) or grass/clover (Lolium perenne/Trifolium repens; IGC), to a single challenge infection. Parasite-naive lambs, grazing grass/clover, were also challenged with the single infection dose providing the uninfected control (UGC) group. Trickle infection significantly reduced worm establishment, inhibited larval development and increased mucosal mast cell (MMC) and globule leucocyte (GL) cells. Grazing treatment (chicory, sulla or grass/clover) significantly affected adult worm (P<0·05), late-L4 (P<0·01) and mid-L4 (P<0·01) larval-stage recoveries of the trickle-infected lambs, with IGC group always carrying higher worm burdens than either IC or IS lambs. MMC and GL cells of trickle-infected lambs were positively correlated with the proportion of early-L4 worms recovered and negatively correlated with both the proportion of adult worms recovered and the total worm establishment, suggesting that the observed effects were due to an enhanced immune response. The results suggest elevated immune responses against T. circumcincta infections in growing lambs grazing on either sulla or chicory compared to those grazing on grass/clover, probably due to differences in forage nutritional values.

A study of the anterior adhesive apparatus (head organs) of Bravohollisia gussevi Lim, 1995 was carried out using light and electron microscopy. The anterior adhesive apparatus or head organs in B. gussevi comprise 6 circular openings or apertures in the antero-lateral region, associated pits lined with specialized microvillous tegument that differ from the general body tegument, a bundle of ducts, and uninucleate gland cells located lateral to the pharynx. The uninucleate glands of the anterior adhesive apparatus (head organs) comprise 2 types of cells, one kind of cell producing rod-like bodies (S1) and the other oval bodies (S2). The S1 bodies are filled with numerous, less electron-dense vesicles in an electron-dense matrix, while S2 bodies have no vesicles but contain a more homogeneous electron-dense matrix. Interlinking band-like structures were observed between S1 bodies. Similar band-like structures were found between S2 bodies. The formation of S1 bodies was followed by transmission electron microscopy. However, the formation of S2 bodies was unclear and could not be resolved. Uniciliated structures were also observed around the openings of the anterior adhesive apparatus. Each uniciliated structure is usually associated with an opening of a gland cell producing granular, electron-dense, secretory bodies, which differ from the secretions produced by the lateral gland cells of the anterior adhesive apparatus.

Many issues concerning the taxonomy of Echinococcus have been resolved in recent years with the application of molecular tools. However, the status of Echinococcus maintained in transmission cycles involving cervid intermediate hosts remains to be determined. The recent characterization of the parasite from cervids in Finland has highlighted the paucity of data available, particularly that from North America. In this study, we have characterized a large number of Echinococcus isolates from cervids from Western Canada on the basis of morphology and molecular genetic techniques. Our results support earlier studies suggesting that Echinococcus of cervid origin is phenotypically and genetically distinct to Echinococcus maintained in domestic host assemblages, and also confirms that Echinococcus of cervid origin does not constitute a genetically homogeneous group. However, our data do not support the existence of 2 distinct genotypes (strains/subspecies) with separate geographical distributions. Our data appear to support the existence of only 1 species in cervids, but additional isolates from cervids and wolves in other endemic regions should be characterized before a final decision is made on the taxonomic status of Echinococcus in cervids.