Fifty-two Leishmania strains, obtained from human patients and dogs in a visceral leishmaniasis focus in Sudan, were characterized by isoenzyme electrophoresis (15 enzymes). The phylogenetic analysis showed that the 7 Leishmania zymodemes obtained hold ancestral positions on the phylogenetic tree, supporting the hypothesis of an East African origin of visceral leishmaniasis.

On the basis of partial amplification of a cloned fragment of kDNA of Leishmania infantum which is specific for this species, we developed a PCR–ELISA technique which avoids the problems associated with classical diagnostic techniques. This technique was tested on 33 L. infantum strains from 19 different zymodemes, which were recognized equally. It was also used on human and canine clinical samples. PCR–ELISA has a higher sensitivity than the other techniques used (IFAT, parasite cultures, optical microscopy of stained samples) and permits detection of a minimum of 0.1 promastigotes or 1 fg of genomic DNA. PCR–ELISA can be used to diagnose human cutaneous leishmaniasis using material obtained by scraping the lesion margin, and human visceral leishmaniasis in HIV(+) individuals and canine leishmaniasis with peripheral blood samples. The presence of L. infantum in dogs with low antibody titres with IFAT technique (20 and 40) was demonstrated indicating that seroprevalence data from epidemiological studies underestimate the true rates of infection.

Intrapredatorus barri, a microsporidian parasite of the predator mosquito Culex fuscanus Wiedemann has been shown to be heterosporous. In many features this species has been reported to be similar to Amblyospora trinus, another microsporidian parasite of a different predator mosquito which was thus proposed for reassignation to the genus Intrapredatorus. In this report small subunit rDNA has been sequenced from I. barri and phylogenetic analysis of 42 microsporidia sequences has been performed.Nucleotide sequence data reported in this paper are available in the Gen Bank™ database under the accession number AY013359. The molecular data show that I. barri can not be a member of the genus Amblyospora. Furthermore, the phylogenetic analysis shows, with high confidence, that the Amblyospora spp. sequences in the present study are not monophyletic.

African trypanosomes of the subgenera Nannomonas and Pycnomonas have been recorded from both wild and domestic suids. However, complete descriptions of some of these trypanosomes with regard to host range, pathogenicity, transmission and distribution are still lacking. Neither the recently described Trypanosoma (Nannomonas) godfreyi nor Trypanosoma (Nannomonas) congolense Tsavo have been isolated from mammalian hosts, while Trypanosoma (Pycnomonas) suis remains the rarest of the Salivarian trypanosomes. The only isolate presumed to be of the latter species is maintained at the Kenya Trypanosomiasis Research Institute, Nairobi. We present here the results of characterization of this isolate by morphology, tsetse transmission, the use of species-specific DNA probes and DNA sequence analysis. Morphology in stained blood smears revealed a small trypanosome with a free flagellum. Experimental transmission through Glossina morsitans morsitans showed a developmental cycle typical of subgenus Nannomonas. A positive identification was obtained with species-specific PCR primers for T. congolense Tsavo; moreover, the sequence of the SSU rRNA gene was almost identical to that of T. congolense Tsavo on database. In phylogenetic analysis of the SSU rRNA genes of Salivarian trypanosomes, T. congolense Tsavo grouped with T. simiae rather than T. congolense, suggesting that the name T. simiae Tsavo is more appropriate.

To dissociate the influence of host age from length of exposure on acquisition of filarial infection, we examined the development of microfilaraemia and anti-filarial IgG4 in all ages of a naive population that became suddenly exposed to Brugia malayi as a result of transmigration. Responses in 247 transmigrants, who had settled for periods of several months up to 6 years in their new homesteads, were compared with those of 133 life-long residents. As shown in earlier studies, anti-filarial IgG4 increased with age in the indigenous population, whose age is equivalent to length of exposure. However, by examining transmigrants, it became clear that development of specific IgG4 was influenced by age, since levels of this antibody were consistently higher in transmigrant adults than in transmigrant children, despite an equal length of exposure to filarial infection. Examining microfilaraemia, it was confirmed that infection establishes more rapidly in adults than in children.

Filarial infections of humans are chronic diseases. Despite an ongoing immune response, adult filariae continuously produce their offspring, the microfilariae (Mf), which are able to persist in sufficient numbers to ensure transmission. In this study, host- and parasite-derived factors, which contribute to persistence of Mf, were investigated using the filariasis model of Litomosoides sigmodontis in mice. Different strains of mice were found to differ widely in their capability to eliminate circulating Mf. Studies of congenic mouse strains showed that early and rapid clearance of Mf was mediated by activation pathways relevant to innate immunity, whereas late or delayed clearance of Mf was pre-determined by MHC-related factors. Genetic knock-out of genes for the MHC class-II molecules totally abrogated resistance. Most interestingly, the presence of only 1 adult female, but not male worms, renders all mice susceptible, irrespective of the genetic background, enabling Mf to circulate for extended periods of time. Such prolonged microfilaraemia was also observed in L. sigmodontis-infected animals challenged with heterologous Mf of Acanthocheilonema viteae. The use of cytokine gene knock-out mice showed that persistence of L. sigmodontis Mf was facilitated by IL-10, but not by IL-4 or IFN-γ. In conclusion, irrespective of a resistant or susceptible host genetic background, survival of Mf of L. sigmodontis in mice is decisively regulated by the presence of adult female L. sigmodontis which will skew and exploit immune responses to facilitate the survival and persistence of their offspring in the infected host.

In contrast to the growing knowledge accumulated on plant resistance to pathogens, mechanisms of parasite resistance largely remain to be elucidated in animal species. In the present study we investigated mechanisms underlying resistance/susceptibility in the snail–trematode system Biomphalaria glabrata–Echinostoma caproni. In particular, we compared the effect of the parasite excretory–secretory (E–S) products on the defence functions of haemocytes from 2 susceptible and 2 resistant snail strains. In vitro experiments showed that E. caproni E–S products inhibit adhesion and phagocytosis of haemocytes from susceptible snails. A partial biochemical characterization also suggested that the interfering factor(s) is (are) heat-labile glycosylated polypeptides of molecular mass between 10 and 30 kDa. Interestingly, haemocytes from resistant snails remained unaffected by the parasite E–S products, suggesting that a constitutive difference results in their refractoriness to the parasite's immunosuppressive factor(s).

We hypothesized that expression of immunity to gastrointestinal nematodes and its relationship with dietary metabolizable protein (MP) supply in ewes depended on body protein- but not on body fat reserves. Twin-bearing ewes were trickle infected with Teladorsagia circumcincta. Three mid-pregnancy feeding treatments were used, calculated to maintain body reserves (HH), maintain body protein- and lose body fat reserves (HL), or lose body reserves (LL), and followed by 2 iso-energetic, periparturient feeding treatments, calculated to provide either scarce (LP) or adequate (HP) amounts of MP. At the end of the mid-pregnancy feeding treatment, HL- and LL-ewes had a smaller backfat depth (∼body fat reserves) than HH-ewes, and LL-ewes had a smaller muscle depth (∼ body protein reserves) and higher faecal egg counts (FEC) than HH- and HL-ewes. Overall, LL-ewes had higher periparturient FEC than HH- and HL-ewes, and LP-ewes had higher periparturient FEC than HP-ewes. However, LL-ewes offered the LP feed had higher periparturient FEC than ewes on any of the other combinations of feeding treatments. Feeding treatments affected plasma pepsinogen but not plasma IgA. The results support the view that body protein- but not body fat reserves may overcome consequences of dietary MP scarcity on periparturient breakdown of immunity to gastrointestinal nematodes.

Stability of trichostrogylid populations indicates that some form of density-dependent regulation occurs which could act through fecundity. We present evidence for intraspecific density-dependent effects in 1 of 2, dominant, abomasal nematodes species (Ostertagia gruehneri) of Svalbard reindeer (Rangifer tarandus platyrhynchus). We found evidence in O. gruehneri, for density-dependent regulation of female worm length in April, July and October 1999. However, it is only in July that female worm length explains the variation in the number of eggs in utero which is also related to egg production per female worm only in this month and not at other times of the year. The seasonal pattern in faecal egg output in this species focuses egg production in the summer months when conditions are favourable to transmission. In contrast, we found no evidence in the other common species (Marshallagia marshalli) for density-dependent regulation of female worm length during or the number of eggs in utero. Faecal egg output in M. marshalli was positively related to worm burden but not to the mean number of eggs in utero. Neither inter-specific interactions nor host body condition appeared to influence worm fecundity. The contrasting patterns of density-dependent regulation of fecundity provides further evidence for divergent life-histories in this nematode community.

One α- and 2 β-tubulin isotypes (isotypes 1 and 2) from the parasitic nematode Haemonchus contortus were artificially expressed in E. coli and purified to obtain tubulin that was capable of polymerizing into microtubules. Binding of [14C] mebendazole (MBZ), a benzimidazole compound, to each individual unpolymerized isotype and to microtubules polymerized from recombinant α- and β-tubulin was assessed and Kd and Bmax values determined. Mebendazole bound to the individual tubulin isotypes with a stoichiometry of 1:1. Binding occurred with highest affinity to α-tubulin followed by β-tubulin isotype 2 and β-tubulin isotype 1 indicating that α-tubulin may play a role in benzimidazole binding to microtubules. Upon polymerization of α- and β-tubulin isotype 2 into microtubules the stoichiometry of binding increased to 2:1 (mebendazole:tubulin) while binding affinity remained the same. Mebendazole binding to α/β-isotype 1 microtubules remained unchanged following polymerization. The increase in the number of benzimidazole receptors on α/β-isotype 2 microtubules suggests the formation of a new benzimidazole receptor upon polymerization.

The possibility that much of the spatial and temporal variation in helminth communities in freshwater fish can be explained in terms of metapopulation theory was tested by examining helminth component communities of eels sampled from 3 localities in the River Exe and 5 of its tributaries on 2 occasions at an interval of 3 years. Four specific predictions of metapopulation theory in relation to community dynamics were tested. The prediction that parasite species could be recognized as common and rare and that this was sensitive to scale was met. The prediction that core species would be more widely distributed and have a higher dispersal ability was met only in part. The prediction that similarity between communities would increase with connectivity and decrease with distance apart was not met. The final prediction that species would go locally extinct and be replaced by rescue effects was not met. There was evidence of local extinctions; not of eel specialists but of species using other fish species as their principal definitive hosts. Eel helminth communities were heavily dominated by 3 species of eel specialists which were relatively stable and responsible for most of the similarity and its changes in space and time. It was concluded that metapopulation theory did not provide a very satisfactory explanation for the changes in the helminth communities in eels and reasons for this included the inherent difficulty of applying metapopulation theory to parasites and the fact that the helminth communities in eels in the River Exe were atypical in being dominated by eel specialists.

Resistance to Ascaris suum infections was investigated in 8- and 15-week-old Iberian pigs. Groups of 3 or 5 pigs were immunized weekly for 6 weeks with antigens of adult A. suum: a 97 kDa body wall (BW) fraction, a 42 kDa fraction of pseudocoelomic fluid (PF) or a 14 kDa PF-fraction; or were inoculated with increasing doses of infective eggs (500–20000), with or without abbreviation by pyrantel pamoate. All immunized pigs and unimmunized control pigs, were challenged with 10000 infective eggs 7 days after the last immunization. The number of liver lesions and lung larvae was substantially lower in the older pigs than in the younger ones 7 days after challenge, but the resistance in immunized pigs of both age groups was similar in comparison to the challenge controls of the same age. The highest degree of resistance against lung larvae was observed in pigs immunized with A. suum eggs (97–99%). The pigs immunized with the 14 kDa and 42 kDa PF-fractions were also well protected (67–93%), while no protection was produced by the 97 kDa BW fraction (0–49%). The reduction of white spots following immunization was less evident, with a maximum of 82% reduction in egg-inoculated young pigs.