Plasmodium ookinetes are elongate, motile and invasive while inside the mosquito gut but promptly metamorphose into spherical immobile oocysts upon coming in contact with the basement membrane surrounding the midgut. There they begin a prolonged growth period characterized by massive DNA synthesis for the production of sporozoites. Living Plasmodium gallinaceum ookinetes attached avidly to the murine extracellular matrix proteins, laminin and collagen type IV. In ELISA-type assays, the main ookinete surface protein, Pgs28 was implicated as a mediator of parasite attachment to these basement membrane constituents. Laminin and collagen IV adhered to ookinete and oocyst lysates spotted onto nitrocellulose membranes. Receptor–ligand blot assays demonstrated that Pgs28 and an oocyst-specific antigen recognized by the mAb 10D6 interact with murine collagen IV and laminin. 10D6 antigen was also recognized by monospecific antiserum against the human epidermal growth factor receptor. Mosquito-derived laminin was incorporated into oocyst capsules of P. gallinaceum growing in Aedes aegypti. We hypothesize that contact with the mosquito basement membrane triggers the transformation of ookinetes into oocysts. Coalescence of basement membrane proteins onto the capsules masks developing oocysts from the mosquito's immune system and facilitates their prolonged extracellular development in the mosquito body cavity.

A cross-sectional sero-epidemiological study was performed in Magoda, Tanzania, an area where malaria is holoendemic. Blood samples were collected from children (1–4 years) and tested for IgG antibody reactivity against 2 recombinant protein fragments of Plasmodium falciparum Rhoptry-Associated Protein-1 (rRAP-1). The data were related to the prevalence of malarial disease and single P. falciparum or mixed Plasmodium infections. Fever ([ges ]37·5 °C) in combination with parasite densities >5000/μl were used to distinguish between children with asymptomatic malaria infections and those with acute clinical disease. Furthermore, C-reactive protein (CRP) was applied as a surrogate marker of malaria morbidity. The prevalence of Plasmodium infections was 96·0%. Eleven children were defined as clinical malaria cases, all with single P. falciparum infections. The density of P. falciparum was significantly lower in children with mixed Plasmodium infections compared to those with single P. falciparum infections. Children with asymptomatic P. falciparum infections had higher IgG reactivities to rRAP-1, compared to IgG reactivities of children with malarial disease. Children with mixed Plasmodium infections generally showed elevated IgG reactivity to rRAP-1, when compared to children with single P. falciparum infections. The possible relationship between mixed species infections, clinical outcome of the disease and antibody responses to RAP-1 is discussed.

This study examines polymorphisms in 2 genes (pfmdr1 and cg2), which have been associated with resistance to chloroquine in Plasmodium falciparum, to determine their value as predictors of resistance status. Among field samples from children in Zaria, northern Nigeria, the Tyr-86 polymorphism in pfmdr1 and Ala-281 and the Dd2κ repeat of cg2, were significantly associated. In 8 samples classified resistant by the micro-in vitro test, or, where this failed, by in vivo trial, 7 showed the cg2 Dd2 type κ repeat, and 6 of these had both the Ala-281 allele and the pfmdr1 Tyr-86 allele. In 26 chloroquine-sensitive samples, none had this combination of 3 polymorphisms (P = 0·00002). This indicates 75% sensitivity and 100% specificity in detection of resistance and shows a positive predictive value for resistant infections of 100%. The negative predictive value, because of sensitivity less than 100%, would depend on the prevalence of resistance. Where prevalence of resistance is approx. 21% as in Zaria, the negative predictive value would be 94%, while in Gabon, with a prevalence of ca 73% it would be 60%. The use of (cg2: Ala-281, Dd2κ. pfmdr1: Tyr-86) genotype detection as a predictive epidemiological tool to examine the distribution of chloroquine-resistance in parts of Africa is therefore possible. The sensitivity of detection of resistant strains still requires improvement.

Eimeria arizonensis, E. albigulae and E. onychomysis, morphologically similar species from closely related murid rodents, were distinguished using nuclear rDNA ITS1 sequences obtained from multiple isolates of each taxon. ITS1 sequences were also obtained from 6 other species parasitizing murid rodents: E. falciformis, E. langebarteli, E. nieschulzi, E. papillata, E. separata and E. sevilletensis, and from E. reedi, a parasite of heteromyid rodents. Under parsimony and maximum likelihood analyses, the isolates of E. arizonensis, E. albigulae and E. onychomysis were differentiated as closely related, monophyletic lineages. Maximum likelihood pairwise distances between the latter species ranged from 7 to 12%, and distances within each species ranged from <1 to 5%; thus it is suggested that ITS1 genetic distances may be used to facilitate taxonomic differentiation of Eimeria spp. Against expectation, phylogenetic procedures placed E. reedi within the phylogeny of the Eimeria of murid rodents. ITS1 sequencing appears to provide data that can be used for taxonomic and phylogenetic studies on the speciose genus Eimeria, and may be especially useful when samples contain insufficient numbers of oocysts for other molecular-based methods, e.g. RAPD–PCR.

A 482 base pair gene fragment from samples of amoebae E. histolytica and E. dispar was amplified by PCR. The amplification products of fragments from the 2 species of amoebae presented differences in mobility in non-denaturing polyacrylamide gel, probably due to sequence-dependent conformational alterations in the DNA fragments. The method described here permits E. histolytica and E. dispar to be distinguished with greater sensitivity and rapidity.

The internal transcribed spacer (ITS-1 and ITS-2) regions and the 5·8S ribosomal RNA gene of 2 Perkinsus spp. (G117 and H49) originating from the softshell clam, Mya arenaria, of the Chesapeake Bay were cloned and sequenced to obtain evidence for their genetic divergence. A high level of heterogeneity in both regions, probably resulting from deletions, insertions, and base substitutions, was evident from alignments of the sequences of the 2 isolates with published sequences of other Perkinsus spp. The isolate G117 and other Perkinsus spp. were highly divergent (13–26% and 19–20% sequence divergence in ITS-1 and ITS-2, respectively). These regions in the isolate H49 and Perkinsus marinus were similar (99·07% and 99% for ITS-1 and ITS-2, respectively). Evidence obtained from a phylogenetic analysis using the aligned sequences suggests that G117 and H49 belong to 2 distinct species of Perkinsus. The isolate G117 possibly belongs to an as yet undescribed species of Perkinsus, and H49 belongs to the species P. marinus. The conclusions drawn from the genetic analysis of H49 and G117 are supported by previously reported morphological characteristics (McLaughlin & Faisal, 1998b). Isolates H49 and G117 originated from the same molluscan species demonstrating that at least 2 different species of Perkinsus can co-exist in 1 host.

The geographical distance between conspecific host populations is no doubt a key determinant of the likelihood that exchanges of parasite species occur between these populations. This variable must therefore be taken into account in studies that compare parasite species richness or similarity among host populations. This paper presents a multivariate approach, based on the permutation of matrices, that allows all pairwise geographical distances between host populations to be included as independent variables. The method is illustrated with 3 separate data sets on parasite communities of conspecific fish from different lakes. In 2 of 3 cases, geographical distances among lakes had a significant influence on the similarity of their parasite communities. The effect of geographical distance on species richness of parasite communities also proved important in 2 of the 3 case studies. These examples demonstrate the pervasive influence of distances among host populations on their parasite communities, and the need to properly control for them in statistical analyses.

Schistosoma japonicum from the People's Republic of China is considered to represent a single species comprising either 1 or 4 ‘strains’. We conducted an allozyme electrophoretic study to examine the extent of genetic variation in S. japonicum from mainland China. The allelic profiles of S. japonicum from 7 provinces were established at 16 enzyme loci. S. japonicum from Sichuan had 3–5 (19–31%) fixed differences compared with those from Zhejiang, Anhui, Jiangxi, Hunan, Hubei and Yunnan, suggesting that S. japonicum in mainland China represents a species complex. In addition, genetic markers were also established for different laboratory-maintained populations of S. japonicum which has significant implications for studying the biology of these organisms in human and animal hosts, and for the control and surveillance of human schistosomiasis in China.

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).

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.

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.

Psoroptic sheep scab and psoroptic otoacariasis of domestic rabbits occur in the same geographical regions of Switzerland. To address the question as to whether Psoroptes mites are naturally transmitted between sheep and rabbits, we determined the sequences of the rDNA second internal transcribed spacer (ITS-2) from psoroptic mites from sheep (5 field isolates from Switzerland and 2 laboratory isolates from Ireland and the UK) and from rabbits (8 field isolates from Switzerland). The ITS-2 sequences for all Psoroptes mites originating from sheep were identical and differed at 1 nucleotide position from all the sequences of the rabbit-derived isolates. A statistically significant difference between a rabbit-derived isolate and isolates originating from sheep was also obtained by morphometric analysis of the lengths of the outer opisthosomal setae. For comparative purposes, the ITS-2 sequences from Chorioptes and Sarcoptes collected in Switzerland were also determined. No intraspecies variation was found in 6 sarcoptic isolates from red foxes, with a sequence identity of 41% as compared to Psoroptes. The ITS-2 sequences of 3 chorioptic isolates differed by 24–29% from the Psoroptes sequence. Identical sequences were found for the Chorioptes isolates from sheep and a camel, which differed by 18% from the sequence of an isolate from a cow. These genetic data of psoroptic mites originating from sheep and rabbits from the same geographical area suggest the existence of epidemiologically separated populations.