The diversity in a 925 bp portion of the Plasmodium vivax MSA1 gene in isolates from the Philippines, China, the Solomon Islands and Papua New Guinea was investigated. A total of 74 base pair changes was found in the amplified fragment from 18 isolates. Most of these changes were single or double base pair substitutions. In several regions, these point changes were tightly linked with one set always present or always absent in the different isolates. Seven such blocks were identified. These blocks were present in different combinations in the different isolates indicating that extensive intragenic recombination has occurred.

The polymerase chain reaction (PCR) has been used to amplify in vitro a semi-conserved region of a gene encoding an Mr 68–72000 surface antigen of Giardia intestinalis trophozoites. Using primers specific for conserved nucleotide sequences identified within the promoter-distal portion of two homologous genes (tsp11 and tsa417) cloned previously from the G. intestinalis isolates Ad-1 (from Australia) and WB (from Afghanistan), a single PCR-amplified DNA fragment of the expected size (0·52 kilobases) was obtained in high yield from either purified DNA or whole trophozoites of the Ad-1 isolate and from every 1 of 9 other axenic G. intestinalis isolates belonging to genetic groups I and II (defined previously on the basis of allozyme electrophoresis data—Andrews et al. 1989). Discernible product was recovered from as few as 2–4 trophozoites. In contrast, 6 G. intestinalis isolates that were assigned by allozymic analysis to genetic groups III/IV yielded small amounts of a 0·37-kilobase (kb) amplification product (with evidence in some samples of an additional 0·4 or 0·18 kb fragment) but no 0·52 kb product. Two animal-derived isolates of G. duodenalis (one from an Australian native rodent, Notomys alexis, the other from a domestic cat) also yielded a single 0·37 kb PCR-amplified fragment, whereas an isolate from another cat produced a 0·34 kb fragment. No product was recovered from G. muris, a morphologically distinct species of Giardia. The results demonstrate that different genotypes of G. duodenalis can be distinguished using this assay and that it is diagnostic for isolates belonging to two major clusters (groups I/II and III/IV) of G. intestinalis. The amplified DNA segment appears to be relatively conserved among group I and group II isolates of G. intestinalis. A related but clearly distinct sequence seems to be conserved among group III/IV isolates of G. intestinalis and some isolates of G. duodenalis.

An incomplete ring of electric nets was placed around uninfected cattle and cattle infected with Trypanosoma congolense. The numbers of fed and unfed Glossina pallidipes caught on the nets were used to estimate the attractiveness of infected and uninfected cattle to tsetse, and the feeding success of tsetse on the cattle. There was no difference in the attractiveness of infected and uninfected cattle to G. pallidipes. However, the feeding success of G. pallidipes on infected cattle was 75 % greater than on uninfected cattle. This suggests that certain effects of T. congolense on cattle behaviour or physiology act to increase the probability of transmission of the parasite by increasing the feeding success of the vector. The nature of the effects of T. congolense on cattle which caused this result is unknown, but several possibilities are discussed.

The effects of exposure to infective larvae on the antibody response to a cocktail of specific recombinant antigens of Onchocerca volvulus and to a worm extract were evaluated by comparing the responses of individuals from a vector controlled area with those from an area of continuing transmission by ELISA. Individuals from the vector controlled areas were found to have reduced responses to both antigen preparations. Amicrofilerdermic (mf–) individuals from the area of vector control exhibited significantly lower total and subclass IgG responses to the worm extract. In contrast, the responses to the cocktail of specific recombinants were significantly reduced in individuals from the area of vector control who were still microfilerdermia positive (mf+). The distribution of IgG subclass specific responses was similar to both antigen preparations, both dominated by the IgG4 and IgG1 subclasses. IgG1 responses to the worm extract remained elevated in the vector controlled individuals but IgG4 was significantly reduced in the mf – individuals. Both subclasses reflected the total IgG response to the cocktail of recombinants and were significantly reduced in individuals from the vector controlled area, when compared to individuals from the hyperendemic area. IgG1 responses to the cocktail of recombinants are significantly lower than IgG4 in all individuals and virtually absent in individuals from the vector-controlled area. Measuring total IgG and IgG4 is more sensitive than IgG1 in detecting infection, 100 or 97% respectively, but they remain elevated in the individuals from the vector controlled areas even after 8–10 years interruption of transmission. These results have important implications for the serological monitoring of control programmes in individuals who have previously been infected.

Variation in the immunogenicity of 3 isolates of Trichinella spiralis was assessed by the parameters of adult worm recovery, mast cell, eosinophil and antibody responses in mice of defined response phenotype. The levels of the protective, inflammatory and immune responses induced by infection differed between the isolates. Isolates showed considerable variation in the capacity to elicit mast cell and eosinophil responses. All induced increases in parasite-specific antibody, levels of total (IgGAM) antibody and of IgM and IgG isotypes rose steadily after infection, but there were significant differences in levels of response. The IgGAM response was correlated with the number of worms present, i.e. the greatest response was seen in low responder (C57BL/10) mice infected with the longest-surviving isolates. All isolates elicited specific IgG1 and IgG2a antibodies after infection, although, again, there were isolate-specific differences in the levels and kinetics of response. Levels of these isotypes were always higher, although not significantly so, in high-responder NIH mice. Low-responder mice showed higher IgE serum levels than high-responder mice after infection, one isolate giving much higher IgE values than the other two.

Reinfection with hookworm (Necator americanus) following chemotherapy was studied over 2 years in a rural village in Madang Province, Papua New Guinea. The prevalence of hookworm infection had returned to pre-treatment levels after 2 years, and the geometric mean hookworm burden had returned to 58 % of the pre-treatment value. The rate of acquisition of adult worms was independent of host age, and was estimated as a geometric mean of 2·9–3·3 worms/host/year (arithmetic mean 7·9–8·9 worms/host/year). There was significant predisposition to hookworm infection; the strength of this predisposition did not vary significantly between age or sex classes.

In the hybrid zone of the two mouse subspecies Mus musculus musculus and Mus musculus domesticus, mice with hybrid genotypes harbour, on the average, more helminth parasites (cestodes and nematodes) than mice of the two parental taxa. In order to determine the roles played by genetic parameters in this phenomenon, mice with recombined and parental genotypes were experimentally infected with the intestinal pinworm Aspiculuris tetraptera, a natural parasite of the house mouse. The results showed that the high susceptibility of the hybrid zone mice is genetically determined. In addition, this study shows the occurrence of variability among resistant parental populations.

The degradation of several protein substrates, including the blood proteins haemoglobin, albumin and fibrinogen, by proteinases present in extracts of adult Haemonchus contortus was examined over a broad pH range. These proteinases were further characterized on the basis of substrate specificity, inhibitor sensitivity and molecular size by spectrophotometric and substrate gel analysis. The majority of the proteinases capable of degrading the blood proteins tested were active at acidic pH and could be ascribed to the cysteine proteinase class. In addition, evidence is presented that these proteinases are differentially recognized and inhibited by immune sera and that parasites capable of withstanding protective host immune responses exhibit modified expression of proteinases.

Indirect immunofluorescence studies using a rabbit polyclonal antiserum have been used to localize the presence of a 32 kDa glycoprotein in the region of the amphids of 2nd-stage juveniles of the root-knot nematode, Meloidogyne incognita. Similar immunoreactivity was also demonstrated in 5 other Meloidogyne species but was not found in representatives of 8 other nematode genera including the closely related cyst nematodes (Globodera and Heterodera). Immunoelectron microscopical studies have shown that the immunoreactivity in M. incognita is associated with the secretory material filling the amphidial channel and probably with the sheath cell.

A low molecular weight (15 kDa) surface antigen of the cattle filarial nematode, Setaria digitata, was earlier shown to be specifically recognized by the antibodies from human bancroftian filarial (Mf positive) patients' sera (Theodore & Kaliraj, 1990). The filarial specific antibodies bound to a 15 kDa peptide in preparative Western blots were eluted and employed in screening of candidate antigens expressed in the genomic library of Wuchereria bancrofti at the IgG4 subclass antibody level. A recombinant clone (λ WbG7) reacting strongly with filarial sera but poorly with sera from patients infected with non-filarial helminths was selected for further studies. The 2 kb DNA insert of the clone λ WbG7 was recloned into a pMAL vector and the recombinant clone pGT7 thus obtained was over-expressed and affinity purified. The purified 105 kDa fusion protein of the clone pGT7 was specific and was not recognized by the non-filarial sera at the IgG4 level. All microfilaraemic individuals were positive by IgG4 assay. However, similar attempts to diagnose by filarial-specific IgE assays failed to recognize microfilaraemic individuals. Moreover, by filarial-specific IgG4 assays, the endemic normals were distinctly divided into two groups, showing higher and lower recognition for this antigen indicating current and past/no infection. Among the filarial-IgG4 (assay)-positive ‘endemic normals’, 14% showed ‘microfilariae’ during repeated peripheral night blood examination, confirming the validity of the recombinant antigen, pGT7 based assay.

Ivermectin has been shown to increase chloride conductances of invertebrate cells. On the muscle cells of the parasitic nematode Ascaris, ivermectin acts as both a GABA receptor antagonist and a chloride channel opener. In this study, ion-sensitive microelectrodes were used to investigate the effect of ivermectin on intracellular C1− concentration of the somatic muscle bag cells of Ascaris suum. Incubation of muscle cells with ivermectin (10 μM in 1% dimethyl sulphoxide vehicle for 60 min) increased intracellular C1− by 2·9 mM or 15% compared to controls (P > 0·01, n = 6).

The central (CNS) and peripheral (PNS) nervous systems of the cyclophyllidean tapeworm, Moniezia expansa, were examined for the presence of cholinergic, serotoninergic and peptidergic elements using enzyme cytochemical and immunocytochemical techniques in conjunction with light and confocal scanning laser microscopy. Cholinesterase activity and 5-hydroxytryptamine- and regulatory peptide-immunoreactivities (IRs) were localized to the nerve fibres and cell bodies of all of the major neuronal components in the CNS of the worm, including the cerebral ganglia and connecting commissure, the 10 longitudinal nerve cords and associated transverse ring commissures. Although each of the 3 systems appeared well developed and comprised a significant portion of the nervous system, the serotoninergic constituent was the most highly developed, consisting of a vast array of nerve fibres and cell bodies distributed throughout the strobila of the worm. A close association of cholinesterase reactivity and peptide-IRs was evident throughout the CNS, indicating the possible co-localization of acetylcholine and neuropeptides. Within the PNS, cholinergic activity and serotoninergic- and peptidergic-IRs occurred in the subtegumental network of nerve fibres and somatic musculature. Although all 3 neuro-chemical elements were present in the acetabula, they were found in different nerve fibres; only cholinergic and peptidergic cell bodies were found. The common genital opening, vagina and ootype regions of the reproductive system displayed a rich innervation of all 3 types of neuronal populations. Within the peptidergic system, immunostaining with antiseraraised to the C-terminus of the neuropeptide Y superfamily of peptides and the invertebrate peptides, neuropeptide F (M. expansa) and FMRFamide was the most prevalent. Limited positive-IR for substance P and neurokinin A were also recorded in the CNS of the worm.