Mass-scale production of Plasmodium vivax sporozoites in Anopheles stephensi was achieved using the chimpanzee (Pan troglodytes) as a source of infective blood. Membrane feeding was as successful as feeding mosquitoes directly on the animal so long as the time between drawing the blood and feeding was restricted to 45 min. Longer delays such as 2–3 h resulted in loss of infectivity in terms of oocyst production. The selected strain of A. stephensi was highly susceptible to P. vivax (Chesson strain). A strain of A. stephensi relatively refractory to P. falciparum showed no cross-refractoriness to P. vivax. Mixed infections of P. falciparum and P. vivax did not interfere with each other in their development in A. stephensi. A second normal blood meal to mosquitoes infected with P. vivax increased the yield of salivary gland sporozoites.

Despite some years of enquiry, the mechanism that leads to intra-erythrocytic death of malarial parasites during the host's response to infection has not been elucidated. We report here that serum from mice undergoing a successful immune response to Plasmodium chabaudi does not inhibit Plasmodium falciparum unless the Pl. chabaudi is virulent enough to rise to at least 50% parasitaemia and to cause illness. This appears to be true of the 556 KA and DS strains of Pl chabaudi, and also other murine malaria parasites. In mice infected with Pl. chabaudi 556 KA inhibitory activity coincided with the presence of TNF in their serum. Exogenous TNF generated inhibitory activity in the serum of mice only if the animals were pretreated with Proprionobacterium acnes, implying a role for activated macrophages downstream from TNF in this process. The difference in inhibitory activity against Pl. falciparum in serum from mice infected with Pl. chabaudi of more or less virulence may be one of degree. Alternatively two distinct mechanisms may operate, the second coming into operation only in ill mice, with higher parasite burdens.

Leishmania turanica n.sp., found infecting the desert rodent Rhombomys opimus in the southern territories of the USSR and the Mongolian People's Republic, is described. This parasite exists sympatrically with L. major and L. gerbilli in R. opimus and is the predominant species. A total of 284 isolates of L. turanica from R. opimus, 3 from naturally infected Phlebotomus andrejevi and 1 from P. papatasi were characterized and found to be clearly distinguishable on isoenzyme and nuclear DNA characteristics from all other Old World taxa of Leishmania.

Development and forward migration of Leishmania parasites in the sandfly gut is accompanied by morphological transformation to highly motile, non-dividing ‘metacyclic’ forms. Previous studies in vitro have demonstrated that this metacyclogenesis is associated with developmentally regulated changes in expression of two major surface glycoconjugates of Leishmania, the lipophosphoglycan (LPG) and the glycoprotein protease GP63. Studies presented here are the first to examine in situ the changes in expression of these two important surface molecules which occur during amastigote-initiated development of L. major in its natural vector Phlebotomus papatasi. Immunocytochemical analysis using a GP63-specific monoclonal (3.8). and others recognizing metacyclic-specific (3F12) and common (WIC79.3) epitopes of LPG on logarithmic and metacyclic promastigotes, demonstrates: (1) clear expression of LPG and GP63 from 2 and 7 days post-bloodfeeding, respectively; (2) developmental modification of the LPG molecule as parasites undergo forward migration and morphological changes associated with metacyclogenesis; and (3) striking deposition of large amounts of parasite-free excreted LPG on/in the epithelial cells of the gut wall.

The completed life-cycle of Cyclospora talpae, an extra-intestinal coccidian (Apicomplexa) of the liver of the mole (Talpa europaea). is reported. Oocysts sporulated within 12–14 days at room temperature (20–22 °C). Both macro- and micro-gametocytes developed independently within the nucleus of epithelial cells lining the bile duct as demonstrated by both light and electron microscopy. Merogony was seen in the mononuclear cells in the capillary sinusoids of the liver. Merozoites showed the typical ultrastructural features of the Apicomplexa. The life-cycle of C. talpae and the ultra-structure of the gametogonic and merogonic stages are reported; the ultrastructure of the merozoites is reported as the first description of this stage for the genus Cyclospora.

An in vitro bioassay was used to examine [14C]glucose incorporation into polysaccharides in albumen glands (AGs) of susceptible M-line Biomphalaria glabrata infected with the NMRI strain of Schistosoma mansoni. Polysaccharide and galactogen synthesis were unaffected by larval trematode infection in AGs of snails at days 14, 21, and 28 post-infection (p.i.) when compared to uninfected controls. Further experiments were conducted to determine if daughter sporocysts, hypothesized to be primary mediators of parasitic castration in this system, were able to exert direct effects on synthetic activity of uninfected AGs via haemolymph-borne molecules or in vitro culture-generated larval excretory–secretory (ES) products. When AGs were incubated in the presence of infected snail haemolymph, significant differences in quantities of polysaccharides and galactogen were detected only in test organs incubated in day 28 p.i. haemolymph. Daughter sporocyst ES products generated during the first 48 h of culture caused a significant reduction in polysaccharide and galactogen synthesis in test organs. When ES products from days 3 to 6 of in vitro culture were tested similarly, no significant differences in either polysaccharide or galactogen synthesis were observed between control and test organs. These data demonstrate that daughter sporocysts are able to modulate a specific aspect of the reproductive activity of the snail host through haemolymph-borne molecules of host or parasite origin, or directly through in vitro culture-generated ES products.

The metacyclic form of African trypanosomes is the first to express genes for the Variant Surface Glycoprotein (VSG) and it uses an unusually predictable subset of the VSG gene repertoire. We have developed a model system for the analysis of metacyclic VSG (M-VSG) gene expression and have used this to demonstrate that, for two M-VSG genes, different modes of expression operate in the insect and mammalian phases of the life-cycle. In metacyclic-derived clones, these genes are expressed in situ, whereas they are routinely activated by duplication in bloodstream trypanosomes. The expression loci for both M-VSG genes studied are structurally simple and we present a model, based on this, for the maintenance of a separate M-VSG repertoire and expression system.

A midgut trypanolysin and an agglutinin from Glossina palpalis subspecies were isolated and partially characterized using anion-exchange chromatography and polyacrylamide gel electrophoresis. FPLC fractions of midgut extracts of Glossina palpalis palpalis caused agglutination and lysis of two trypanosome species (Trypanosoma congolense and Trypanosoma brucei brucei). although Glossina palpalis gambiensis caused only agglutination. The trypanolysin and agglutinin were active only in the posterior midguts, were heat labile above 50%C, had a periodic cycle of ‘activity’ in response to bloodmeal intake and were not affected by protease inhibitors or trypsin but were inactivated by pronase. The lytic substance contained two proteins with approximate molecular weights (Mr) of 12000 and 10000 Da respectively. The agglutinin had an approximate Mr of 67000 Da. Gamma-irradiation of the two subspecies caused a temporary inhibition of trypanolytic and agglutinin activities in midgut extracts.

All pairwise combinations of three cloned stocks of Trypanosoma brucei (STIB 247L, STIB 386AA and TREU 927/4) were co-transmitted through tsetse flies (Glossina morsitans) and screened for the production of hybrid trypanosomes. Clones of metacyclic and bloodstream trypanosomes from flies harbouring mature infections containing hybrid trypanosomes were established and screened for several isoenzyme and restriction fragment length polymorphisms. For each of the three combinations of parents, some progeny clones were observed to be of a phenotype and genotype indicating that genetic exchange had occurred during development of the trypanosomes in flies. These hybrid clones shared three salient features: (1) where the parents were homozygous variants the progeny were heterozygous, (2) where one of the parents was heterozygous, allelic segregation was observed and (3) the progeny clones were shown to be recombinant when two or more markers for which one of the parents was heterozygous were examined. These results are consistent with the progeny being an F1 in a diploid mendelian genetic system involving meiosis and syngamy. Our observations show that all possible combinations of the three stocks may undergo genetic exchange. A marker analysis of a series of clones each derived from single metacyclic trypanosomes showed that individual flies transmit a mixture of trypanosome genotypes corresponding to F1 progeny and to parental types, indicating that genetic exchange was a non-obligatory event in the life-cycle of the trypanosome. In addition, a preliminary analysis of the phenotype of procyclic stage trypanosomes derived from flies infected with two stocks, indicates that genetic exchange is unlikely to occur at this stage.

Oxytetracycline (OTC) significantly inhibited cytochrome c oxidase activity in bovine lymphocytes infected with Theileria parva and in uninfected mitogen-stimulated lymphocytes. The inhibitory effect was detected in vitro within 24 h of treatment with drug concentrations as low as 1 µg/ml. Following mitogen stimulation of lymphocytes, concentrations of 3 and 10 µg/ml OTC completely inhibited an increase in cytochrome c oxidase activity for 48–72 h. This inhibitory activity was considered to be due to a direct effect on lymphoblast mitochondrial protein synthesis. As a consequence, adenosine triphosphate activity was significantly reduced in lymphocytes stimulated either by infection with T. parva sporozoites or by mitogen and then treated with OTC. The results also indicated that parasite mitochondrial protein synthesis was inhibited by OTC. The activity of OTC reported in this study could explain the suppression of disease following ‘infection and treatment’ immunization against East Coast fever and the in vitro drug-inhibition of schizont development.

Spermatogenesis and the fine structure of the mature spermatozoon of Fasciola hepatica have been studied by transmission electron microscopy. The primary spermatogonia display a typical gonial morphology and occupy the periphery of the testis. They undergo 3 mitotic divisions to give rise to 8 primary spermatocytes forming a rosette of cells connected to a central cytophore. The primary spermatocytes undergo 2 meiotic divisions, resulting in 32 spermatids that develop into spermatozoa. Intranuclear synaptonemal complexes in primary spermatocytes confirm the first meiotic division. The onset of spermiogenesis is marked by the formation of the zone of differentiation which contains 2 basal bodies and a further centriole derivative, the central body. The zone extends away from the spermatid cell to form the median process; into this migrates the differentiated and elongate nucleus. Simultaneously, 2 axonemes develop from the basal bodies. During development, they rotate through 90° to extend parallel to the median process. The migration of the nucleus to the distal end of the median process coincides with the fusion of the axonemes to the latter to form a monopartite spermatozoon. The mature spermatozoon possesses 2 axonemes of the 9 + ‘1’ pattern typical of parasitic platyhelminths, 2 elongate mitochondria and a variable array of peripheral microtubules. The nuclear region of the spermatozoon is immotile. The value of sperm ultrastructure as a taxonomic tool in platyhelminth phylogeny is discussed.

A great deal of published experimental evidence shows that parasite-induced changes in host choice, and biting and survival rates of bloodsucking insects are an important source of bias in estimates of Vectorial Capacity. I argue here that, because of the difficulties of handling these and other biases in practice, indices (based on the Vectorial Capacity) which purport to measure transmission rate are best used in a comparative rather than an absolute way. They should attempt to measure, not transmission rate, but changes in transmission rate. In this case, the less restrictive assumption that parasites are responsible for constant bias (rather than no bias) leads to relatively small errors. The results encourage a more optimistic view of the performance of classical entomological indices such as the parous rate. They should also help to redirect the principal epidemiological question about vector–parasite interactions: future studies should be concerned with the impact of parasites, not directly on Vectorial Capacity, but on the comparative measures which are actually used to evaluate transmission rate.

This paper uses simple mathematical models and statistical estimation techniques to analyse the frequency distribution of microfilariae (mf) in blood samples from human populations which are endemic for lymphatic filariasis. The theoretical analysis examines the relationship between microfilarial burdens and the prevalence of adult (macrofilarial) worms in the human host population. The main finding is that a large proportion of observed mf-negatives may be ‘true’ zeros, arising from the absence of macrofilarial infections or unmated adult worms, rather than being attributable to the blood sampling process. The corresponding mf distribution should then follow a Poisson mixture, arising from the sampling of mf positives, with an additional proportion of ‘true’ mf-zeros. This hypothesis is supported by analysis of observed Wuchereria bancrofti mf distributions from Southern India, Japan and Fiji, in which zero-truncated Poisson mixtures fit mf-positive counts more effectively than distributions including the observed zeros. The fits of two Poisson mixtures, the negative binomial and the Sichel distribution, are compared. The Sichel provides a slightly better empirical description of the mf density distribution; reasons for this improvement, and a discussion of the relative merits of the two distributions, are presented. The impact on observed mf distributions of increasing blood sampling volume and extraction efficiency are illustrated via a simple model, and directions for future work are identified.

This paper examines the effects of host age and sex on the frequency distribution of Wuchereria bancrofti infections in the human host. Microfilarial counts from a large data base on the epidemiology of bancroftian filariasis in Pondicherry, South India are analysed. Frequency distributions of microfilarial counts divided by age are successfully described by zero-truncated negative binomial distributions, fitted by maximum likelihood. Parameter estimates from the fits indicate a significant trend of decreasing overdispersion with age in the distributions above age 10; this pattern provides indirect evidence for the operation of density-dependent constraints on microfilarial intensity. The analysis also provides estimates of the proportion of mf-positive individuals who are identified as negative due to sampling errors (around 5% of the total negatives). This allows the construction of corrected mf age–prevalence curves, which indicate that the observed prevalence may underestimate the true figures by between 25% and 100%. The age distribution of mf-negative individuals in the population is discussed in terms of current hypotheses about the interaction between disease and infection.

Jirds (Meriones libycus) were infected with various numbers of Acanthocheilonema viteae L3 stage parasites. During the course of the ensuing 16 weeks, blood samples were collected at 2 weekly intervals and the amount of the major parasite excretory–secretory product (E–S 62) and antibodies directed against it measured. After 16 weeks, animals were sacrificed and the size of the mature worm burden established. In spite of interaction between E–S 62 and host antibody, a statistically significant relationship was found to exist between the amount of E–S 62 present in the bloodstream and the size of the parasite load. It is suggested that the detectable antigen level is more influenced by the size of the worm burden than the presence of antibody and that antibody is only likely to affect adversely antigen measurement in situations where the amount released is relatively low. Examples of this are early in infection and in low-level infections. These ideas are discussed in relation to the development and assessment of serological assays which attempt to predict parasite burden in human filarial infections.

The capacity of Acanthocheilonema viteae to metabolize fructose was investigated in vitro. In common with other filarial species A. viteae oxidized fructose to lactate but its rate of consumption was only 40% of the glucose-containing control value. Fructose was not incorporated into glycogen. Release of 14CO2 from [U-14C]fructose was not detected in the presence of glucose and was about 40% of the glucose-containing value under conditions where fructose was the sole hexose substrate. Fructose consumption and lactate excretion increased in proportion to the external concentration of fructose. However, worm viability was not maintained in fructose over a 120 h in vitro incubation. In the presence of fructose, protein synthesis (measured incorporation of [35S]methionine into acid-insoluble material) was reduced compared to the glucose-containing control group; but was significantly greater than the value obtained under glucose-free conditions.

In and around the British Isles, three biologically specialized strains of the fish acanthocephalan Pomphorhynchus laevis have been recognized, each differing in geographical distribution and both final and intermediate host utilization. Six restriction endonuclease digests of the genomic DNA of the English freshwater and marine strains were compared by genomic cross-hybridization using the randomly sheared and radio-isotope labelled genomic DNA of freshwater P. laevis, and also by using a heterologous isotope labelled ribosomal DNA probe. Autoradiographs of hybridized Southern blots provided evidence for considerable homogeneity in the total repetitive DNA of the two strains as well as some preliminary evidence for possible restriction fragment length polymorphisms using EcoR I and Pvu II.

Hymenolepis diminuta liberated measurable amounts of ‘Lowry-positive material’ (LPM) and protein during incubation for 2 h in vitro. When tapeworms were incubated in the presence of bovine trypsin (BT), or when BT was added to the medium after removing the tapeworms, the enzyme's proteolytic activity was inhibited significantly. Centrifugation of the medium at 30 000 g yielded a pellet composed of tegumental elements, but this fraction did not inhibit BT. The 30 000 g supernatant fraction contained a chemical(s) that inhibited the proteolytic enzymes of the rodent host's intestinal contents (IC). The inhibitor(s) was stable following repeated freeze-thaw cycles, heat labile, and not degraded by BT or IC, and it inhibited the amidase activity of BT in a non-competitive manner.

Cyclosporin A (CsA), administered in 5 daily subcutaneous doses of 50 mg/kg to MF1 mice immediately following infection with Hymenolepis diminuta enhanced parasite growth relative to controls. Drug administered at 24 h intervals for 10 days, and thereafter every 48 h to MF1 and CBA/Ca mice infected with H. diminuta, increased worm survival and growth, delayed host-mediated expulsion of the parasite and enabled some worms to develop to patency. Worm survival and weight both increased in a dose-dependent manner following daily CsA treatment of infected CBA/Ca and BALB/c mice (0–150 mg/kg CsA/day). Delay in parasite elimination was accompanied by increased frequency of worm-attachment in the anterior small intestine (MF1 mice given 5 daily doses of CsA [0–150 mg/kg] following infection); posteriad migration of worms was restricted in a dose-dependent manner. The data presented contrast markedly with the action of the same drug on H. microstoma in mice. Thus CsA treatment acts in opposing ways on two closely related parasites in the same host; this possibly reflects the mechanistic antagonism between immunosuppression and anthelmintic activity. This paper reports the first use of a specific T cell-suppressive drug on H. diminuta in the mouse, implicating the role of T cells in protective immunity to this parasite.

Immunodiagnostic tests for Taenia-specific faecal antigen based on polyclonal rabbit antisera against Taenia saginata or Taenia solium proglottid extracts in capture-type ELISA assays have been developed. Taenia-specific antigen was detected in detergent-solubilized faecal extracts from T. solium- and T. saginata-infected hosts. Coproantigen from T. solium-infected hamsters did not cross-react with faeces from rodents infected with Hymenolepis diminuta, H. citelli, H. micro-stoma, Necator americanus, Strongyloides ratti or Nematospiroides dubius and faeces from uninfected animals. When the T. saginata-capture ELISA was tested with faecal samples positive for T. solium antigen, no cross-reactions were obtained. However, faecal samples from humans infected with T. solium or T. saginata, including some with extremely low egg counts, were cross-reactive by either test. Nevertheless, considerably higher O.D. values were obtained with stool samples from Taenia patients compared to Hymenolepis nana-infected or uninfected individuals. Two individuals, infected with Taenia sp. and positive for coproantigens by ELISA, became antigen-negative 6 days after treatment with Niclosamide. The possibility of developing species-specific immunodiagnostic tests for human taeniasis through coproantigen detection is discussed.