Showing papers in "Parasitology in 1990"

Parasite-host coevolution.

Abstract: In this paper we wish to develop three themes, each having to do with evolutionary aspects of associations between hosts and parasites (with parasite defined broadly, to include viruses, bacteria and protozoans, along with the more conventionally defined helminth and arthropod parasites). The three themes are: the evolution of virulence; the population dynamics and population genetics of host–parasite associations; and invasions by, or ‘emergence’ of, new parasites.

Commitment of the malaria parasite Plasmodium falciparum to sexual and asexual development.

Abstract: Blood-stage malaria parasites in the vertebrate host can develop either into the asexual, multiplying forms, called schizonts, or into gametocytes, the sexual stages of the parasite. In the present work we studied the differentiation into asexual parasites or gametocytes of the progeny of single, isolated schizonts of the clone 3D7A of Plasmodium falciparum, using monoclonal antibodies specific for the sexual or asexual stages of the parasite. We observed that schizonts obtained from a continuous culture undergoing serial cycles of growth and dilution with fresh red blood cells produced either only gametocytes or only asexual parasites, showing a high degree of commitment to one or the other developmental pathway. The relative proportion of schizonts which produced gametocytes was very low at low parasite densities in culture, while at high parasite densities a much greater proportion of schizonts produced gametocytes. Nevertheless, at both low and high parasite densities individual schizonts were almost always fully committed to producing only gametocytes or only asexual parasites.

Immunodiagnosis of taeniasis by coproantigen detection.

Abstract: 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. microstoma, 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.

Glutathione transferase in helminths.

Abstract: The helminth glutathione (GSH) transferases are present as isoenzymes but fail to show a clear biochemical homology to any of the three mammalian GSH transferase families. GSH transferase is one of the major detoxification systems found in helminths, particularly high levels being found in cestodes and digeneans. Helminth GSH transferases bind a range of anthelmintics but there is limited evidence that the enzymes can conjugate anthelmintics with glutathione. Other natural substrates of helminth GSH transferase may be secondary products of lipid peroxidation including lipid hydroperoxides and reactive carbonyls. Lipid peroxidation can arise via free radicals produced by host immuno-effector cells and helminth GSH transferase may help form a defence system against immune-mediated damage. GSH transferase has also been identified as a protective antigen in schistosomiasis.

Selection and evolution of virulence in bacteria: an ecumenical excursion and modest suggestion.

Abstract: Why do parasites kill their hosts? During this past decade, research in three different areas; evolutionary ecology, medical microbiology, and population genetics has provided theory and data that address this and related questions of selection and the evolution and maintenance of parasite virulence. A general theory of parasite-host coevolution and the conditions for selection to favour parasite virulence has been put forth. Considerable advances have been made in elucidating the mechanisms of pathogenicity and inheritance of virulence in bacteria. The population genetic structure and the relationship between pathogenic and non-pathogenic forms has been determined for a number of species of bacteria. We critically review these developments and their implications for questions of selection and the evolution and maintenance of virulence in bacteria. We postulate how selection may operate on specific types of bacterial virulence and present a general protocol to experimentally test hypotheses concerning selection and the evolution of virulence in bacteria.

Epidemiology and immunology of Necator americanus infection in a community in Papua New Guinea: humoral responses to excretory-secretory and cuticular collagen antigens.

Abstract: Baseline data from an immuno-epidemiological study of hookworm infection in a rural village in Madang Province, Papua New Guinea are reported. Necator americanus was found to be the commonest helminth infection, with a prevalence of near 100% and intensity of 40 worms per host in adults. Enterobius vermicularis, Ascaris lumbricoides and Trichuris trichiura were also present, at prevalences of 53, 10 and 3% respectively; Ancylostoma duodenale was absent. The frequency distribution of N. americanus was highly over-dispersed, and was well described by a negative binomial distribution with aggregation parameter, k, of 0.370. Intensity of infection was significantly related to host age, but did not differ between the sexes. Haemoglobin levels and haematocrit values were indicative of anaemia in the community, but were unrelated to hookworm infection. Levels of antibodies (IgG, IgA and IgM combined) against adult Necator cuticular collagen and excretory-secretory (ES) products were determined. Serum concentrations of the two types of antibody were significantly correlated with each other. Significant positive correlations were found between anti-ES antibody levels and hookworm egg production, and between anti-collagen antibody levels and host age. It is suggested that the level of anti-collagen antibodies may reflect cumulative exposure to infection, whereas levels of anti-ES antibodies may be more dependent on current worm burden. No evidence was found to suggest that either antibody response is important in regulating parasite population growth. Similarly, the presence of a positive correlation between eosinophil concentration and infection intensity in adults indicates that eosinophilia reflects, rather than determines, the host's worm burden.

A rhoptry antigen of Plasmodium falciparum is protective in Saimiri monkeys.

Abstract: A non-polymorphic antigen associated with the rhoptry organelles of Plasmodium falciparum has been purified by immunoaffinity chromatography. The antigen, RAP-1 (rhoptry associated protein-1), which is defined by monoclonal antibodies which inhibit parasite growth in vitro, is a multi-component antigen consisting of four major proteins of 80, 65, 42 and 40 kDa and two minor proteins of 77 and 70 kDa. These proteins were electro-eluted from preparative sodium dodecyl sulphate polyacrylamide gels and protected Saimiri sciureus monkeys from a lethal blood-stage infection of P. falciparum malaria. Sera from the protected animals recognized only proteins of the RAP-1 antigen when used to probe a Western blot of total parasite protein extract, confirming that RAP-1 is responsible for eliciting the protective immune response.

The relationship between the frequency distribution of Ascaris lumbricoides and the prevalence and intensity of infection in human communities.

Abstract: Observed field data from a range of geographically distinct human communities suggest a consistent non-linear relationship between prevalence and mean intensity of Ascaris lumbricoides infection. Utilizing the negative binomial distribution as a description of observed aggregation, maximum-likelihood analysis reveals that the degree of aggregation is a negative linear function of mean worm burden. The factors responsible for this relationship in human populations require further study but may involve some combination of (i) density-dependent reduction in worm numbers within individuals, (ii) density-dependent parasite-induced host mortality or (iii) self-treatment by heavily infected hosts. Variability in the degree of aggregation appears dependent on the level of infection in a community and independent of geographical differences in the host or parasite populations.

Occurrence of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in British soils

Abstract: Soil from 403 hedgerow, roadside verge, woodland, heathland or field sites was assessed for presence of entomopathogenic rhabditid nematodes with aGalleria larva baiting technique. Steinernematids were recovered from 48 % of the sites but only one site yielded Heterorhabditis sp. The species recovered most frequently was Steinernema bibionis (Bovien). Another unidentified Steinernema sp. was also isolated. Prevalence of steinernematids in the different types of habitat formed continuum so that roadside verges harboured them most often while heathland sites yielded them least often. Mean soil temperatures at 5 cm were about 15 °C in fields and verges, 13 °C in hedgerows and heathland and 12 °C in woodland. Prevalence of the nematodes varied in different parts of Britain and was influenced by soil type. They were associated with calcareous soils, especially those with a calcareous subsoil horizon. Soils which are periodically or seasonally water-logged were suitable. Soils enriched by humus and hence high in organic matter frequently harboured the nematodes and some suitable soils had significant clay contents or subsoils high in clay. It appears that the two steinernematids are relatively unspecialized species, occurring in diverse habitats and soil types and are adapted to cooler temperatures.

Evidence that the mechanism of gene exchange in Trypanosoma brucei involves meiosis and syngamy.

Abstract: 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 F 1 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 F 1 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.

Sexual development in malarial parasites: gametocyte production, fertility and infectivity to the mosquito vector.

Abstract: Using cloned lines of Plasmodium berghei producing mixed asexual and sexual (clone 234L) and purely asexual (clone 233L) parasitaemias, the courses of parasitaemia, gametocytogenesis, exflagellation, ookinete production in vitro and mosquito infectivity have been followed. For clone 234L mosquito infectivity is maximal at day 3 and has ceased by day 6 post-infection. Conversely, gametocytogenesis, exflagellation and ookinete production are at minimal levels at day 3 and rise to peaks between days 10 and 15 of infection (in TO mice infected with blood at mechanical blood passage 3). Sexual potential declines progressively with sustained mechanical passage (up to P14). Gametocyte conversion is highest early in infection and declines exponentially; however, upon each mechanical passage conversion is again raised but decreases more rapidly with succeeding passages. For practical mosquito transmission in the laboratory we consider P8 to be the useful limit for mechanical transmission for this parasite clone. Asexual parasite growth (virulence) is more rapid with increased mechanical passage inducing a rapid fall in haematocrit. By P14 the course of infection closely parallels that of the purely asexual clone 233L.

Parasites, desiderata lists and the paradox of the organism.

Abstract: Eavesdrop morning coffee at any major centre of evolutionary theory today, and you will find ‘parasite’ to be one of the commonest words in the language. Parasites are touted as prime movers in the evolution of sex, promising the final solution to that problem of problems, the puzzle that led G. C. Williams to proclaim in 1975 ‘a kind of crisis’ at hand in evolutionary biology (Hamilton, 1980; Tooby, 1982; Seger & Hamilton, 1988). Parasites seem to offer a plausible justification for the otherwise futile effort females put into choosing among posturing males (Hamilton & Zuk, 1982; but see Read, 1990). Frequency-dependent selection exerted by parasites is, according to one admittedly minority view, largely responsible for the high levels of diversity found in gene pools (Clarke, 1979). One might even extrapolate to a time when the entire metazoan body could come to be seen as a gigantic adaptation against microscopic pathogens.

Avermectin and avermectin derivatives are antagonists at the 4-aminobutyric acid (GABA) receptor on the somatic muscle cells of Ascaris; is this the site of anthelmintic action?

Abstract: The mechanism underlying the ability of the anthelmintic avermectin to paralyse the nematode Ascaris is not yet fully understood. Using conventional two-electrode electrophysiological recording techniques we have demonstrated that micromolar concentrations of ivermectin block the inhibitory GABA response on the muscle cells of the parasitic nematode Ascaris. The ability of a number of avermectin derivatives to act as receptor antagonists for the Ascaris muscle GABA receptor has been determined. This provides useful information to compare with the in vivo anthelmintic potency of these compounds. Abamectin, the most potent anthelmintic, was the most potent compound at inhibiting the GABA response whilst octahydroavermectin, a compound which lacks anthelmintic activity, did not block the GABA receptor. This is consistent with the notion that the GABA receptor antagonist properties of the avermectins could contribute to their anthelmintic action.

Expression of LPG and GP63 by different developmental stages of Leishmania major in the sandfly Phlebotomus papatasi.

Abstract: 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.

Rostellar hook morphology of Echinococcus granulosus (Batsch, 1786) from natural and experimental Australian hosts, and its implications for strain recognition.

Abstract: An analysis of the rostellar hooks of Australian isolates of Echinococcus granulosus revealed that there was less variation in larval (metacestode) than adult characters and that metacestode characters could be measured directly from adult worms. A factor analysis indicated that two factors, one representing a contrast between number of hooks and their length, and the other representing blade lengths, were sufficient to account for 87.5% of the variance in metacestode hook measurements. These results indicate that rostellar hook morphology is not useful for discriminating strains of E. granulosus in Australia. The Tasmanian and mainland domestic strains were found to be indistinguishable using rostellar morphology. Although many of the isolates from sylvatic hosts differed from those from domestic hosts, there was not a clear separation as would be expected if two distinct strains existed. Evidence was presented to show that the morphological differences seen in sylvatic hosts could be attributed to host-induced effects, and that the previously accepted existence of two mainland strains should be investigated further.

Characterization of Trichomonas vaginalis haemolysis

Abstract: The haemolytic activity of live Trichomonas vaginalis organisms was investigated. Optimal haemolysis of human erythrocytes was observed at a parasite to erythrocyte ratio of 1:5 during a 2 h incubation period. No haemolytic activity was detected in concentrated culture supernatants after overnight growth of trichomonads or when parasites were separated from erythrocytes by a 3 microns filter, suggesting a contact-dependent mechanism for haemolysis. The haemolytic activity was temperature-dependent and maximal haemolysis occurred at 37 degrees C. Treatment of trichomonads with metronidazole reduced levels of haemolysis by greater than 50%. Maximal haemolysis occurred at the pH range of the vagina during trichomoniasis. N-alpha-tosyl-L-lysyl-chloromethyl ketone and iodoacetamide, inhibitors of trichomonad cysteine proteinases, reduced the haemolytic activity of live parasites.

The influence of genes mapping within the major histocompatibility complex on resistance to Trichuris muris infections in mice.

Abstract: Two panels of H-2 recombinant strains of mice were used in an attempt to map the H-2-linked genes which control resistance to infection with Trichuris muris. Response phenotypes could be related to the presence of 'resistance' (q,b) or 'susceptibility' (k,d) alleles at I-A. The influence of these genes was modulated by other alleles, particularly q or d alleles, at the D end of the H-2. Absence of I-E molecules correlated with resistance to infection in some but not all strains studied. Thus the (B10.BR x B10.G) F1 strain which expressed I-Ek gene products was resistant to infection. A study of the time-course of infection in strains of mice expressing q alleles throughout the H-2 on 4 different genetic backgrounds (NIH, SWR, DBA and B10) revealed that most strains were resistant to infection. However, the DBA/1 strain exhibited differential responsiveness, 4 out of 6 individuals harbouring mature adult parasites on day 35 post-infection.

The serotoninergic, cholinergic and peptidergic components of the nervous system in the monogenean parasite, Diclidophora merlangi: a cytochemical study

Abstract: Confocal scanning laser microscopy has been employed with immunocytochemical techniques to map the distribution of serotoninergic and peptidergic components in the nervous system of the monogenean gill-parasite, Diclidophora merlangi; results are compared with the distribution of cholinergic components, following histochemical staining for cholinesterase activity. While all three neurochemical elements are present in the central and peripheral nervous systems, the cholinergic and peptidergic systems dominate the CNS, whereas the PNS has a majority of serotoninergic nerve fibres. The cholinergic and peptidergic neuronal pathways overlap extensively in staining patterns, suggesting possible co-localization of acetylcholine and neuropeptides. Within the peptidergic nervous system, immunoreactivity to the pancreatic polypeptide family of peptides and FMRFamide were the most prevalent. Gastrin/cholecystokinin (CCK)-, neuropeptide Y-, substance P-, neurokinin A- and eledoisin-like immunoreactivities have been demonstrated for the first time in a monogenean parasite. The gastrin/CCK- and tachykinin-like immunoreactivities had an apparently restricted distribution in the worm.

Schistosoma mansoni and S. japonicum: comparison of levels of ultraviolet irradiation for vaccination of mice with cercariae.

Abstract: When cercariae of Schistosoma mansoni and of S. japonicum were irradiated with various levels of u.v. light at 254 nm, their development to perfusable worms was reduced to below 1% at about 200 microW min cm-2. Cercariae attenuated with about 300 microW min cm-2 induced partial resistance against an homologous challenge infection in mice. No differences were observed between the two schistosome species when the same treatment was given to the cercariae. Thus the same u.v. dose can confer immunizing ability to cercariae of both S. mansoni and S. japonicum.

Virulence of lizard malaria: the evolutionary ecology of an ancient parasite-host association.

Abstract: The negative consequences of parasitic infection (virulence) were examined for two lizard malaria parasite-host associations: Plasmodium agamae and P. giganteum, parasites of the rainbow lizard, Agama agama, in Sierra Leone, West Africa; and P. mexicanum in the western fence lizard, Sceloporus occidentalis, in northern California. These malaria species vary greatly in their reproductive characteristics: P. agamae produces only 8 merozoites per schizont, P. giganteum yields over 100, and P. mexicanum an intermediate number. All three parasites appear to have had an ancient association with their host. In fence lizards, infection with malaria is associated with increased numbers of immature erythrocytes, decreased haemoglobin levels, decreased maximal oxygen consumption, and decreased running stamina. Not affected were numbers of erythrocytes, resting metabolic rate, and sprint running speed which is supported by anaerobic means in lizards. Infected male fence lizards had smaller testes, stored less fat in preparation for winter dormancy, were more often socially submissive and, unexpectedly, were more extravagantly coloured on the ventral surface (a sexually dimorphic trait) than non-infected males. Females also stored less fat and produced smaller clutches of eggs, a directly observed reduction in fitness. Infected fence lizards do not develop behavioural fevers. P. mexicanum appears to have broad thermal buffering abilities and thermal tolerance; the parasite's population growth was unaffected by experimental alterations in the lizard's body temperature. The data are less complete for A. agama, but infected lizards suffered similar haematological and physiological effects. Infected animals may be socially submissive because they appear to gather less insect prey, possibly a result of being forced into inferior territories. Infection does not reduce clutch size in rainbow lizards, but may lengthen the time between clutches. These results are compared with predictions emerging from several models of the evolution of parasite virulence. The lack of behavioural fevers in fence lizards may represent a physiological constraint by the lizards in evolving a thermal tolerance large enough to allow elimination of the parasite via fever. Such constraints may be important in determining the outcome of parasite-host coevolution. Some theory predicts low virulence in old parasite-host systems and higher virulence in parasites with greater reproductive output. However, in conflict with this argument, all three malarial species exhibited similar high costs to their hosts.

The evolutionary biology of molecular parasites.

Abstract: A parasite can be considered to be the device of a nucleic acid which allows it to exploit the gene products of other nucleic acid–the host organisms. In this view, all parasites are ‘molecular parasites’. But it is interesting to restrict our attention to nucleic acids which do not encode organisms, as these live in a purely molecular world which lacks emergent features such as fangs and ovipositors. Viruses and transposons are molecular parasites in this sense. Most viral nucleic acids do code for some proteins, such as replicases and the protein shell in which they travel between their cellular oases. Some, however, do not even have a shell and code for nothing at all–these are the ‘viroids’ (Reisner & Gross, 1985), the smallest parasites in the world.

Differential diagnosis of Taenia saginata and Taenia solium with DNA probes

Abstract: A size selected genomic DNA library was constructed using DNA extracted from Taenia saginata. The DNA was digested using the restriction enzyme EcoR1 under star conditions and the 2-4 kbase fraction, selected following sucrose density-gradient separation, was cloned in the bacteriophage lambda gt 10. A panel of cestode DNAs including Taenia saginata, Taenia solium, Taenia taeniaeformis, Taenia crassiceps, Echinococcus granulosus and DNAs of bovine, porcine and human origin were used in conjunction with hybridization analysis to identify two recombinant bacteriophages. The first probe, designated HDP1, reacted specifically with T. saginata DNA. The second, designated HDP2, reacted with DNA from both T. saginata and T. solium but not the other DNA samples and thus provided a general reagent for positive identification of fragments of Taenia spp. proglottides of human faecal origin. If used in conjunction the two DNA probes allow positive identification of T. saginata. In the clinical situation it is important to be able to distinguish T. saginata and T. solium infections and DNA probes such as these may be useful in such differentiation.

Transmission of taeniid tapeworm eggs via blowflies to intermediate hosts

Abstract: Experiments showed that dead blowflies containing eggs of Taenia hydatigena can transmit infection if ingested by lambs during grazing. Rabbits became infected with T. pisiformis after grazing grass exposed to blowflies that had been in contact with dog faeces containing the eggs of this parasite. When captive blowflies were exposed first to dog faeces containing proglottids of T. hydatigena and then to cooked meat, 100% of pigs fed on this meat became infected. The experiments are intended to model the transmission dynamics that result in human larval tapeworm infections. The implications of the results to the epidemiology of human hydatid diseases caused by Echinococcus granulosus and E. multilocularis as well as of T. solium cysticercosis are discussed.

Genetic diversity and genetic differentiation in Echinococcus granulosus (Batsch, 1786) from domestic and sylvatic hosts on the mainland of Australia.

Abstract: Enzyme electrophoresis was used to examine genetic variation within and between populations of Echinococcus granulosus from domestic and sylvatic hosts in western and eastern Australia. Substantial genetic diversity was found within all populations. There was no evidence, however, of genetic differentiation between populations from different hosts or geographic areas. When isolates were grouped into previously described domestic or sylvatic strains on the basis of rostellar hook morphology, most (94%) of the genetic variation occurred within, rather than between strains. These results conflict with the currently accepted theory of separate domestic and sylvatic strains of E. granulosus on the mainland of Australia.

Spermatogenesis and the fine structure of the mature spermatozoon of the liver fluke, Fasciola hepatica (Trematoda: Digenea)

Abstract: 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 degrees 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.

Bloodmeal digestion by strains of Anopheles stephensi Liston (Diptera: Culicidae) of differing susceptibility to Plasmodium falciparum.

Abstract: Blood digestion was studied in strains of Anopheles stephensi which had been genetically selected for either refractoriness or susceptibility to infection by Plasmodium falciparum. Females of the refractory Pb3-9a strain ingested more blood than selected (Sda-500) and unselected (Punjab) susceptible females and began to degrade the haemoglobin soon after feeding. In susceptible females, haemoglobin degradation started only after a significant post-feeding lag period. Total protein content of the midgut after the bloodmeal was correspondingly higher for refractory than for susceptible females, but absolute and relative rates of protein degradation were not significantly different between the different mosquito strains. Bloodmeal induction of midgut trypsin activity and the maximal trypsin activity were the same for the different strains. The residual aminopeptidase activity and its relative post-feeding activity (enzyme units per midgut) were significantly higher in refractory females. However, when converting to specific aminopeptidase activity, no differences between strains were evident. The results indicate that both the early initiation of haemoglobin degradation and higher aminopeptidase activity in the Pb3-9a refractory strain are important in the limitation of parasite development within the mosquito midgut, whereas trypsin plays no role in this process.

The evolution of inducible defence.

Abstract: Defences against parasites are characterized by inducible, amplifiable responses, often with a memory component. Inducible defences with similar properties are common in a variety of other types of interactions, for example many aquatic invertebrates produce inducible structural defences against their predators and competitors. Most inducible defences have the following properties: (1) a threshold of activation; (2) an amplification of response with increasing stimulus; (3) a memory component. Specificity, amplification and memory are the basis for defining a defence as ‘immune’ (Klein, 1982), and these properties are present in both the vertebrate and invertebrate internal defence responses to pathogens. Invertebrates differ in the absence of immunoglobulins and therefore in reduced specificity. Although the reduced specificity of invertebrate internal defence systems is often viewed as proof of their ‘primitiveness’, the differences in defence systems of vertebrates and invertebrates may be more related to their respective selection regimes than to phylogeny. The syngeneic recognition system of vertebrates functions to recognize small departures from self, such as would arise from neoplasia. Are vertebrates under more intense selection from neoplasia, perhaps due to a greater incidence of hormonal imbalance or hypersensitivity reactions? The invertebrate internal defence systems are all less discriminating than the vertebrate, but there are marked differences in degree of discrimination depending on whether the group is colonial or not. Even the phyla of colonial animals with quite simple body plans, the sponges and cnidarians, have a more discriminating recognition system than the phyla of solitary animals with more complex body plans, such as the molluscs and arthropods. The primary effectors of all invertebrate responses to parasites are encapsulation and phagocytosis, although in some phyla there are specific antibacterial proteins that can also be induced. A primary effector of the vertebrate immune system is also based on phagocytosis, but is mediated largely by immunoglobulins. Although memory is a widespread property of inducible defence systems, the mechanisms, underlying memory components are unknown except in the vertebrate immune response. The function and adaptiveness of many of these accelerated or amplified second-set responses remain unclear, although it is tempting to conclude that a memory component is a beneficial adaptation to multiple, intermittent stimuli. From this review I conclude that the conditions that favour the evolution of inducible defences include: (1) an intermittent, unpredictable selective regime; (2) selection for a ‘camouflaged’ defence, to slow parasite counter-adaptation; (3) reliable cues and (4) high costs of defence.

Survey of 15 sites over 28 months for entomopathogenic nematodes (Rhabditida: Steinernematidae)

Abstract: The presence of entomopathogenic nematodes in soil from 15 sites on 10 sampling occasions over a period of 28 months was documented. Sites were chosen to represent habitats classified as heathland, hedgerow, roadside verge, pasture or woodland. Bioassays utilized Galleria larvae and most of the nematodes obtained were identified asSteinernema (= Neoaplectana) bibionis (Bovien). A sibling species which fails to interbreed with S. bibionis was also found at 2 of the sites. Bioassays were at 15 and 20°C and there was no significant difference in the results at these temperatures. There was no evidence for seasonality in the presence of the nematodes and at 2 sites they were found on every sampling occasion. Otherwise, sites converted unpredictably from positive to negative or negative to positive over the 28-month sampling period.

Schistosoma mansoni: stage-dependent damage after in vivo treatment with praziquantel.

Abstract: The in vivo effects of a single, subcurative dose (200 mg/kg body wt of mouse) of praziquantel on the structure of the tegument and subtegumental tissues of juvenile (21-, 26- and 30-day-old) and adult Schistosoma mansoni are described. In juvenile worms praziquantel caused only moderate damage to both the tegument and subtegumental tissues although the degree of drug-induced damage was related to both the sex of the worms and to the developmental status of each individual worm. In general, male worms exhibited more extensive and longer lasting surface damage than females. However, the level and extent of the changes increased as the worms became more developmentally advanced with adult worms showing extensive damage to the tegument and vacuolization and disruption of the subtegumental tissues. The possible correlation between the stage-dependent changes in susceptibility to praziquantel and age-related changes, particularly in the phospholipid composition of the outer tegumental membrane is discussed.

Frequency distribution of lymphatic filariasis microfilariae in human populations: population processes and statistical estimation.

Abstract: 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.