Showing papers in "Parasitology in 1991"
TL;DR: Examination of M. expansa's primary structure shows that it displays significant sequence homology with the vertebrate neuropeptide Y superfamily, suggesting that this platyhelminth-derived peptide is the phylogenetic precursor.
Abstract: Using a C-terminally directed pancreatic polypeptide (PP) antiserum and immunocytochemical methods, PP-immuno-reactivity (IR) was localized throughout the central (CNS) and peripheral nervous systems (PNS) of the cestode, Moniezia expansa. In the CNS, immunostaining was evident in the paired cerebral ganglia (primitive brain), connecting commissure, and the paired longitudinal nerve cords that are cross-linked by numerous regular transverse connectives. The PNS was seen to consist of a fine anastomosing nerve-net of immunoreactive fibres, many of which were closely associated with reproductive structures. Radioimmunoassay of this peptide IR in acid-alcohol extracts of the worm measured 192·8 ng/g of PP–IR. HPLC analyses of the M. expansa PP–IR identified a single molecular form which was purified to homogeneity. Plasma desorption mass spectrometry (PDMS) of purified parasite peptide resolved a single peptide with a molecular mass of 4599±10 Da. Automated gas-phase Edman degradation identified a 39-amino acid peptide with a C-terminal phenyl-alaninamide. Examination of its primary structure shows that it displays significant sequence homology with the vertebrate neuropeptide Y superfamily, suggesting that this platyhelminth-derived peptide is the phylogenetic precursor. Neuropeptide F (M. expansa) is the first regulatory peptide to be fully sequenced from the phylum Platyhelminthes and may represent a member of an important new class of invertebrate neuropeptide.
181 citations
TL;DR: Respiratory assay showed NADH, alpha-glycerophosphate and succinate to be the substrates with the greatest potential for metabolism, and the apparent absence of a respiratory chain 'Site I' in such mitochondria may mean that NADH-fumarate reductase serves to reoxidize mitochondrial NADH.
Abstract: Highly purified mitochondrial fractions have been isolated from the intraerythrocytic stages of two mammalian Plasmodium spp., Plasmodium yoelii of rodents and Plasmodium falciparum of man. Mitochondria of the former parasite are cristate whereas those of the latter are essentially acristate. Isolated mitochondria from both parasite species were heterogeneous with respect to size, shape, density of matrix staining and extent of internal structure. Respiratory assay, by reduction of exogenous cytochrome c, showed NADH, alpha-glycerophosphate and succinate to be the substrates with the greatest potential for metabolism. Additionally, proline, dihydroorotate and glutamate (P. falciparum only) were oxidized at low rates. A number of NAD(+)-linked substrates were not utilized. The NADH-dependent reduction of cytochrome c was insensitive to rotenone and antimycin A. Fumarate inhibited the NADH-dependent reduction of cytochrome c and stimulated the oxidation of NADH, suggestive of an NADH-fumarate reductase pathway. Oxidation of either alpha-glycerophosphate or succinate was fully inhibited by standard mitochondrial electron transport inhibitors, including a number of Complex III inhibitors, although the concentrations required of such inhibitors (notably myxothiazol) were relatively high compared to mammalian mitochondria. Dithionite-reduced minus oxidized difference spectra indicated the presence of cytochromes aa3, b, c and c1 in mitochondria of both parasite species, but at a higher cytochrome to protein ratio in P. yoelii. Freshly isolated mitochondria from either species exhibited only low respiratory control ratios with alpha-glycerophosphate or succinate as substrates. The apparent absence of a respiratory chain 'Site I' in such mitochondria may mean that NADH-fumarate reductase serves to reoxidize mitochondrial NADH.
168 citations
TL;DR: A statistical analysis of 2-locus genetic diversity of clones within isolates concludes that not all multiple-clone infections result from superinfection, but that some are due to single multiple- clone inoculations.
Abstract: The occurrence of multiple-clone Plasmodium falciparum haploid blood-stage infections is a pre-requisite for cross-fertilization and genetic exchange at the diploid stage in the mosquito. Using monoclonal antibodies against 3 polymorphic blood-stage antigens, a method of two-colour differential immunofluorescence allowed the resolution of between 1 and 4 clones/isolate. A mean of 2 P. falciparum clones was detected in the blood of malaria patients in The Gambia. The mean number of clones/patient showed no correlation with age, parasitaemia, or disease severity. There was a slight difference in mean number between sample periods, probably reflecting temporal differences in transmission intensity. A statistical analysis of 2-locus genetic diversity of clones within isolates concludes that not all multiple-clone infections result from superinfection, but that some are due to single multiple-clone inoculations.
135 citations
TL;DR: There has been a 2.5-fold increase in human infectiousness in the last 25 years and it is argued that chloroquine would be expected to select for increased infectivity in the parasite, and this may also have contributed to the observed increase.
Abstract: In an area of holoendemic malaria in Northern Tanzania, Anopheles gambiae s.l. females were age-graded by Polovodova's method and dissected for sporozoites. Age-specific sporozoite rates implied that mosquitoes acquired new infections at all ages. The extrinsic period lasted just over 3 gonotrophic cycles (9-11 days). Very high sporozoite rates in the oldest females implied the absence or rarity of genetic refractoriness to infection. A method is described for estimating the proportion of bloodmeals which result in mosquito infection. This method makes relatively few assumptions about mosquito behaviour, and could be useful for evaluating transmission-blocking interventions. Overall, it is estimated that about 21% of meals are infectious. This is much higher than previous estimates derived either from experimental mosquito feeding studies or from similar age-grading data collected from the same area in 1962. Various alternative explanations are considered, and it is concluded that there has been a 2.5-fold increase in human infectiousness in the last 25 years. This is partly attributable to suppression of human infectiousness by widespread chloroquine usage during the 1960s, followed by removal of this effect by drug resistance. It is argued that chloroquine would be expected to select for increased infectivity in the parasite, and this may also have contributed to the observed increase.
130 citations
TL;DR: Metabolic labelling and immunoprecipitation showed that G RA 2 and GRA 3 were processed from lower molecular weight precursors, and that GRA 2 andGRA 4 incorporated [3H] glucosamine and are thus likely to be glycosylated.
Abstract: The biosynthesis and fate of 4 different dense granule proteins of Toxoplasma gondii were studied with 3 monoclonal antibodies raised against tachyzoites and 1 polyclonal antibody raised against a recombinant protein. These proteins have the following molecular weights: 27 kDa (GRA 1), 28 kDa (GRA 2), 30 kDa (GRA 3) and 40 kDa (GRA 4). All four proteins were found in dense granules by immunoelectron microscopy; in T. gondii-infected cells, they were found in the vacuolar network but, in addition, GRA 3 was also detected on the parasitophorous vacuole membrane. Therefore, dense granule contents undergo differential targeting when exocytosed in the parasitophorous vacuole. Metabolic labelling and immunoprecipitation showed that GRA 2 and GRA 3 were processed from lower molecular weight precursors, and that GRA 2 and GRA 4 incorporated [3H] glucosamine and are thus likely to be glycosylated.
127 citations
TL;DR: Bloodstream forms of Trypanosoma congolense were continuously cultivated in vitro at 34-36 degrees C in the absence of feeder cell layers, using HMI-93 medium which was modified from Iscove's modified Dulbecco's MEM.
Abstract: Bloodstream forms of Trypanosoma congolense (2 clones: ILNat3.1 and IL3000, and 4 stocks: IL2079, IL2466, IL3266 and CP-81) were continuously cultivated in vitro at 34-36 degrees C in the absence of feeder cell layers, using HMI-93 medium which was modified from Iscove's modified Dulbecco's MEM (Flow Laboratories, Irvine, Scotland). The modification was done by supplementing the medium with 0.05 mM bathocuproine sulphonate, 1.5 mM L-cysteine, 0.5 mM hypoxanthine, 0.12 mM 2-mercaptoethanol, 1 mM sodium pyruvate, 0.16 mM thymidine, 20% (v/v) heat-inactivated young goat serum and 5% (v/v) Serum Plus (Hazleton Biologics, Lenaxa, KS, USA). Trypanosomes obtained from two different sources were used to initiate primary cultures: (1) metacyclic forms which were produced in vitro at 27 degrees C, and (2) bloodstream forms obtained from Balb/c mice which had been infected with the bloodstream forms transformed in vitro from the metacyclic forms. Metacyclic forms placed in 25 cm2 T-type (T-25) flasks rapidly attached to the bottom of the flasks and transformed to bloodstream forms during the initial 24 h and continued to proliferate. The bloodstream forms isolated from the infected mouse blood by means of diethylaminoethyl cellulose (DE52) column chromatography also continued to proliferate in the flasks. Cultures were maintained by replacing the medium every 24 h. Every 4-5 days, the attached bloodstream forms were resuspended in fresh medium by gentle pipetting and then were subcultured. The method was further simplified by initiating primary cultures directly with 10 microliters of the tail blood of infected mice in 24-well culture plates and then by subcultivating either in wells or in T-25 flasks. The shortest population doubling time, 9 h, was achieved by seeding subcultures with 10(6) bloodstream forms/ml. The bloodstream forms propagated in this system were morphologically similar to those seen in infected mouse blood, they were covered with a surface coat as examined by electron microscopy and they were infective to mice.
105 citations
TL;DR: It is concluded that recombination among unlinked loci is a common consequence of sexual reproduction of P. falciparum in The Gambia and slightly lower genotype diversity was observed in each of two villages, which may be a consequence of smaller population size compared with the urban region.
Abstract: Serotyping with monoclonal antibodies was used to estimate the number and frequencies of allelic variants of two merozoite surface proteins, MSP1 and MSP2, and an exported protein Exp-1, in a sample of 344 clinical isolates of Plasmodium falciparum from an urban region of The Gambia. Represented among the isolates were 36, 8 and 2 alleles of the MSP1, MSP2 and Exp-1 loci respectively. Relative frequencies of these alleles remained stable in the parasite population over the 2 years of the study. A computer program was used to calculate from the frequencies of individual alleles at the three loci, the probable number of different genotypes in samples from the population, assuming random assortment among the loci. No significant difference was found between the expected and the observed genotype diversity. It is concluded that recombination among unlinked loci is a common consequence of sexual reproduction of P. falciparum in The Gambia. Slightly lower genotype diversity was observed in each of two villages, which may be a consequence of smaller population size compared with the urban region.
105 citations
TL;DR: The relationship between faecal examination for egg output and worm burden of Opisthorchis viverrini in man of 181 autopsy cases from Northeast Thailand is described and the accuracy of egg counts for estimating worm burden and its relevance to parasite epidemiological research are discussed.
Abstract: The relationship between faecal examination for egg output and worm burden of Opisthorchis viverrini in man of 181 autopsy cases from Northeast Thailand is described. Diagnosis of the parasite infection by stool examination for the presence of eggs was less sensitive than the worm recovery technique. Using Stoll's dilution and formalin-ether technique, no eggs were detected in the faeces of 20 cases harbouring low worm burdens (less than 20 worms). The quantitative faecal egg count by Stoll's dilution technique showed a strikingly close positive correlation with the number of worms recovered (r = 0.96, P less than 0.001) indicating a strong linear association between eggs per gram of faeces (epg) and worm burden. The number of epg per worm was inversely correlated to the worm burden (P less than 0.001), suggesting that density-dependent constraints on fecundity could operate to restrict egg output in heavy infections. The accuracy of egg counts for estimating worm burden and its relevance to parasite epidemiological research are discussed.
104 citations
TL;DR: Preliminary analysis of lipids in a rhoptry fraction purified from Toxoplasma gondii tachyzoites found that significant amounts of phosphatidic acid and lysophospholipids were found, which could be related to functional involvement of the organelle in host-cell invasion.
Abstract: Quantitative and qualitative analysis of lipids has been performed on a rhoptry fraction purified from Toxoplasma gondii tachyzoites. The lipid to protein ratio was estimated to be 0.26. The cholesterol to phospholipid ratio was unusually high at 1.48. Phosphatidylcholine was the major phospholipid; phosphatidylserine, phosphatidylinositol and sphingomyelin were absent whereas significant amounts of phosphatidic acid and lysophospholipids were found. This pelicular composition could be related to functional involvement of the organelle in host-cell invasion.
103 citations
TL;DR: It is hypothesized that vector population dynamics and the susceptibility of the vector population to infection with T. parva are among the most important factors which influence the expression of ECF as a disease entity, and the likelihood of transformation occurring.
Abstract: The protozoan parasite Theileria parva, transmitted by the ixodid tick Rhipicephalus appendiculatus, is the cause of East Coast fever (ECF) and the related syndromes of Corridor disease and January disease in cattle of eastern, central and southern Africa. It is likely that buffalo (syncerus caffer) are the natural host of T. parva. In eastern and southern Africa, there exists both buffalo-adapted and cattle-adapted T. parva. Disease caused by buffalo-adapted parasites is called Corridor disease, and that caused by cattle-adapted parasites is termed East Coast fever. In eastern Africa, it has been shown experimentally that buffalo-adapted T. parva can, after serial passage in cattle, become adapted to cattle, in which it can then be maintained and cause ECF. This adaptation has been termed transformation. The transformation of buffalo-adapted T. parva to a cattle-adapted parasite has not been reported in southern Africa, and ECF, eradicated from South Africa, Swaziland and southern Mozambique by 1960, has not reappeared in the subcontinent. This paper discusses the possible reasons for this, and hypothesizes that vector population dynamics and the susceptibility of the vector population to infection with T. parva are among the most important factors which influence the expression of ECF as a disease entity, and the likelihood of transformation occurring. It also considers the possibility that disappearance of ECF from southern Africa resulted from the extinction, as a result of vigorous control measures and unfavourable climatic conditions, of non-diapausing populations of R. appendiculatus that may have been introduced from eastern Africa with cattle imported in 1901.
102 citations
TL;DR: The relationship of survival and reproduction of Gyrodactylus salaris Malmberg on the Atlantic salmon to water temperature, was studied on the basis of temporal sequence of births and age at death of individual parasites on isolated salmon, and of infrapopulation growth on isolated and grouped salmon.
Abstract: The relationship of survival and reproduction of Gyrodactylus salaris Malmberg on the Atlantic salmon (Salmo salar) to water temperature (2.5-19.0 degrees C), was studied on the basis of temporal sequence of births and age at death of individual parasites on isolated salmon, and of infrapopulation growth on isolated and grouped salmon. Mean life-span of the parasite was negatively correlated with water temperature: 33.7 days at 2.5 degrees C and 4.5 days at 19.0 degrees C. The average number of offspring per parasite peaked between 6.5 and 13.0 degrees C, and was approximately 2.4 at these two temperatures. Both the period between the successive births of the offspring (max 4) and the estimated generation time were negatively correlated with temperature. The innate capacity for increase (rm) was positively correlated with temperature: from 0.02 (/parasite/day) at 2.5 degrees C to 0.22 (/parasite/day) at 19.0 degrees C. Growth of the infrapopulations was positively correlated with water temperature and was higher on isolated fish than on grouped fish, though less than the potential parasite population growth estimated from rm. In the infrapopulations the mean intensity of parasites continued to increase throughout all the experiments on both isolated fish and on grouped fish.
TL;DR: In this article, three techniques for estimating the intensity of Opisthorchis viverrini infection in individuals from a Northeast Thai community are compared, using a quantitative formalin/ethyl acetate technique, worm burdens were estimated by expulsion chemotherapy and antibody levels measured by ELISA.
Abstract: Three techniques for estimating the intensity of Opisthorchis viverrini infection in individuals from a Northeast Thai community are compared. Egg counts were determined using a quantitative formalin/ethyl acetate technique, worm burdens were estimated by expulsion chemotherapy and antibody levels were measured by ELISA. Log-transformed worm and egg counts were closely correlated (r = 0.80), suggesting that both measurements provide good assessments of relative intensity of infection. However, no Opisthorchis worms were recovered from 34 people with high egg counts; probably due to problems with the expulsion technique in some individuals. Examination of egg production per fluke indicated that each fluke contributed an average of 180 eggs per gram (epg) of faeces and fecundity was negatively associated with total worm burden. Serum IgG levels correlated significantly with Opisthorchis egg count (r = 0.61) at two independent assessments. Although significant associations were observed between antibody levels and echinostome infection, analysis suggested that these reflected independent associations between these two variables and Opisthorchis infection and age. We conclude that all three measurements are useful for epidemiological studies.
TL;DR: A preliminary serological evaluation of the Em2(G11) antigen by ELISA revealed identical immunodiagnostic characteristics, compared to Em2 obtained by classical means, thus suggesting the presented method for future isolation of large-scale Em2 antigen.
Abstract: A monoclonal antibody (MAb G11) species-specific to the Em2 antigen of Echinococcus multilocularis was generated for (i) further biological characterization of the Em2 antigen, (ii) easy affinity-purification of Em2 antigen for immunodiagnostic and immunological investigations and (iii) development of a sandwich-ELISA for the detection of Em2 antigen in diagnostic samples and thus species-specific identification of E. multilocularis metacestode material. The MAb G11 was used in an antibody sandwich-ELISA to detect soluble Em2 antigen with a methodical sensitivity of 80 ng E. multilocularis antigen/ml of solution. MAb G11 specifically detected Em2 antigen in all of 15 E. multilocularis-isolates originating from various geographical areas and in none of other helminth isolates (e.g. Echinococcus granulosus, E. vogeli, and others). Further biological analysis by FITC-labelled MAb G11 demonstrated unique binding activity to the laminated layer of the metacestode. Also, oncospheres were binding FITC-labelled MAb G11 on an outer layer synthesized during cultivation in vitro for 13 days after hatching. Application of the MAb G11 antibody sandwich-ELISA for investigation of solubilized oncospheres confirmed the in vitro synthesis of Em2 antigen by oncospheres on day 13 p.i. Adult stages (somatic antigens) and freshly hatched oncospheres were always MAb G11 negative. Solid-phase MAb G11 was used for purification of the corresponding Em2 antigen by affinity chromatography. A preliminary serological evaluation of the Em2(G11) antigen by ELISA revealed identical immunodiagnostic characteristics, compared to Em2 obtained by classical means, thus suggesting the presented method for future isolation of large-scale Em2 antigen.
TL;DR: It is concluded that, in areas of low morbidity such as Kangundo, chemotherapy of schoolchildren only, at intervals of up to 3 years, is a satisfactory way of producing a long-term reduction in both intensity of infection and morbidity.
Abstract: A comparison was made of the long-term impact of different methods of administration of chemotherapy (oxamniquine, 30 mg/kg in divided doses; or praziquantel, 40 mg/kg) on prevalence and intensity of Schistosoma mansoni infection in four areas in Kangundo Location, Machakos District, Kenya. In Area A, treatment was offered in October 1983 and again in April 1985 to all infected individuals. In Area H, treatment was offered in April 1985 to individuals excreting greater than or equal to 100 eggs per gram (epg) of faeces. In Area S, treatment was offered in April 1985 to all infected school children, within the framework of the primary schools. In the witness area, Area W, treatment was given in April 1985, for ethical reasons, to a small number of individuals excreting greater than or equal to 800 epg. Prevalence and intensities of infection were subsequently monitored at yearly intervals for three complete post-treatment years. In the Area S schools, clinical examination was also carried out at yearly intervals. Treatment of all infected individuals on two occasions (Area A) was the most effective and long-lasting way of reducing prevalence and intensity of infection. In this area, however, some earlier interventions had been carried out and pre-treatment intensities were lower than in the other areas. Treatment only of infected schoolchildren (Area S) also had a marked and prolonged effect, comparable to or better than treatment of individuals with heavy infections (Area H). Treatment of infected schoolchildren also caused a persistent reduction in the prevalence of hepatomegaly, and there was suggestive evidence from intensities of infection in community stool surveys (but not from incidence rates) of an effect on transmission. In all study areas, reinfection was most rapid and most intense among children. These findings are discussed in the light of theoretical considerations and of results from other studies, both on schistosomiasis and on intestinal helminths. We conclude that, in areas of low morbidity such as Kangundo, chemotherapy of schoolchildren only, at intervals of up to 3 years, is a satisfactory way of producing a long-term reduction in both intensity of infection and morbidity.
TL;DR: Passive transfer of monospecific anti-omega 1 sera into S. mansoni infected, T cell-deprived mice completely prevented the occurrence of microvesicular hepatocyte damage in these animals.
Abstract: T cell-deprived mice acutely infected with S. mansoni suffer microvesicular hepatocyte damage which is not seen in infected, immunological intact animals. A cationic fraction (CEF6) of the PBS-soluble portion of S. mansoni eggs (SEA) induces antibodies which, on passive transfer, prevent hepatocyte damage. CEF6 contains 2 antigens, omega 1 and alpha 1, and has also been shown to be a useful serodiagnostic reagent. This paper describes the purification and characterization of the 2 antigens present in CEF6. omega 1 is a monomeric glycoprotein with a pI greater than 9.0 and a molecular weight of 31 kDa. Alpha 1 consists of two immunologically cross-reactive dimers, 41 and 36 kDa in non-reducing conditions, each of which consists of one unique and one common glycoprotein subcomponent. In ELISA with mouse and human infection sera omega 1 is shown to be S. mansoni specific and is better able to distinguish S. mansoni infections from other schistosome infections than are unfractionated SEA, CEF6 or alpha 1. Passive transfer of monospecific anti-omega 1 sera into S. mansoni infected, T cell-deprived mice completely prevented the occurrence of microvesicular hepatocyte damage in these animals. Monospecific anti-alpha 1 serum had no hepatoprotective capacity.
TL;DR: Observations of essentially all water contacts made over a two week period by all individuals in a rural community in eastern Zimbabwe provide strong quantitative support for control programmes aimed at heavily used sites or at the minority of individuals making most water contacts.
Abstract: Variations in the amount of water contact made by individuals and in the amount of water contact made at different sites may have significant impacts on patterns of human schistosome infection. Previous studies have reported variations in the rate of water contact and differences in the sites used between age/sex classes, but there is limited information on variations in individual water contact behaviour. In this paper we report and analyse observations of essentially all water contacts made over a two week period by all individuals in a rural community in eastern Zimbabwe. The mean rate of water contact was 0.43 contacts/person/day. These data were over-dispersed, ranging from zero to 3.3 contacts/person/day; 90% of contacts were made by only 37% of the population. Contact rates were related to age (highest in 8 to 10-year-olds) but not sex, with substantial variation unaccounted for by these variables. Age and sex classes differed in types of water-related activities and the time of day of contact. A greater diversity of sites was used by children than by adults and by males than by females. Individual contact rates were correlated with intensities of infection, although the risk of infection per contact was estimated to be highest in 2 to 4-year-old children and higher for males than females. Five contact sites were used during the study period, with more than 50% of contacts occurring at just 2 sites. Different age and sex classes used different sites and there were additional site-related differences in types of activity and the time of day of use. The implications of these water contact patterns for schistosome epidemiology are discussed. In particular the results provide strong quantitative support for control programmes aimed at heavily used sites (e.g. focal mollusciciding) or at the minority of individuals making most water contacts (e.g. targeted chemotherapy).
TL;DR: It is concluded that infections of E. cuniculi are common in the tropics and that reactivations of these infections might be a hazard to AIDS patients.
Abstract: The enzyme-linked immunosorbent assay (ELISA) was used to survey human sera for antibodies to Encephalitozoon cuniculi using spores obtained from in vitro cultures as antigen. Sera were obtained from patients with tropical diseases, neurological and renal disorders, patients who were HIV positive and those who had been tested for HIV but found to be negative. Sera from inhabitants of the village of Jali, The Gambia and from healthy blood donors were also examined. Numerous sera from all groups except the blood donors gave positive ELISA reactions at dilutions of 1:400. On titration, those with titres of 1:400 were reclassified as negative. Antibody titres of 1:800 and above were considered to be indicative of past or present infections with E. cuniculi. Many of these ELISA seropositives were also positive by IFAT or PAP. When examined by Western blotting of SDS-PAGE protein profiles of E. cuniculi spores, sera from many patients who had a tropical association reacted with the characteristic profiles shown by known positive mouse and rabbit sera. Others in the tropical group showed antibody binding to some but not all of the immunodominant polypeptides and yet others were negative in spite of their reactivity in the ELISA, IFAT or PAP test. Less agreement between ELISA and Western blotting results was obtained with the other groups of patients, although reactivity with one or more of the major polypeptide bands was sometimes seen. Serum from one blood donor, examined by ELISA and Western blotting, was positive. Differences in the methods of antigen preparation and of epitopes recognized by individuals may account for different reactivities in the tests. It is concluded that infections of E. cuniculi are common in the tropics and that reactivations of these infections might be a hazard to AIDS patients.
TL;DR: Most of the successful anti-nematode drugs currently available affect the nematode locomotory system, presumably due to their interactions with molecules associated with the main neuro-transmitters of the motor nervous system, such as acetylcholine and GABA as discussed by the authors.
Abstract: Most of the successful anti-nematode drugs currently available affect the nematode locomotory system. Their success is due to their interactions with molecules associated with the main neuro-transmitters of the motor nervous system, acetylcholine and GABA. These drugs tend to have a relatively broad spectrum of action, affecting a wide variety of nematodes, presumably because nematode motor nervous systems are conservative in their use of these transmitters.
TL;DR: The asymmetric distribution of phospholipids in the erythrocyte membrane during the intracellular development of the human malaria parasite Plasmodium falciparum was studied and changes already present in trophozoite-infected cells were studied.
Abstract: The asymmetric distribution of phospholipids in the erythrocyte membrane during the intracellular development of the human malaria parasite Plasmodium falciparum was studied. Infected cells of high parasitaemia were treated with phospholipase A2 or sphingomyelinase C, followed by isolation of the host red cell membrane using the Affigel (731) bead method. Additionally, phosphatidylserine on the surface of infected cells was probed using a phosphatidylserine-sensitive prothrombinase assay. Trophozoite-infected cells showed an increase in phosphatidylethanolamine and phosphatidylserine and a decrease in phosphatidylcholine in the outer leaflet. In addition to the changes already present in trophozoite-infected cells, schizont-infected cells showed a decrease in sphingomyelin as well as a further increase in phosphatidylserine in the outer leaflet. The results are discussed with respect to possible mechanisms and consequences of these changes.
TL;DR: Up to 200 laboratory reared Anopheles dirus mosquitoes were fed on each of 496 symptomatic Thai men who had patent, naturally acquired Plasmodium vivax gametocytaemia, suggesting that each oocyst contributed about 850 sporozoites to a gland infection.
Abstract: Up to 200 laboratory reared Anopheles dirus mosquitoes were fed on each of 496 symptomatic Thai men who had patent, naturally acquired Plasmodium vivax gametocytaemia. Mean gametocyte densities were 455/mm3 (range: 0–3281), geometric mean oocyst number was 9 (0–142), mean frequency of infection was 43% (0–100%), and mean sporozoite number in salivary glands was 9525 (0–285 000). There was little relation between gametocyte density and either oocyst number or frequency of mosquito infection. There were, however, statistically strong positive correlations between oocyst numbers and frequency of infection, and between number of oocysts and number of salivary gland sporozoites. The data suggest that each oocyst contributed about 850 sporozoites to a gland infection.
TL;DR: Quantitative estimates of reinfection rates suggested that individuals aged 12 years or less acquired substantially more infection (measured as egg output) than individuals more than 12 years old.
Abstract: Infection with Schistosoma haematobium was studied in a rural community of approximately 500 persons in eastern Zimbabwe. The overall prevalence of infection, as determined by urine egg counts, was 40.1%, and of heavy infections (greater than or equal to 50 eggs/10 ml urine) was 11.0%. The prevalence of both heavy and all infections was highest in the 8 to 10-year-old age class. During 1987-88 data were obtained from 102 individuals on intensity of reinfection 14 weeks after treatment with praziquantel, the efficacy of treatment having been determined after 4 weeks. The water contact made by these individuals during 2-week periods immediately following treatment was recorded. The relative abundance of patent infected intermediate host snails, Bulinus globosus, was also monitored. An index of exposure was developed which weighted each water contact by its duration, the type of activity, the time of day, and the abundance of infected snails at the site used. The relationships between rates of reinfection, rates of exposure, and age were examined. Although only 13 individuals showed positive rates of reinfection, there were statistically significant and independent effects of both exposure and age on reinfection rate. Quantitative estimates of reinfection rates suggested that individuals aged 12 years or less acquired substantially more infection (measured as egg output) than individuals more than 12 years old.
TL;DR: These data indicate that, overall, male and female S. vulgaris respond differently to infection and that intestinal helminths normally thought to be of little or no pathological consequence to the host are factors that should be addressed in future studies regarding animal energetics, ecology, and behaviour.
Abstract: Although the relationship between intestinal parasitism, the ingestion and use of energy, and host survival is expected, little work has been done to outline the effect of such organisms upon their host's nutritional requirements in an ecological context. This study is the first to demonstrate that an intestinal helminth previously reported to be of little or no histopathological consequence, Plagiorhynchus cylindraceus, has a significant detrimental impact upon the flow of food energy through a definitive host, the European starling, Sturnus vulgaris. Within both male and female adult European starlings reductions in standard metabolic rates occurred as the result of initial infection, indicating that the host's basal metabolism/thermal regulatory abilities were altered. Moreover, initially infected male starlings, but not females, had an increased consumption and excretion of energy and maintained lower average daily body weights versus controls when temperature stressed. These results appear to be due to either a parasite-mediated alteration in host activity and/or to the disruption of host-digestive abilities. Additionally, these data indicate that, overall, male and female S. vulgaris respond differently to infection and that intestinal helminths normally thought to be of little or no pathological consequence to the host are factors that should be addressed in future studies regarding animal energetics, ecology, and behaviour.
TL;DR: Positive interactions are more common than negative interactions in determining the species richness of helminth infracommunities of bats, and positive associations become even more important as the community becomes more complex.
Abstract: Twelve populations of bats were examined to determine the extent of interspecific associations in determining the species richness of intestinal helminth infracommunities. The pool of helminth species which was available to individual bats ranged from 2 to 21. The 'summed binomial' distribution was determined to underlie the host frequency distribution of the number of helminth species per host. Overall covariation in occurrences of species in replicated communities can be detected by testing for the equality of the observed variance of the host frequency distribution to the variance expected when species are allocated to hosts at random. Where statistically significant the covariance was indicative of a majority of positive rather than negative interspecific associations. As the mean number of species per host in a host population increases not only does the number of positive associations increase but so does the proportion of species pairs which exhibit positive associations. Although there is an increase in the proportion of species pairs which exhibit positive associations as the number of species increases, the magnitude of the associations (as indicated by the mean positive or the mean negative pairwise covariances) does not. Therefore, we concluded that positive interactions are more common than negative interactions in determining the species richness of helminth infracommunities of bats. Further, positive associations become even more important as the community becomes more complex. However, the increased importance is derived from the number rather than the strength of the associations.
TL;DR: The possibility that the antigen detected in the cytoplasm and nuclei of the nurse cell is produced by the stichocytes of the nematode and that it is controlling genes of the altered muscle fibre, either directly or indirectly, is discussed.
Abstract: Monoclonal antibodies raised against antigens present in the excretions/secretions (E/S) of larval Trichinella spiralis , polyclonal antibodies raised against E/S and antisera from rabbits and pigs infected with T. spiralis were used in conjunction with immunocytochemical techniques to detect antigens in sections of muscle from mice that had been infected with T. spiralis for 15 or 30 days. The antibodies recognized epitopes in the stichocytes, on the surface of the cuticle, in the lumen of the oesophagus and in the lumen of the intestine of encysted larvae. Monoclonal antibodies 7C 2 C 5 and 1H7 and the polyclonal antibodies recognized epitopes in the cavity occupied by the larva, in the cytoplasm of the nurse cell, and in the hypertrophic nuclei of the nurse cell, but did not recognize material in the smaller nuclei of the nurse cell, in the cyst wall or in the surrounding muscle. Monoclonals 3B2E6 and 1D11G8B2, which recognized epitopes in the stichocytes and on the surface of the cuticle of the larvae, gave negative results with the cytoplasm and nuclei of the nurse cell. A polyclonal antibody raised against Trichuris suis recognized epitopes in the muscle and hypodermis of encysted T. spiralis but gave negative results with material in the nurse cell and nurse cell nuclei. The possibility that the antigen detected in the cytoplasm and nuclei of the nurse cell is produced by the stichocytes of the nematode and that it is controlling genes of the altered muscle fibre, either directly or indirectly, is discussed.
TL;DR: The neuropeptide story began in 1928 with the description by Ernst Scharrer of gland-like nerve cells in the hypothalamus of the minnow, Phoxinus laevis, which were overwhelmingly specialized for secretory activity, overshadowing other neuronal properties, and were termed ‘neurosecretory neurons’.
Abstract: The neuropeptide story began in 1928 with the description by Ernst Scharrer of gland-like nerve cells in the hypothalamus of the minnow, Phoxinus laevis. Because these nerve cells were overwhelmingly specialized for secretory activity, overshadowing other neuronal properties, Scharrer termed them ‘neurosecretory neurons’. What was even more remarkable about the cells was that their products were released into the bloodstream to act as hormones, specifically neurohormones. Neurosecretory cells were identified largely on morphological grounds. That is, they could be stained with special techniques, such as chrome-haematoxylin and paraldehyde-fuchsin, although the techniques are far from specific, staining non-neurosecretory cells as well. However, the basis for the ‘special’ neurosecretory techniques is the demonstration of sulphur-containing proteins – so they are indicative of peptide-producing neurones. An alternative characteristic of neurosecretory cells is the presence of large (> 100 nm), dense-cored vesicles at the electron microscope level; these are the so-called elementary granules of neurosecretion, or ENGs. However, implicit in the concept of neurosecretion is that the prime function of the neurosecretory cell is in endocrine regulation, exerting a hormone-like control over some aspect of the organism's metabolism, by controlling endocrine glands and other effector organs. To satisfy this criterion, evidence had to be obtained of cycles of secretory activity within the cell that could be correlated with a change in the physiological condition of the organism.
TL;DR: Bicuculline, securinine, pitrazepine, SR95531 and RU5135 are potent vertebrate GABAa antagonists but have little effect on GABA receptors, and the potency order of the arylaminopyridazine GABA antagonists is compared.
Abstract: The organization of Ascaris motoneurones and nervous system is summarized. There is an anterior nerve ring and associated ganglia, main dorsal and ventral nerve cords which run longitudinally, and a small set of posterior ganglia. Cell bodies of motoneurones are found in the ventral nerve cord and occur in 5 repeating 'segments'; each contains 11 motoneurones. Seven morphological types of excitatory or inhibitory motoneurone are recognized. Each Ascaris somatic muscle cell is composed of the contractile spindle; the bag region, containing the nucleus; the arm; and the syncytial region, the location of neuromuscular junctions. The resting membrane potential of muscle is approximately -30 mV and shows regular depolarizing, Ca-dependent 'spike potentials' superimposed on smaller Na(+)- and Ca2(+)-dependent 'slow waves' and even slower 'modulation waves'. The membrane shows high Cl- permeability. Adjacent cells are electrically coupled so that electrical activity in the cells is synchronized. Acetylcholine (ACh) and gamma-aminobutyric acid (GABA) affect the electrical activity. Bath-applied ACh increases membrane cation conductance, depolarizes the cells, alters the frequency and amplitude of spike potentials and produces contraction. Bath-applied GABA increases Cl- conductance, decreases spike activity and causes hyperpolarization and muscle relaxation. The extra-synaptic ACh receptors on the bag region of Ascaris muscle can be regarded as a separate subtype of nicotinic receptor. ACh and anthelmintic agonists (pyrantel, morantel, levamisole) produce a dose-dependent increase in cation conductance and membrane depolarization which is blocked by tubocurarine, mecamylamine but not by hexamethonium. The potency of GABA agonists, with the exception of sulphonic acid derivatives, correlates with the vertebrate GABAa receptor. The potency of antagonists does not. Thus, bicuculline, securinine, pitrazepine, SR95531 and RU5135 are potent vertebrate GABAa antagonists but have little effect on GABA receptors. The potency order of the arylaminopyridazine GABA antagonists: SR95103, SR95132, SR42666, SR95133, SR95531, SR42627 and SR42640 at the Ascaris GABA receptors contrasts with that at vertebrate GABAa receptors. It has been suggested that the receptor is referred to as a GABAn receptor. Patch-clamp studies show that ACh activates a non-selective cation channel which has a main conductance of 40-50pS and apparent mean open time of 1.3 ms; a smaller channel of 20-30 pS with a similar open-time is also activated. Pyrantel and levamisole also produce openings with similar conductances and open-times.(ABSTRACT TRUNCATED AT 400 WORDS)
TL;DR: The intensity of Trichuris trichiura and hookworm infections among the villagers before and after intervention were not observed to have changed significantly regardless of selective, targeted or mass treatment.
Abstract: SUMMARY A study to compare effects of mass, targeted and selective chemotherapy with levamisole (Ketrax) as an action for the control of Ascaris lumbricoides was carried out in three communities in rural Oyo State, Nigeria. Selective treatment was applied in one village by treating the most heavily infected 20 % of the inhabitants, targeted treatment in the second village involved children aged 2-15 years, while mass treatment was offered to all inhabitants excluding infants under 1 year and pregnant women in the third village. Recommended doses of levamisole were given in the villages, as described above, at 3-monthly intervals during a period of 1 year. Prevalence and intensity (epg) of A. lumbricoides infection were determined immediately before and 3 months after the period of intervention using a modified Kato-Katz technique. In the selective treatment village, no significant differences were found between the pre- and post-treatment egg counts (mean( + s.D.) epg 6776+10791 versus 4259 + 10909 respectively) of A. lumbricoides in the total population. In the targeted treatment village, significant differences were recorded in pre- and post-treatment egg counts for the total population (9057 + 15 797 versus 2579±6529) among the children alone (10935 + 20094 versus 992 + 3175) and among the untreated adults (7742 + 9782 versus 4561 ± 8798). In the mass treatment village, significant differences in pre- and post-treatment egg count values were also recorded (11907 + 17220 versus 1489 + 5165). The intensity of Trichuris trichiura and hookworm infections among the villagers before and after intervention were not observed to have changed significantly regardless of selective, targeted or mass treatment.
TL;DR: The results of treatment of the 65 kDa protein with trypsin and chymotrypsin are consistent with the 65KDa protein being a truncated and covalently modified band 3 molecule which consists of the first 540 amino acids of human band 3.
Abstract: A monoclonal antibody, 1C4, was produced which recognizes a 65 kDa protein that is localized to the plasma membrane of human erythrocytes infected with Plasmodium falciparum. By immunofluorescence the antigen was visualized as dots on the surface of the infected cell. The 65 kDa protein was present in 4 strains of diverse geographical origin, and in erythrocytes infected with a knobless strain. The 65 kDa protein was insoluble in non-ionic detergents, but was partly soluble in SDS and some high (1 M) salt solutions. The 65 kDa protein is recognized by antibodies specific for the cytoplasmic domain and the N-terminal side of the membrane-spanning region of human band 3, but was not recognized by an antibody specific to the C-terminal side of the membrane-spanning region. The results of treatment of the 65 kDa protein with trypsin and chymotrypsin are consistent with the 65 kDa protein being a truncated and covalently modified band 3 molecule which consists of the first 540 amino acids of human band 3.
TL;DR: Analysis of L3 excretory-secretory products revealed enzymes of Mr 51, 58, 79, approximately 150 and approximately 250 kDa, indicating that the major 51 kDa protease is a metallo-enzyme, and incubation at 100 degrees C generated a previously unobserved activity at 20 kDa.
Abstract: Proteases from infective larval (L3) and adult stages of Nippostrongylus brasiliensis were investigated with a combination of techniques involving gelatin degradation and cleavage of fluorogenic substrates. Analysis of L3 excretory-secretory (ES) products revealed enzymes of Mr 51, 58, 79, approximately 150 and approximately 250 kDa. Inhibition profiles indicate that the major 51 kDa protease is a metallo-enzyme. Significantly, little activity was present in larval somatic extracts, suggesting the synthesis of zymogens or precursor forms prior to secretion. Adult ES contained a distinct enzyme, of 50 kDa, and a number of other proteases were detected in somatic extracts of this stage, ranging from 51 to greater than 300 kDa. The largest of these adult somatic enzymes is also a putative metallo-protease. While nearly all enzymes from both L3 and adult are heat labile, incubation at 100 degrees C generated a previously unobserved activity at 20 kDa. Furthermore, a protease of similar size may be found in uninfected rat intestinal tissue, suggesting specific uptake of a host-associated enzyme by the parasite in the form of an inactive, heat-labile complex.
TL;DR: Although ligand-gated receptors differ markedly in terms of the endogenous ligands they recognize and the ion channels that they gate, the structural similarities suggest a strong evolutionary relationship, and the study of the pharmacology of receptor proteins for neurotransmitters in invertebrates is now gathering momentum.
Abstract: Classical transmitters are present in all phyla that have been studied; however, our detailed understanding of the process of neurotransmission in these phyla is patchy and has centred on those neurotransmitter receptor mechanisms which are amenable to study with the tools available at the time, for example, high-affinity ligands, tissues with high density of receptor protein, suitable electrophysio-logical recording systems. Studies also clearly show that many neurones exhibit co-localization of classical transmitters and neuropeptides. However, the physiological implications of this co-localization have yet to be elucidated in the vast majority of examples.The application of molecular biological techniques to the study of neurotransmitter receptors (to date mainly in vertebrates) is contributing to our understanding of the evolution of these proteins. Striking similarities in the structure of ligand-gated receptors have been revealed. Thus, although ligand-gated receptors differ markedly in terms of the endogenous ligands they recognize and the ion channels that they gate, the structural similarities suggest a strong evolutionary relationship. Pharmacological differences also exist between receptors that recognize the same neurotransmitter but in different phyla, and this may also be exploited to further the understanding of structure-function relationships for receptors. Thus, for instance, some invertebrate GABA receptors are similar to mammalian GABAa receptors but lack a modulatory site operated by benzodiazepines. Knowledge of the structure and subunit composition of these receptors and comparison with those that have already been elucidated for the mammalian nervous system might indicate the functional importance of certain amino acid residues or receptor subunits. These differences could also be exploited in the development of new agents to control agrochemical pests and parasites of medical importance.The study of the pharmacology of receptor proteins for neurotransmitters in invertebrates, together with the application of biochemical and molecular biological techniques to elucidate the structure of these molecules, is now gathering momentum. For certain receptors, e.g. the nicotinic receptor, we can expect to have fundamental information on the function of this receptor at the molecular level in both invertebrates and vertebrates in the near future.