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At least 19 recordsLinked to original sources

PubMed1994

Characterization of a high molecular weight antigen of Cryptosporidium parvum micronemes possessing epitopes that are cross-reactive with all parasitic life cycle stages.

Crossreacting antigens between life cycle stages of Cryptosporidium parvum (Protozoa, Apicomplexa) were detected using monoclonal antibodies (mAbs). Shared epitopes were demonstrated by immunoelectron microscopy, at the level of micronemes of the sporozoite and merozoite stages; some dense granules were also labelled but not so intensively. The parasitophorous vacuole membranes of all intracellular stages, the wall-forming bodies of macrogametes and the outer oocyst walls all shared these epitopes. The antigens that bear these epitopes were characterized using the whole oocyst and sporozoite stages as sources of antigenic material. Complex labelling patterns were observed on Western blots. However, all the mAbs used in this study recognized an antigen of more than 500 kDa. The glycoproteinic nature of this antigen was demonstrated by its sensitivity to pronase and periodate treatments. The expression of this high molecular weight immunoreactive antigen in the intracellular stages of C parvum was not investigated and remains to be found.

Animals↗

PubMed1997

Brugia pahangi: differential induction and regulation of jird inflammatory responses by life-cycle stages.

It has been hypothesized that different life-cycle stages of filarial nematodes induce different host responses. This concept was examined in the Brugia pahangi-jird model of lymphatic filariasis by measuring the kinetics of inflammatory responses to parasite antigens following intraperitoneal inoculation of different life-cycle stages. For this purpose, viable female or male worms, L3, L4, or microfilarial stage, were used. Dead worms served as controls. Worm and microfilarial burdens, pulmonary granulomatous inflammation (PGRN) to soluble adult worm antigen (SAWA)-coated beads, and peritoneal eosinophil and macrophage numbers were assessed at different days post-inoculation. All jirds inoculated with any of these life-cycle stages developed an early PGRN to SAWA which was later significantly reduced. Only viable worms induced down-regulation of the PGRN response. These results indicate that the hyporesponsive state is induced and maintained by all life-cycle stages. Also, the degree of granulomatous response was influenced by worm burden, with larger worm burdens inducing lower initial levels of PGRN to SAWA. Peritoneal inflammatory responses differed from the systemic response in that numbers of macrophages increased with time and microfilarial accumulation. No correlation was observed between peritoneal inflammatory responses measured by eosinophil and macrophage numbers and PGRN to SAWA.

Animals↗

PubMed1991

Endopeptidase variations among different life-cycle stages of African trypanosomes.

Lysates of different life-cycle stages of Trypanosoma congolense, Trypanosoma vivax and Trypanosoma brucei were analysed for endopeptidase activity, using reaction conditions which permitted a distinction to be made between lysosomal and non-lysosomal activity [Lonsdale-Eccles, J. D. & Grab, D. J. (1987) Eur. J. Biochem. 169, 467-475]. Hydrolysis of Z-Arg-Arg-NHMec (Z = benzyloxycarbonyl, NHMec = 7-amino-4-methylcoumaryl) and Z-Gly-Gly-Arg-NHMec occurred predominantly at alkaline pH and was observed in lysates of both insect and mammalian infective forms of T. brucei and T. congolense. Compared to their other life-cycle stages, procyclic forms of T. brucei and epimastigote forms of T. congolense exhibited enhanced hydrolysis of these substrates. Low levels of hydrolysis of Z-Arg-Arg-NHMec were observed in the bloodstream and epimastigote forms of T. vivax. The hydrolysis of Z-Gly-Gly-Arg-NHMec in each of the life-cycle stages of T. vivax was generally below detectable levels. In lysates of T. congolense, proteolytic and Z-Phe-Arg-NHMec-hydrolytic activity in bloodstream forms greater than metacyclic greater than epimastigote greater than procyclic forms. In T. vivax Z-Phe-Arg-NHMec-hydrolytic activity differed slightly according to the origin of the parasite but, in general, followed the same pattern (i.e. bloodstream forms greater than epimastigote forms, with metacyclic forms usually intermediate between these two). In T. brucei, Z-Phe-Arg-NHMec-hydrolytic activity in bloodstream forms greater than procyclic forms. Upon differentiation of the long, slender bloodstream forms into short, stumpy forms the Z-Phe-Arg-NHMec-hydrolytic activity was elevated even further. Thus, during their life cycle, each of these African trypanosomes exhibits complex changes of endopeptidase activity, suggestive of an induction of lysosomal activity between the insect and mammalian forms.

Amino Acid Sequence↗

PubMed2000

Life-cycle stage morphology of Psoroptes mange mites.

Detailed life-cycle stage descriptions for the ectoparasitic mite Psoroptes ovis (Hering) (Acari: Psoroptidae) from rabbit hosts (syn. Psoroptes cuniculi) are presented. The results resolve a number of contradictions in the literature relating to the recognition of the life cycle stages of these mites. This study supports the view that there are two distinct male nymphal stages, both lacking dorsoposterior tubercles. The male tritonymph is significantly larger than the protonymph and has five pairs of metapodosomal setae rather than three. In addition, male tritonymphs have two pairs of cuticular pits on the central metapodosoma rather than the single pair of the protonymph. The results also show that the female protonymph can be distinguished from the male nymphal stages and the female tritonymph. Both female nymphal stages possess dorsoposterior tubercles, but the protonymph is significantly smaller than the tritonymph. In addition, the protonymph possesses three pairs of metapodosomal setae rather than five, one pair of cuticular pits rather than two, and a pulvillus on leg IV which is absent in the female tritonymph. The presence of dorsoposterior tubercles enables the female nymphs to be distinguished easily from the males. By contrast, distinguishing between the nymphal stages of the same sex relies on the identification of both the number of metapodosomal setae and cuticular pits. These descriptions are used to produce a key, which allows the various stages of both sexes to be distinguished.

Animals↗

PubMed1992

Antigenic differences between the life cycle stages of Cooperia oncophora.

The antigenic differences among the life cycle stages of Cooperia oncophora were studied by SDS-gel electrophoresis and Western blotting. Sodium dodecylsulphate polyacrylamide gel electrophoresis of the different life cycle stages of C oncophora revealed a complex protein pattern with a decreasing number of protein bands towards the adult stages. Several bands of the fourth stage larvae were in common with both the third stage and the adult nematode. Western blotting with sera from C oncophora monoinfected calves showed that the antigens of the fourth stage larvae were recognised predominantly and the presence of stage specific antigens in all stages. Strong cross reactivity was demonstrated when serum from Ostertagia ostertagi infected calves was used.

Animals↗

PubMed2006

An approach to revealing blood fluke life cycles, taxonomy, and diversity: provision of key reference data including DNA sequence from single life cycle stages.

Revealing diversity among extant blood flukes, and the patterns of relationships among them, has been hindered by the difficulty of determining if specimens described from different life cycle stages, hosts, geographic localities, and times represent the same or different species. Persistent collection of all available life cycle stages and provision of exact collection localities, host identification, reference DNA sequences for the parasite, and voucher specimens eventually will provide the framework needed to piece together individual life cycles and facilitate reconciliation with classical taxonomic descriptions, including those based on single life cycle stages. It also provides a means to document unique or rare species that might only ever be recovered from a single life cycle stage. With an emphasis on the value of new information from field collections of any available life cycle stages, here we provide data for several blood fluke cercariae from freshwater snails from Kenya, Uganda, and Australia. Similar data are provided for adult worms of Macrobilharzia macrobilharzia and miracidia of Bivitellobilharzia nairi. Some schistosome and sanguinicolid cercariae that we recovered have peculiar morphological features, and our phylogenetic analyses (18S and 28S rDNA and mtDNA CO1) suggest that 2 of the new schistosome specimens likely represent previously unknown lineages. Our results also provide new insights into 2 of the 4 remaining schistosome genera yet to be extensively characterized with respect to their position in molecular phylogenies, Macrobilharzia and Bivitellobilharzia. The accessibility of each life cycle stage is likely to vary dramatically from one parasite species to the next, and our examples validate the potential usefulness of information gleaned from even one such stage, whatever it might be.

Animals↗

PubMed1986

Antigenic differences among the life cycle stages of Oesophagostomum radiatum.

Maturation of Oesophagostomum radiatium is characterised by production of different proteins in all the life cycle stages that occur within the bovine host. One-dimensional sodium dodecyl sulphate-polyacrylamide gels stained for total protein, showed that fourth stage larvae (L4) and adult worms have many proteins in common but L4 has a major additional protein (82,000 D); third stage larvae have many different proteins. Analysis of the proteins of each life cycle stage by Western blotting with sera from naturally infected calves showed that each life cycle stage had specific antigenic proteins. The major protein unique to L4 was identified as a developmentally regulated antigen.

Animals↗

PubMed2006

Variation in the intensity of inbreeding depression among successive life-cycle stages and generations in gynodioecious Silene vulgaris (Caryophyllaceae).

Inbreeding depression is one of the hypotheses explaining the maintenance of females within gynodioecious plant populations. However, the measurement of fitness components in selfed and outcrossed progeny depends on life-cycle stage and the history of inbreeding. Comparative data indicate that strong inbreeding depression is more likely to occur at later life-cycle stages. We used hermaphrodite individuals of Silene vulgaris originating from three populations located in different valleys in the Swiss Alps to investigate the effect of two generations of self- and cross-fertilization on fitness components among successive stages of the life cycle in a glasshouse experiment. We detected significant inbreeding depression for most life-cycle stages including: the number of viable and aborted seeds per fruit, probability of germination, above ground biomass, probability of flowering, number of flowers per plant, flower size and pollen viability. Overall, the intensity of inbreeding depression increased among successive stages of the life cycle and cumulative inbreeding depression was significantly stronger in the first generation (delta approximately 0.5) compared with the second generation (delta approximately 0.35). We found no evidence for synergistic epistasis in our experiment. Our finding of more intense inbreeding depression during later stages of the life cycle may help to explain the maintenance of females in gynodioecious populations of S. vulgaris because purging of genetic load is less likely to occur.

Crosses, Genetic↗

PubMedDate not supplied

Isolation and characterization of glutathione reductase from Physarum polycephalum and stage-specific expression of the enzyme in life-cycle stages with different oxidation-reduction levels.

Physarum polycephalum has a life cycle with several distinct phases that have different oxidation-reduction requirements. To investigate the relationship between the life cycle and the oxidation-reduction state, we isolated glutathione reductase (GR; EC 1.6.4.2) from Physarum microplasmodia. The enzyme was found to be a homodimer with a subunit M(r) of 49,000, and K(m) values for oxidized glutathione and NADPH of 40 and 28.6 microM, respectively. We then constructed a cDNA library from microplasmodium mRNA and cloned GR cDNA from the library. The isolated cDNA consisted of 1,475 bp encoding a polypeptide of 452 amino acids. The amino acid sequence similarity was about 50% with GRs of other organisms, and several conserved sequence motifs thought to be necessary for activity are evident in the Physarum enzyme. Escherichia coli transformed with an expression vector containing the cDNA synthesized the active GR. Genomic Southern blot analysis indicated that the GR gene is present as a single copy in the Physarum genome. Immunoblot analysis and RT-PCR analysis detected GR mRNA expression in the microplasmodium, plasmodium, and sclerotium, but not in the spore or flagellate. GR activity was low in the spore and flagellate. These results suggest that the glutathione oxidation-reduction system relates to the Physarum life cycle.

Amino Acid Sequence↗

PubMedDate not supplied

Ultrastructural observations on life cycle stages of Pneumocystis carinii.

An ultrastructural study of life cycle stages of Pneumocystis carinii in infected rat lungs in situ was undertaken utilizing 8 different modes of fixation. Three of the fixatives employed gave good fixation of cysts and intracystic bodies, but for the trophic forms fixation was only fair. Both the trophic forms and intracystic bodies have nuclear pores. The mitochondria of the organism have cristae that appear lamellar. One of the fixation modes revealed a thin, electron-dense layer on the outer surface of the cell wall, a "fuzzy coat" that had not been described previously. This material appears to mediate tight adhesion of trophic forms with other trophic forms, cysts, and with pneumocytes of the lung alveolus.

Animals↗

PubMed1995

The life cycle stages of three Diplostomum species maintained in the laboratory.

The taxonomy of Diplostomum species is in a highly confused state due to synonomy and descriptions based on only one life cycle stage. The objective of this study was to establish and maintain life cycles of several Diplostomum species in the laboratory so that accurate and detailed descriptions could be made. Metacercariae taken from the eyes of fresh water fish were identified using the key of Shigin (1986). Adopting the method of Field, McKeown & Irwin (1994) life cycles of Diplostomum spathaceum, Diplostomum parviventosum and Diplostomum volvens were established and each stage was described, measured and photographed or illustrated as required. The results of this work will provide standard descriptions for the life cycle stages of these Diplostomum species. It will relieve some of the identification difficulties experienced when only one stage in the life cycle is encountered.

Animals↗

PubMed2000

Novel species specific antigens of trypanosoma congolense and their different localization among life-cycle stages.

Seven monoclonal antibodies (mAbs) were raised against Trypanosoma congolense procyclic form (PCF). Localization of the antigens recognized by the mAbs was determined in bloodstream form (BSF), PCF, epimastigote form (EMF) and metacyclic form (MCF) by confocal laser scanning microscopy (CLSM). Two mAbs (10F9 and 20H12) showed different fluorescent patterns among different life-cycle stages of the parasite. The 10F9 recognized a 76 kDa antigen of all life-cycle stages of the parasite and the antigen localization corresponded with that of a mitochondrion. While the 20H12 recognized 119 and 122 kDa antigens of all the life-cycle stages and the antigen localization corresponded with a flagellum in BSF and MCF, tip of a flagellum in PCF, and part of cytoplasm in EMF. Moreover, the 20H12 did not react to T. brucei gambiense, T. b. rhodesiense and T. evansi antigens in both CLSM and immunoblotting. Therefore, the antigens recognized by the 20H12 seem to be T. congolense specific. Although, further studies will be required for a full characterization of the T. congolense specific 119 and 122 kDa antigens, the mAb 20H12 and the specific antigens may be useful in not only establishment of T. congolense specific diagnosis methods but also studies on molecular mechanisms regulating differentiation of the parasite during life-cycle.

Animals↗

PubMed1998

[Genotypic variability in populations of the mound-builder mouse Mus spicilegus Pet., 1882, at various life cycle stages].

Genotypic variability in two Moldovian populations of moundbuilder mice Mus spicilegus Pet. was examined at different stages of their life cycle--at the beginning of overwintering in mounds and in the middle of summer in agrocenoses. Thirty allozymes were assayed. Significant differences in allele frequencies and heterozygosity between overwintered and mice born in the year of the experiment were recorded only for Idh-1. This fact was related to the presence of differential mortality among overwintered mice in the populations by the middle of summer. The increase in heterozygosity for Idh-1 in the populations occurred because of "old" mice that lived until the middle of summer, at which point they still have not contributed to the gene pool of the population. The genetic effect of the annual separation of generations of mound-builder mice due to their overwintering in mounds is analyzed. Overwintering has a bottleneck effect on the population. The relatively low genetic variability in mound-builder-mice populations is considered with regard to their biological and ecological features. Based on obtained data, a conclusion on the high genetic stability of moundbuilder-mice populations during their entire life cycle is made.

Animals↗

PubMed2004

The Schistosoma mansoni soluble proteome: a comparison across four life-cycle stages.

Differential analysis of immune responses to schistosomes has routinely been performed using complex mixtures of soluble proteins from various life-cycle stages, on the assumption that these differed significantly in composition. Proteomic techniques now allow us to characterise and compare such mixtures. The soluble proteins from cercariae, lung-schistosomula, adult worms and eggs of Schistosoma mansoni were separated by high-resolution two-dimensional electrophoresis and the resulting images analysed using appropriate software. A high degree of quantitative and qualitative similarity in spot pattern was revealed across the life-cycle, greatest between adjacent stages. To initiate mapping of these soluble proteomes, the 40 most abundant spots in each preparation, accounting for 21-46% of the total protein, were subjected to peptide fingerprinting by mass spectrometry. On average 55% of the spots were identified, but overall, these comprised only 32 different protein species. With one exception all proteins originated in the cytosol and 24 of the 32 had previously been pinpointed by virtue of their immunoreactivity, including four of the WHO priority vaccine candidates. The similarity in composition between the four preparations means that they are unlikely to discriminate adequately between immune responses to different life-cycle stages and argues strongly for the need to identify true stage-specific marker proteins. Equally, it is difficult to reconcile the abundance and immunogenicity of such cytosolic proteins with their status as vaccine candidates, as it is unlikely they will be accessible to the immune system in an intact parasite.

Animals↗

PubMed1996

Immunization against Trichinella spiralis using antigens from different life-cycle stages experimental study in mice.

Crude antigen preparations from different life-cycle stages of T. spiralis using 10 micrograms and 100 micrograms antigen/dose, were tested for their ability to immunize mice against homologous challenge infection. Immunization was assessed by accelerated expulsion of adult worms, reduction in female worm fecundity and decreased yield of muscle larvae. The dose of 100 micrograms proved to be more protective in all antigen preparations. The AA provided highly significant protection against adult stages (89%) and muscle larvae (80%). NLA produced highly significant protection against muscle larvae only (76%). MLA provided highly significant protection against adult stages (74%), female fecundity (74%) and muscle larvae (80%). Combination of AA and MLA produced highly significant protection against adult stages (96%), female fecundity (73%) and muscle larvae (86). The results show that all antigen preparations provided variable, considerable but partial protection against challenge infection. The variations in immunization potential are discussed.

Animals↗

PubMed2006

Vaccination against filarial nematodes with irradiated larvae provides long-term protection against the third larval stage but not against subsequent life cycle stages.

Sustainable control of human filariasis would benefit enormously from the development of an effective vaccine. The ability to vaccinate experimental animals, with reductions in worm burden of over 70%, suggests this aim is possible. However, in experimental vaccinations the challenge is usually administered 2 weeks after the immunisation phase and thus the protection obtained is likely to be biased by persisting inflammation. Using the murine model Litomosoides sigmodontis, we increased the time between immunisation with irradiated larvae and challenge with fully infective L3 to 5 months. Significant protection was achieved (54-58%) and the reduced worm burden was observed by 10 days p.i. The developmental stage targeted was the L3, since no nematodes died once they reached the pleural cavity of vaccinated mice, as has been previously shown in short-term protocols. However, larval developmental rate was faster in vaccinated than in primary-infected mice. Immunological assessments were made prior to challenge and then from 6 h to 34 days post-challenge. Samples were taken from the subcutaneous tissue where the larvae were inoculated, the lymph nodes through which they migrate and the pleural cavity in which they establish. Eosinophils were still present although scarce in the subcutaneous tissue of vaccinated mice before challenge. Cytokine and specific antibody production of vaccinated and challenged mice were L3-specific and Th2-biased and greatly exceeded the response of primary-infected mice. The heightened Th2 response may explain the faster development of the filarial worms in vaccinated mice. Thus, long-term vaccination protocols generated a strong memory response that led to significant but incomplete protection that was limited to the infective larval stage suggesting alternative vaccination strategies are needed.

Animals↗

PubMed1986

Ultrastructural localization of the circulating cathodic antigen in the digestive tract of various life-cycle stages of Schistosoma mansoni.

In the present study the ultrastructural localization of the schistosome-derived circulating cathodic antigen (CCA) within the digestive tract of various life-cycle stages of the blood fluke Schistosoma mansoni has been determined. Use was made of CCA-specific monoclonal antibodies in a two-step gold-labeling procedure. The following results were obtained: In cercariae gold label was found in the cytoplasm and in the luminal surface coat of the gut epithelium. The oesophagus of this life-cycle stage of the parasite showed minimal CCA reactivity, as gold label was limited to the luminal surface coat, locating proximally to the gut. In 3 1/2-week-old worms and in adult male and female worms CCA was demonstrable in the Golgi apparatus, in cytoplasmic vesicles and in the luminal surface coat; in the caudal quarter of the gut of adult worms CCA was also present in lysosome-like bodies. The oesophagus of these worm preparations did not show any CCA reactivity. In miracidia CCA was not present.

Animals↗

PubMed1987

Interaction of the plant toxin ricin with different life cycle stages of Schistosoma mansoni.

The binding of the plant toxin ricin to various life cycle stages of Schistosoma mansoni has been studied. Fluorescein isothiocyanate (FITC)-ricin exhibited binding to schistosomula and adult worms, but not to cercariae or to freshly transformed schistosomula. Binding was specifically inhibited by the presence of galactose in the medium. Analysis of protein synthesis in treated parasites illustrated that ricin failed to intoxicate schistosomula or adult worms. Indeed, fluorescence quenching studies suggested that the fluorescent toxin was confined to the outer bilayer and was not internalised.

Animals↗