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E Schein

Publications and source records attributed to E Schein.

At least 37 records · Page 2Linked to original sources

DNA measurements and ploidy determination of developmental stages in the life cycles of Theileria annulata and T. parva.

The relative DNA levels of different developmental stages of Theileria annulata and T. parva in the cow and the tick were measured by the cytophotometric DNA technique using the fluorochrome Hoechst 33258 as a staining dye. The results revealed that sporozoites, merozoites, gamonts, and gametes were haploid, whereas multinucleated intralymphocytic schizonts were polyploid. No difference was observed between T. parva and T. annulata in these stages. For both Theileria species, the DNA measurements revealed that fusion of gametes occurred in the gut of the final host, thus providing evidence of sexual reproduction. However, differences were observed between the two parasites in the tick. Whereas T. parva zygotes underwent a two-step meiotic division, a comparable reduction division could not be unequivocally detected in T. annulata. Differences could also be detected in the further development of kinetes, indicating that Theileria species are not characterized by only one life cycle, which is specific for this genus.

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DNA measurements reveal differences in the life cycles of Babesia bigemina and B. canis, two typical members of the genus Babesia.

The relative DNA levels of different developmental stages of Babesia bigemina and B. canis were measured by cytophotometry using the fluorochrome Hoechst 33258 as a staining dye. The DNA measurements provided direct proof of sexual reproduction, i.e., fusion of gametes resulting in the formation of zygotes followed by a meiotic division. Both Babesia species are considered to be typical members of the genus Babesia; however, the DNA measurements revealed important differences in the life cycle of these parasites, indicating that Babesia species are not characterized by a life cycle, which is specific for this genus.

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New findings in the development of Babesia (Theileria) equi (Laveran, 1901) in the salivary glands of the vector ticks, Hyalomma species.

The development of the piroplasm Babesia equi was studied by light microscopy in the salivary glands of three different Hyalomma species during and after the engorgement of nymphs on experimentally infected horses and after adults had fed on a vertebrate host following ecdysis. The stock of B. equi used was isolated from a horse imported from Turkmenistan (CIS) in 1991. The findings, being identical in all three Hyalomma species, differ with regard to the chronological order of the development stages in several respects from the results of previous studies based upon light or electron microscopy. A first sporogony phase of B. equi was found to develop in the salivary glands of the engorged nymphs before the ticks moulted to adults. Beginning at day 6 postinfestation (p. infest.) of the nymphs, spindle-shaped sporozoites appeared to be formed by both rapid sequential fission of a multinucleated complex and a process of radial budding from multiple fission bodies. Sporozoites isolated from the salivary glands of the engorged nymphs proved to be infectious when they were injected into a susceptible horse. After the nymphs had moulted, a second sporogony phase similar to the first one observed in the salivary glands of engorged nymphs could also be initiated in the salivary glands of adults when they were attached to a vertebrate host. Sporozoites produced in the salivary glands of adults were equally infectious for horses. Thus, two completely separate sporogony phases in B. equi seem to develop successively in the salivary glands of Hyalomma species during a transstadial transmission.

Acari↗

The development of Babesia (Theileria) equi (Laveran, 1901) in the gut and the haemolymph of the vector ticks, Hyalomma species.

The development of the piroplasm Babesia equi was studied by light microscopy in the gut and the haemolymph of three different Hyalomma species during and after the nymphs had engorged on parasitaemic horses. The stock of B. equi used was isolated from a horse imported from Turkmenistan (CIS) in 1991. The existence of gamogony was identified by the occurrence of gamonts and gametes in the gut contents of the nymphs at between 3 and 4 days after infestation of the nymphs, before the ticks dropped off the experimentally infected horses. Zygotes and kinetes were observed in the intestinal cells from day 4 until day 7 after infestation i.e. 2 days prior to engorgement until repletion of the nymphs. Simultaneously, kinetes could also be seen in the haemolymph of engorged nymphs. The morphology and the sequence of the developmental stages of B. equi identified in the gut and the haemolymph of the vector ticks were identical in all three Hyalomma species. They resembled typical developmental stages of bovine Theileria species. Because of the close similarity of the developmental cycle of B. equi in Hyalomma species to the life cycle of Theileria species and due to the differences to the other Babesia species, a discussion about classifying B. equi into the family of Theileriidae now seems to be justified.

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Transmission of the agent of Lyme disease on a subtropical island.

To determine whether the agent of Lyme disease (Borrelia burgdorferi) perpetuates on a subtropical island, we recorded prevalence of spirochetal infection in Ixodes ricinus ticks collected on Madeira Island and identified local reservoir hosts of both pathogen and vector tick. Spirochetes infect at least 1.3% of the nymphal ticks collected in a particular site. Subadult ticks frequently parasitized Norway as well as black rats on the island, and Lyme disease spirochetes were found in these hosts. Each was competent as reservoir host. Canaries, lizards and domestic ungulates, on the other hand, were incompetent for Madeiran spirochetal isolates. Madeiran spirochetes stimulated rodents to recognize the same antigens as did spirochetes isolated on the European continent. The polymerase chain reaction amplified identical specific sequences of Madeiran spirochetes as of European-derived spirochetes. Rats appear to be the main reservoir hosts of the agent of Lyme disease on Madeira Island, and cattle and sheep serve as definitive hosts of the vector tick. The agent of Lyme disease is enzootic on this subtropical island some 10 degrees of latitude south of the northern Mediterranean coast. Because I. ricinus ticks frequently attack people on Madeira Island, Lyme disease should be considered as a cause of locally acquired human illness.

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Production of interferon by Theileria annulata- and T. parva-infected bovine lymphoid cell lines.

Theileria annulata and T. parva-infected lymphoblastoid cells were examined for their capacity to produce interferon (IFN). Supernatants of such cells were tested in biological assay for their antiviral activity. Only T. parva-infected cells of T-cell origin were capable of producing IFN-gamma. Supernatants of some but not all T. annulata-infected cells showed also antiviral activity, which was greatly reduced after exposure to a pH of 2. Northern-blot analysis of the cells using an IFN-gamma cDNA probe confirmed the results obtained for T. parva-infected cells in a biological assay. No IFN-gamma mRNA was detected in T. annulata-infected cells. The importance of IFN for the pathogenesis of theileriosis is discussed.

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Effect of buparvaquone on the expression of interleukin 2 receptors in Theileria annulata-infected cells.

Theileria annulata-infected cells were cultured in the presence or absence of human recombinant interleukin 2 (hrIL-2). This growth factor proved to be capable of enhancing the growth of the infected cells: its effect was marked, particularly when the cells were seeded at low densities, and it varied from cell line to cell line. The infected cells produced a factor that possessed the biological activities of IL-2, since their supernatants could enhance the proliferation of concanvalin A-stimulated (Con A) blasts. The reactivity of the parasitized cells to hrIL-2 was abolished following their treatment with the antitheilerial drug buparvaquone. In addition, the drug inhibited the binding of 125I-IL-2 to T. annulata-infected cells but failed to suppress its binding to Con A blasts. Northern blot analysis revealed that the drug had no effect on the expression of the alpha chain of the IL-2 receptor (IL-2R). Therefore, it is possible that buparvaquone interferes with the expression of the beta chain of the IL-2R. The role of IL-2 and the IL2R in the permanent proliferation of T. annulata-infected cells is discussed.

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Borna disease virus-specific antigens. II. The soluble antigen is a protein complex.

Borna disease virus-specific soluble antigen from persistently infected rat brains was purified to homogeneity using preparative sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The soluble antigen is a complex of three proteins with apparent molecular weights of 35 kDa, 38 kDa and 24 kDa. The 35/38 kDa antigen double band was separated into its two components. The 24 kDa protein has no common epitopes with the 35/38 kDa protein.

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[Parasite fauna of red foxes in Berlin (West)].

The infestation with ecto- and endoparasites of red foxes in Berlin (West) was investigated. As shown by post mortems of 100 animals (47 males and 53 females), none of them was found to be free of parasites. With regard to ectoparasites, 3 species of ixodes ticks, 7 flea species, one species of biting lice and mite species were demonstrated. The sarcoptic mange is often inducing the death of the foxes and is considered to be a limiting factor of the population increment. Concerning endoparasites, cestodes (6 species), trematodes (7 species), nematodes (10 species) and 3 coccidian species were found. The infestation rate with different species of hook worms, ascarids and mesocestoides was remarkably high. However, none of the foxes were infected with Trichinella spiralis or Echinococcus multilocularis. As shown by ELISA, in 15% of foxes antibodies against Borrelia burgdorferi were found. Therefore, the red fox must be considered as a reservoir host for Lyme-borreliosis.

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Effect of cyclosporin A on the proliferation of bovine lymphocytes to concanavalin A and on the growth of Theileria annulata-infected bovine cells.

After being inoculated by Hyalomma ticks, the sporozoites of Theileria annulata invade bovine lymphocytes, where they subsequently differentiate to schizonts. The infected cells are induced to a continuous proliferation which can be enhanced by human recombinant interleukin 2 (hrIL-2). In the present study, we examined the influence of cyclosporin A (CsA) on the growth of schizont-containing cells and compared with its effect on bovine peripheral blood lymphocytes (PBL) responding to Concanavalin A (ConA). In both cell types, the proliferation was inhibited in a dose dependent manner, which was not restorable in T. annulata-infected cells even after addition of hrIL-2. In contrast, ConA-blasts were able to undergo a proliferative response provided they were treated with high doses of CsA. Both, T. annulata-infected cells and bovine ConA-blasts express IL-2 receptors (IL-2R). The binding of radiolabelled hrIL-2 to ConA-blasts and T. annulata-infected cells was only partially inhibited after treatment with CsA. CsA was not toxic for the parasites, since the treated cells still contained schizonts which did not show any morphological abnormality.

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Sexual cycle of Babesia divergens confirmed by DNA measurements.

The DNA content of the developmental stages within the life cycle of Babesia divergens was measured by means of fluorescence microscopy using the DNA-specific bisbenzimide Hoechst 33258. By comparing the mean relative fluorescence intensities, the assumption of sexual reproduction in the gut of the tick vector (Ixodes ricinus) was confirmed. However, no proof of a pre- or postzygotic meiosis was found.

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Buparvaquone but not cyclosporin A prevents Theileria annulata-infected bovine lymphoblastoid cells from stimulating uninfected lymphocytes.

The influence of Buparvaquone on the morphology, proliferation, and stimulation with T and B cell mitogens of Theileria annulata-infected cells was studied. In addition, the stimulatory capacity of the infected cells before and after treatment with Buparvaquone or cyclosporin A (CsA) was also examined and compared to that of ConA-stimulated bovine peripheral blood cells (PBL). After incubation of the cells for 4 days with Buparvaquone only few schizonts were detectable in the cells. Prolongation of the incubation time to 8, 12, or 14 days eliminated completely the parasites. Despite the elimination of the parasites, the cells were still unable to undergo a proliferative response to Con A or PWM. However, the drug did not interfere with the response of normal PBL to these mitogens. Furthermore, Buparvaquone but not CsA inhibits the generation of mixed lymphocyte reaction (MLR). None of the drugs could prevent ConA-blasts from stimulating autologous PBL. These results suggest that the antigen expressed by the infected cells and recognised by the responder PBL was induced by the schizonts.

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In vitro proliferative and cytotoxic responses of PBL from Theileria annulata-immune cattle.

Peripheral blood lymphocytes were isolated from healthy calves and were subsequently infected with sporozoites of Theileria annulata in vitro. The infected cells were passaged for 50 times and thereafter inoculated into animals from which they were previously isolated. Within 4-5 days, schizont-containing cells were demonstrable in the lymph nodes of all animals. Few days later, merozoites were detected in erythrocytes. A slight decrease in the counts of lymphocytes and leucocytes was also found. After 2 months these animals and a group of uninfected calves were heavily infected by tick-infestation and showed severe symptoms of theileriosis with 60% schizont-containing cells in the lymph nodes and a parasitaemia of about 35%. Because of the severity of the infection, all control calves were treated with Halofuginone. In contrast, the initially immunized cattle (by inoculation of culture cells), survived the infection without chemotherapy. Less than 10% of their lymph node cells contained schizonts, whereas less than 1% of their erythrocytes were found to be infected with merozoites. In all immunized animals, specific cytotoxic PBL, with the capacity to lyse autologous but not allogeneic infected cells, were demonstrated. In addition, a population of PBL were found to be able to inhibit the growth of T.annulata-infected culture cells in vitro. However, in comparison to PBL of immune animals, PBL of acute infected calves were superior in their capacity to inhibit the proliferation of schizont-containing cells. In mixed lymphocyte reactions, T. annulata-infected cells could induce a more pronounced proliferative response in PBL from immune than in PBL of uninfected animals.

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The effects of drugs on the formation of Theileria annulata merozoites in vitro.

The effects of colchicine, taxol, dinitrophenol, sodium azide, TLA-144, and obioactin on Theileria annulata schizonts were tested in vitro and studied by means of light and electron microscopy. Colchicine (0.001-5 microM) and taxol (1 microM) completely inhibited the mitosis of the host cell. This resulted in higher numbers of schizont nuclei after 96 h, since the division of the parasites was apparently not affected. Increasing numbers of schizont nuclei were also obtained after incubation of parasitized lymphocytes for 7 day at 41 degrees C with a daily medium exchange.

Alkaloids↗

Effect of temperature on transovarial transmission of Babesia bigemina (Smith and Kilborne, 1893) in Boophilus annulatus (Say, 1821).

The effect of the temperature on the transovarial passage of Babesia bigemina has been studied in female Boophilus annulatus during their oviposition. Kinetes of Babesia were present in eggs laid at the temperatures of 16, 20, 26, 28, 30 and 35 degrees C. The rate of infection was temperature dependent. It reached at least 50%, even at low temperatures. The average infection level at 26 and 30 degrees C was 270 kinetes per egg. However, between 20 and 35 degrees C the eggs laid during the first 3 days were parasite free. At 16 degrees C, no kinetes were detected during the first 13 days of oviposition.

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The role of antibodies in immunity against Theileria annulata infection in cattle.

The role of antibodies in protecting cattle against Theileria annulata infection was investigated. It was found that serum samples from immune cattle had a neutralizing effect on the sporozoites preventing in this way the invasion of lymphocytes. However, the same antiserum neither caused lysis of macroschizont- infected lymphoblastoid cells nor inhibited their proliferation in vitro. Free merozoites but not the infected erythrocytes were opsonized by immune serum. In addition, complement alone, without antibody participation, was able to lyse free merozoites. The importance of humoral factors in immunity to T. annulata infection is discussed.

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The role of interleukin 2 (IL 2) in the proliferation of Theileria annulata-infected bovine lymphocytes.

The role of interleukin 2 (IL 2) in the proliferation of Theileria annulata-infected lymphoblastoid cells was studied. It was found that these cells neither require nor produce IL 2. This was based on the following findings: (a) The growth of T. annulata-infected cells was not enhanced by addition of IL 2 to the culture medium. (b) IL 2-dependent Con A-stimulated bovine peripheral blood lymphocytes (Con A-blasts) no longer required IL 2 after being infected with sporozoites of T. annulata. (c) The supernatants of T. annulata-infected cells did not support growth of IL 2-dependent Con A-blasts.

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