[Issues in nursing research of body image].
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Adult↗
Biomedical subjects
K Fujisaki
Publications and source records attributed to K Fujisaki.
Theileria sergenti and T. buffeli: polymerase chain reaction-based marker system for differentiating the parasite species from infected cattle blood and infected tick salivary gland.
Benign Theileria species in cattle. Theileria sergenti and T. buffeli, are morphologically indistinguishable. The polymerase chain reaction (PCR) was used to amplify the genes encoding the 33- and 34-kDa major piroplasm antigens (p33/34) of T. sergenti and T. buffeli from cattle blood infected with these parasites and tick salivary gland infected with T. sergenti. Following amplification, the p33 gene from T. sergenti and the p34 gene from T. buffeli were clearly differentiated using the restriction enzyme sites that were not shared between them. The oligonucleotide primer set, designed from the p33/34 genes, was specific for these Theileria species, since no amplification was detected with DNA from Babesia ovata, B. bovis, Anaplasma marginale, A. centrale, Eperythrozoon wenyoni, bovine white blood cells, and uninfected tick salivary glands. One tenth vol of the template prepared from either 25 microliters of blood with 0.5% parasitemia or individual tick salivary glands with six infected acini allowed sufficient amplification for differentiation of the two parasite species by restriction enzyme digestion. In addition, this system could be used to demonstrate the simultaneous, experimentally induced infection of cattle with T. sergenti and T. buffeli. The PCR-based marker system therefore provides a means to differentiate T. sergenti from T. buffeli in infected cattle blood and infected tick salivary glands. This system may also be useful for the characterization of other benign Theileria species in cattle.
Animals↗
Expression of a 32 kilodalton Theileria sergenti piroplasm surface protein by recombinant baculoviruses.
Previous studies detected a single amino acid substitution (Ala196 to Gly196) between cDNA clones encoding a 32 kDa antigen (p32) of Theileria sergenti (Chitose stock) obtained from a persistently infected calf. In this study, 2 different recombinant baculoviruses (pAc/p32-Ala196 and pAc/p32-Gly196) were constructed for the expression of p32. Molecular masses of the polypeptides produced in Spodoptera frugiperda cells infected with the recombinant baculoviruses were the same as that of authentic p32. pAc/p32-Ala196 produced additional polypeptides, with molecular masses higher than 32 kDa, which resulted from differential N-glycosylation as revealed by endo N-glycosidase treatment. The results indicate that a single amino acid substitution may lead to a conformational change in p32 which affected post-translational modification of recombinant products.
Alanine↗
Detection of Hepatozoon canis oocyst from ticks collected from the infected dogs.
We collected 220 ticks from 58 dogs infected with Hepatozoon canis from 1990 to 1992 in Yamaguchi and Fukuoka prefectures, the western provinces of Japan. These ticks were identified as Haemaphysalis campanulata, H. longicornis, H. flava, Ixodes ovata, and I. nipponensis. By histopathological examination of these ticks, two oocysts which seemed to be those of H. canis were observed in the haemocoele of an adult female H. longicornis and a nymph H. flava tick. The oocyst, about 300 x 150 microns in size, was composed of 50 to 70 sporocysts with 10 to 16 banana-shaped sporozoites each.
Animals↗
Morphological, serological and antigenic characteristics, and protein profile of newly isolated Japanese bovine Babesia parasite with particular reference to those of B. ovata.
An intraerythrocytic large protozoan, tentatively designated Babesia sp. 1, was recently isolated from cattle in Hokkaido Prefecture, Japan. This parasite closely resembled B. ovata in shape of piroplasms, but was distinguishable by other morphological, immunological, and biochemical characters. The paired pyriform piroplasm of B. sp. 1 was larger than that of B. ovata. The results from serological and antigenic examination by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis showed that there were cross- but distinguishable-reaction between B. sp. 1 and B. ovata. Protein profiles of both Babesia parasites piroplasms analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) were apparently different from each other. Several major proteins revealed by 2D-PAGE and the immunodominant proteins resolved by Western blot analysis (40 kDa for B. sp. 1 and 29 kDa for B. ovata) were unique to each parasite. The results of the present study indicate the possibility that B. sp. 1 is a species different from B. ovata.
Animals↗
The taxonomy of the bovine Theileria spp.
The classification of the benign species of Theileria of cattle is very confusing Representatives of this group of parasites appear worldwide, and are known as T. sergenti in Japan, T. buffeli in Australia and T. orientalis elsewhere. Consequently, these parasites are frequently referred to as the T. sergenti/buffeli/orientalis group. Kozo Fujisaki, Shin-ichiro Kawazu and Tsugihiko Kamio here argue that the Australian T. buffeli and British T. orientalis belong to one and the same species, and that the Japanese T. sergenti is a separate species.
Journal Article↗
Development in vitro of free-living infective larvae to the parasitic stage of Strongyloides venezuelensis by temperature shift.
Free-living infective larvae Strongyloides venezuelensis were cultured in Dulbecco's modified Eagle's medium at 25 and 37 degrees C, and development to the parasitic stage was evaluated using morphological, protein and antigenicity criteria. Few larvae cultured at 25 degrees C showed development whereas, in most of the larvae cultured at 37 degrees C, there appeared characteristic changes such as a bulb-like head and droplets under the cuticle with an increase of body width of the larvae. The results obtained from two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) revealed that the protein spot patterns of the larvae cultured at 25 and 37 degrees C were differentiated by 17 specific spots. In addition, Western blot analysis combined with 2D-PAGE for reaction with serum obtained from an infected rat revealed that protein spots showing immunodominant antigen at 37 degrees C were almost the same as those of the larvae recovered from the rats rather than those of the larvae at 25 degrees C. These results strongly suggested that a temperature shift from 25 to 37 degrees C has an important role in the development of free-living infective larvae to the parasitic stage of S. venezuelensis. The culture system established in the present study was useful for biological and biochemical studies in the development from/of the free-living to the parasitic stage of Strongyloides species.
Animals↗
Protein analysis of Anaplasma marginale and Anaplasma centrale by two-dimensional polyacrylamide gel electrophoresis.
Protein compositions of Anaplasma marginale and A. centrale were analyzed by two-dimensional gel electrophoresis. Both species had a major protein which was composed of 3-4 spots. The molecular weights of these two proteins were approximately 39 kDa. However, the position of these proteins in the gels were slightly different when 2 gel maps were superimposed. Five other protein spots were shared by A. marginale and A. centrale, whereas all the other protein spots were appeared to be unique to each of the species.
Anaplasma↗
Development of Theileria sergenti schizonts in the lymph node of experimentally infected cattle.
Schizogony of Japanese Theileria sergenti of cattle was studied by light and electron microscopy. Schizonts were detected in the draining lymph node between 4 and 8 days after sporozoite inoculation. Macroschizonts (the phase of nuclear division having invaginations) were formed 6 days after inoculation. Subsequently, microschizonts (the phase of merozoite formation displaying rosette-like appearance) were observed 8 days after inoculation. Multiple infections of a host cell with sporozoites were suggested to occur since different stages of schizonts were simultaneously detected in the same cell. Host cells of schizonts were considerably enlarged by parasitism. However, morphological characteristics of the developmental stages of T. sergenti schizonts resembled those of malignant Theileria species (e.g. T. parva). Schizogony of T. sergenti observed in this study seems to be the primary generation.
Animals↗
Seroepidemiological studies of bovine babesiosis caused by Babesia ovata, B. bigemina and B. bovis in Peninsular Malaysia.
Cattle in Peninsular Malaysia were examined for evidence of infection with Babesia ovata, B. bigemina and B. bovis by an enzyme-linked immunosorbent assay for detection of antibody to the three Babesia species. All of the test samples when assayed with B. ovata antigen, resulted in low value indicating low probability of cattle infected with B. bigemina, 74.4% were positive for B. bovis and 72.6% were positive for both Babesia species. In addition, a serological survey with regard to age difference was carried out on a milk production farm. High reactivity antibody to B. bigemina and B. bovis was detected in calves less than 1 month of the age. The reactivity decreased in calves 1-3 months of the age. Then, the reactivity increased for both Babesia species in 6 months old calves. These results suggested that cattle infected with B. bigemina and B. bovis were widespread throughout Peninsular Malaysia and that both parasites might exist as an enzootical parasite.
Age Factors↗
Theileria sergenti: experimental transmission by the long-nosed cattle louse, Linognathus vituli.
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Animals↗
Histological observations on the schizonts in cattle infected with Japanese Theileria sergenti.
Histological observations were performed on the schizonts of Japanese Theileria sergenti in three calves necropsied 8 and 10 days after application of nymphal Haemaphysalis longicornis infected with T. sergenti. In all the three calves, schizonts were observed in the cytoplasm of huge cells formed in the drainage lymph nodes, liver and spleen. The huge cells were 50 to 200 microns in diameter. Schizonts had granular appearance and an irregular shape and were 1 to 7 microns in diameter. Ultrastructurally, schizonts had more than one nuclei and were formed in the unit enlarged cell. Schizonts showed a specific reaction against anti-T. sergenti anti serum, therefore, it was concluded that the schizonts were those of T. sergenti histologically.
Animals↗
Theileria sergenti: transformation of zygotes into kinetes in vector ticks, Haemaphysalis (Kaiseriana) longicornis and H. (K.) mageshimaensis.
Invagination or transformation of a zygote of Theileria sergenti into a kinete was examined with Giemsa-stained smears of the midgut of vector ticks, Haemaphysalis longicornis and H. mageshimaensis. Zygotes invaginated almost simultaneously with the moulting of host ticks. They were round and oval, and measured 5.4 to 6.7 microns in diameter on average. The kinetes developing in the zygotes were situated in the periphery of cytoplasm and were spindle- or club-shaped.
Animals↗
Protein analysis of Strongyloides venezuelensis by two-dimensional polyacrylamide gel electrophoresis.
Infective larvae, larvae in the lung and adult-stage worms in the small intestine of Strongyloides venezuelensis were analysed for protein by two-dimensional gel electrophoresis. The infective larvae were differentiated from the other two stages of parasite with 13 stage-specific spots, whereas the larvae in the lung and the adult-stage worms were identical to each other in spot patterns except for 6 spots.
Animals↗
Antigenic differences between Japanese Theileria sergenti and other benign Theileria species of cattle from Australia (T. buffeli) and Britain (T. orientalis).
Serological comparisons among piroplasm antigens of the benign Theileria species of cattle from Japan, Australia and Britain, which are frequently referred to as T. sergenti, T. buffeli and T. orientalis, were carried out. The results obtained from comparative enzyme-liked immunosorbent assay (ELISA) using sera from infected cattle suggest that T. sergenti could be differentiated from both T. buffeli and T. orientalis by their serological dissimilarities. Western blotting combined with two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) revealed that both the 33-kDa piroplasm protein of T. sergenti and the similar 34-kDa protein of T. buffeli and T. orientalis corresponded to immuno-dominant antigens against cattle. The other 32-kDa proteins of . T. buffeli and T. orientalis also represented immunodominant antigens. Cross-reactivities of the 32-and 34-kDa proteins were observed between T. buffeli and T. orientalis, whereas the 33-kDa protein of T. sergenti could be differentiated from the similar 34-kDa proteins of T. buffeli/orientalis. The present study suggests that T. sergenti should be separated from T. buffeli and T. orientalis on the basis of their serological dissimilarities.
Animals↗
Analysis of the genes encoding immunodominant piroplasm surface proteins of Theileria sergenti and Theileria buffeli by nucleotide sequencing and polymerase chain reaction.
The nucleotide sequences of the cDNAs encoding a 33-kDa piroplasm protein of Theileria sergenti (p33) and a similar protein of Theileria buffeli (p34) were determined. Both of the genes contained an open reading frame of 849 base pairs. The predicted amino acid sequence of p33 and p34, consisting of 283 residues, showed 82% similarity. A transmembrane hydrophobic domain and signal peptides were predicted. The polymerase chain reaction was used to amplify p33/34 genes from the piroplasm DNA of T. sergenti, T. buffeli and Theileria orientalis. Following amplification, p33 and p34 genes were clearly differentiated using the restriction enzymes sites that were not shared between them. These results indicated that p33 and p34 were conserved molecules among these Theileria species, and the genes that encode p33/34 proteins were suitable for discrimination of T. sergenti from T. buffeli/T. orientalis.
Amino Acid Sequence↗
Preliminary biochemical characterization of 'veil' structure purified from Theileria sergenti-, T. buffeli- and T. orientalis-infected bovine erythrocytes.
A structure called 'veil' in Theileria sergenti-, T. buffeli- or T. orientalis-infected bovine erythrocytes was purified from erythrocyte lysates by Percoll density-gradient centrifugation. On electron microscopical examination, the veils consisted of electron-dense material showing a periodicity of striations and were not surrounded by membranes. Analyses of veil proteins by one- and two-dimensional polyacrylamide gel electrophoresis revealed that the veils contain haemoglobin and several other basic proteins of molecular weight ranging from 15.5 to 46 kDa. By comparing the protein patterns of the veil with those of purified piroplasms and uninfected bovine erythrocytes, these basic proteins appeared to be of parasite origin. It would appear likely that the veils formed by intra-erythrocytic precipitation of haemoglobin and proteins excreted by the parasites. Differences in veil constituents were found between T. sergenti, T. buffeli and T. orientalis.
Animals↗
Plasma high density lipoprotein in anemic cattle infected with Theileria sergenti.
Changes in plasma concentration of lipid composition were analysed in cattle with anemia due to Theileria sergenti infection. Plasma levels of phospholipids, cholesterol, free cholesterol, high density lipoprotein cholesterol, and vitamin E decreased to 40-67% of the pre-infection levels, corresponding to the decrease of PCV due to the infection. However, no definite changes were detected in plasma level of triglyceride. By gradient centrifugation, it was confirmed that lipid components, other than triglyceride, occur in high density lipoprotein (HDL) and these decreases lowered the HDL value. There was no correlation between this phenomenon and liver function. As similar changes in lipid composition were also observed in phlebotomized calves, it was considered that this phenomenon might partially depend on the acceleration of erythropoiesis as a reaction to anemia caused by T. sergenti infection.
Anemia↗