Showing papers by "Chihiro Sugimoto published in 2007"
TL;DR: Loop-mediated isothermal amplification (LAMP) is a rapid and simple technique since it can be carried out in 1 h and requires only a simple heating device for incubation, which has great potential of being used for diagnosis of trypanosomosis in the laboratory and the field.
138 citations
TL;DR: Loop-mediated isothermal amplification could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis by using primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections.
80 citations
TL;DR: Results indicated that AMA1 is a cross-reactive antigen between N. caninum and T. gondii and a potential common vaccine candidate to control two parasites.
68 citations
TL;DR: It is suggested that a phylogenetic analysis of the hsp70 gene sequence might be helpful in classifying Babesia and Theileria species, and that canine babesial isolates might be closely related to each other, indicating their evolution from the same ancestry.
35 citations
TL;DR: An unknown trypanosome species isolated from naturally infected Haemaphysalis hystricis ticks is reported on and a phylogenetic tree constructed with the 18S rRNA gene indicates that Trypanosoma KG1 is a member of the stercorariantrypanosomes.
Abstract: Common arthropod vectors for trypanosomes are flies, fleas and bugs. This study reports on an unknown trypanosome species isolated from naturally infected Haemaphysalis hystricis ticks, hereby, referred to as Trypanosoma KG1 isolate. The parasite has been successfully cultured in vitro with L929 or HEK 293T cell line as feeder cells. This trypanosome cannot survive in vitro without feeder cells. Following experimental infections of ticks, the trypomastigote-like and the epimastigote-like forms of this trypanosome could be detected by Giemsa-stained smears of the midgut and salivary glands of Ornithodoros moubata ticks which were made to feed on a culturing medium containing Trypanosoma KG1 isolate through an artificial membrane. Trypanosoma KG1 isolate could also be detected from Giemsa-stained smears of the haemolymph up to 30 days post-inoculation into the O. moubata haemocoel. Trypanosoma KG1 isolate cannot be propagated in laboratory animals including mice, rats, rabbits and sheep. A phylogenetic tree constructed with the 18S rRNA gene indicates that Trypanosoma KG1 is a member of the stercorarian trypanosomes.
32 citations
TL;DR: Results indicate that P0 is a cross-reactive antigen between N. caninum and T. gondii and a potential common vaccine candidate to control both parasites.
31 citations
TL;DR: This is the first report of tick myosin alkali light chain and the function of this protein is discussed, and it is proved Hq02 protein was a "concealed" antigen.
21 citations
TL;DR: In this article, a primary cDNA library with a size of 1.34 × 106 PFU was constructed from Haemaphysalis qinghaiensis eggs and was immunoscreened with rabbit anti-Hq22 serum.
Abstract: A primary cDNA library with a size of 1.34 × 106 PFU was constructed from Haemaphysalis qinghaiensis eggs and was immunoscreened with rabbit anti-H. qinghaiensis serum. One clone (Hq22, named following those clones obtained from adult Haemaphysalis qinghaiensis cDNA library which we constructed before) screened from the cDNA library was selected randomly for sequencing. The entire sequence of the clone was subsequently obtained using rapid amplification of the cDNA ends (RACE). A search of the cloned sequence against GenBank revealed that it related to ribosomal protein L23a (Rpl23a) and had a high percentage similarity to this protein from different species. Conserved domains for Rpl23a were also identified in the cloned sequence. Expression analysis by RT-PCR showed that this gene is expressed in salivary glands, midguts, other tissues and different developmental stages of H. qinghaiensis. Based on the H. qinghaiensis Rpl23a sequence, open reading frames (ORF) of Rpl23a of Heamaphysalis longicornis and Boophilus microplus were also cloned and were performed for comparison with Rpl23a of H. qinghaiensis and other organisms as well. Vaccine based on Rpl23a recombinant protein cannot protect sheep against H. qinghaiensis.
20 citations
TL;DR: LAMP is introduced as an alternative molecular diagnostic tool that could be used in large-scale epidemiological surveys and is shown to be effective in detecting Theileria spp.
Abstract: The sensitivity of LAMP, PCR and microscopy to detect Theileria spp. and Trypanosoma congolense in field-derived bovine blood samples from Tanzania was evaluated and compared. No parasites were detected by microscopy. Furthermore, no bovine Theileria spp. were detected by LAMP and PCR from all the 24 samples collected from Arusha. Four and one out of 24 samples were positive for Theileria congolense infection by LAMP and PCR respectively while, 18 and nine out of 40 samples from Dar es Salaam were positive by LAMP and PCR for Theileria spp. Infection, respectively. Although all samples from Dar es Salaam were negative for Trypanosoma congolense infections by PCR, 12 out of 40 samples were LAMP positive. Whilst PCR is an established gene amplification method for the detection of Theileria and trypanosome parasites, this study introduces LAMP as an alternative molecular diagnostic tool that could be used in large-scale epidemiological surveys.
15 citations
TL;DR: Exogenous TGF-β1 is immunostimulative, inducing partial protection against T. congolense infection, possibly through mechanisms involving innate immune responses, as well as activating macrophages that may contribute to parasite control.
Abstract: The socioeconomic implications of trypanosomosis in sub-Saharan Africa and the limitations of its current control regimes have stimulated research into alternative control methods. Considering the pro- and anti-inflammatory properties of transforming growth factor β1 (TGF-β1) and its potential to enhance immunity against protozoan parasites, we examined the effects of intraperitoneally delivered TGF-β1 in C57BL/6 mice infected with Trypanosoma congolense, the hemoprotozoan parasite causing nagana in cattle. A triple dose of 10 ng TGF-β1 significantly reduced the first parasitemic peak and delayed mortality of infected mice. Furthermore, exogenous TGF-β1 significantly decreased the development of trypanosome-induced anemia and splenomegaly. The apparent TGF-β1-induced antitrypanosome protection, occurring mainly during the early stage of infection, correlated with an enhanced parasite antigen-specific Th1 cell response characterized by a skewed type I cytokine response and a concomitant stronger antitrypanosome immunoglobulin G2a antibody response. Infected TGF-β1-pretreated mice exhibited a significant reduction in the trypanosome-induced hyperexpansion of B cells. Furthermore, evidence is provided herein that exogenous TGF-β1 activates macrophages that may contribute to parasite control. Collectively, these data indicate that exogenous TGF-β1 is immunostimulative, inducing partial protection against T. congolense infection, possibly through mechanisms involving innate immune responses.
12 citations
01 Jan 2007
TL;DR: In this article, the authors used loop-mediated isothermal amplification (LAMP) primers for detection of Theileria equi and Babesia caballi infections, and the results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis.
Abstract: Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10 6 dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T. equi was amplified by a T. equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T. equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T. equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis. # 2006 Elsevier B.V. All rights reserved.
TL;DR: The present study conducted by using suppressive subtractive hybridization (SSH) technique to seek the highly expressed genes especially in host indicates that no trypanosome genes could be differentially expressed either the in vivo or in vitro propagatedtrypanosomes.
Abstract: With the hypothesis that African trypanosomes could have in vivo specific genes for adaptation to host’s environment, the present study was conducted by using suppressive subtractive hybridization (SSH) technique to seek the highly expressed genes especially in host. A total of 328 clones from the in vivo SSH library and that of 160 clones from the in vitro SSH library were analyzed in order to determine their expression levels, but none of the above-mentioned genes showed differential expression. This indicates that no trypanosome genes could be differentially expressed either the in vivo or in vitro propagated trypanosomes. Alternatively, there might be limitation for detecting specifically expressed genes in African trypanosomes using this method, because of their polycistronic gene expression. Key Words : differential expression; infection; Trypanosoma brucei The development of axenic culture methods for both bloodstream form (BSF) and procyclic form (PCF) of African trypanosomes (Hirumi and Hirumi 1989; Hirumi and Hirumi 1991) has contributed to further investigation of African trypanosomes. However, this invention has given rise to a question of whether biological characteristics of