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1.
锥虫抗药性机制的研究进展   总被引:1,自引:0,他引:1  
为了较全面地了解锥虫抗药性机制的研究进展,综述了国内外有关报道。锥虫抗药性形成的机制主要表现在三方面,即锥虫体内酶的活性(或含量)发生变化,抗药株虫体的苹果酸酶、锥虫硫酮还原酶或鸟氨酸脱羧酶等酶的活性明显升高;锥虫细胞膜上的特异载体,即细胞膜上转运腺苷酸和腺苷的P2载体发生改变,药物进入虫体受阻;虫体的抗药基因被激活或基因发生重组、突变或扩增。本文还总结了锥虫抗药性机制研究中存在的问题及今后的研究  相似文献   

2.
A direct card agglutination test for Trypanosoma evansi, CATT/T. evansi based on the predominant variable antigen-type (pVAT) RoTat 1.2 was evaluated previously in the field in Isiolo District, Kenya. Sixteen out of 51 (31.4%) parasitologically positive camels were negative by the antibody detection test. In the present study, trypanosomes isolated from the camels were analysed in an attempt to determine the cause of the false negative results of CATT/T. evansi. A total of 20 field isolates comprised 16 stocks from camels that were negative by CATT/T. evansi, and 4 from CATT/T. evansi-positive camels. In addition, 15 known T. evansi and four T. brucei were used as reference. Purified DNA samples were tested using an established RoTat 1.2-based polymerase chain reaction (PCR) that yields a 488 bp product for the specific detection of T. evansi. Antibodies to RoTat 1.2 variant surface glycoprotein (VSG) were used in Western blotting to detect RoTat 1.2 VSG linear epitopes. Results of PCR and Western blot showed that the 16 stocks isolated from CATT/T. evansi-negative camels fell into three groups. In Group 1, both the RoTat 1.2 VSG gene and the VSG were absent in three stocks. In five trypanosome stocks in Group 2, the RoTat 1.2 VSG gene was detected, but Western blot was negative indicating absence of the expressed VSG. Five other stocks containing the RoTat 1.2 VSG gene were also in this group. The RoTat 1.2 VSG gene was detected and Western blot was positive in all four trypanosome stocks in Group 3. All four stocks from CATT/T. evansi-positive camels contained the RoTat 1.2 VSG gene and the expressed VSG. The reference T. evansi KETRI 2479 lacked the RoTat 1.2 VSG gene and there was no immune reactivity detected by Western blot. The rest of the reference T. evansi stocks examined contained the RoTat 1.2 VSG gene. All the four T. brucei samples examined were negative by PCR and Western blot. In conclusion, this study showed that the RoTat 1.2 VSG gene was absent from some T. evansi trypanosomes in Kenya.  相似文献   

3.
The first outbreak of trypanosomosis caused by Trypanosoma evansi in camels in France was reported on a farm in the Aveyron Department. Five camels were imported from the Canary Islands to the farm in early July 2006, and trypanosomes were observed on a stained blood smear from one of them, which died in October. On further investigations, trypanosomes were observed in the blood of five camels, three of them indigenous to the farm and two that had been imported. On the basis of microscopical examination (morphological criteria and measurements) and serological results based on the card agglutination T evansi test and PCR typing, the parasites were identified as T evansi. After treatment with melarsomine, the infected camels rapidly became negative by parasitological tests and were negative two to four months later by serological tests. The parasite was probably transmitted by tabanids and Stomoxys calcitrans, which were abundant in July to September 2006. No parasites were observed in other animals on the farm or on neighbouring farms, but some of the sheep on these farms were positive by PCR or serology.  相似文献   

4.
Ten isolates of Trypanosoma evansi from the Pantanal region of Brazil, recently derived from coati (Nasua nasua, carnivora, Procyonidae), horses and dogs, were characterized on the basis of biological (experimental infections in Wistar rats) and biochemical (multilocus enzyme eletrophoresis) data. Biological data were analyzed by Nested analysis of variance and Kruskal-Wallis. Marked heterogeneity in virulence was observed in the isolates. Some of the isolates showed an undulating parasitaemia, typical for African trypanosomes. This biological heterogeneity did not correspond with the biochemical homogeneity observed in the T. evansi isolates. T. evansi has one of the widest distributions and greatest range of mammalian hosts and is widely recognized to have evolved from Trypanosoma brucei. Adaptability of T. evansi was not reflected in the variability of biochemical and molecular parameters studied to date. The variability in virulence was very significant, but not correlated with the host from which it was derived. These data suggested that, in the region studied, T. evansi is transmitted among both domestic and sylvatic animals in one single transmission cycle.  相似文献   

5.
The development and distribution of Trypanosoma congolense, T vivax and T brucei in the skin of goats was examined after the animals were bitten by infected Glossina morsitans centralis. Following the tsetse bite, the trypanosomes in the skin multiplied, reaching maximum numbers when the skin reaction (chancre) of the host attained its maximum size. In goats infected with T vivax and T brucei, trypanosomes were observed circulating in the blood before the peak of the chancre, while in T congolense-infected goats microscopically detectable parasites were found in blood only during the decline of the chancre. In contrast to T vivax, large numbers of T congolense and T brucei parasites were found in the skin following tsetse-transmitted infection. Ultrastructural differences were observed in T congolense and T brucei indicating an intracutaneous transformation from metacyclic to blood stream forms. T congolense forms in the skin reactions had a well developed secretory reticulum, small mitochondria and lacked large lipid inclusions compared to metacyclic and blood stream forms. The intracutaneous forms of T brucei had smaller mitochondria, the glycosomes were of more uniform size and the rough endoplasmic reticulum was less developed than in metacyclic or blood stream forms.  相似文献   

6.
In Venezuela, two non-tsetse transmitted trypanosomes, Trypanosoma evansi and Trypanosoma vivax, are the major etiological agents of animal trypanosomosis. Rodents can be experimentally infected with T. evansi in order to obtain enough parasites to prepare antigens for serological tests. On the contrary, the production of T. vivax antigens is a limiting factor in most laboratories. Since T. evansi and T. vivax have exhibited a very high immunological cross-reactivity, we have focused on the identification of antigens from T. evansi responsible for this phenomenon. The predominant 64 kDa glycosylated cross-reacting antigen was recently purified from the TEVA1 T. evansi Venezuelan isolate [Parasitology 124 (2002) 287]. Here, we purified two additional cross-reacting antigens with molecular masses of approximately 51 and 68 kDa from the cytosolic fraction of the same T. evansi isolate, by sequential chromatography on DEAE-sepharose and sephacryl S-300. Sera obtained from animals infected with T. evansi or T. vivax recognized both purified proteins, suggesting their potential use as diagnostic reagents.  相似文献   

7.
伊氏锥虫和布氏锥虫动基体DNA酶切电泳比较   总被引:2,自引:0,他引:2  
限制性内切酶MboI,DdeI,Hinfi和TaqI对布氏锥虫KDNA进行酶切后,电泳中均显示出多条DNA区带,其总Kb数约等于20kb,而各限制酶对伊氏锥虫KNDA消化后均显示出1至2条区带,总和约1kb明显区别于布氏锥虫。  相似文献   

8.
伊氏锥虫同工酶、蛋白质和抗原组分的比较研究   总被引:2,自引:0,他引:2  
本文采用生化技术对八个中国伊氏锥虫株及一个布氏锥虫株的同工酶、蛋白质和抗原组分进行比较研究。根据同工酶电泳结果,可将伊氏锥虫和与其形态上不能区分的布氏锥虫区别开来,亦可将伊氏锥虫分为两个酶株群(Z1、Z2)。根据SDS-聚丙烯酰胺凝胶电泳、等电聚焦电泳和免疫印迹试验的结果,可将酶株群Z1分成五个不同多肽群(株)。本研究结果表明,中国伊氏锥虫遗传变异程度较低,是一个相对稳定的种群。  相似文献   

9.
Blood stream forms of drug-resistant and sensitive Trypanosoma brucei brucei, Trypanosoma brucei evansi and Trypanosoma vivax were incubated in a liquid medium for 24 h at 37 degrees C in the presence of various concentrations of diminazene aceturate (Berenil) or isometamidium chloride (Samorin), and assayed for infectivity in mice. Whereas the infectivity to mice of all Samorin-sensitive trypanosomes was decreased after incubation with 1 ng Samorin ml-1, the Samorin-resistant stocks remained infective for mice. Two of the Samorin-resistant stocks remained infective after incubation with Samorin concentrations of up to 50 ng ml-1. The infectivity of Berenil-resistant trypanosome stocks were also retained after incubation with drug concentrations (0.5 or 1.0 micrograms ml-1) which otherwise inhibited the infectivity of Berenil-sensitive trypanosome stocks. In addition, differences in infectivity were observed when Berenil-resistant and sensitive trypanosome stocks were incubated in medium supplemented with serum from goats previously treated with Berenil. Thus, drug-resistant and sensitive trypanosomes can be clearly distinguished using the drug incubation infectivity test.  相似文献   

10.
作者观测不同保护剂、稀释液、PH值、降温方法、复苏温度、冻融次数、液相气相交替以及解冻后在普通冰瓶中存放时间对伊氏锥虫浙江虫浙江虫株的感染性及致病力的影响。在此试验基础上,又对伊氏锥虫的6个不同虫株、媾疫锥虫、铡果锥虫、布氏锥虫等4个种的9个早株,进行了超低温保藏试验和长期保藏效果观察。已测定的有效保藏期伊氏锥虫达574-3200天,媾疫锥虫达2866天,则果锥虫达763天,布氏锥虫783天。通过  相似文献   

11.
A card agglutination test (CATT/T. evansi) was evaluated for detection of antibodies against Trypanosoma evansi (T. evansi) in experimentally and naturally infected buffaloes. Four calves were inoculated with a strain of T. evansi isolated from a dromedary camel. Parasitological examination of the calves revealed trypanosomes in the blood from days 4 to 9 post-inoculation (PI). General emaciation appeared from day 26 PI and aggravated until the end of the experiment (day 88 PI). Antibodies against T. evansi were detectable from day 8 PI till the end of the experiment. Parasitological examination of 200 water buffalo blood samples obtained from slaughterhouses revealed negative results. Serological examination of these animals showed that 48 (24%) water buffaloes had anti-T. evansi antibodies.  相似文献   

12.
The variable surface glycoprotein of Trypanosoma evansi RoTat 1.2 variable antigen type (VAT) is used as an antigen in different antibody detection assays for T. evansi. To obtain more information on the predominant character of RoTat 1.2 and its diagnostic potential in antibody detection tests, we checked its expression in 10 different T. evansi stocks and clones from different parts of the world. Cryostabilates were injected into mice and the trypanosomes of the first peak parasitaemia were screened for the presence of RoTat 1.2 by VAT specific immunofluorescence. To monitor the appearance of RoTat 1.2 specific antibodies during infection, rabbits were infected and serologically tested at different time intervals with VAT specific immune trypanolysis, CATT/T. evansi, LATEX/T. evansi and ELISA/T. evansi.Test results confirm the predominant character of RoTat 1.2.  相似文献   

13.
Teneral tsetse flies infected with either Trypanosoma brucei or T. vivax were fed on healthy cattle. Blood samples collected daily from the cattle were examined by microscopy for the presence of trypanosomes, in thick smear, thin smear and in the buffy coat (BC). All the cattle fed upon by infected tsetse developed a fluctuating parasitaemia. DNA was extracted from the blood of these cattle and subjected to polymerase chain reaction (PCR) using oligonucleotide primers specific for T. brucei or T. vivax. The PCR products unique to either T. brucei or T. vivax were identified following amplification of DNA from the blood samples of infected cattle, whereas none was detectable in the DNA from the blood of the cattle exposed to non-infected teneral tsetse. In a concurrent set of experiments, one of the oligonucleotide primers in each pair was biotinylated for use in PCR-ELISA to examine all the blood samples with this assay. Both the PCR and the PCR-ELISA revealed trypanosome DNA in 85% of blood samples serially collected from the cattle experimentally infected with T. brucei. In contrast, the parasitological assays showed trypanosomes in only 21% of the samples. In the blood samples from cattle experimentally infected with T. vivax, PCR and PCR-ELISA revealed trypanosome DNA in 93 and 94%, respectively. Microscopy revealed parasites in only 63% of the BCs prepared from these cattle. Neither PCR nor PCR-ELISA detected any trypanosome DNA in blood samples collected from the animals in the trypanosome-free areas. However, both assays revealed the presence of trypanosome DNA in a number of blood samples from cattle in trypanosomosis-endemic areas.  相似文献   

14.
Trypanosoma evansi was diagnosed for the first time in camels in the Canary Islands in 1997. Several sanitary measures including treatment of infected animals were taken; however, nowadays a little area is still infected. In order to determine possible reservoirs 138 wild rodents were trapped, 64 of them in the infected farms and the remaining 74 in other areas. The captured species were Rattus rattus (24), Rattus norvegicus (69) and Mus musculus domesticus (45). Serological (CATT/T. evansi), parasitological (micro-Hematocrit Centrifugation technique and stained smears) and molecular (PCR) methods for T. evansi and T. lewisi were used as diagnostic methods. None of the examined rodents was positive for T. evansi; 18, however, showed motile trypanosomes at micro-Hematocrit Centrifugation technique and resulted positive for T. lewisi by PCR. The results would suggest that the studied rodent species would not play a relevant role in the epidemiology of T. evansi infection in Canaries.  相似文献   

15.
The role played by domestic animals in the transmission of gambiense Human African Trypanosomosis remains uncertain. Northwest Uganda is endemic for Trypanosoma brucei gambiense. Of the 3267 blood samples from domestic animals in four counties examined by hematocrit centrifugation technique (HCT), 210 (6.4%) were positive for trypanosomes. The prevalence of animal trypanosomosis was estimated at 13.8% in Terego County, 4.2% in East Moyo County, 3.1% in Koboko County, and zero in West Moyo County. The trypanosome infection rates varied from 0.2% in goats, 3.5% in dogs, 5.0% in sheep, 7.5% in cattle, to 15.5% in pigs. DNA was extracted from the blood samples by Chelex method, Sigma and Qiagen DNA extraction Kits. A total of 417(12.8%) DNA samples tested positive by polymerase chain reaction (PCR) using T. brucei species specific primers (TBR) indicating that the DNA was of Trypanozoon trypanosomes while 2850 (87.2%) samples were TBR-PCR negative. The T. brucei infection rates based on TBR-PCR were highest in pigs with 21.7%, followed by cattle (14.5%), dogs (12.4%), sheep (10.8%), and lowest in goats with 3.2%, which indicated that pigs were most bitten by infected tsetse than other domestic animals. TBR-PCR detected 6.3% more infected domestic animals that had been missed, and confirmed the 6.4% cases detected by HCT in the field. Statistical analysis done using one-way ANOVA Kruskal-Wallis test (Prism version 5.0) showed no significant difference in trypanosome infections among domestic animals using both HCT and TBR-PCR techniques in the different counties (Confidence Interval of 95%, p-values >0.05). All the 417 trypanosome DNA samples were negative by PCR using two sets of primers specific for the T. b. gambiense specific glycoprotein gene and serum resistance associated gene of T. b. rhodesiense, indicating that they were probably not from the two human infective trypanosomes. Polymerase chain reaction using primers based on ribosomal internal transcribed spacer-1 region (ITS-PCR) resolved the 417 DNA of trypanosome samples into 323 (77.5%) as single trypanosome infections due to T. brucei and 39 (9.4%) mixed infections but missed detecting 55 (13.1%) samples, possibly because of the low sensitivity of ITS-PCR as compared to TBR-PCR. The 31 mixed infections were due to T. brucei (T.b) and T. vivax (T.v); while 8 mixed infections were of T. congolense (T.c) and T. brucei but no mixed trypanosome infections with T. congolense, T. brucei, and T. vivax were detected. Statistical analysis done using one way ANOVA Kruskal-Wallis test (Prism version 5.0) to compare single and mixed trypanosome infections showed no significant difference in trypanosome infections due to single (T.v, T.b, T.c) and mixed (T.v+T.b; T.v+T.c; T.b+T.c; T.v+T.b+T.c) trypanosome species among domestic animals in the different counties using ITS-PCR technique (Confidence Interval of 95%, p-values >0.05). It was concluded that domestic animals in northwest Uganda were probably not reservoirs of T. b. gambiense and there was no infection, as yet, with T. b. rhodesiense parasites.  相似文献   

16.
According to several authors, Trypanosoma evansi is a monomorphic trypanosome found exclusively in slender intermediate forms, although additional studies have revealed that many strains present stumpy forms on rare occasions. In a recent T. evansi outbreak in mainland Spain, several atypical forms were observed in blood smear examinations. Molecular procedures were then necessary to confirm the causal agent. Morphological and biometric measures were taken to characterize the different forms of T. evansi. In contrast to published information, the results of this study would indicate that biometrically distinct T. evansi could also be found in the same farm and even in the same animal species. These data could be useful for many trypanosomes endemic areas of the world where molecular methods are not commonly available.  相似文献   

17.
伊氏锥虫TR酶的初步研究   总被引:4,自引:3,他引:1  
国外研究者已报道在布氏锥虫、枯氏锥早、刚果锥虫中发现了存在TR酶,此酶是锥虫代谢的重要酶之一。伊氏锥虫TR酶研究的尚属空白。本研究目的是为了确定伊氏锥虫中是存在TR酶。我们利用分光光度法测出了伊氏锥虫中有TR酶活性,但无GR酶活性,测出了TR酶的Km和Vm值;发现了治疗锥虫药物安锥赛和盐酸锥双对TR酶有抑制作用,抑制率分别达28%和31%;比较了七株中国伊氏锥虫和布氏锥虫敏毫克蛋白中TR酶活力,其  相似文献   

18.
Blood samples from 3000 Somali camels (Camelus dromedarius) were examined for trypanosome infection. Of these, 160 (5.33%) were infected with Trypanosoma evansi, one (0.03%) with T. congolense and one (0.03%) with T. brucei. Camel trypanosomiasis occurred in most areas of tabanid infestation throughout the country. The tabanids Philoliche zonata and P. magretti are incriminated as the major vectors of the disease.  相似文献   

19.
用小鼠治疗试验和体外药敏试验观察了4个布氏锥虫伊氏亚种虫株在长期体外培养条件下药敏性的稳定性.各虫株的原始群体、连续培养30d和90d的群体对贝尼尔、苏拉灭、安锥赛和硫胂聚氰胺的敏感性基本相同,说明连续培养90d各虫株对上述4种抗锥虫药的敏感性无明显改变.  相似文献   

20.
以家畜伊氏锥虫弱毒株免疫家兔12只,然后进行攻虫试验。其中11只被保护,1只发病死亡。以BA-ELISA检测家兔免疫后及攻虫后特异性IgG、IgM的变化,发现免疫后IgG、IgM均于第21d达到峰值;攻虫后IgM于第6d达到峰值,IgG于第9d达到峰值。攻虫后,随着免疫组家兔体内特异性IgG、IgM的不断上升,其血液内的锥虫也同时被清除。体外中和试验也表明,特异性抗体对锥虫的感染力具有明显的抑制作用。  相似文献   

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