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Phosphoenolpyruvate carboxylase(PEPC)is essential for the glycolytic pathway and parasite proliferation in Babesia gibsoni
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作者 Dong‑Fang Li Ling‑Na Wang +7 位作者 Yi‑Dan Bai Yu‑Xin Yu Xing Lu Xing‑Ai Guan Fang‑Jie Li Sen Wang Lan He Jun‑Long Zhao 《Animal Diseases》 2025年第3期343-354,共12页
Apicomplexan parasites predominantly generate ATP and lactic acid through glycolysis and anaerobic glucose metabolism,incorporating CO_(2) into glycolysis via a stage-dependent phosphoenolpyruvate carboxylase(PEPC)mec... Apicomplexan parasites predominantly generate ATP and lactic acid through glycolysis and anaerobic glucose metabolism,incorporating CO_(2) into glycolysis via a stage-dependent phosphoenolpyruvate carboxylase(PEPC)mechanism.Although the role of PEPC in plant and bacterial carbon fixation is well documented,its function within Babesia remains largely unexplored.This study employs reverse genetics to probe the biological role of PEPC in Babesia gibsoni,noting its conservation across similar protozoa,suggesting a pivotal and conserved biological function.Western blotting and immunofluorescence(IFA)experiments using the BgPEPC-3×Flag strain revealed that the BgPEPC protein has a molecular weight of 105 kDa and localizes predominantly to the cytoplasm.Attempts to knock out the PEPC gene in BgPEPC-3×Flag strains failed under standard media conditions,succeeded only with the addition of 5 mM malate,an upstream metabolite of oxaloacetic acid(OAA).In addition to malate,the downstream metabolite of OAA can also partially compensate for the phenotypic defects caused by PEPC deficiency.This intervention alleviated severe growth deficits,underscoring the critical role of aspartate in the parasite lifecycle.Moreover,metabolic inhibitors such as L-cycloserine and triazamidine,which target aspartate aminotransferase and mitochondrial functions,respectively,demonstrated increased efficacy against BgPEPC knockout strains.The lack of a compensatory response to malic acid supplementation underscores the integral role of BgPEPC in intermediary carbon metabolism and its necessity in providing aspartate as a precursor to pyrimidine synthesis.Collectively,these findings suggest that PEPC could be a potential target for future drug development against B.gibsoni infections. 展开更多
关键词 babesia gibsoni PHOSPHOENOLPYRUVATE Phosphoenolpyruvate carboxylase MALATE ASPARTATE
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Pyronaridine combined with diminazene aceturate inhibits Babesia in vitro and in vivo
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作者 Shimaa Abd El-Salam El-Sayed Mohamed Z.Sayed-Ahmed +5 位作者 Shaimaa Ahmed Awad Ali Nourah Alsadaan Nawazish Alam Mahmoud S.Alkhoudary Ikuo Igarashi Mohamed Abdo Rizk 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2024年第3期93-99,共7页
Objective:To evaluate the combination therapy of pyronaridine tetraphosphate and diminazene aceturate against Babesia in vitro and in vivo.Methods:Bioinformatic analysis was performed using atom pair fingerprints.An i... Objective:To evaluate the combination therapy of pyronaridine tetraphosphate and diminazene aceturate against Babesia in vitro and in vivo.Methods:Bioinformatic analysis was performed using atom pair fingerprints.An in vitro combination test was performed against Babesia bovis and Theileria equi.Moreover,the in vivo chemotherapeutic efficacy of pyronaridine tetraphosphate in combination with diminazene aceturate was investigated against the growth of Babesia microti in mice using a fluorescence inhibitory assay.Results:Pyronaridine tetraphosphate and diminazene aceturate exhibited nearly similar molecular weights.The in vitro combination of pyronaridine tetraphosphate and diminazene aceturate was synergistic on Babesia bovis and additive on Theileria equi.In addition,5 mg/kg pyronaridine tetraphosphate combined with 10 mg/kg diminazene aceturate inhibited Babesia microti growth significantly compared with those observed after treatment with 25 mg/kg diminazene aceturate alone from day 6 post treatment to day 12 post treatment.The combination therapy also normalized the hematological parameters of infected mice.Conclusions:An oral dose of pyronaridine tetraphosphate combined with a subcutaneous dose of diminazene aceturate inhibits Babesia in vitro and in mice,suggesting it might be a new paradigm for the treatment of babesiosis. 展开更多
关键词 babesia Pyronaridine tetraphosphate Diminazene aceturate BABESIOSIS THEILERIA
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在中国新发现的一种羊的巴贝斯虫(Babesia)未定种 被引量:5
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作者 连灿 何虎成 +2 位作者 白启 韩根凤 刘光远 《中国兽医学报》 CAS CSCD 北大核心 1997年第2期116-119,共4页
在中国甘肃某地自然感染发病绵羊的血液涂片中,观察到一种大型的巴贝斯虫(Babesia)。虫体形态具多形性,有双梨籽形、单梨籽形、圆环形、棒状、不规则形、逗点形和三叶形。典型双梨籽形虫体的大小为1.8~2.5μm×... 在中国甘肃某地自然感染发病绵羊的血液涂片中,观察到一种大型的巴贝斯虫(Babesia)。虫体形态具多形性,有双梨籽形、单梨籽形、圆环形、棒状、不规则形、逗点形和三叶形。典型双梨籽形虫体的大小为1.8~2.5μm×0.9~1.8μm,平均(2.21±0.12)μm×(1.17±0.18)μm。个别红细胞中寄生有4个或8个虫体。该虫种特征性的形态是三叶形。它是相连的双梨籽形虫体两尖端的核质延伸(出芽),并生出原生质,发育成一个完整的梨籽形虫体,与母体共同排列形成三叶形的。该虫种可引起绵羊严重发病和死亡。本研究对自然感染发病羊的临床症状和死亡羊的病理变化进行了观察。该虫种的形态大小和致病力,与莫氏巴贝斯虫(B.motasi)和粗糙巴贝斯虫(B.crassa)明显不同。 展开更多
关键词 巴贝斯虫 绵羊 形态学 临床症状 病理变化
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Inhibitory effects of methanolic Olea europaea and acetonic Acacia laeta on growth of Babesia and Theileria 被引量:2
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作者 Amany Magdy Beshbishy Gaber El-Saber Batiha +2 位作者 Oluyomi Stephen Adeyemi Naoaki Yokoyama Ikuo Igarashi 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2019年第9期425-434,共10页
Objective:To evaluate the antipiroplasmic activities of methanolic extract of Olea europaea(MOE)and acetonic extract of Acacia laeta(AAL)against Babesia and Theileria parasites in vitro and evaluate the chemotherapeut... Objective:To evaluate the antipiroplasmic activities of methanolic extract of Olea europaea(MOE)and acetonic extract of Acacia laeta(AAL)against Babesia and Theileria parasites in vitro and evaluate the chemotherapeutic effects of these extracts against Babesia(B.)microti in vivo.Methods:Fluorescence assay using SYBR Green 1 nucleic acid stain was used to detect inhibitory effects of the two extracts as well as the combination effects of the two extracts with diminazene aceturate and atovaquone on four Babesia species and Theileria equi in vitro while for in vivo experiments,8-weekold female BALB/c mice were injected intraperitoneally with 1× 107 B.microti-iRBCs and treated orally at a dose of 150 mg/kg of both extracts.Results:The half maximal inhibitory concentration(IC50)values of AAL against B.bovis,B.bigemina,B.divergens,B.caballi,and Theileria equi were lower than those of MOE extracts.Toxicity assay on Madin-Darby bovine kidney,mouse embryonic fibroblast(NIH/3T3),and human foreskin fibroblast cell lines showed that MOE and AAL affected only the viability of Madin-Darby bovine kidney cell line with half maximal effective concentrations(EC50)of(794.7±41.9)and(873.9±17.5)μg/mL,respectively.The oral treatments of MOE and AAL at 150 mg/kg inhibited the growth of B.microti in mice by 80.4% and 64.4%,respectively.The MOE and diminazene aceturate combination showed a higher chemotherapeutic effect than that of monotherapy.Conclusions:MOE and AAL have the potential to be an alternative remedy for treating piroplasmosis.Furthermore,the combination therapy of MOE + DA was more potent against B.microti infection in mice than their monotherapies. 展开更多
关键词 OLEA europaea ACACIA laeta babesia THEILERIA In VITRO In vivo Inhibition
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Differential IL10 mRNA Profiles Associated to <i>Babesia bovis</i>and <i>B. bigemina</i>Infection Levels in Persistently Infected Animals 被引量:2
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作者 Talita Barban Bilhassi Rodrigo Giglioti +4 位作者 Cintia Hiromi Okino Wilson Malagó Júnior Henrique Nunes de Oliveira Cíntia Righetti Marcondes Márcia Cristina de Sena Oliveira 《Open Journal of Veterinary Medicine》 2019年第11期161-169,共9页
This work aimed to find quantitative phenotypic traits that can be used to discriminate the levels of resistance/susceptibility to B. bovis and B. bigemina in two groups of cattle presenting the highest (H) or lowest ... This work aimed to find quantitative phenotypic traits that can be used to discriminate the levels of resistance/susceptibility to B. bovis and B. bigemina in two groups of cattle presenting the highest (H) or lowest (L) infection levels and Rhipicephalus microplus ticks count. The animals were selected from a previous study of 50 Canchim (5/8 Charolais/zebu) heifers raised in an endemic area for these parasites. These animals were evaluated regarding their TNFα, IL10, IFN-γ, IL12 and iNOS mRNA levels. No differences were found between these groups regarding TNFα, IFN-γ, IL12β or iNOS transcripts. However, the IL10 transcripts were significantly higher in the H group compared to the L group. Moreover, significant correlation coefficients were observed between B. bovis loads and both IL10 and IFN-γ transcripts, while no correlations were found for B. bigemina loads and all tested immune-related transcripts, suggesting that differential IL10 mRNA profiles were closely associated to B. bovis loads. Our results have contributed to a better understanding of the immune responses against Babesia infection, as we demonstrated that the IL10 cytokine levels might also influence or be influenced by parasitemia levels in persistently infected animals. 展开更多
关键词 babesia BOVIS Cytokines Gene Expression IL10 RT-qPCR
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Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab 被引量:1
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作者 Samreen Zulfiqar Sadia Shahnawaz +9 位作者 Muhammad Ali Arif Mahmood Bhutta Shahid Iqbal Sikandar Hayat Shazia Qadir Muhammad Latif Nazia Kiran Ali Saeed Muhammad Ali Furhan Iqbal 《Asian Pacific Journal of Tropical Biomedicine》 SCIE CAS 2012年第2期104-108,共5页
Objective:To determine the presence of Babesia bovis(B.bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals.Methods:Blood samples were collected fr... Objective:To determine the presence of Babesia bovis(B.bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals.Methods:Blood samples were collected from 144 large ruminants,including 105 cattle and 39 buffaloes,from six districts in southern Punjab including Multan,Layyah,Muzaffar Garh,Bhakar,Bahawalnagar and Vehari.Data on the characteristics of animals and herds were collected through questionnaires.Different blood(hemoglobin,glucose) and serum(ALT,AST,LDH,cholesterol)parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B.bovis on the blood and serological profile of infected animals.Results:27 out of 144 animals.from 5 out of 6 sampling districts,produced the541-bp fragment specific for B.bovis.Age of animals(P=0.02).presence of ticks on animals(P=0.04)and presence of ticks on dogs associated with herds(P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area.ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle.Conclusions:This study has reported for the first time the presence of B.bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output. 展开更多
关键词 babesia bovis PCR amplification CATTLE HEMATOLOGY Serum biochemistry RUMINANT BABESIOSIS Serological profile CATTLE
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Inhibition of <i>Ehrlichia canis</i>and <i>Babesia canis</i>transmission among ticks fed together on dogs vaccinated with Bm86 antigen 被引量:1
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作者 Alina Rodríguez-Mallon Gervasio H. Bechara +5 位作者 Rosangela M. Zacarias Efrain Benavides-Ortiz Jose Luis Soto-Rivas Arlen Patricia Gómez-Ramírez Javier Andrés Jaimes-Olaya Mario Pablo Estrada-García 《Open Journal of Animal Sciences》 2013年第3期24-32,共9页
GAVAC (Heber Biotec S.A, Havana, Cuba) is a commercially available vaccine developed with the Rhipicephalus (Boophilus) microplus Bm86 recombinant antigen. Bm86 is a “concealed” antigen that is present in the plasma... GAVAC (Heber Biotec S.A, Havana, Cuba) is a commercially available vaccine developed with the Rhipicephalus (Boophilus) microplus Bm86 recombinant antigen. Bm86 is a “concealed” antigen that is present in the plasmatic membrane of tick gut epithelial cells with unknown function so far. It is well known that after vaccination in the last fifteen years in Cuba, there was a significant decrease of babesiosis (Babesia bovis and Babesia bigemina) and anaplasmosis (Anaplasma marginale) in cattle. A reduced transmission capacity of ticks fed on tick-immune animals and humans has been reported for several tick-borne pathogens. Recent experiments have demonstrated that an anti-tick vaccine may contribute to the control of tick-borne pathogens not only by decreasing the exposure of susceptible hosts to ticks, but also by reducing the vector capacity of ticks. In this study, the potential of Bm86 vaccination to interfere with pathogen transmission among ticks was evaluated by using as experimental model the brown dog tick Rhipicephalus sanguineus and the tickborne Babesia canis and Ehrlichia canis pathogens. Dogs, vaccinated and not vaccinated, were infested with pathogen-infected ticks and noninfected nymphs of R. sanguineus. After feeding, the pathogen transmission to newly molted adults from co-feeding uninfected nymphs was studied by conventional PCR and qPCR. Results suggest that the anti-Bm86 antibodies could be able to block the transmission of B. canis and/or E. canis from infected to non-infected ticks. 展开更多
关键词 RHIPICEPHALUS sanguineus EHRLICHIA CANIS babesia CANIS Tick-Borne Diseases DOGS Bm86
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田鼠巴贝虫(Babesia mocroti)的超微结构观察
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作者 陈韶红 蔡玉春 +5 位作者 陈家旭 卢艳 俞彰 张永年 艾琳 周晓农 《中国人兽共患病学报》 CAS CSCD 北大核心 2013年第11期1072-1075,共4页
目的观察田鼠巴贝虫的超微结构,了解田鼠巴贝虫在宿主红细胞中发育的形态变化。方法运用扫描电镜观察田鼠巴贝虫侵入宿主红细胞的过程,运用透射电镜观察田鼠巴贝虫发育的形态变化过程及宿主红细胞形态变化情况。结果扫描电镜下观察到... 目的观察田鼠巴贝虫的超微结构,了解田鼠巴贝虫在宿主红细胞中发育的形态变化。方法运用扫描电镜观察田鼠巴贝虫侵入宿主红细胞的过程,运用透射电镜观察田鼠巴贝虫发育的形态变化过程及宿主红细胞形态变化情况。结果扫描电镜下观察到田鼠巴贝虫裂殖子大小在406~981nm之间,虫体发育过程分为裂殖子、分裂中的裂殖子、滋养体3个阶段,裂殖子是通过红细胞膜上微孔进入宿主红细胞,分裂中的裂殖子可使红细胞变形,滋养体是通过溶解红细胞膜游离红细胞。透射电镜观察到田鼠巴贝虫裂殖子的核膜为双层膜结构,虫体中有核糖体、微管、内质网、线粒体和溶酶体等完整的细胞器,分裂中的裂殖子在侵入后期可见食物空泡,滋养体的外膜和核不规则,胞浆中含有颗粒和空泡,宿主红细胞的电子密度随着虫体的发育而逐渐变稀疏。结论田鼠巴贝虫是含有完整细胞器的单细胞有机体,具有一般细胞所有的基本结构,它能完成多细胞动物所具有的生命机能,并以宿主红细胞中的血红蛋白作为生存氧料。 展开更多
关键词 田鼠巴贝虫 红细胞 超微结构
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田鼠巴贝西虫Babesiamicroti截短型分泌抗原的重组表达及其在诊断上的应用 被引量:4
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作者 黄艳丽 曹杰 +4 位作者 蔺智兵 张厚双 周勇志 胡永浩 周金林 《寄生虫与医学昆虫学报》 CAS 2012年第4期213-219,共7页
田鼠巴贝西虫分泌抗原l(Babesiamicrotisecretedantigen1,BmSA1)是一个重要的诊断抗原候选分子,但全长蛋白的表达量少,也不宜纯化。本研究在此基础上,通过生物信息学分析,选取N-末端信号肽后,C-末端跨膜区前,包含CPSF73-100_C... 田鼠巴贝西虫分泌抗原l(Babesiamicrotisecretedantigen1,BmSA1)是一个重要的诊断抗原候选分子,但全长蛋白的表达量少,也不宜纯化。本研究在此基础上,通过生物信息学分析,选取N-末端信号肽后,C-末端跨膜区前,包含CPSF73-100_C功能区的228个氨基酸所编码基因片段,重组高效表达了一个可溶性截短型抗原,解决了完整蛋白重组表达纯化的困难。纯化的截短型抗原作为酶联免疫试验(ELISA)诊断抗原,可清晰地区分阴性及阳性鼠血清,并与其他病原感染无交叉反应,显示其诊断的特异性。鼠人工感染田鼠巴贝西虫系列血清检测表明,该抗原可检测早期感染(5天)和感染(55天)以后的样本,显示其对不同时期感染的宿主均具有诊断价值。结果表明,重组B.microti截短型抗原可作为一种诊断制剂检测田鼠巴贝西虫抗体。 展开更多
关键词 田鼠巴贝西虫 截短型抗原BmSAl 克隆 表达 诊断
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Babesia infection in cattle and dogs in Suizhou City,Hubei Province,China
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作者 Guandu Wu Xiaofan Zhou +11 位作者 Fang Guo Jiao Xu Jingjing Song Zhen Jin Huijie Cao Ju Tang Huiya Lu Zezheng Jiang Tianmei Yu Xiaoyong Zhang Xiaohui Liu Xue-jie Yu 《Infectious Medicine》 2025年第1期43-50,共8页
Background Babesiosis is a zoonotic disease caused by the intraerythrocytic parasite Babesia,which poses a serious threat to public health.Currently,the prevalence of babesiosis in domestic animals and the genetic div... Background Babesiosis is a zoonotic disease caused by the intraerythrocytic parasite Babesia,which poses a serious threat to public health.Currently,the prevalence of babesiosis in domestic animals and the genetic diversity of Babesia in Central China have not been comprehensively studied.Methods In this study,we collected 1093 ticks,including 95.24%(1041/1093)Haemaphysalis longicornis,4.67%(51/1093)Rhipicephalus microplus,and 0.09%(1/1093)Ixodes sinensis.Blood samples from 216 goats,56 cattle,and 25 dogs were collected from Suizhou City,Hubei Province,China,and animal blood DNA was extracted for the detection of Babesia with PCR.Results PCR results showed that 50.00%(28/56)of cattle and 32.00%(8/25)of dogs were Babesia-positive,including for Babesia bovis 3.57%(2/56),B.bigemina 3.57%(2/56),and B.ovata 42.86%(24/56)in cattle and B.gibsoni 32.00%(8/25)in dogs.All goats(216)and ticks(1093)were Babesia-negative.Conclusions Our findings showed that Babesia infections are prevalent in cattle and dogs in Central China,indicating that babesiosis should be monitored in animals and humans in Central China. 展开更多
关键词 babesia CATTLE Goats DOGS TICKS
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Transcriptome study reveals tick immune genes restrict Babesia microti infection
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作者 Tingting Feng Hao Tong +6 位作者 Feihu Zhang Qianqian Zhang Heng Zhang Xia Zhou Hang Ruan Qihan Wu Jianfeng Dai 《Insect Science》 2025年第2期457-470,共14页
A systems biology approach was employed to gain insight into tick biology and interactions between vectors and pathogens.Haemaphysalis longicornis serves as one of the primary vectors of Babesia microti,significantly ... A systems biology approach was employed to gain insight into tick biology and interactions between vectors and pathogens.Haemaphysalis longicornis serves as one of the primary vectors of Babesia microti,significantly impacting human and animal health.Obtaining more information about their relationship is crucial for a comprehensive un-derstanding of tick and pathogen biology,pathogen transmission dynamics,and potential control strategies.RNA sequencing of uninfected and B.microti-infected ticks resulted in the identification of 15056 unigenes.Among these,1051 were found to be differentially expressed,with 796 being upregulated and 255 downregulated(P<0.05).Integrated tran-scriptomics datasets revealed the pivotal role of immune-related pathways,including the Toll,Janus kinase/signal transducer and activator of transcription(JAK-STAT),immunod-eficiency,and RNA interference(RNAi)pathways,in response to infection.Consequently,3 genes encoding critical transcriptional factor Dorsal,Relish,and STAT were selected for RNAi experiments.The knockdown of Dorsal,Relish,and STAT resulted in a substantial increase in Babesia infection levels compared to the respective controls.These findings significantly advanced our understanding of tick–Babesia molecular interactions and pro-posed novel tick antigens as potential vaccine targets against tick infestations and pathogen transmission. 展开更多
关键词 babesia immune signaling pathway TICK TRANSCRIPTOME vector-pathogen interactions
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Babesia microti(Babesiidae,Piroplasmida)infection in a Chinese traveler returning from the United States of America
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作者 Xin-An Huang Rong Xiang +10 位作者 Ru-He Liao Yu-Bo Luan Yi-Lin Zhao Ji-Hu Yang Chun-Feng Luo Lin Huang Luo-Yuan Xia Dai-Yun Zhu Yi Sun Lei Wang Jia-Fu Jiang 《Infectious Diseases of Poverty》 2025年第3期92-97,共6页
Background Human babesiosis,caused by intraerythrocytic protozoa of the genus Babesia(Piroplasmida,Babesiidae),is a globally emerging zoonosis transmitted primarily through Ixodes spp.ticks.Babesia microti,which is en... Background Human babesiosis,caused by intraerythrocytic protozoa of the genus Babesia(Piroplasmida,Babesiidae),is a globally emerging zoonosis transmitted primarily through Ixodes spp.ticks.Babesia microti,which is endemic particularly in the northeastern and midwestern United States,accounts for the majority of globally reported human cases.Recent studies highlight its spread to non-traditional regions and cross-border transmission,driven by climate change,blood transfusions and increased human mobility.Despite increasing reports of autochthonous B.microti infections in certain areas of China,imported cases remain critically underrecognized due to overlapping clinical manifestations with malaria and limited diagnostic awareness.Case presentation We report a diagnostically challenging case of acute B.microti infection in a 52-year-old Chinese woman,presenting with a sudden recurrent fever(39.0–41.0℃),hemolytic anemia(hemoglobin 104 g/L),thrombocytopenia(platelet 78×10^(9)/L)and splenic hypodense lesions on July 11,2023,seven days after returning from a 14-day visit to rural Wisconsin,United States.Peripheral blood smears demonstrated characteristic intraerythrocytic ring forms(parasitemia:7800 organisms/μl)and pathognomonic"Maltese cross"tetrads.Polymerase chain reaction(PCR)targeting the 18S rRNA gene confirmed B.microti infection(GenBank No.PP087232),showing 99.8%identity with the US-type strain Gray(AY693840)and the sequence obtained from a US travel-acquired case in Singapore(MK609547).The patient received intravenous clindamycin(600 mg twice daily),oral dihydroartemisinin(80 mg twice daily),packed red blood cell transfusions,and supportive care,ultimately achieving full recovery after 17 days.Conclusions This study documented the first imported cases of human babesiosis in China,emphasizing the need for heightened clinical and public health vigilance.Screening travelers from endemic areas presenting with fever or hemolytic anemia for Babesia,bolstering molecular diagnosis,improving transfusion safety,and intensifying regional surveillance are crucial in reducing underdiagnosis and preventing transmission.These measures are essential for controlling babesiosis in China. 展开更多
关键词 Human babesiosis babesia microti HEMOPHAGOCYTOSIS Travel-related diseases
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天冬氨酸蛋白酶在卵形巴贝斯虫侵染长角血蜱过程中的作用研究
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作者 罗金 张高峰 +9 位作者 崔晶晶 柳志成 赵帅阳 任巧云 胡东生 曹立华 常国勤 殷宏 罗建勋 刘光远 《中国兽医科学》 北大核心 2025年第2期163-170,共8页
本试验以天冬氨酸蛋白酶(AP)为目标基因,探究其表达抑制后对卵形巴贝斯虫侵染长角血蜱的能力。利用5′/3′-RACE和PCR扩增天冬氨酸蛋白酶基因的全长和CDS,然后根据获得的序列设计siRNA干扰引物。通过颈静脉接种卵形巴贝斯虫的方式构建... 本试验以天冬氨酸蛋白酶(AP)为目标基因,探究其表达抑制后对卵形巴贝斯虫侵染长角血蜱的能力。利用5′/3′-RACE和PCR扩增天冬氨酸蛋白酶基因的全长和CDS,然后根据获得的序列设计siRNA干扰引物。通过颈静脉接种卵形巴贝斯虫的方式构建动物感染模型,将通过RNA干扰(RNAi)AP基因表达后的长角血蜱置于感染卵形巴贝斯虫牛体表,使蜱叮咬后检测蜱及蜱卵中卵形巴贝斯虫的感染率。结果显示,长角血蜱AP基因全长为1497 bp,编码区序列为1176 bp。该基因的RNAi可使AP的表达丰度降低81.91%(P=0.003<0.01)具有显著性差异。感染动物过程中,相较于空白对照组和阴性对照组,qRT-PCR结果表明RNAi组的巴贝斯虫感染率下降。试验结果表明,干扰长角血蜱AP能有效抑制卵形巴贝斯虫的垂直传播,虽然该基因在蜱种间有较高保守性,但是其核酸序列仍然存在一定的差异。该基因第626~644位或第257~275位定点结合可作为潜在的阻断卵形巴贝斯虫传播的疫苗设计靶点,该位点可以发挥抑制/干扰AP表达的作用。该作用下可有效阻止卵形巴贝斯虫的垂直传播,而且可导致巴贝斯虫的感染水平的显著降低。因此,本研究不仅有助于揭示巴贝斯虫的生物学特性,也为疾病防控提供了潜在的新策略,为公共卫生提供更加有效的干预措施。 展开更多
关键词 长角血蜱 卵形巴贝斯虫 天冬氨酸蛋白酶 RNA干扰 RACE
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福建省1例人感染田鼠巴贝虫的实验诊断
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作者 林耀莹 陈云虹 +2 位作者 谢汉国 欧阳榕 肖丽贞 《海峡预防医学杂志》 2025年第2期1-3,21,共4页
目的分析福建省2024年1例人感染田鼠巴贝虫的实验诊断结果,促进提升各级实验室的巴贝虫病诊断筛查能力,及时诊断和治疗有关病例,降低疾病危害。方法收集福建省少见的1例人感染巴贝虫病例的抗凝全血及病例相关信息,用姬姆萨染色镜检、巴... 目的分析福建省2024年1例人感染田鼠巴贝虫的实验诊断结果,促进提升各级实验室的巴贝虫病诊断筛查能力,及时诊断和治疗有关病例,降低疾病危害。方法收集福建省少见的1例人感染巴贝虫病例的抗凝全血及病例相关信息,用姬姆萨染色镜检、巴贝虫虫种核酸18S rRNA鉴定方法等对病例血样进行检测。结果姬姆萨染色镜检结果初步判定该病例感染巴贝虫;经NCBI-BLAST线上比对,显示该病例血样18S rRNA片段扩增产物(GenBank accession number:PQ999821)与田鼠巴贝虫(KF410827)的一致性为99.5%,判断该病例为田鼠巴贝虫感染。结论人感染巴贝虫病是一种新发寄生虫病,应加强巴贝虫镜检和核酸筛查等常态化检测,以准确了解人感染巴贝虫病的流行情况,为疾病防控制定对应的策略。 展开更多
关键词 田鼠巴贝虫 巴贝虫病 新发寄生虫病 实验诊断 疾病控制
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我国部分茶园蜱种类及其携带的两种新发病原的调查研究
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作者 石涵 周勇志 +5 位作者 曹杰 王亚楠 张厚双 夏宣保 徐前明 周金林 《中国动物传染病学报》 北大核心 2025年第3期145-151,共7页
为了保障茶叶主产区茶农的生命健康,查清新发人兽共患重要蜱传病的本底,本研究对我国中东部的河南信阳、安徽六安、浙江金华三地区的茶园中蜱种类及其携带的蜱传田鼠巴贝斯虫和发热伴血小板减少综合征病毒开展了调查研究。在2022—2023... 为了保障茶叶主产区茶农的生命健康,查清新发人兽共患重要蜱传病的本底,本研究对我国中东部的河南信阳、安徽六安、浙江金华三地区的茶园中蜱种类及其携带的蜱传田鼠巴贝斯虫和发热伴血小板减少综合征病毒开展了调查研究。在2022—2023年,共采集了963只蜱样本,发现有长角血蜱、微小扇头蜱、褐黄血蜱、龟形花蜱、微形血蜱5种蜱,分别占总数的72.90%、16.72%、9.76%、0.42%、0.21%。应用荧光定量PCR和巢式PCR方法对三地蜱样本中的田鼠巴贝斯虫和发热伴血小板减少综合征病毒的进行检测。结果显示,田鼠巴贝斯虫在河南信阳、安徽六安、浙江金华三地的蜱样本中阳性率分别为25.17%、13.98%、4.99%。长角血蜱的阳性率占比最高(83.33%),褐黄血蜱和龟形花蜱中没有检测出携带田鼠巴贝斯虫。发热伴血小板减少综合征病毒在河南信阳、安徽六安、浙江金华三地的阳性率分别为20.07%、5.77%、7.84%,5种蜱均检测到阳性,其中长角血蜱的阳性率占比最高(60.82%),微形血蜱阳性率占比最低(1.03%)。在长角血蜱和微形血蜱中发现有田鼠巴贝斯虫和发热伴血小板减少综合征病毒共感染情况,其中长角血蜱若蜱共感染最为显著。茶园是新发蜱传病高风险场景,应加强相关防控研究。 展开更多
关键词 蜱传病 田鼠巴贝斯虫 发热伴血小板减少综合征病毒 分子检测
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云南省金沙江流域小型兽类感染巴贝虫的分子流行病学调查
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作者 王帆 张云 +7 位作者 邵宗体 李玉琼 浦恩念 何志海 姚明国 别双双 江佳富 杜春红 《中国人兽共患病学报》 北大核心 2025年第7期767-774,共8页
目的了解云南省金沙江流域小型兽类感染巴贝虫的种类及其分布,掌握该地区小型兽类携带巴贝虫情况,为巴贝虫病的防控提供科学依据。方法选取云南省金沙江流域的其中10个县(市),在农耕地、林区等不同环境,捕获分属5目10科25属54种的小型兽... 目的了解云南省金沙江流域小型兽类感染巴贝虫的种类及其分布,掌握该地区小型兽类携带巴贝虫情况,为巴贝虫病的防控提供科学依据。方法选取云南省金沙江流域的其中10个县(市),在农耕地、林区等不同环境,捕获分属5目10科25属54种的小型兽类1493只,取其肝、脾组织提取DNA后,运用分子生物学方法检测巴贝虫18S rRNA的150 bp目的片段作初筛,阳性者进一扩增18S rRNA 1600 bp目的片段,通过序列比对及系统进化分析进行虫种特征分析。结果在云南省金沙江流域的6个县(市)小型兽类中检测出感染巴贝虫的小型兽类14只,总阳性率为0.93%(14/1493)。经序列分析与比对为田鼠巴贝虫的Otsu型和Kobe型,与人感染巴贝虫的病例患者检出的序列具有较高的相似度和较近的进化关系。6个县(市)的阳性率分别为:巧家县2.34%(3/128)、永善县2.06%(2/97)、元谋县1.88%(4/213)、德钦县1.03%(3/291)、香格里拉市0.95%(1/105)和水富县0.78%(1/128)。阳性小兽分属1目2科6属8种,检出巴贝虫阳性率分别是白尾松田鼠5.56%(1/18)、斯氏家鼠3.36%(4/119)、巢鼠3.33%(1/30)、西南绒鼠2.94%(1/34)、社鼠2.65%(3/113)、针毛鼠2.35%(2/85)、大耳姬鼠1.16%(1/86)、中华姬鼠0.98%(1/102)。巢鼠中检出巴贝虫为首次报道。在3种生境和海拔均检出小兽感染巴贝虫,其中1500~2500 m林区和高海拔居民区捕获的小兽巴贝虫的感染率较高。结论云南省金沙江沿岸部分县(市)的多种小型兽类存在巴贝虫感染,这些地区巴贝虫的流行情况值得关注。 展开更多
关键词 云南省 金沙江流域 小型兽类 巴贝虫 调查
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牛巴贝斯虫SBP2蛋白生物信息学分析及多克隆抗体制备
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作者 冯秀娟 任冀超 +7 位作者 崔泽云 赵雪晴 李佳欣 张杨 张伟 巴音查汗·盖力克 郭庆勇 李永畅 《中国畜牧兽医》 北大核心 2025年第8期3847-3856,共10页
【目的】探究牛巴贝斯虫(Babesia bovis)SBP2蛋白(spherical body protein 2,SBP2)的生物信息学特征并进行原核表达及多克隆抗体制备,为牛巴贝斯虫疫苗和诊断抗原筛选提供理论依据。【方法】对牛巴贝斯虫SBP2基因进行扩增和克隆,运用Meg... 【目的】探究牛巴贝斯虫(Babesia bovis)SBP2蛋白(spherical body protein 2,SBP2)的生物信息学特征并进行原核表达及多克隆抗体制备,为牛巴贝斯虫疫苗和诊断抗原筛选提供理论依据。【方法】对牛巴贝斯虫SBP2基因进行扩增和克隆,运用Mega7.0构建牛巴贝斯虫SBP2蛋白系统进化树,利用IEDB Analysis Resource等生物信息学方法对牛巴贝斯虫SBP2蛋白的磷酸化位点和B细胞抗原表位进行预测分析;对SBP2蛋白与弓形虫致密颗粒蛋白(GRA)的氨基酸序列比对分析;构建原核表达载体p ET-28a-SBP2,诱导表达SBP2重组蛋白并进行蛋白纯化,通过Western blotting验证SBP2重组蛋白反应原性;以SBP2重组蛋白为免疫原,免疫BALB/c小鼠制备多克隆抗体,并利用间接ELISA方法检测其效价。【结果】系统进化树显示,本研究牛巴贝斯虫SBP2蛋白与泰国株(OM46855)的氨基酸序列亲缘关系最近。生物信息学分析显示,SBP2蛋白包含17个B细胞抗原表位和31个磷酸化位点,其中包括12个丝氨酸位点、14个苏氨酸位点及5个酪氨酸位点。牛巴贝斯虫SBP2蛋白氨基酸序列与弓形虫GRA蛋白之间有很强的相关性。成功构建了重组质粒p ET-28a-SBP2;Western blotting结果显示,SBP2重组蛋白分子质量大小约为35ku,具有良好的反应原性;制备的多克隆抗体效价为1∶204800。【结论】本研究通过预测牛巴贝斯虫SBP2蛋白生物学特性,加深了对SBP2蛋白的认识,利用原核表达系统成功获得牛巴贝斯虫SBP2重组蛋白,并获得其小鼠源多克隆抗体,为进一步研究SBP2功能及其在牛巴贝斯虫致病机制中的作用奠定了基础。 展开更多
关键词 牛巴贝斯虫 球状体蛋白2 生物信息学分析 原核表达 多克隆抗体
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甘肃省犬巴贝斯虫病分子流行病学调查
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作者 李娇 王锦明 +3 位作者 关贵全 王桂荣 叶得河 马永华 《中国动物传染病学报》 北大核心 2025年第3期188-194,共7页
本研究从甘肃兰州、陇南、天水等14个地区共收集宠物犬抗凝血1628份,采用吉姆萨染色和PCR方法进行检测,并对阳性样本进行18S rRNA基因测序,将测序结果与GenBank中收录的巴贝斯虫基因序列进行同源性分析,构建分子系统进化树。结果显示,... 本研究从甘肃兰州、陇南、天水等14个地区共收集宠物犬抗凝血1628份,采用吉姆萨染色和PCR方法进行检测,并对阳性样本进行18S rRNA基因测序,将测序结果与GenBank中收录的巴贝斯虫基因序列进行同源性分析,构建分子系统进化树。结果显示,全省感染率为3.13%(51/1628)。以18S rRNA为靶基因的遗传分析发现,甘肃地区流行的虫株与陕西分离株、河南分离株、日本分离株、印度分离株的B.gibsoni处于同一分支,表明甘肃地区犬感染的巴贝斯虫种类为B.gibsoni。通过本次调查研究发现甘肃省犬巴贝斯虫的感染情况和流行优势虫种。可为甘肃省的犬巴贝斯虫病的防治提供流行病学数据。 展开更多
关键词 18S rRNA 吉氏巴贝斯虫 巢氏PCR扩增技术 分子生物学诊断
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巴贝斯虫TRAP基因敲除虫株的构建及鉴定
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作者 叶雨欣 王锦明 +7 位作者 年月丽 刘泽恩 梁勤东 张玉婷 拜娅楠 刘培琪 殷宏 关贵全 《中国兽医科学》 北大核心 2025年第9期1227-1232,共6页
为阐明巴贝斯虫TRAP在入侵宿主细胞中的作用,采用同源重组技术敲除羊巴贝斯虫未定种新疆株的TRAP基因,通过PCR、间接免疫荧光及Western-blot等方法进行验证。结果显示,羊巴贝斯虫未定种新疆株的TRAP基因被成功敲除,这为靶向TRAP的药物... 为阐明巴贝斯虫TRAP在入侵宿主细胞中的作用,采用同源重组技术敲除羊巴贝斯虫未定种新疆株的TRAP基因,通过PCR、间接免疫荧光及Western-blot等方法进行验证。结果显示,羊巴贝斯虫未定种新疆株的TRAP基因被成功敲除,这为靶向TRAP的药物及疫苗的研发奠定了基础。 展开更多
关键词 巴贝斯虫 TRAP基因 基因敲除 入侵
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核糖体蛋白S6激酶在卵形巴贝斯虫侵染长角血蜱过程中的作用
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作者 车丽妍 罗金 +15 位作者 高仙 张高峰 赵帅阳 任巧云 胡东生 李元飞 曹立华 林向鑫 郑立东 常国勤 李星 王希伟 樊亮 殷宏 罗建勋 刘光远 《中国兽医科学》 北大核心 2025年第8期1033-1040,共8页
卵形巴贝斯虫(Babesia ovata)是一种寄生于牛血液中的梨形虫,可引起患畜卵形巴贝斯虫病。长角血蜱(Haemaphysalis longicornis)是其主要的生物媒介。核糖体蛋白S6激酶(RpS6K)作为一个重要的细胞信号传导分子,在病原侵染蜱的过程中扮演... 卵形巴贝斯虫(Babesia ovata)是一种寄生于牛血液中的梨形虫,可引起患畜卵形巴贝斯虫病。长角血蜱(Haemaphysalis longicornis)是其主要的生物媒介。核糖体蛋白S6激酶(RpS6K)作为一个重要的细胞信号传导分子,在病原侵染蜱的过程中扮演着关键角色。本研究利用PCR扩增长角血蜱RpS6K基因的全长序列,并根据获得的序列设计RpS6K的小干扰RNA(siRNA)。试验中,牛颈静脉接种卵形巴贝斯虫,建立动物感染模型。通过siRNA抑制RpS6K基因表达,应用方差分析方法(ANOVA)对Sp6K在卵形巴贝斯虫侵染长角血蜱过程中的生理指标数据进行分析。结果显示,长角血蜱RpS6K基因全长为2922 bp。抑制试验使RpS6K基因的表达丰度降低至31.54%(P<0.05)。动物感染试验表明,相较于对照组,试验组卵中巴贝斯虫的感染率显著下降。干扰长角血蜱RpS6K可以有效阻断卵形巴贝斯虫在蜱体中的垂直传播。因此,本研究为防治巴贝斯虫病提供了新的方向。 展开更多
关键词 长角血蜱 卵形巴贝斯虫 核糖体蛋白S6激酶 RNAI
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