Objective:To green synthesize and characterize copper nanoparticles(Cu NPs)using Astragalus sinicus,as well as evaluate the acaricidal,larvacidal,and repellent activities of Cu NPs against Hyalomma anatolicum(H.anatol...Objective:To green synthesize and characterize copper nanoparticles(Cu NPs)using Astragalus sinicus,as well as evaluate the acaricidal,larvacidal,and repellent activities of Cu NPs against Hyalomma anatolicum(H.anatolicum),one of the most prevalent ticks infesting cattle in Saudi Arabia.Methods:Cu NPs were green synthesized by adding the Astragalus sinicus extract to a copper sulfate solution.The acaricidal,larvicidal,and repellent activities of Cu NPs against H.anatolicum were assessed via the adult immersion test,the larval packet test,and the vertical movement behavior of tick larvae,respectively.The effects of Cu NPs on acetylcholinesterase as well as oxidative enzyme activities were examined.Results:The green synthesized Cu NPs displayed a spherical form with a size range of 15-75 nm.After exposure of adult H.anatolicum to different concentrations of Cu NPs,the viability rate of adult H.anatolicum and the mean number,weight,and hatchability of eggs were noticeably reduced,in comparison to the control group(P<0.001).In addition,the viability rate of larvae considerably declined(P<0.001)with the LC_(50)and LC_(90)values of 11.30 and 20.34μg/m L,respectively.The maximum repellent activity of Cu NPs was observed at 50,100,and 200μg/m L with complete repellent activity after 60,120,and 180 min of exposure,respectively.Cu NPs,mainly at 1/2 LC_(50)and LC_(50)concentrations,markedly suppressed the acetylcholinesterase activity of the larval stage of H.anatolicum(P<0.001).Moreover,Cu NPs,mainly at LC_(50)dose,significantly elevated malondialdehyde level while declining glutathione-S-transferase level in H.anatolicum larvae(P<0.001).Conclusions:Cu NPs show potent acaricidal,larvicidal,and repellent activities against adults and larvae of H.anatolicum.However,further studies must be performed to clarify the precise mechanisms and the efficacy of Cu NPs in practical use.展开更多
Hyalomma anatolicum, Hyalomma asiaticum and Hyalomma detritum are wide-spread tick vectors in China. They could transmit a great variety of serious animal and human pathogens, which are great threats to the health of ...Hyalomma anatolicum, Hyalomma asiaticum and Hyalomma detritum are wide-spread tick vectors in China. They could transmit a great variety of serious animal and human pathogens, which are great threats to the health of human beings and the safety of stockbreeding. Most of them are distributed in Xinjiang and Inner Mongolia, and they share similar morphologies. This study is to establish a method for identifying H. anatolicum, H. asiaticum and H. detritum with molecular markers and to revealing the phylogenetic relationship of these ticks. Ticks were collected from domestic animals in Xinjiang and Inner Mongolia autonomous regions and classified by morphological characters. 16S rRNA and mitachondrial cytochrome oxidase subunit I gone (CO1) of ticks were amplified by PCR and subsequently sequenced. Phylogenetic trees were constructed by MEGA 5.0 and Mrbayes 3.2. On the 16S rRNA-based phylogenetic tree, H. anatolicum and H. aisaticum were clustered together with their respective classes. H. detritum was clustered with their respective class and the H. anatolicum, H. asiaticum formed distinct branches on the phylogenetic trees based on the COL The method based on morphology that combined with molecular 16S rRNA and COl seemed a simple and accurate method for species identification of H. anatolicum, H. asiaticum and H. detritum. n展开更多
A long-term blood feeder, like the <i><span>Hyalomma</span></i><span> </span><i><span>dromedarii</span></i><span> tick, requires extended con...A long-term blood feeder, like the <i><span>Hyalomma</span></i><span> </span><i><span>dromedarii</span></i><span> tick, requires extended control over all hemostatic defense mechanisms generated by the host during feeding, including blood coagulation. To overcome this, ticks have evolved numerous molecules that target proteases in the blood coagulation cascade. New insights into the role of clotting factors in the development and progression of cancer have identified anticoagulant treatment as a potential therapeutic approach. In this context, the present work assessed the anticoagulation activities of crude and fractionated salivary gland extract (SGE) prepared from semi-fed </span><i><span>H</span></i><span>. </span><i><span>dromedarii</span></i><span> females. Additionally, the antitumor effects of the potent anti-thrombin fractions were determined against colon cancer (Caco-2) and normal skin (HFB4) cells. Crude SGE significantly prolonged clotting time in prothrombin time (PT), activated partial thromboplastin time (aPTT) and thrombin time (TT) assays and inhibited thrombin in FII-activity assay. Using anion-exchange chromatography, the fractions that strongly inhibited thrombin (3.A4 and 3.A5) were eluted. Both fractions prolonged the aPTT and TT clotting times and reduced the activity of FII significantly. The protein profiles of both fractions indicated the presence of a single polypeptide band of about 99 kDa. Regarding anti-cancer potential of the tested fractions, Caco-2 cells showed reduced viability with obvious morphological changes, induced apoptosis and a reduced level of vascular endothelial </span><span>growth factor (VEGF). G2/M cell cycle arrest was observed only in 3.A5-treated</span><span> cells. No cytotoxic effects were observed in HFB4 cells. These results demonstrated the potential of tick-derived anticoagulants, specifically thrombin inhibitors, as effective tools in colorectal cancer treatment. Further purification of the effector molecule(s) is required to fully characterize their structures and mechanisms of action.</span>展开更多
Hyalomma dromedarii ticks are important disease vectors to camels in the UAE and worldwide. Ticks can be identified using DNA-based techniques. In addition, such techniques could be utilized to study the intraspecific...Hyalomma dromedarii ticks are important disease vectors to camels in the UAE and worldwide. Ticks can be identified using DNA-based techniques. In addition, such techniques could be utilized to study the intraspecific genetic diversity in tick populations. In this study, the genetic diversity of four H. dromedarii populations was investigated using mitochondrial cytochrome c oxidase subunit I (COI) gene and randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). The results showed that both of the aforementioned techniques produced similar grouping patterns. Moreover, they revealed that the four tick populations had high levels of genetic similarity. However, one population was slightly different from the three other populations. The current study demonstrated that H. dromedarii ticks in the UAE are very similar at the genetic level and that investigating more locations and screening larger numbers of ticks could reveal larger genetic differences.展开更多
自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环...自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环形泰勒虫。将此饱血成蜱经实验室饲喂条件下,孵化出的幼虫分别饲喂于牛与家兔两种不同的动物,结果在饱血若虫阶段所有的蜱均从两种动物体表脱落,不再叮咬,饥饿成蜱又更换另一宿主,证明其是二宿主蜱。展开更多
文摘Objective:To green synthesize and characterize copper nanoparticles(Cu NPs)using Astragalus sinicus,as well as evaluate the acaricidal,larvacidal,and repellent activities of Cu NPs against Hyalomma anatolicum(H.anatolicum),one of the most prevalent ticks infesting cattle in Saudi Arabia.Methods:Cu NPs were green synthesized by adding the Astragalus sinicus extract to a copper sulfate solution.The acaricidal,larvicidal,and repellent activities of Cu NPs against H.anatolicum were assessed via the adult immersion test,the larval packet test,and the vertical movement behavior of tick larvae,respectively.The effects of Cu NPs on acetylcholinesterase as well as oxidative enzyme activities were examined.Results:The green synthesized Cu NPs displayed a spherical form with a size range of 15-75 nm.After exposure of adult H.anatolicum to different concentrations of Cu NPs,the viability rate of adult H.anatolicum and the mean number,weight,and hatchability of eggs were noticeably reduced,in comparison to the control group(P<0.001).In addition,the viability rate of larvae considerably declined(P<0.001)with the LC_(50)and LC_(90)values of 11.30 and 20.34μg/m L,respectively.The maximum repellent activity of Cu NPs was observed at 50,100,and 200μg/m L with complete repellent activity after 60,120,and 180 min of exposure,respectively.Cu NPs,mainly at 1/2 LC_(50)and LC_(50)concentrations,markedly suppressed the acetylcholinesterase activity of the larval stage of H.anatolicum(P<0.001).Moreover,Cu NPs,mainly at LC_(50)dose,significantly elevated malondialdehyde level while declining glutathione-S-transferase level in H.anatolicum larvae(P<0.001).Conclusions:Cu NPs show potent acaricidal,larvicidal,and repellent activities against adults and larvae of H.anatolicum.However,further studies must be performed to clarify the precise mechanisms and the efficacy of Cu NPs in practical use.
基金Supported by National Key Technology Research and Development Program ofChina(2012BAK11B04)
文摘Hyalomma anatolicum, Hyalomma asiaticum and Hyalomma detritum are wide-spread tick vectors in China. They could transmit a great variety of serious animal and human pathogens, which are great threats to the health of human beings and the safety of stockbreeding. Most of them are distributed in Xinjiang and Inner Mongolia, and they share similar morphologies. This study is to establish a method for identifying H. anatolicum, H. asiaticum and H. detritum with molecular markers and to revealing the phylogenetic relationship of these ticks. Ticks were collected from domestic animals in Xinjiang and Inner Mongolia autonomous regions and classified by morphological characters. 16S rRNA and mitachondrial cytochrome oxidase subunit I gone (CO1) of ticks were amplified by PCR and subsequently sequenced. Phylogenetic trees were constructed by MEGA 5.0 and Mrbayes 3.2. On the 16S rRNA-based phylogenetic tree, H. anatolicum and H. aisaticum were clustered together with their respective classes. H. detritum was clustered with their respective class and the H. anatolicum, H. asiaticum formed distinct branches on the phylogenetic trees based on the COL The method based on morphology that combined with molecular 16S rRNA and COl seemed a simple and accurate method for species identification of H. anatolicum, H. asiaticum and H. detritum. n
文摘A long-term blood feeder, like the <i><span>Hyalomma</span></i><span> </span><i><span>dromedarii</span></i><span> tick, requires extended control over all hemostatic defense mechanisms generated by the host during feeding, including blood coagulation. To overcome this, ticks have evolved numerous molecules that target proteases in the blood coagulation cascade. New insights into the role of clotting factors in the development and progression of cancer have identified anticoagulant treatment as a potential therapeutic approach. In this context, the present work assessed the anticoagulation activities of crude and fractionated salivary gland extract (SGE) prepared from semi-fed </span><i><span>H</span></i><span>. </span><i><span>dromedarii</span></i><span> females. Additionally, the antitumor effects of the potent anti-thrombin fractions were determined against colon cancer (Caco-2) and normal skin (HFB4) cells. Crude SGE significantly prolonged clotting time in prothrombin time (PT), activated partial thromboplastin time (aPTT) and thrombin time (TT) assays and inhibited thrombin in FII-activity assay. Using anion-exchange chromatography, the fractions that strongly inhibited thrombin (3.A4 and 3.A5) were eluted. Both fractions prolonged the aPTT and TT clotting times and reduced the activity of FII significantly. The protein profiles of both fractions indicated the presence of a single polypeptide band of about 99 kDa. Regarding anti-cancer potential of the tested fractions, Caco-2 cells showed reduced viability with obvious morphological changes, induced apoptosis and a reduced level of vascular endothelial </span><span>growth factor (VEGF). G2/M cell cycle arrest was observed only in 3.A5-treated</span><span> cells. No cytotoxic effects were observed in HFB4 cells. These results demonstrated the potential of tick-derived anticoagulants, specifically thrombin inhibitors, as effective tools in colorectal cancer treatment. Further purification of the effector molecule(s) is required to fully characterize their structures and mechanisms of action.</span>
文摘Hyalomma dromedarii ticks are important disease vectors to camels in the UAE and worldwide. Ticks can be identified using DNA-based techniques. In addition, such techniques could be utilized to study the intraspecific genetic diversity in tick populations. In this study, the genetic diversity of four H. dromedarii populations was investigated using mitochondrial cytochrome c oxidase subunit I (COI) gene and randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). The results showed that both of the aforementioned techniques produced similar grouping patterns. Moreover, they revealed that the four tick populations had high levels of genetic similarity. However, one population was slightly different from the three other populations. The current study demonstrated that H. dromedarii ticks in the UAE are very similar at the genetic level and that investigating more locations and screening larger numbers of ticks could reveal larger genetic differences.
文摘自新疆伊宁地区黄牛体表采集的盾糙璃眼蜱饥饿或半饱血成虫,带回实验室感染除脾牛。经血涂片检查发现,于感染后第21天,病牛血液中出现一种以圆环形与卵圆形为主的泰勒虫。经形态学观察和18 S rRNA基因测序和进化关系分析证明,它们为环形泰勒虫。将此饱血成蜱经实验室饲喂条件下,孵化出的幼虫分别饲喂于牛与家兔两种不同的动物,结果在饱血若虫阶段所有的蜱均从两种动物体表脱落,不再叮咬,饥饿成蜱又更换另一宿主,证明其是二宿主蜱。