作者： 李中波;尧国民;侯强红;程天印

期刊： 中国畜牧兽医,2021年48(02):607-618 ISSN：1671-7236

作者机构： [侯强红; 李中波; 尧国民] 怀化职业技术学院;湖南农业大学动物医学院;[程天印] 怀化职业技术学院<&wdkj&>湖南农业大学动物医学院

关键词： 液相色谱-串联质谱;褐黄血蜱;中肠蛋白

摘要： 本研究旨在探明4个时间段褐黄血蜱（采自刺猬体表）中肠蛋白质成分,揭示参与血餐消化的蛋白种类及其含量变化规律。采用液相色谱-串联质谱法（LC-MS/MS）对采集于刺猬体表的4个时间段褐黄血蜱的中肠蛋白组成分进行检测。基于该蜱的唾液、中肠转录组翻译文库及Uniprot数据库,利用软件Mascot 2.2对所获肽段及蛋白质进行鉴定。结果显示:在褐黄血蜱中肠蛋白提取液中共检测出特异性肽段3 046条,鉴定303种蛋白,其中271种为高可信蛋白;在所有的高可信蛋白中,23种含量较为丰富,125种在后期消化阶段（第2至第4阶段）的含量为0,123种的含量发生不同程度的变化（24种蛋白的含量变化明显,其中12种含量上升明显,12种含量下降明显）。确定148种高可信蛋白来自于刺猬血清,推断24种可信蛋白可能参与蜱虫对血餐的消化。

语种： 中文

作者： Liu, L.;He, X. -M.;Feng, L. -L.;Duan, D. -Y.;Zhan, Y.;...

期刊： Medical and Veterinary Entomology,2020年34(2):192-200 ISSN：0269-283X

通讯作者： Cheng, T. -Y.

作者机构： [He, X. -M.; Duan, D. -Y.; Liu, L.; Cheng, T. -Y.; Feng, L. -L.; Zhan, Y.] Hunan Agr Univ, Coll Vet Med, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha, Hunan, Peoples R China.;[Cheng, T. -Y.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, T. -Y.] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： complementary DNA;contig;heat shock protein 70;heat shock protein A2A;heat shock protein A2B;heat shock protein A5;heat shock protein A9;unclassified drug;arthropod protein;heat shock protein 70;antigen;cell organelle;cytoplasm;DNA;gene expression;protein;RNA;tick;alpha helix;amino acid sequence;Article;computer simulation;cytoplasm;DNA synthesis;endoplasmic reticulum;gene amplification;gene expression;Haemaphysalis;Haemaphysalis flava;insect genome;mitochondrion;molecular cloning;multigene family;nonhuman;open reading frame;protein expression;protein localization;real time polymerase chain reaction;RNA extraction;salivary gland;transcriptomics;animal;chemistry;female;genetics;Ixodidae;metabolism;molecular cloning;Acari;Haemaphysalis flava;Ixodida;Ixodidae;Amino Acid Sequence;Animals;Arthropod Proteins;Cloning, Molecular;Female;HSP70 Heat-Shock Proteins;Ixodidae;Multigene Family

摘要： The heat shock protein 70 (HSPA) family and their genes have been studied in ticks and are considered as possible antigen candidates for the development of anti-tick vaccines. However, knowledge about their members, structure and function in ticks is incomplete. Based on our transcriptomic data, the full length of four HSPA genes in Haemaphysalis flava (Acari: Ixodidae) was cloned via rapid amplification of cDNA ends. The open reading frame of HSPA2A, HSPA2B, HSPA5 and HSPA9 was 1920, 1911, 1983 and 2088 bp in length, respectively. Three family signatures and one localization motif were in the encoding proteins. HSPA2A and HSPA2B were predicted to be located at cytoplasm/nucleus, whereas HSPA5 and HSPA9 were at endoplasmic reticulum and mitochondria, respectively. In silico simulation demonstrated that those proteins had distinct numbers of α-helixes, extended strands and coils, and different antigenic epitopes. Expression of HSPA5 and HSPA9 in the salivary gland was significantly higher in partially-engorged female adult ticks than the fully-engorged (P ' 0.01) as shown by a quantitative polymerase chain reaction. Our data indicated that H. flava ticks had at least four HSPA genes encoding proteins with different cellular locations, structures and expression profiles, suggesting their diverse roles in tick biology. © 2019 The Royal Entomological Society

语种： 英文

作者： Duan, D. -Y.;Zhou, H. -M.;Cheng, T. -Y.*

期刊： Medical and Veterinary Entomology,2020年34(2):215-224 ISSN：0269-283X

通讯作者： Cheng, T. -Y.

作者机构： [Duan, D. -Y.; Zhou, H. -M.; Cheng, T. -Y.] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, T. -Y.] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： bacterial DNA;RNA 16S;bacterial DNA;RNA 16S;adult;bacterium;coliform bacterium;digestive system;ectoparasite;female;female headed household;microbial activity;microbial community;relative abundance;Article;Bartonella;comparative study;controlled study;ectoparasite;Escherichia coli;female;Melophagus ovinus;microbial community;midgut;nonhuman;phylum;Proteobacteria;sample;sheep;Streptococcus;Wolbachia;animal;bacterium;China;Diptera;feeding behavior;intestine flora;isolation and purification;microbiology;microflora;China;Gansu;Jiuquan;Arsenophonus;Bacteria (microorganisms);Bartonella;Bartonella chomelii;Escherichia coli;Melophagus ovinus;Ovis aries;Proteobacteria;Streptococcus hyointestinalis;Wolbachia;Animals;Bacteria;China;Diptera;DNA, Bacterial;Feeding Behavior;Female;Gastrointestinal Microbiome;Microbiota;RNA, Ribosomal, 16S

摘要： Melophagus ovinus is a type of ectoparasite infesting sheep. Data regarding the comprehensive bacterial community associated with the whole body and midgut of M. ovinus under different engorged statuses are required. Melophagus ovinus were collected from the city of Jiuquan, China. Bacterial DNA was extracted from the whole body and midgut of fully engorged female adults, or newly hatched and unfed adult female M. ovinus. The 16S rRNA gene V3–V4 hypervariable regions were sequenced using the IonS5™XL platform (Thermo Fisher Scientific, Waltham, MA, U.S.A.). The whole body bacterial diversity of the newly hatched, unfed adult females was greater compared with that of the other three samples. Proteobacteria was the dominant bacterial phylum in all of the samples. Of the 42 total bacterial genera present in all of the experimental samples, Arsenophonus, Bartonella and Wolbachia were the dominant genera. The relative abundance of Arsenophonus in midgut was greater than that in the whole body. The relative abundance of Bartonella in fully engorged adults was far greater than those in newly hatched, unfed adults. The relative abundance of Wolbachia was highest in the whole body of newly hatched, unfed adults. Seventeen bacterial species were identified in all experimental samples. Bartonella chomelii, Streptococcus hyointestinalis and Escherichia coli were the first species reported in M. ovinus. © 2019 The Royal Entomological Society

语种： 英文

作者： Duan, D-Y;Tang, J-M;Chen, Z.;Liu, G-H;Cheng, T-Y

期刊： Medical and Veterinary Entomology,2020年34(2):229-235 ISSN：0269-283X

通讯作者： Liu, GH;Cheng, TY

作者机构： [Tang, J-M; Chen, Z.; Duan, D-Y; Cheng, T-Y; Liu, G-H; Cheng, TY] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Liu, GH; Cheng, TY] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： genomic DNA;mitochondrial DNA;ribosome RNA;transfer RNA;ectoparasite;endangered species;genome;mammal;mitochondrial DNA;molecular ecology;population genetics;taxonomy;tick;agar gel electrophoresis;Amblyomma americanum;Amblyomma fimbriatum;Amblyomma javanense;Article;controlled study;female;gene structure;male;Manis javanica;maximum likelihood method;mitochondrial genome;nonhuman;Pholidota (animal);phylogeny;RNA gene;Sanger sequencing;tick;animal;China;genetics;insect genome;Ixodidae;California;San Diego;United States;Amblyomma;Amblyomma americanum;Ixodidae;Manis javanica;Animals;China;Female;Genome, Insect;Genome, Mitochondrial;Ixodidae;Male

摘要： Amblyomma javanense is an important ectoparasite of Manis javanica, although the population genetics, molecular biology and systematics of A. javanense remain poorly understood. In the present study, the mitochondrial genome of A. javanense was sequenced using the Illumina HiSeq sequencing platform (Illumina, San Diego, CA, U.S.A.) and compared with the genomes of two closely related species: Amblyomma fimbriatum and Amblyomma americanum. The intraspecies and interspecies relationships of A. javanense and another 21 selected species were investigated by constructing a maximum-likelihood tree and a neighbour-joining tree. The mitochondrial genome of A. javanense was 14 780 bp in length and contained 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and two control regions. The results of the comparisons indicate that there is great similarity among these three species, and both trees indicate that A. javanense is a member of the Amblyomminae. The study of A. javanense of pangolins also indicates the premise and foundation of the relationship between the parasite and other species. © 2019 The Royal Entomological Society

语种： 英文

作者： Feng, Li-Li;Cheng, Tian-Yin*

期刊： Experimental and Applied Acarology,2020年80(2):269-287 ISSN：0168-8162

通讯作者： Cheng, Tian-Yin

作者机构： [Cheng, Tian-Yin; Feng, Li-Li] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-Yin] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.

关键词： Proteome;Midgut content;Blood meal;High-confidence proteins

摘要： Tick blood meals are stored and digested in their midguts. Blood digestion is complex, and many proteins are involved. Study of the tick-derived proteins in the midgut content may aid in the discovery of active molecules that would be useful for anti-tick vaccines. We analyzed the midgut content proteomes of partially engorged female Haemaphysalis flava, fully engorged female H. flava, and hedgehog serum using liquid chromatography tandem-mass spectrometry and label-free quantitation. In this study, high-confidence protein profiling of tick midgut content was determined. Based on the search against our in-house transcriptome database, the 28 high-confidence proteins were identified. Of these, 17 were identified as tick-derived, and the rest were of unspecified origin (proteins that could not be differentiated as host-derived or tick-derived proteins). The function of these midgut content proteins identified here may involve nutrient transportation, anti-coagulation, erythrocyte lysis, detoxification, lipid metabolism, and immunization. The presence of hemoglobin suggested that the red blood cells were lysed in the gut lumen. The midgut contents contain a large amount of fibrinogen and it has the ability to clot immediately. The midgut contained mostly host-derived proteins, and these host proteins provide rich nutrients for tick development and reproduction. However, some intracellular proteins were also identified, suggesting the possibility of shedding of the midgut epithelium and ingestion of saliva during feeding. This finding advances our understanding of the digestive mechanism and will be useful in the screening of vaccine antigens.

语种： 英文

作者： Yi, J. N.;Jin, Y. C.;Liu, J. H.;Li, F.;Li, Z. B.;...

期刊： TROPICAL BIOMEDICINE,2019年36(1):297-303 ISSN：0127-5720

通讯作者： Cheng, T. Y.

作者机构： [Yi, J. N.; Li, Z. B.; Jin, Y. C.; Li, F.; Liu, J. H.; Cheng, T. Y.] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Li, F.; Liu, J. H.; Cheng, T. Y.] Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, T. Y.] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

摘要： The cattle tick, Rhipicephalus microplus (formerly Boophi/us microp lus), is the most important blood-feeding ectoparasite of cattle in tropical and subtropical regions of the world. In this study, we examined sequence variability in three mitochondrial (mt) DNA (cox1, nad1, nad4) among cattle tick R. microplus originating from four provinces of China. A portion of earl (pcox1), nad1 (pnad1) and nad4 (pnad4) genes were amplified by polymerase chain reaction (PCR) separately from adult R. microplus individuals and the amplicons were subjected to sequence from both directions. The sequence of mt cox1, nad1, nad4 genes was 817 bp, 350 bp, and 794 bp in size, respectively. The intraspecific sequence variations within R. microplus were 0-8.6% for coxl, 0-4.9% for nad1 and 0-10.3% for nad4. However, the interspecific sequence differences among the members of the Rhipicephalus [R. sanguineus (JX416325) and R. turanicus (NC035946)] were significantly higher, being 16.9-20.5%, 18-22.8%, 22.8-25.3% for pcox1, pnad1 and pnad4, respectively. In addition, genetic differences were 7.9-8.6% for cox1, 4.3-4.9% for nad1 and 10-10.3% for nad4 between the two detected lineages (R. microplus clade A and clade B). Phylogenetic analyses indicated that all the Rhipicephalus isolates from the present study represents R. microplus, supporting that R. microplus represents species complex. Our result provided an additional genetic evidence for the existence of species complex within R. microplus in China.

语种： 英文

作者： He, Xiao-ming;Liu, Lei*;Cheng, Tian-yin*

期刊： Ticks and Tick-borne Diseases,2019年10(1):170-175 ISSN：1877-959X

通讯作者： Liu, Lei;Cheng, Tian-yin

作者机构： [He, Xiao-ming; Cheng, Tian-yin; Liu, Lei] Hunan Agr Univ, Coll Vet Med, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Liu, Lei; Cheng, TY] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Liu, L; Cheng, TY] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： Tick;HSC70;Heat shock protein;Anticoagulation

摘要： Ticks and tick-borne diseases are major global health threats. During blood feeding, ticks insert their hypostomes into hosts and inject an array of anticoagulant molecules to maintain fluidity of the blood-meal. These anticoagulant molecules may provide insights into understanding the feeding biology of ticks and to develop vaccines against infestations. In Haemaphysalis flava, the heat shock cognate 70 (HSC70), a member of the heat shock protein (HSP) family, is differentially expressed in salivary glands at different levels of engorgement during blood feeding. However, its function in ticks is largely not known. The present study was designed to explore the possible effects of HSC70 on the plasma. The open reading frame (ORF) of HSC70 was expressed in a prokaryotic system, and recombinant HSC70 (rHSC70) was purified and characterized. The anticoagulation activity of rHSC70 was estimated by measuring prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen (FIB) with/without its inhibitor, VER155008. The results demonstrated that rHSC70 from H. flava extended TT (P < 0.001) and FIB clotting times (>300 s), but showed little effect on PT and APTT. Adding an inhibitor reversed anticlotting effects of rHSC70 on TT and FIB. These data indicate that rHSC70 is an anticoagulant agent, and the anticlotting activity likely attributes to the inhibition of thrombin and the transformation of fibrinogen into fibrin. © 2018 Elsevier GmbH

语种： 英文

作者： 段德勇;刘国华;程天印

期刊： 中国兽医科学,2019年49(8):990-997 ISSN：1673-4696

作者机构： 湖南农业大学动物医学院,湖南长沙,410128;[刘国华; 段德勇; 程天印] 湖南农业大学

关键词： 波斯锐缘蜱;形态学;分子标记;特征

摘要： 为了精准鉴定波斯锐缘蜱,从新疆、甘肃、河北等地采集寄生蜱,借助体视显微镜超景深系统对其形态学特征进行观察;经PCR扩增、测序获得其16S rDNA和cox1基因序列,结合GenBank数据库中参考序列,比对分析同源性并构建系统进化树。结果显示,波斯锐缘蜱幼蜱形态与其他发育阶段差异较大,其体型较小,背、腹表面光滑,无盘窝,生殖孔未形成,足3对。若蜱除生殖孔不显著外,其他形态结构均与成蜱相似。雌、雄成蜱形态学差异主要为:雌蜱生殖孔呈横裂形,雄蜱生殖孔呈半圆形。同时,在16S rDNA和cox1基因序列的邻位相接系统进化树中,波斯锐缘蜱分布于两大进化枝之一的软蜱科,同种聚类良好。结果提示:波斯锐缘蜱具有显著、易辨识、易鉴定的形态学和分子标记特征。

语种： 中文

作者： 张海;程天印

期刊： 湖南畜牧兽医,2019年(04):3-4 ISSN：1006-4907

作者机构： 湖南农业大学动物医学院,湖南长沙410128;湘西自治州兽医局,湘西吉首416000;湖南农业大学动物医学院,湖南长沙,410128;[张海; 程天印] 湖南农业大学

关键词： 畜牧养殖;家庭牧场;发展对策

摘要： 为了促进湘西自治州畜牧养殖业发展,特别是家庭牧场这种新型农业经营主体的发展,文章对湘西自治州家庭牧场发展现状进行了调查,提出了推动家庭牧场规范、有序发展的相应对策.

语种： 中文

作者： Xu, Lv;Liu, Lei;Cheng, Tian-yin*

期刊： JOURNAL OF MEDICAL ENTOMOLOGY,2019年56(2):569-575 ISSN：0022-2585

通讯作者： Cheng, Tian-yin

作者机构： [Xu, Lv; Cheng, Tian-yin; Liu, Lei] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-yin] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.

关键词： tick;GAPDH;RACE;real-time PCR;salivary gland

摘要： Haemaphysalis flava (Acari: Ixodidae) harbors pathogenic microorganisms and transfers these to hosts during blood feeding. Proteomic analysis in the midgut contents of H. flava detected glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and contig 1683 was retrieved as a GAPDH gene fragment by searching our previous transcriptomic library. In the study, the 5' and 3' ends of contig 1683 were cloned by rapid amplification of cDNA ends (RACE) and a full length, 1340 bp cDNA of Hf-GAPDH was obtained. The open-reading frame had 999 bp and coded for 333 amino acids. Hf-GAPDH was predicted to have an N-terminal NAD binding domain and a C-terminal glyceraldehyde dehydrogenase catalytic domain. The molecular structure of Hf-GAPDH was analyzed and the evolutionary relationship also established. The GAPDH protein sequence was conserved among ticks. The expression pattern of Hf-GAPDH, analyzed by real-time PCR, significantly differed among life phases, feeding stages, and tissues. As the ticks grew, the expression level of Hf-GAPDH was up-regulated. The expression levels of Hf-GAPDH in salivary glands and midguts from half-engorged ticks were lower than the same tissues from engorged ticks. This study will provide reference data for the follow-up verification of the GAPDH-related function and the feasibility as a potential anti-tick vaccine.

语种： 英文

作者： Li, Zhong-Bo;Fu, Yi-Tian;Cheng, Tian-Yin;Yao, Guo-Min;Hou, Qiang-Hong;...

期刊： Acta Parasitologica,2019年64(2):360-366 ISSN：1230-2821

通讯作者： Liu, Guo-Hua;Zou, Feng-Cai

作者机构： [Li, Zhong-Bo; Zhao, Yu; Fu, Yi-Tian; Cheng, Tian-Yin; Liu, Guo-Hua; Li, Fen] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Cheng, Tian-Yin; Liu, Guo-Hua; Li, Fen] Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.;[Li, Zhong-Bo; Hou, Qiang-Hong; Yao, Guo-Min] HuaiHua Vocat & Tech Coll, Coll Anim Sci & Technol, Huaihua 418000, Hunan, Peoples R China.;[Zhao, Yu] Xinyang Agr & Forestry Univ, Coll Anim Sci & Vet Med, Xinyang 464000, Henan, Peoples R China.;[Zou, Feng-Cai] Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan, Peoples R China.

通讯机构： [Liu, Guo-Hua] H;[Zou, Feng-Cai] Y;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.;Yunnan Agr Univ, Coll Anim Sci & Technol, Kunming 650201, Yunnan, Peoples R China.

关键词： Haemaphysalis longicornis;Genetic variation;Mitochondrial DNA;Phylogenetic analysis

摘要： Introduction: Haemaphysalis longicornis is an important ectoparasite of domestic and wild animals that can transmit many pathogens including viruses, fungi, bacteria and protozoa. Materials and methods: In this study, we examined genetic variation and population genetics in three mitochondrial (mt) genes [cox1 (cytochrome c subunit 1), rrnL (large subunit ribosomal RNA) and nad5 (NADH dehydrogenase 5)] among four H. longicornis populations from China. Results: The sizes of the partial sequences of cox1, rrnL and nad5 were 776bp, 409bp, 510bp, respectively. Among the obtained sequences, we identified 22 haplotypes for cox1, 2 haplotypes for rrnL and 17 haplotypes for nad5. Low gene flow and significant genetic differentiation (66.2%) were detected among H. longicornis populations. There was no rapid expansion event in the demographic history of four H. longicornis populations in China. In addition, phylogenetic analyses confirmed that all the Haemaphysalis isolates were H. longicornis which were segregated into two major clades. Conclusion: The mt DNA genes provide a potential novel genetic marker for molecular epidemiology of H. longicornis and assist in the control of tick and tick-borne diseases in humans and animals. © 2019, Witold Stefański Institute of Parasitology, Polish Academy of Sciences.

语种： 英文

作者： 郭鹏宇;程天印

期刊： 畜牧兽医科技信息,2019年(8):43-45 ISSN：1671-6027

作者机构： 湖南农业大学动物医学院,湖南长沙,410128;[郭鹏宇; 程天印] 湖南农业大学

关键词： 鸡;盐酸莱克多巴胺;残留消除规律

摘要： 为建立盐酸莱克多巴胺（RAC）在鸡血清和组织中残留的检测方法,探究其消除规律,本研究以1mg/kg、3mg/kg和9mg/kg的添加剂量将R AC混入饲料,连续饲喂21d,在用药后第1、3、7、14、21d和休药后1、3、7d,分别采集实验组和对照组鸡血液、肌肉、肝脏和胃组织样品,酶联免疫吸附试验（ELISA）检测各样品RAC含量。实验结果为实验组鸡血清中RAC残留量在第7d达到高峰,第7～14天持续下降,第21d出现第2个残留高峰;肌肉中残留量在第7d达到高峰,后逐渐降低;肝脏中比较容易达到残留高峰,第21天后,RAC残留量迅速下降;胃中残留量第14d达到高峰。用ELISA对组织和血清中残留RAC的检测发现,RAC残留浓度基本遵循肝脏>胃>肌肉>血清,休药7d后为胃>肝脏>肌肉>血清。

语种： 中文

作者： 徐律;程天印

期刊： 畜牧兽医学报,2019年50(3):627-636 ISSN：0366-6964

作者机构： 湖南农业大学动物医学院,长沙410128;湖南畜禽安全生产协同创新中心,长沙410128;兽用蛋白质工程疫苗湖南省重点实验室,长沙410128;[徐律; 程天印] 湖南农业大学

关键词： 褐黄血蜱;卵;液相色谱-串联质谱;卵黄蛋白原;特异性肽段

摘要： 本研究旨在探明褐黄血蜱卵内蛋白组分,揭示蜱胚胎发育中的关键营养物质,为筛选可干扰胚胎发育的抗原分子奠定基础。采用液相色谱-串联质谱法对新鲜褐黄血蜱的卵蛋白成分进行分析,基于该蜱的唾液腺转录组翻译文库、中肠转录组翻译文库和Uniprot数据库对各蛋白成分进行鉴定。结果显示:从褐黄血蜱卵黄蛋白提取液中检出特异性肽段221条,由此鉴定蛋白53种,其中高可信蛋白12种,能够确定的功能蛋白有肌动蛋白(actin)和卵黄蛋白原Ⅱ(vitellogenin 2,Vg-2),且Vg-2与卵黄蛋白(vitellins,Vn)一级结构相同。蜱卵原生质所含蛋白较少;Vg-2是蜱卵内存在的唯一一种卵黄蛋白原。

语种： 中文

作者： 周鸿铭;程天印;段德勇

期刊： 中国兽医科学,2019年49(3):317-323 ISSN：1673-4696

作者机构： 湖南农业大学动物医学院,湖南长沙,410128;[段德勇; 周鸿铭; 程天印] 湖南农业大学

关键词： 草原革蜱;森林革蜱;形态学;分子标记;差异分析

摘要： 为了比较草原革蜱(采自内蒙古呼伦贝尔)和森林革蜱(采自宁夏固原)的形态结构与分子标记异同点,借助体视显微镜超景深系统观察了这2种蜱的形态结构;通过分子生物学技术,测定了这2种蜱COX1、ITS2及16S rDNA基因序列,并通过比对这3种基因的同源性、构建系统发生树,分析了这2种蜱的分子标记差异。结果显示,这2种蜱均具有珐琅斑,须肢粗短,缘垛整齐等革蜱属的典型特征。草原革蜱珐琅彩浓厚且色浅呈银白色,刻点小且分布均匀、夹杂少数大刻点,背突直而短,缘垛分布均匀;森林革蜱珐琅彩覆盖大部分盾板且色深呈深褐色,刻点粗细混杂、分布较为稠密,背突向背方弯曲,尾部缘垛较其他缘垛窄;两者具有显著的形态学差异。草原革蜱COX1、ITS2、16S rDNA基因序列与GenBank中已知草原革蜱基因序列的相似度分别为99.88%、99.90%、97.58%;森林革蜱COX1、ITS2、16S rDNA基因序列与GenBank中已知森林革蜱基因序列的相似度分别为98.64%、98.60%、99.53%。进一步对比这2种蜱的COX1、ITS2、16S rDNA基因序列,发现其相似度分别为99.76%、99.75%、98.29%。结果提示:虽然草原革蜱和森林革蜱形态学差异显著,但COX1、ITS2、16S rDNA基因序列相似度极高,两者可能为同一种的不同亚种或COX1、ITS2、16S rDNA基因不适用于革蜱属的种间鉴定。

语种： 中文

作者： Feng, Li-Li;Liu, Lei;Cheng, Tian-Yin*

期刊： Experimental and Applied Acarology,2019年78(3):443-460 ISSN：0168-8162

通讯作者： Cheng, Tian-Yin

作者机构： [Cheng, Tian-Yin; Feng, Li-Li; Liu, Lei] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Cheng, Tian-Yin; Feng, Li-Li; Liu, Lei] Hunan Agr Univ, Hunan Colaborat Innovat Ctr Safety Prod Livestock, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-Yin] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;Hunan Agr Univ, Hunan Colaborat Innovat Ctr Safety Prod Livestock, Changsha 410128, Hunan, Peoples R China.

关键词： Rhipicephalus microplus;Salivary proteins;Proteome;iTRAQ

摘要： Rhipicephalus microplus salivary gland secretes a number of complex bioactive proteins during feeding. These components are important in feeding and affect anti-coagulation, anti-inflammation and also have anti-microbial effects. In this study, tick saliva was collected from partially engorged female (PEF) and fully engorged female (FEF) ticks. Liquid chromatography tandem–mass spectrometry (LC–MS/MS) and isobaric tags for relative and absolute quantification (iTRAQ) were used to identify and quantify R. microplus salivary proteins. A total of 322 unique peptides were detected and 151 proteins were characterized in both PEF and FEF. Of these, 41 proteins are considered as high-confidence proteins. Fifteen high-confidence proteins were upregulated and six high-confidence proteins were downregulated (p < 0.05; PEF:FEF ratio ≥ 1.2 or PEF:FEF ratio ≤ 0.83); 17 high-confidence proteins are slightly changed (PEF:FEF ratio > 0.83 and < 1.2). These high-confidence proteins are involved in several physiological roles, including egg development, transportation of proteins, immunity and anti-microorganism, anti-coagulant, and adhesion. In comparison with PEF, the number of upregulated proteins exceeded the number of proteins downregulated. Salivary protein may be induced by the blood-meal and these proteins contribute to successful feeding. © 2019, Springer Nature Switzerland AG.

语种： 英文

作者： Liu, Lei;Liu, Yi-song;Liu, Guo-hua;Cheng, Tian-yin*

期刊： Parasites & Vectors,2018年11(1):1-7 ISSN：1756-3305

通讯作者： Cheng, Tian-yin

作者机构： [Cheng, Tian-yin; Liu, Guo-hua; Liu, Yi-song; Liu, Lei] Hunan Agr Univ, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-yin] H;Hunan Agr Univ, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： Haemaphysalis flava;Tick;Faeces;Proteome;Blood digestion

摘要： Background: Ticks and tick-borne diseases are of major public health concern. Currently, development of vaccines against ticks is considered crucial for their control. A critical step in this process is the screening of viable antigens. Faeces are byproducts of digestion and blood meal utilization, and partly reflect the vitality and vector potential of ticks. However, an integrated analysis of proteins in tick faeces is lacking. The present study explored the protein components in the faeces of the tick Haemaphysalis flava, by liquid chromatography-tandem mass spectrometry (LC/MS-MS) to identify potential protein antigens for vaccine development against ticks. Methods: Faeces from adult H. flava engorged females were collected. Proteins were extracted from faeces, and the trypsin-digested peptides were analyzed by LC/MS-MS. High confidence proteins were identified based on unique peptides revealed by MS. Potential faecal protein genes, as well as their sources, were also characterized by searching previous transcriptome datasets from the salivary glands and midgut of H. flava. Results: In total, 21 were recognized with confidence. Amongst these, 18 were of likely tick origin, while three proteins (serum albumin, haemoglobin α and β subunits) were likely from hosts. Seventeen unigenes corresponding to these proteins were retrieved by searching our previous H. flava salivary glands and midgut transcriptomic datasets. Some proteins were reported to prevent blood clotting, play a role in immunity and antibiosis, and formation of musculature. The functions of the remaining proteins are unknown. Conclusions: Identifying antigens for tick vaccine development is feasible by analyzing the faecal proteome as well as the transcriptomes of salivary glands and midguts. The vast number of proteins detected in tick faeces highlights the complexity of blood digestion in ticks, a field that needs more investigation. © 2018 The Author(s).

语种： 英文

作者： Liu, Lei;Cheng, Tian-yin*;He, Xiao-ming

期刊： Ticks and Tick-borne Diseases,2018年9(3):490-495 ISSN：1877-959X

通讯作者： Cheng, Tian-yin

作者机构： [He, Xiao-ming; Cheng, Tian-yin; Liu, Lei] Hunan Agr Univ, Hunan Collaborat Innovat Ctr Safety Prod Livestoc, Coll Vet Med, Changsha, Hunan, Peoples R China.;[Cheng, Tian-yin] Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-yin] H;Hunan Agr Univ, Coll Vet Med, Changsha 410128, Hunan, Peoples R China.

关键词： *Blood meal;*Digestion;*Haemaphysalis flava;*Midgut contents;*Proteome

摘要： Scant information is available regarding the proteins involved in blood meal processing in ticks. Here, we aimed to highlight the midgut proteins involved in preventing blood meal coagulation, and in facilitating intracellular digestion in the tick Haemaphysalis flava. Proteins were extracted from the midgut contents of fully engorged and partially engorged ticks. We used liquid chromatography tandem-mass spectrometry (LC–MS/MS) analysis to identify 131 unique peptides, and 102 proteins. Of these, 15 proteins, each with at least two unique peptides, were recognized with high confidence. We also retrieved 18 unigenes from our previous published transcriptomic libraries of the midguts and salivary glands of H. flava, and inferred the primary structures of nine proteins and fragments of five proteins. There were 23 and 21 unique proteins in the midgut contents of fully engorged and partially engorged ticks, respectively. We detected 58 shared proteins in the midgut contents of both fully engorged and partially engorged ticks. Of these, seven were significantly differentially expressed between fully engorged and partially engorged ticks: actin, calmodulin, elongation factor-1α, hsp90, multifunctional chaperone, tubulin α, and tubulin β. Our results demonstrated that the proteome of the midgut contents, combined with the transcriptome of the midgut, was a viable method for the reinforcement of protein identification. This method will facilitate further study of blood meal processing by ticks, as well as the identification of clues for tick infestation control. The existence of numerous proteins detected in the midgut contents also highlight the complexity of blood digestion in ticks; this area is in need of further investigation. © 2018 Elsevier GmbH

语种： 英文

作者： Li, Zhong-Bo;Liu, Guo-Hua;Cheng, Tian-Yin*

期刊： Experimental and Applied Acarology,2018年74(2):171-176 ISSN：0168-8162

通讯作者： Cheng, Tian-Yin

作者机构： [Li, Zhong-Bo; Cheng, Tian-Yin; Liu, Guo-Hua] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Cheng, Tian-Yin; Liu, Guo-Hua] Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Cheng, Tian-Yin] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

关键词： Haemaphysalis longicornis;ITS regions;China;Hedgehog;Goat

摘要： In the present study, the entire first and second internal transcribed spacer (ITS-1 and ITS-2) regions of nuclear ribosomal DNA (rDNA) of Haemaphysalis longicornis from China were amplified by polymerase chain reaction. The 45 representative amplicons were sequenced, and sequence variation in the ITS was examined. The ITS sequences of H. longicornis were 3644bp in size, including the part of 18S rDNA, 28S rDNA sequences and the complete ITS-1, 5.8S rDNA and ITS-2 sequences. Sequence analysis revealed that the ITS-1, 5.8S rDNA and ITS-2 of this hard tick were 1582, 152, and 1610bp in size, respectively. The intra-specific sequence variations of ITS-1 and ITS-2 within H. longicornis were 0–2 and 0–2.2%; however, the inter-specific sequence differences among members of the genus Haemaphysalis were significantly higher, being 35.1–55.2 and 37–52% for ITS-1 and ITS-2, respectively. The molecular approach employed in this study provides the foundation for further studies of the genetic variation of H. longicornis from different hosts and geographical origins in China. © 2018, The Author(s).

语种： 英文

作者： He, X. -M.;Cheng, T. -Y.*

期刊： JOURNAL OF APPLIED ENTOMOLOGY,2018年142(1-2):173-180 ISSN：0931-2048

通讯作者： Cheng, T. -Y.

作者机构： [He, X. -M.; Cheng, T. -Y.] Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.

通讯机构： [Cheng, T. -Y.] H;Hunan Agr Univ, Coll Vet Med, Changsha, Hunan, Peoples R China.

关键词： bacteria;Haemaphysalis flava;microbiome;saliva

摘要： <jats:title>Abstract</jats:title><jats:p>The tick <jats:italic>Haemaphysalis flava</jats:italic> is one of the most significant blood‐feeding arthropod parasites and is a vector for numerous human and animal pathogens. However, a comprehensive investigation of the microbial communities found in the saliva of this tick species is lacking. This study used 16S <jats:styled-content style="fixed-case">rRNA</jats:styled-content> Illumina sequencing to characterize the compositions of microbiomes present in saliva and whole tick samples isolated from engorged and partially fed adult <jats:italic>H.flava</jats:italic> females. This revealed that the bacterial diversity present in tick saliva increased after a prolonged blood meal, and that the species diversity found in saliva was significantly higher than that of whole ticks. Three bacteria phyla, in particular, made up more than 80% of the microbial community across all samples—<jats:italic>Proteobacteria, Firmicutes</jats:italic> and <jats:italic>Actinobacteria</jats:italic>. Furthermore, some of the genera identified in this study had not previously been reported in ticks before, such as <jats:italic>Facklamia, Vagococcus, Ruminococcus, Lachnospira, Bradyrhizobium, Peptostreptococcus, Jeotgalicoccus, Roseburia, Brachybacterium, Sporosarcina, u114, Megamonas</jats:italic> and <jats:italic>Dechloromonas</jats:italic>. Finally, we found that many of the isolated bacteria were opportunistic pathogens, indicating a potential risk to humans and livestock exposed to <jats:italic>H.flava</jats:italic>. These results will contribute to fully understanding the transmission of tick‐borne pathogens.</jats:p>

语种： 英文

作者： Tang, Jia-Min;Li, Fen;Cheng, Tian-Yin;Duan, De-Yong*;Liu, Guo-Hua*

期刊： Parasitology Research,2018年117(8):2677-2683 ISSN：0932-0113

通讯作者： Duan, De-Yong;Liu, Guo-Hua

作者机构： [Duan, De-Yong; Liu, Guo-Hua; Tang, Jia-Min; Cheng, Tian-Yin; Li, Fen] Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;[Duan, De-Yong; Liu, Guo-Hua; Cheng, Tian-Yin; Li, Fen] Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

通讯机构： [Duan, DY; Liu, GH] H;Hunan Agr Univ, Coll Vet Med, Hunan Prov Key Lab Prot Engn Anim Vaccines, Changsha 410128, Hunan, Peoples R China.;Hunan Coinnovat Ctr Anim Prod Safety, Changsha 410128, Hunan, Peoples R China.

关键词： amino acid;genomic DNA;mitochondrial DNA;molecular marker;nucleotide;Article;China;Diptera;DNA extraction;ectoparasite;geographic distribution;Melophagus ovinus;microbial genome;mitochondrial genome;molecular biology;nonhuman;nucleotide sequence;paraphyly;parasite genome;population genetics;priority journal;sequence analysis;sheep;start codon;Xinjiang;animal;comparative study;Diptera;female;genetics;geography;male;mitochondrial genome;parasitology;phylogeny;sheep disease;Animals;China;Diptera;Female;Genome, Mitochondrial;Geography;Male;Phylogeny;Sheep;Sheep Diseases

摘要： The sheep ked Melophagus ovinus is mainly found in Europe, Northwestern Africa, and Asia. Although M. ovinus is an important ectoparasite of sheep in many countries, the population genetics, molecular biology, and systematics of this ectoparasite remain poorly understood. Herein, we determined the mitochondrial (mt) genome of M. ovinus from Gansu Province, China (MOG) and compared with that of M. ovinus Xinjiang Uygur Autonomous Region, China (MOX). The mt genome sequence (15,044bp) of M. ovinus MOG was significantly shorter (529bp) than M. ovinus MOX. Nucleotide sequence difference in the whole mt genome except for non-coding region was 0.37% between M. ovinus MOG and MOX. For the 13 protein-coding genes, comparison revealed sequence divergences at both the nucleotide (0–1.1%) and amino acid (0–0.59%) levels between M. ovinus MOG and MOX, respectively. Interestingly, the cox1 gene of M. ovinus MOX is predicted to employ unusual mt start codons AAA, which has not been predicted previously for any parasite genome. Phylogenetic analyses showed that M. ovinus (Hippoboscoidea) is related to the superfamilies Oestroidea + Muscoidea. Our results have also indicated the paraphylies of the four families (Anthomyiidae, Calliphoridae, Muscidae, and Oestridae) and two superfamilies (Oestroidea and Muscoidea). This mt genome of M. ovinus provides useful molecular markers for studies into the population genetics, molecular biology, and systematics of this ectoparasite. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.

语种： 英文