摘要：
Bioactive components in the midgut of ticks play a key role in tick blood digestion, feeding and pathogen transmission. The study of protein and gene targets in midgut provides opportunities to explore novel tick control strategies. Only a few nucleotide sequences are available in public databases for Haemaphysalis flava, an important disease vector for humans and animals. Knowledge of the process of blood digestion by the ticks and protein expression in the digestive tract is limited. Here, we utilize high-throughput sequencing to characterize the mid-gut transcriptome of fully engorged (FE, average length of 10 mm) and partially engorged (PE, average length of 5 mm) female H. flava. 6.8 GB and 8.3 GB of high-quality sequence datawere obtained using Illumina sequencing technology. 54,357,490 and 66,116,050 reads were finally assembled into 76,556 unigenes with mean length of 704 bp. The transcripts involved in bloodmeal digestion were classified into eight large categories, including peptidase inhibitor, peptidase (serine-, metallo-, cysteine-, aspartic-peptidase), phospholipase, carbohydrate digestion/hydrolases, lipid binding, immunity-related proteins, iron/heme metabolism and secreted proteins. A total of 5508 differentially expressed genes (DEGs) were identified between FE and PE. To confirm the DEG results, ten genes involved in blood digestion, feeding and defending from pathogens, were validated using qPCR. Our results not only contribute to better understanding of the changes inmidgut transcript expression during different blood feeding stages, but also provide a valuable resource for identifying targets for future tick control studies. (C) 2015 Elsevier B.V. All rights reserved.
摘要：
Trichuris ovis parasitizes the caeca and colon of hosts, causing trichuriasis. Despite this parasite being of animal health significance, causing considerable socio-economic losses globally, little is known of the molecular characteristics of T. ovis. In the present study, the entire first and second internal transcribed spacer (ITS-1 and ITS-2) regions of nuclear ribosomal DNA (rDNA) of T. ovis from goats in Hunan province, China were amplified by polymerise chain reaction (PCR), and the representative amplicons were subjected to sequencing in order to estimate sequence variation. The ITS rDNA sequences for the T. ovis samples were 1289-1303 bp in length. Sequence analysis revealed that the ITS-1 rDNA, 5.8S and ITS-2 rDNA of these whipworms were 725-734 bp, 163 bp and 401-406 bp in size, respectively. Sequence variation in ITS rDNA within and among T. ovis was also examined. Excluding nucleotide variations in the simple sequence repeats, the intra-species sequence variation in the ITS-1 was 0-2.7%, and ITS-2 was 0-4.1%. The inter-species sequence variation among the T. ovis and other whipworms were more than 45.7% for ITS-1 rDNA and 51.3% for ITS 2 rDNA. These results demonstrated that the ITS rDNA sequences provide additional genetic markers for the identification and differentiation of the T. ovis. These data should be useful for studying the epidemiology, population genetics of T. ovis, as well as for the diagnosis of trichuriasis in sheep and goats.
摘要：
Saliva plays an important role in feeding and pathogen transmission, identification and analysis of tick salivary gland (SG) proteins is considered as a hot spot in anti-tick researching area. Herein, we present the first description of SG transcriptome of Haemaphysalis flava using next-generation sequencing (NGS). A total of over 143 million high-quality reads were assembled into 54,357 unigenes, of which 20,145 (37.06%) had significant similarities to proteins in the Swiss-Prot database. 13,513 annotated sequences were associated with GO terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that 14,280 unigenes were assigned to 279 KEGG pathways in total. Reads per kb per million reads (RPKM) analysis showed that there were 3035 down-regulated unigenes and 2260 up-regulated unigenes in the engorged ticks (ET) compared with the semi-engorged one (SET). Several important genes are associated with blood feeding and ingestion as secreted salivary proteins, concluding cysteine, longipain, 408, calreticulin, metalloproteases, serine protease inhibitor, enolase, heat shock protein and AV422 in SG, were identified. The qRT-PCR results confirmed that patterns of these genes (except for the longipain gene) expression were consistent with RNA-seq results. This de novo assembly of SG transcriptome of H.flava not only provides more chance for screening and cloning functional genes, but also forms a solid basis for further insight into the changes of salivary proteins during blood-feeding. (C) 2015 Elsevier B.V. All rights reserved.
摘要：
蜱类系统分类学研究已经进入分子阶段，与传统的形态学、生物学鉴定相比，无疑更加客观准确，建立系统进化树更能全面反映其亲缘关系，极大地提高了鉴定效率与可信度。论文总结了近年来应用分子生物学技术鉴定硬蜱的主要方法，包括核酸序列分析、随机扩增 DNA 多态性分析、限制性片段长度多态性分析。随着分子生物学和现代统计学的广泛应用，蜱类的鉴定将更加依赖现代分类学方法。
摘要：
The mir-155 family is not only involved in a diversity of cancers, but also as a regulator of the immune system. However, the evolutionary history of this family is still unclear. The present study indicates that mir-155 evolved independently with lineage-specific gain of miRNAs. In addition, arm switching has occurred in the mir-155 family, and alternative splicing could produce two different lengths of ancestral sequences, implying the alternative splicing can also drive evolution for intragenic miRNAs. Here we screened validated target genes and immunity-related proteins, followed by analyzation of the mir-155 family function by high-throughput methods like the gene ontology (GO) and Kyoto Eneyclopedin of Genes and Genemes (KEGG) pathway enrichment analysis. The high-throughput analysis showed that the CCND1 and EGFR genes were outstanding in being significantly enriched, and the target genes cebpb and VCAM1 and the protein SMAD2 were also vital in mir-155-related immune reponse activities. Therefore, we conclude that the mir-155 family is highly conserved in evolution, and CCND1 and EGFR genes might be potential targets of mir-155 with regard to progress of cancers, while the cebpb and VCAM1 genes and the protein SMAD2 might be key factors in the mir-155 regulated immune activities.
