To obtain fusion protein of Mycobacterium bovis with high purity, the recombinant prokaryotic expression vector for Mb2277 gene was constructed and the immunogenicity of its products was initially investigated in the present study.A pair of primer was designed according the gene sequence Mb2277 from the genomic DNA of M.Bovis in GenBank. and was amplified by PCR using DNA of M.Bovis 93006 strain as template. The PCR product and pET-28a(+) was then digested by BamHⅠ and EcoR Ⅰdouble enzyme. To constructed a prokaryotic expression plasmid, the purified Mb2277 was cloned to pET28a(+). Then the recombinant plasmid was transformed into competent cell of E.coli BL21(DE3).The bacteria were induced by IPTG and its lysates were analyzed by SDS-PAGE and Western-blotting. In this way, the prokaryotic expression plasmid for M. bovis Mb 2277 protin was obtained, and a expression band with molecular of 25 ku could be found in SDS-PAGE analysis. As demonstrated by Western blotting this expression product showed excellent reactivity with rabbit immune sera against M. bovis.

HEV is a positive strand RNA virus containing 3 open reading frames(ORFs), while the function of its binding protein pORF3 is poorly understood. In our study, the interactive protein of the pORF3 from human liver cDNA library was screened by the SOS recruitment system, and the construction of the pSOS bait-ORF3 plasmid was identified. The expression of the pORF3 was analyzed by Western blotting, and the detection result of the self activation of bait protein was obtained as expected. Totally, 8 putative interacting proteins had been successively screened. Some of these cellular binding proteins of pORF3, especially the important immunological pathway kinase p110δ, may advance our understanding of the role in ORF3 protein of HEV, and the possible implications of these candidate proteins for the further research are discussed.

Objective To evaluate the influence of Eperythrozoon infection on human and mouse erythrocytes and to explore the pathogenesis of Eperythrozoonosis. Methods The specific gene fragment of Eperythrozoon was detected by polymerase chain reaction (PCR) from the venous blood samples of five patients infected with Eperythrozoon. The complement receptor type I (CD35) expression on erythrocytes of these five patients was determined by flow cytometry. Thereafter, the Eperythrozoons were purified from human samples and injected into mice through the tail veins. Blood smear microscopy, PCR and transmission electron microscopy were used to assure the successful infection. The hematological indicators of human and mice, such as red blood cell (RBC) count,hemoglobin (Hb) content, hematocrit and superoxide dismutase (SOD) were evaluated. All results were analyzed by t test. Results More than 80% of treated mice were confirmed to be infected with Eperythrozoon successfully. A fragment of 801 bp specific gene of Eperythrozoon was detected by PCR in samples from both infected patients and infected mice, which were not detected in samples from healthy control people or control mice. CD35 was highly expressed on the erythrocytes of infected patients, but not expressed on the erythrocytes of infected mice. Both RBC counts and Hb content dramatically decreased in infected patients and infected mice. Hematocrit and the activity of SOD also slightly decreased in infected patients and infected mice. Conclusions Eperythrozoon can spread between human and mice and destroy erythrocyte structure. Eperythrozoon can upregulate CD35 expression in human, but there is no CD35 expression in mice.

Objective To study the genomic molecular organization and genotype of human astro-virus infected infants in Shanghai of China. Methods Based on the published genomic sequence of HAstV (GenBank), the whole genome of one isolate human astrovirus was sequenced by specific primers. The PCR-products were cloned to pMD18-T vector and sequenced, phylogenetic tree was constructed by the Neighbor-Joining method with MEGA 4 software. Results The genome of HAstV-SH is 6807 hp, contains three ORFs: ORF1a and 1b encode the non-structural protein (from 83 nt to 4372 nt), ORF2 encodes the structural protein (from 4364 nt to 6727 nt). Compared with the ORF2 gene of those eight astrovirus sero-types in GenBank, revealed that the highest homology is with genotype 1 (97%). Homology with other gen-otypes ranged between 63% and 70%. Conclusion HAstV-SH belonged to genotype 1 and closely clus-tered with a strain of Japan (AB009985).

Background: Ticks are one of the most common external parasites in dogs, and are associated with the transmission of a number of major zoonoses, which result in serious harm to human health and even death. Also, the increasing number of pet dogs and pet owners in China has caused concern regarding human tick-borne illnesses. Accordingly, studies are needed to gain a complete understanding of the bacterial composition and diversity of the ticks that parasitize dogs. Objectives: To date, there have been relatively few reports on the analysis of the bacterial community structure and diversity in ticks that parasitize dogs. The objective of this study was to investigate the microbial composition and diversity of parasitic ticks of dogs, and assessed the effect of tick sex and geographical region on the bacterial composition in two tick genera collected from dogs in China. Methods: A total of 178 whole ticks were subjected to a 16S ribosomal RNA (rRNA) next generation sequencing analysis. The Illumina MiSeq platform targeting the V3–V4 region of the 16S rRNA gene was used to characterize the bacterial communities of the collected ticks. Sequence analysis and taxonomic assignment were performed using QIIME 2 and the GreenGene database, respectively. After clustering the sequences into taxonomic units, the sequences were quality-filtered and rarefied. Results: After pooling 24 tick samples, we identified a total of 2,081 operational taxonomic units, which were assigned to 23 phyla and 328 genera, revealing a diverse bacterial community profile. The high, moderate and low prevalent taxa include 46, 101, and 182 genera, respectively. Among them, dominant taxa include environmental bacterial genera, such as Psychrobacter and Burkholderia. Additionally, some known tick-associated endosymbionts were also detected, including ,Coxiella, Rickettsia, and Ricketssiella. Also, the potentially pathogenic genera Staphylococcus and Pseudomonas were detected in the tick pools. Moreover, our preliminary study found that the differences in microbial communities are more dependent on the sampling location than tick sex in the tick specimens collected from dogs. Conclusions The findings of this study support the need for future research on the microbial population present in ticks collected from dogs in China.

Background: Ticks are one of the most common external parasites in dogs, and are associated with the transmission of a number of major zoonoses, which result in serious harm to human health and even death. Also, the increasing number of pet dogs and pet owners in China has caused concern regarding human tick-borne illnesses. Accordingly, studies are needed to gain a complete understanding of the bacterial composition and diversity of the ticks that parasitize dogs. Objectives: To date, there have been relatively few reports on the analysis of the bacterial community structure and diversity in ticks that parasitize dogs. The objective of this study was to investigate the microbial composition and diversity of parasitic ticks of dogs, and assessed the effect of tick sex and geographical region on the bacterial composition in two tick genera collected from dogs in China. Methods: A total of 178 whole ticks were subjected to a 16S ribosomal RNA (rRNA) next generation sequencing analysis. The Illumina MiSeq platform targeting the V3–V4 region of the 16S rRNA gene was used to characterize the bacterial communities of the collected ticks. Sequence analysis and taxonomic assignment were performed using QIIME 2 and the GreenGene database, respectively. After clustering the sequences into taxonomic units, the sequences were quality-filtered and rarefied. Results: After pooling 24 tick samples, we identified a total of 2,081 operational taxonomic units, which were assigned to 23 phyla and 328 genera, revealing a diverse bacterial community profile. The high, moderate and low prevalent taxa include 46, 101, and 182 genera, respectively. Among them, dominant taxa include environmental bacterial genera, such as Psychrobacter and Burkholderia. Additionally, some known tick-associated endosymbionts were also detected, including ,Coxiella, Rickettsia, and Ricketssiella. Also, the potentially pathogenic genera Staphylococcus and Pseudomonas were detected in the tick pools. Moreover, our preliminary study found that the differences in microbial communities are more dependent on the sampling location than tick sex in the tick specimens collected from dogs. Conclusions The findings of this study support the need for future research on the microbial population present in ticks collected from dogs in China.