| Objective:The objective in this study is to conduct a survey on the distribution and prevelance characteristics of Bartonella spp.isolated from rodents in different provinces in China,and to explore their phylogenetic relationship and genetic diversity.This study will provide the scientific foundation for the prevention and provide basic information for control of the Bartonellosis.Methods:1.Rodents were captured between 2013 and 2016 on the basis of zoogeographic division of China(including fifteen provinces).Rodents were live-trapped using the trap-at-night method.Tissue samples were taken from liver and kidney under the aseptic condition.2.Bacterial isolation was performed by inoculating spleen onto Tryptic Soy Agar medium in which the 5%defibrinated sheep blood was minxed,then put the plates into incubator at 35℃ in 5%CO2 for 20-30 days.Bartonella-like colonies were identified by the citrate synthase gene(gltA)and visualizing the target gene fragment by gel electrophoresis.For all identified isolates,amplify the following four genes:16S rRNA gene、cell division protein-encoding gene(ftsZ)、the RNA polymerase β subunit-encoding gene(rpoB).And then their sequences were obtained3.By the nucleotide BLAST program(National Center for Biotechnology Information),each gene sequences obtained were compared with sequences deposited in Genbank.Alignment and phylogenetic analysis of individual gene sequences and concatenated sequences(gltA-16S rRNA-ftsZ-rpoB)were performed using MEGA 7.0(Molecular Evolutionary Genetics Analysis software).The phylogeny tree was construted using the Neighbor-joining method and Maximum likelihood method.4.Nucleotide diversity and the haplotype diversity were analysed by DnaAP 5.10.1(DNA Sequence Polymorphism software).Results:1.A total of 2826 liver tissues were inoculated in this study,and 26.97%(757/2826)of culture-medium were PCR positive for Bartonella spp.The total of 757 Bartonella-infected animals with a total 62 species belonging to 3 orders,9 families,25 genera and 62 species.There were differences about Bartonella infection rate in rodents.And the species of rodents in study province was different.2.Bartonella were detected in rodents from all the study province.There were significant differences about the positive rates for Bartonella infection between different study provinces.3.According to the comparison for the homology of nucleotide and the analysis of molecular evolutionary genetics,12 Bartonella genetypes were detected in the different species of rodents from zoogeographic regions.It includes B.taylorii、B.grahamii、B.rattimassiliensis、B.elizabethae、B.japonica、B.tribocorum、B.silvatica、B.queenslandensis、B.jaculi、B.washoensis、B.phoceensis、B.heixiaziensis,and new potential genetypes.It has been found that Bartonella genetype in different rigions were different.Conclusion:This study confirmed that Bartonella were tested in a wild rage of rodents,and widely spread in different regions in China.It was characterized by broad adaptability and host diversity.The results of genetic evolution analysis revealed that it possess genetic diversity and there may be new potential species.Furthermore,there may be possible that some unknown diseases might be associated with Bartonella infection because of the exist of pathogenic Bartonella,potential or new one.Hence,this subject provides scientific references for prevention and control,and epidemiology of Bartonellosis in China. |
