广斑虻3个部位携带菌群种类及结构分析

Species and structure of bacterial flora carried by three parts of Chrysops Vanderwulpi

目的调查广斑虻中肠、马氏管和卵巢中携带菌群种类以及结构差异。方法在黑龙江省大庆市某牛场附近捕捉广斑虻(Chrysops vanderwulpi),无菌采集其中肠、马氏管和卵巢3个部位进行16S V3-V4区测定和分析,并进行细菌分离培养。结果研究样本25份,共获得1331个操作分类单元(OTUs),中肠、马氏管和卵巢分别为1211、1204和1257个OTUs,其中1051个OTUs为3种样本所共有。3个部位在门水平中,厚壁菌门、变形菌门、拟杆菌门最丰富,合计超过80.00%。在属水平上,不同部位细菌组成情况不同,毛螺菌属在中肠相对丰度最高,占菌群总含量的8.38%;乳酸菌属在马氏管中相对丰度最高,占菌群总含量的12.13%;肠杆菌属在卵巢中相对丰度最高,占菌群总含量的10.60%。对所有序列进行Blast分析,部分序列与致病菌人型支原体(CP055150)、索氏梭菌(MT539084)、副流感嗜血杆菌(LT674777)、类志贺邻单胞菌(MT573123)、伯克霍尔德菌(MN150518)、微小脲原体(CP041199)、螺旋杆菌(MF440359)同源性均达到100.00%。在培养结果中,需氧条件培养出37株5属9种,厌氧条件下共培养出20株6属7种,未培养出致病菌。结论本研究首次对虻类进行了细菌多样性分析,初步了解广斑虻中肠、马氏管和卵巢的细菌组成及差异情况;在虻类中检测到多种潜在致病菌,并对部分细菌进行了分离鉴定,未培养出致病菌,研究结果为虻及虻传播疾病的深入研究提供了可靠的数据。

Objective In order to clarify the species and structure differences of bacteria in the gut,Malpighia's duct and ovary of the Chrysops.vanderwulpi.Methods C.vanderwulpi was captured near a cattle farm in Daqing city,Heilongjiang province.The 16S V3-V4 region of the midgut,Malpighenian duct and ovary of 25 C.vanderwulpi flies was detected and analyzed,and the bacteria were isolated and cultured.Results A total of 1331 OTUs were obtained,with 1211,1204 and 1257 OTUs in the midgut,Malpigsian tubule and ovary,respectively,of which 1051 OUTs were shared by all three samples.At the phylum level of the three sites,Firmicutes,Proteobacteria,and Bacteroidetes were the most abundant,totaling more than 80.00%.At the genus level,the composition of bacteria in different parts was different.The relative abundance of Lachnospiraceae was the highest in the midgut,accounting for 8.38%of the total content of bacteria.The relative abundance of Lactobacillus was the highest in Malwinia tubule,accounting for 12.13%of the total content of bacteria.Enterobacter was the most abundant genus in the ovary,accounting for 10.60%of the total flora.Blast analysis of all sequences showed that some sequences were similar to those of the pathogenic bacteria Metamycoplasma hominis,(CP055150),Paeniclostridium sordellii,(MT539084),Haemophilus parainfluenzae(LT674777),Plesiomonas shigelloides(MT57312),Paraburkholderia terrae(MN150518),Ureaplasma parvum(CP041199),Helicobacter cinaedi,Paraburkholderia terrae(MF440359)were 100.00%homology.In the culture results,37 strains,5 genera and 9 species were cultured under aerobic condition,while 20 strains,6 genera and 7 species were cultured under anaerobic condition,and no pathogenic bacteria were cultured.Conclusion In this study,the bacterial diversity of Tabanus was analyzed for the first time,and the composition and differences of bacteria in the gut,Malpiglet's midduct and ovary of C.vanderwulpi were analyzed.A variety of potential pathogens have been detected in the Horseflies,and some bacteria were isolated and identified,but no pathogenic bacteria were cultured.The results provide reliable data for the further study of the flies and the diseases transmitted by them.