家蝇发育过程中肠道可培养共生细菌的分离与鉴定

发布时间：2018-06-27 03:48

本文选题：家蝇 + 变态发育　；参考：《中国寄生虫学与寄生虫病杂志》2017年02期

【摘要】：目的研究家蝇(Musca domestica)不同发育阶段肠道中可培养共生细菌的组成。方法在无菌条件下,对不同发育阶段家蝇虫体进行体表消毒。对卵和孵化1 d的幼虫,取5枚卵或5条幼虫,加1 ml无菌水研磨;对孵化2 d以上的幼虫、蛹和成蝇,各虫期分别取5条,解剖出肠道,加1 ml无菌水研磨。取研磨液按传统分离培养法分离和鉴定家蝇肠道的共生细菌,挑取形态有差异的单菌落,置于LB培养基中小量培养,提取菌体DNA,PCR扩增16S r DNA基因,测序后在GenBank中进行比对,鉴定到属。计数家蝇不同虫期肠道中分离的细菌菌属数的变化情况。结果通过培养分离出家蝇各阶段肠道共生细菌26属,其中,卵期未分离到共生细菌,幼虫期分离到11属,蛹期分离到19属,成蝇分离到14属。不同虫期之间存在重复,幼虫期和蛹期共有的细菌有6属,分别为变形杆菌属(Proteus)、库特氏菌属(Kurthia)、博德特氏菌属(Bordetella)、微杆菌属(Microbacterium)、产碱杆菌属(Alcaligenes)和明串球菌属(Leuconostoc);蛹期和成蝇共有的细菌有3属,分别为鞘氨醇杆菌属(Sphingobacterium)、肠球菌属(Enterococcus)、苍白杆菌属(Ochrobactrum);幼虫期和成蝇共有的细菌有1属,为克雷伯菌属(Klebsiella);3个虫态共有的细菌有4属,分别为普罗威登斯菌属(Providencia)、葡萄球菌属(Staphylococcus)、香味菌属(Myroides)和假单胞菌属(Pseudomonas)。幼虫期、蛹期和成蝇革兰阳性菌的比例分别为4/11、6/19和3/14。结论在家蝇由幼虫经蛹发育到成蝇的过程中,其肠道中可培养细菌菌属数呈先上升后下降趋势,革兰阳性菌的比例逐渐下降。
[Abstract]:Objective to study the composition of culturable symbiotic bacteria in the intestine of Musca domestica at different developmental stages. Methods the body surface of Musca domestica at different developmental stages was disinfected under aseptic condition. Five eggs or five larvae were taken from eggs and larva hatched for one day, and 1 ml sterile water was added to grind them, and 5 larvae, pupae and adult fly hatched more than 2 days were taken from each stage, and intestinal tract was dissected and ground with 1 ml aseptic water. The symbiotic bacteria in the intestinal tract of Musca domestica were isolated and identified by the traditional isolation and culture method. The single colony with different morphology was picked up and cultured in a small amount in LB medium. The 16s r DNA gene was amplified by PCR from the DNA of the bacteria, and compared with that in GenBank after sequencing. The genus was identified. The number of bacterial genera isolated from intestinal tract of Musca domestica at different stages was counted. Results 26 genera of intestinal symbiotic bacteria were isolated from housefly in different stages. Among them, no symbiotic bacteria were isolated in egg stage, 11 genera in larval stage, 19 genera in pupal stage and 14 genera in adult fly. There are 6 genera of bacteria in larval and pupa stages. They are Proteus, Kurthia, Bordetella, Microbacterium, (Alcaligenes) and Leuconostoc. They are Sphingobacterium, Enterococcus and Ochrobactrum, respectively. They are Providencia, Staphylococcus, Myroides and Pseudomonas. The percentages of gram-positive bacteria in larval stage, pupa stage and adult fly were 4 / 11 / 6 / 19 and 3 / 14, respectively. Conclusion during the development of housefly from larva through pupa to adult fly, the number of culturable bacteria in the intestinal tract of Musca domestica increased first and then decreased, and the proportion of Gram-positive bacteria decreased gradually.
【作者单位】：泰山医学院新发传染病溯源及防控协同创新中心;泰山医学院基础医学院;
【基金】：国家自然科学基金(No.81401693,81572028,81271874) 山东省科技发展计划项目(No.2014GSF121007)~~
【分类号】：R384.2

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