论文部分内容阅读
目的了解进食对家蝇成虫肠道共生菌数量和种类的影响。方法刚羽化的家蝇随机分为正常喂食组和未喂食组,每天分雌、雄取样,解剖肠道后,分离其肠道内细菌,直到全部死亡。家蝇肠道共生细菌采用传统方法分离培养。挑取形态有差异的单菌落,于LB培养基中摇菌过夜,提取DNA,进行16srDNA基因扩增,扩增产物测序,并在NCBI中进行序列比对,鉴定到属,计数家蝇不同虫态肠道中分离到的细菌菌属数,分析其变化。结果在家蝇成蝇肠道内共分离到共生菌17属,其中正常进食成蝇分离到12属,未进食成蝇肠离到10属,两组共有细菌6属,分别为普罗威登斯菌属、葡萄球菌属、香味菌属、克雷伯菌属、假单胞菌属和不动杆菌属。正常喂食的家蝇肠道中特有的细菌有7属,分别为苍白杆菌属、鞘氨醇杆菌属、肠球菌属、寡养单胞菌属、土壤杆菌、代尔夫特菌属和漫游球菌属;未喂食家蝇肠道中特有的细菌有4属,分别为白色杆菌属、肠杆菌属、短状杆菌属和微杆菌属。正常喂食组不同日龄雌虫和雄虫肠道中分离到的共生细菌菌属数量差异有统计学意义(F值分别为5.57和3.57,P<0.05或P<0.01);未进食组不同日龄雌虫和雄虫肠道中分离到的共生细菌菌属数量差异无统计学意义(F值分别为0.17和0.92,P>0.05)。结论进食不但可影响家蝇的寿命,还可影响家蝇成虫体内的肠道共生细菌的组成,推测其肠道共生细菌可能部分来自于食物和生活环境。
Objective To understand the effects of feeding on the numbers and types of intestinal commensal bacteria in housefly adults. Methods The newly emerged houseflies were randomly divided into normal feeding group and non-feeding group. Female and male samples were taken daily to dissect the intestine and the bacteria in the intestine were isolated until all died. Housefly gut symbiotic bacteria were isolated and cultured by traditional methods. Single colonies with different morphology were picked and shaken overnight in LB medium. DNA was extracted and 16srDNA gene was amplified. The amplified products were sequenced and sequenced in NCBI. State bacteria intestinal bacteria isolated from the number, analysis of its changes. Results In the flies of Musca domestica, a total of 17 genera of commensal bacteria were isolated. Among them, 12 genera were isolated from normal feeding adult flies and 10 genera were isolated from non-fed adult flies. There were 6 genera of bacteria belonging to Providencia Staphylococcus, Flavobacterium, Klebsiella, Pseudomonas and Acinetobacter. There are 7 genera in the gut of housefly that are normally fed, which are Corynebacterium, Sphingomonas, Enterococcus, Stenotrophomonas, Agrobacterium, Delfisteria, ; Non-fed Housefly intestinal specific bacteria have 4 genera, respectively, white bacillus, Enterobacter, Bacillus and Microbacterium genus. There were significant differences in the number of commensal bacteria isolated from the gut of male and female in the normal feeding group (F = 5.57 and 3.57 respectively, P <0.05 or P <0.01) There were no significant differences in the number of commensal bacteria isolated from the gut and male gut (F = 0.17 and 0.92, P> 0.05). Conclusions Feeding can not only affect the life span of house flies, but also affect the composition of intestinal symbionts in housefly adults. It is speculated that intestinal commensal bacteria may come partly from food and living environment.