本文選題：家蠅 + 變態(tài)發(fā)育��； 參考：《中國寄生蟲學(xué)與寄生蟲病雜志》2017年02期

【摘要】：目的研究家蠅(Musca domestica)不同發(fā)育階段腸道中可培養(yǎng)共生細(xì)菌的組成。方法在無菌條件下,對不同發(fā)育階段家蠅蟲體進(jìn)行體表消毒。對卵和孵化1 d的幼蟲,取5枚卵或5條幼蟲,加1 ml無菌水研磨;對孵化2 d以上的幼蟲、蛹和成蠅,各蟲期分別取5條,解剖出腸道,加1 ml無菌水研磨。取研磨液按傳統(tǒng)分離培養(yǎng)法分離和鑒定家蠅腸道的共生細(xì)菌,挑取形態(tài)有差異的單菌落,置于LB培養(yǎng)基中小量培養(yǎng),提取菌體DNA,PCR擴(kuò)增16S r DNA基因,測序后在GenBank中進(jìn)行比對,鑒定到屬。計數(shù)家蠅不同蟲期腸道中分離的細(xì)菌菌屬數(shù)的變化情況。結(jié)果通過培養(yǎng)分離出家蠅各階段腸道共生細(xì)菌26屬,其中,卵期未分離到共生細(xì)菌,幼蟲期分離到11屬,蛹期分離到19屬,成蠅分離到14屬。不同蟲期之間存在重復(fù),幼蟲期和蛹期共有的細(xì)菌有6屬,分別為變形桿菌屬(Proteus)、庫特氏菌屬(Kurthia)、博德特氏菌屬(Bordetella)、微桿菌屬(Microbacterium)、產(chǎn)堿桿菌屬(Alcaligenes)和明串球菌屬(Leuconostoc);蛹期和成蠅共有的細(xì)菌有3屬,分別為鞘氨醇桿菌屬(Sphingobacterium)、腸球菌屬(Enterococcus)、蒼白桿菌屬(Ochrobactrum);幼蟲期和成蠅共有的細(xì)菌有1屬,為克雷伯菌屬(Klebsiella);3個蟲態(tài)共有的細(xì)菌有4屬,分別為普羅威登斯菌屬(Providencia)、葡萄球菌屬(Staphylococcus)、香味菌屬(Myroides)和假單胞菌屬(Pseudomonas)。幼蟲期、蛹期和成蠅革蘭陽性菌的比例分別為4/11、6/19和3/14。結(jié)論在家蠅由幼蟲經(jīng)蛹發(fā)育到成蠅的過程中,其腸道中可培養(yǎng)細(xì)菌菌屬數(shù)呈先上升后下降趨勢,革蘭陽性菌的比例逐漸下降。
[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.
【作者單位】： 泰山醫(yī)學(xué)院新發(fā)傳染病溯源及防控協(xié)同創(chuàng)新中心;泰山醫(yī)學(xué)院基礎(chǔ)醫(yī)學(xué)院;
【基金】：國家自然科學(xué)基金(No.81401693,81572028,81271874) 山東省科技發(fā)展計劃項目(No.2014GSF121007)~~
【分類號】：R384.2

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