Bacterial diversity analysis of larvae and adult midgut microflora using culture-dependent and culture-independent methods in lab-reared and field-collected Anopheles stephensi-an Asian malarial vector.
Bacterial diversity analysis of larvae and adult midgut microflora using culture-dependent and culture-independent methods in lab-reared and field-collected Anopheles stephensi-an Asian malarial vector.
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DOI：
10.1186/1471-2180-9-96
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发表时间：
2009-05-19
期刊：
影响因子：
4.2
通讯作者：
Bhatnagar RK
中科院分区：
文献类型：
--
作者：
Rani A;Sharma A;Rajagopal R;Adak T;Bhatnagar RK
蚊子是许多致病生物的中间宿主。媒介控制是研究最多的抑制蚊媒疾病的策略之一。斯氏按蚊是间日疟原虫的传播媒介之一。该寄生虫在蚊子中肠内经历主要的发育和成熟步骤，并且对按蚊相关的中肠微生物群知之甚少。蚊子中肠植物群的鉴定和表征可能有助于更好地了解蚊子生物学，包括寿命、繁殖和蚊子-病原体相互作用，这些对于发展病媒控制机制的战略至关重要。实验室饲养和野外采集的A.通过“培养依赖和培养非依赖”方法筛选斯氏雄虫、雌虫和幼虫。构建了5个实验室和野外采集的16 SrRNA基因文库。进一步分析了斯氏蚊和来自两个样品的总共115个可培养分离物。从中肠鉴定出68个属。stephensi，53只来自野外捕获，15只来自实验室饲养的蚊子。在田间和实验室饲养的A中，分别检测到171个和44个不同的基因型，与数据库中最接近的基因型相似性为85%至99%。stephensi中肠。这些OTU的Shannon多样性指数值在1.74-2.14之间（实验室饲养），在2.75-3.49之间（野外捕获）。斯蒂芬蚊子。高物种均匀度值为0.93至0.99，在现场收集的成人和幼虫中肠植物群表明，通过这些方法检索的微生物多样性的广阔。现场捕获的成年雄虫A.实验室饲养的蚊虫中，粘质沙雷氏菌和脑膜败血冷杆菌含量丰富。50%以上的细菌类型与未培养的细菌有关。有趣的是，发现的几种细菌与其他昆虫中已知的共生体有关。在我们的研究中发现的分离株中，很少有是γ-变形菌中的新种，这些新种不能在遗传学上归入已知的种类。据我们所知，这是第一次尝试研究A。从实验室饲养的和现场收集的成虫和幼虫使用“培养依赖和独立的方法”。
Mosquitoes are intermediate hosts for numerous disease causing organisms. Vector control is one of the most investigated strategy for the suppression of mosquito-borne diseases. Anopheles stephensi is one of the vectors of malaria parasite Plasmodium vivax. The parasite undergoes major developmental and maturation steps within the mosquito midgut and little is known about Anopheles-associated midgut microbiota. Identification and characterization of the mosquito midgut flora is likely to contribute towards better understanding of mosquito biology including longevity, reproduction and mosquito-pathogen interactions that are important to evolve strategies for vector control mechanisms. Lab-reared and field-collected A. stephensi male, female and larvae were screened by "culture-dependent and culture-independent" methods. Five 16S rRNA gene library were constructed form lab and field-caught A. stephensi mosquitoes and a total of 115 culturable isolates from both samples were analyzed further. Altogether, 68 genera were identified from midgut of adult and larval A. stephensi, 53 from field-caught and 15 from lab-reared mosquitoes. A total of 171 and 44 distinct phylotypes having 85 to 99% similarity with the closest database matches were detected among field and lab-reared A. stephensi midgut, respectively. These OTUs had a Shannon diversity index value of 1.74–2.14 for lab-reared and in the range of 2.75–3.49 for field-caught A. stephensi mosquitoes. The high species evenness values of 0.93 to 0.99 in field-collected adult and larvae midgut flora indicated the vastness of microbial diversity retrieved by these approaches. The dominant bacteria in field-caught adult male A. stephensi were uncultured Paenibacillaceae while in female and in larvae it was Serratia marcescens, on the other hand in lab-reared mosquitoes, Serratia marcescens and Cryseobacterium meninqosepticum bacteria were found to be abundant. More than fifty percent of the phylotypes were related to uncultured class of bacteria. Interestingly, several of the bacteria identified are related to the known symbionts in other insects. Few of the isolates identified in our study are found to be novel species within the gammaproteobacteria which could not be phylogenetically placed within known classes. To the best of our knowledge, this is the first attempt to study the midgut microbiota of A. stephensi from lab-reared and field-collected adult and larvae using "culture-dependent and independent methods".
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影响因子：
4
作者：
Beard, CB;Dotson, EM;Durvasula, RV
通讯作者：
Durvasula, RV
影响因子：
5.8
作者：
Huber, T;Faulkner, G;Hugenholtz, P
通讯作者：
Hugenholtz, P
影响因子：
2.1
作者：
BEIER, MS;PUMPUNI, CB;DAVIS, JR
通讯作者：
DAVIS, JR
DOI：
10.4269/ajtmh.1996.54.219
发表时间：
1996-02-01
影响因子：
3.3
作者：
DeMaio, J;Pumpuni, CB;Beier, JC
通讯作者：
Beier, JC
DOI：
10.1073/pnas.0610451104
发表时间：
2007-05-22
影响因子：
11.1
作者：
Favia, Guido;Ricci, Irene;Daffonchio, Daniele
通讯作者：
Daffonchio, Daniele