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A metagenomic assessment of the bacteria associated with Lucilia sericata and Lucilia cuprina (Diptera: Calliphoridae)
Authors:Baneshwar Singh  Tawni L. Crippen  Longyu Zheng  Andrew T. Fields  Ziniu Yu  Qun Ma  Thomas K. Wood  Scot E. Dowd  Micah Flores  Jeffery K. Tomberlin  Aaron M. Tarone
Affiliation:1.Department of Entomology,Texas A&M University,College Station,USA;2.Department of Forensic Science,Virginia Commonwealth University,Richmond,USA;3.Southern Plains Agricultural Research Center,Agricultural Research Service, USDA,College Station,USA;4.State Key Laboratory of Agricultural Microbiology, National Engineering Research Center of Microbe Pesticide,Huazhong Agricultural University,Wuhan,China;5.School of Marine and Atmospheric Sciences,Stony Brook University,Stony Brook,USA;6.Tianjin Institute of Industrial Biotechnology,Chinese Academy of Science,Tianjin,China;7.Department of Chemical Engineering,The Pennsylvania State University,University Park,USA;8.MR DNA Molecular Research LP,Shallowater,USA;9.Walter Reed Army Institute of Research,Silver Spring,USA
Abstract:Lucilia Robineau-Desvoidy (Diptera: Calliphoridae) is a blow fly genus of forensic, medical, veterinary, and agricultural importance. This genus is also famous because of its beneficial uses in maggot debridement therapy (MDT). Although the genus is of considerable economic importance, our knowledge about microbes associated with these flies and how these bacteria are horizontally and trans-generationally transmitted is limited. In this study, we characterized bacteria associated with different life stages of Lucilia sericata (Meigen) and Lucilia cuprina (Wiedemann) and in the salivary gland of L. sericata by using 16S rDNA 454 pyrosequencing. Bacteria associated with the salivary gland of L. sericata were also characterized using light and transmission electron microscopy (TEM). Results from this study suggest that the majority of bacteria associated with these flies belong to phyla Proteobacteria, Firmicutes, and Bacteroidetes, and most bacteria are maintained intragenerationally, with a considerable degree of turnover from generation to generation. In both species, second-generation eggs exhibited the highest bacterial phylum diversity (20 % genetic distance) than other life stages. The Lucilia sister species shared the majority of their classified genera. Of the shared bacterial genera, Providencia, Ignatzschineria, Lactobacillus, Lactococcus, Vagococcus, Morganella, and Myroides were present at relatively high abundances. Lactobacillus, Proteus, Diaphorobacter, and Morganella were the dominant bacterial genera associated with a survey of the salivary gland of L. sericata. TEM analysis showed a sparse distribution of both Gram-positive and Gram-negative bacteria in the salivary gland of L. sericata. There was more evidence for horizontal transmission of bacteria than there was for trans-generational inheritance. Several pathogenic genera were either amplified or reduced by the larval feeding on decomposing liver as a resource. Overall, this study provides information on bacterial communities associated with different life stages of Lucilia and their horizontal and trans-generational transmission, which may help in the development of better vector-borne disease management and MDT methods.
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