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Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis

BACKGROUND: Microbial ecology of phlebotomine sand flies is not well understood although bacteria likely play an important role in the sand fly biology and vector capacity for Leishmania parasites. In this study, we assessed the significance of the microbial community of rabbit feces in oviposition...

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Autores principales: Peterkova-Koci, Kamila, Robles-Murguia, Maricela, Ramalho-Ortigao, Marcelo, Zurek, Ludek
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3414784/
https://www.ncbi.nlm.nih.gov/pubmed/22827861
http://dx.doi.org/10.1186/1756-3305-5-145
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author Peterkova-Koci, Kamila
Robles-Murguia, Maricela
Ramalho-Ortigao, Marcelo
Zurek, Ludek
author_facet Peterkova-Koci, Kamila
Robles-Murguia, Maricela
Ramalho-Ortigao, Marcelo
Zurek, Ludek
author_sort Peterkova-Koci, Kamila
collection PubMed
description BACKGROUND: Microbial ecology of phlebotomine sand flies is not well understood although bacteria likely play an important role in the sand fly biology and vector capacity for Leishmania parasites. In this study, we assessed the significance of the microbial community of rabbit feces in oviposition and larval development of Lutzomyia longipalpis as well as bacterial colonization of the gut of freshly emerged flies. METHODS: Sterile (by autoclaving) and non-sterile (control) rabbit feces were used in the two-choice assay to determine their oviposition attractiveness to sand fly females. Bacteria were identified by amplification and sequencing of the 16S rRNA gene with universal eubacterial primers. Sterile, control (non-sterile), and sterilized and inoculated rabbit feces were used to assess the significance of bacteria in L. longipalpis development. Newly emerged adult flies were surface-sterilized and screened for the bacterial population size and diversity by the culturing approach. The digestive tract of L4 sterile and control larvae was incubated with Phalloidin to visualize muscle tissues and DAPI to visualize nuclei. RESULTS: Two-choice behavioural assays revealed a great preference of L. longipalpis to lay eggs on rabbit feces with an active complex bacterial community (control) (85.8 % of eggs) in comparison to that of sterile (autoclaved) rabbit feces (14.2 %). Bioassays demonstrated that L. longipalpis larvae can develop in sterile rabbit feces although development time to adult stage was greatly extended (47 days) and survival of larvae was significantly lower (77.8 %) compared to that of larvae developing in the control rabbit feces (32 days and 91.7 %). Larval survival on sterilized rabbit feces inoculated with the individual bacterial isolates originating from this substrate varied greatly depending on a bacterial strain. Rhizobium radiobacter supported larval development to adult stage into the greatest extent (39 days, 88.0 %) in contrast to that of Bacillus spp. (76 days, 36.0 %). From the complex natural bacterial community of rabbit feces, R. radiobacter survived pupation and colonized the newly emerged females most successfully (82.6 % of all bacteria cultured); however, only 25 % of females were positive for bacteria in the digestive tract upon emergence. Immunohistochemistry did not reveal any obvious differences in anatomy of the digestive tract between control and axenic larvae. CONCLUSIONS: The bacterial community in the sand fly larval habitat affects oviposition and larval development although bacteria are not essential for successful development of L. longipalpis. Different bacteria contribute to larval development to various degrees and some, e.g. Rhizobium radiobacter, survive pupation and colonize the digestive tract of newly emerged females. With the establishment of the axenic rearing system, this study opens new venues to study the effect of bacteria on the gut epithelial immunity and vector competence of sand flies for Leishmania parasites with a goal to develop paratransgenic approaches for Leishmania control.
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spelling pubmed-34147842012-08-10 Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis Peterkova-Koci, Kamila Robles-Murguia, Maricela Ramalho-Ortigao, Marcelo Zurek, Ludek Parasit Vectors Research BACKGROUND: Microbial ecology of phlebotomine sand flies is not well understood although bacteria likely play an important role in the sand fly biology and vector capacity for Leishmania parasites. In this study, we assessed the significance of the microbial community of rabbit feces in oviposition and larval development of Lutzomyia longipalpis as well as bacterial colonization of the gut of freshly emerged flies. METHODS: Sterile (by autoclaving) and non-sterile (control) rabbit feces were used in the two-choice assay to determine their oviposition attractiveness to sand fly females. Bacteria were identified by amplification and sequencing of the 16S rRNA gene with universal eubacterial primers. Sterile, control (non-sterile), and sterilized and inoculated rabbit feces were used to assess the significance of bacteria in L. longipalpis development. Newly emerged adult flies were surface-sterilized and screened for the bacterial population size and diversity by the culturing approach. The digestive tract of L4 sterile and control larvae was incubated with Phalloidin to visualize muscle tissues and DAPI to visualize nuclei. RESULTS: Two-choice behavioural assays revealed a great preference of L. longipalpis to lay eggs on rabbit feces with an active complex bacterial community (control) (85.8 % of eggs) in comparison to that of sterile (autoclaved) rabbit feces (14.2 %). Bioassays demonstrated that L. longipalpis larvae can develop in sterile rabbit feces although development time to adult stage was greatly extended (47 days) and survival of larvae was significantly lower (77.8 %) compared to that of larvae developing in the control rabbit feces (32 days and 91.7 %). Larval survival on sterilized rabbit feces inoculated with the individual bacterial isolates originating from this substrate varied greatly depending on a bacterial strain. Rhizobium radiobacter supported larval development to adult stage into the greatest extent (39 days, 88.0 %) in contrast to that of Bacillus spp. (76 days, 36.0 %). From the complex natural bacterial community of rabbit feces, R. radiobacter survived pupation and colonized the newly emerged females most successfully (82.6 % of all bacteria cultured); however, only 25 % of females were positive for bacteria in the digestive tract upon emergence. Immunohistochemistry did not reveal any obvious differences in anatomy of the digestive tract between control and axenic larvae. CONCLUSIONS: The bacterial community in the sand fly larval habitat affects oviposition and larval development although bacteria are not essential for successful development of L. longipalpis. Different bacteria contribute to larval development to various degrees and some, e.g. Rhizobium radiobacter, survive pupation and colonize the digestive tract of newly emerged females. With the establishment of the axenic rearing system, this study opens new venues to study the effect of bacteria on the gut epithelial immunity and vector competence of sand flies for Leishmania parasites with a goal to develop paratransgenic approaches for Leishmania control. BioMed Central 2012-07-24 /pmc/articles/PMC3414784/ /pubmed/22827861 http://dx.doi.org/10.1186/1756-3305-5-145 Text en Copyright © 2012 Peterkova-Koci et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Peterkova-Koci, Kamila
Robles-Murguia, Maricela
Ramalho-Ortigao, Marcelo
Zurek, Ludek
Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title_full Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title_fullStr Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title_full_unstemmed Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title_short Significance of bacteria in oviposition and larval development of the sand fly Lutzomyia longipalpis
title_sort significance of bacteria in oviposition and larval development of the sand fly lutzomyia longipalpis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3414784/
https://www.ncbi.nlm.nih.gov/pubmed/22827861
http://dx.doi.org/10.1186/1756-3305-5-145
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