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Escherichia coli expressing a green fluorescent protein (GFP) in Anopheles stephensi: a preliminary model for paratransgenesis

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Abstract

Issues, like emerging insecticide resistance in Anopheles mosquitoes, have led to a breakdown in many vector control programs. In this study, a recombinant Escherichia coli with plasmid expressing a green fluorescent protein (E.coli-GFP) was used as a paratransgenesis model to determine: the possibility of E. coli-GFP trans-stadial transmission. The effect of the water microflora, of bacteria-impregnated sugar solutions, and of blood-feeding on E. coli-GFP colonization and localization within An. stephensi tissues, were studied. The results demonstrated the persistence of E. coli-GFP during molting and metamorphosis events and its trans-stadial transmission. Also the efficacy of bacteria-impregnated sugar solutions for transferring the bacteria to the adult mosquito’s midgut was shown. A blood meal dramatically increased the number of bacteria within 24–48 h post feeding. In addition to fluorescence microscope evaluation, GFP gene PCR amplification showed the presence of the bacteria in the midgut of larvae, pupae, and adults up to 13 days after eclosion. Massive colonization of bacteria was observed in the larvae and in the adult mosquito’s malpighian tubules which may play a role in retaining bacteria in adult mosquitos. The results of this study showed that E. coli could be used as a laboratory model in paratransgenesis studies for the evaluation of various effector molecules as anti-parasite agents for malaria and filariasis.

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Acknowledgment

Authors would like to thank Eng. Abaei, MR. and Eng. Rafie, F. for their assistance in An. stephensi rearing. This study was financially supported by the Tehran University of Medical Sciences (TUMS). The authors also thank the anonymous reviewers for their helpful comments and Professor David Richardson for assistance with editing and improving the English.

Competing interests

All authors declare that they have no competing interests.

Authors’ contributions

ARC did the experiments, analyzed the results and wrote the draft of manuscript, MAO and HV designed the study and coordinated the team works, BY and FZ assist the bacterial experiments and AR reviewed the manuscript and finalized the study design.

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Authors and Affiliations

  1. Department of Medical Entomology and Vector Control, School of Public Health, Urmia University of Medical Sciences (UMSU), Urmia, Iran

    Ali Reza Chavshin

  2. Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences (TUMS), Tehran, Iran

    Ali Reza Chavshin, Mohammad Ali Oshaghi & Hasan Vatandoost

  3. Department of Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran

    Bagher Yakhchali & Fahimeh Zarenejad

  4. The malaria office, Center for Diseases Control (CDC), Ministry of Health, Tehran, Iran

    Ahmad Raeisi

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  1. Ali Reza Chavshin
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  2. Mohammad Ali Oshaghi
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  3. Hasan Vatandoost
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  4. Bagher Yakhchali
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  5. Ahmad Raeisi
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  6. Fahimeh Zarenejad
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Correspondence to Mohammad Ali Oshaghi.

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Chavshin, A.R., Oshaghi, M.A., Vatandoost, H. et al. Escherichia coli expressing a green fluorescent protein (GFP) in Anopheles stephensi: a preliminary model for paratransgenesis. Symbiosis 60, 17–24 (2013). https://doi.org/10.1007/s13199-013-0231-5

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