Assessing Survival of Transgenic Bacteria, Serratia AS1 and Enterobacter cloacae, in Sugar Bait, White Saxaul Plant (Haloxylon persicum) and Rodent Barrow’s Soil, A Contained-Field Study for Paratransgenesis Approach

Marzieh Ghassemi; Amir Ahmad Akhavan; Alireza  Zahraei-Ramezani; Bagher Yakhchali; Mohammad Reza Zarean; Reza Jafari; Mohammad Ali Oshaghi

doi:10.18502/jad.v18i1.15668

Marzieh Ghassemi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Amir Ahmad Akhavan Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Alireza Zahraei-Ramezani Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Bagher Yakhchali Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran
Mohammad Reza Zarean Esfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Reza Jafari Esfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Ali Oshaghi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

DOI: https://doi.org/10.18502/jad.v18i1.15668

Keywords: Paratransgenesis; Symbionts; Sand fly; Leishmaniasis; Vector-borne diseases

Abstract

Background: The viability and persistence of engineered bacterium candidates in field conditions is one of the considerable challenges in the paratransgenesis approach to fighting vector-borne diseases.

Methods: In this study two engineered bacterium candidates to produce paratransgenic sand flies, Serratia AS1 and Enterobacter cloacae expressing m-Cherry fluorescent were applied on the leaves of the white saxaul plant (Haloxylon persicum), sugar bait, and rodent burrow soil and their persistent time was tested in desert condition, Matin Abad County, Isfahan, August 2022. A PBS suspension of 10⁹ cells/ml was used for sugar bait, spraying on plant leaves (~10 cm²) and 10 cm² of rodent burrow soil. Sand fly samples were taken daily and were plated on LB Agar and the fluorescent cells were counted after 24 hours.

Results: Time course in general caused a decrease in the number of bacteria for both strains. The two strains were persistent in sugar bait and on plant leaves for four days and on soil for two days. Although there were slight differences between the number of the bacteria in sugar baits, which was not significant (P< 0.05). The number of E. cloacae surviving on plant and in soil were significantly (P< 0.0001 and P= 0.046) higher than Serratia AS1.

Conclusion: This study shows that plants or sugar bait are useful routes for delivery of the transformed bacteria for the paratransgenesis approach, although, the bacteria ought to be sprayed on plants or sugar baits should be replaced with new ones in four days intervals.