Ghassemi Marzieh, Akhavan Amir Ahmad, Zahraei-Ramezani Alireza, Yakhchali Bagher, Zarean Mohammad Reza, Jafari Reza, Oshaghi Mohammad Ali
Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Institute of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
J Arthropod Borne Dis. 2024 Mar 31;18(1):12-27. doi: 10.18502/jad.v18i1.15668. eCollection 2024 Mar.
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.
In this study two engineered bacterium candidates to produce paratransgenic sand flies, AS1 and expressing m-Cherry fluorescent were applied on the leaves of the white saxaul plant (), 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 10 cells/ml was used for sugar bait, spraying on plant leaves (∼10 cm) and 10 cm 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.
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 surviving on plant and in soil were significantly (P< 0.0001 and P= 0.046) higher than AS1.
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.
工程菌候选菌株在野外条件下的生存能力和持久性是抗媒介传播疾病的共生转基因方法中一个相当大的挑战。
在本研究中，两种用于生产共生转基因白蛉的工程菌候选菌株AS1和表达m-樱桃荧光蛋白的菌株，被应用于白梭梭植物的叶片、糖饵和啮齿动物洞穴土壤上，并于2022年8月在伊斯法罕省马廷阿巴德县的沙漠条件下测试它们的持续时间。将10个细胞/毫升的磷酸盐缓冲盐水（PBS）悬浮液用于糖饵、喷洒在植物叶片（约10厘米）和10厘米深的啮齿动物洞穴土壤上。每天采集白蛉样本，接种在LB琼脂平板上，24小时后对荧光细胞进行计数。
总体而言，时间进程导致两种菌株的细菌数量减少。这两种菌株在糖饵和植物叶片上持续存在四天，在土壤上持续存在两天。虽然糖饵中细菌数量之间存在细微差异，但不显著（P<0.05）。在植物上和土壤中存活的数量显著高于AS1（P<0.0001和P=0.046）。
本研究表明，植物或糖饵是共生转基因方法中递送转化细菌的有用途径，不过，细菌应该每隔四天喷洒在植物上或更换糖饵。
