Evaluating the role of plant growth-promoting bacteria in biological control of Salmonella typhimurium pathogen

Document Type : Research Paper

Authors

1 Department of Soil science, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

2 Department of Soil Science, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran

3 Department of Nanotechnology, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran

Abstract

Human pathogens such as Salmonella enter agricultural soils from various sources. Salmonella is one of the most common food-borne microorganisms and is a causative agent of infections shared between humans and animals. In the current study, some plant growth-promoting bacteria were used to control the pathogen Salmonella typhimurium. The results showed that, in order, 1) Bacillus rugosus CS5, 2) Bacillus vallismortis AS4, 3) Priestia aryabhattai CL1 and 4) Bacillus sp. SS4 had the highest inhibitory effect on the grrowth of Salmonella in liquid medium. However, in solid medium, the bacteria 1) B. vallismortis AS4, 2) B. rugosus CS5, 3) P. aryabhattai CL1 and 4) Bacillus sp. SS4 exhibited the greatest inhibition of this pathogen's growth. The ability to produce siderophores, proteases, lipases, and hydrogen cyanide was investigated in these four bacteria. All four bacteria were capable of producing siderophore. The highest protease production index, with values of 2.5 and 2.9, belonged to P. aryabhattai CL1 and B. rugosus CS5, respectively. B. vallismortis AS4 was capable of producing lipase. Both P. aryabhattai strain CL1 and B. rugosus strain CS5 had the ability to produce HCN, with values of 0.48 mg/mL and 0.2 mg/mL, respectively. The simple impact of four separate bacteria and a microbial consortium on the Salmonella population in soil was investigated. The results showed that the Salmonella population decreased 15 days after inoculation due to the treatments. The greatest reduction in the the population of this pathogen was related to the microbial consortium and the Bacillus sp. strain SS4 treatment, with a 92 % and 98 % reduction in population compared to the control, respectively. The findings indicate a positive effect of plant growth-promoting bacteria in soil for controlling S. typhimurium, which could reduce the negative impacts of the pathogen's presence in soil, thereby enhancing human health and food security.

Keywords

Main Subjects

EXTENDED ABSTRACT

Introduction

Human pathogens, such as Salmonella, pose a threat to public health and food security by contaminating soil and agricultural products. This contamination can lead to salmonellosis in consumers. Recognizing the role of plant growth-promoting bacteria in enhancing soil quality and suppressing pathogens, this study investigated their effects on controlling Salmonella typhimurium in soil. The aim was to mitigate the spread of this pathogen in crops, the food chain, and groundwater.

Materials and Methods

The study involved both laboratory and pot experiments. In the laboratory, we measured the optical density of Salmonella in broth medium and the colony diameter on solid medium to screen the antagonistic properties of plant growth-promoting bacteria. We evaluated their ability to produce siderophores, lipase, protease enzymes, and hydrogen cyanide, which are known to inhibit pathogen growth. In the pot experiments, we tested the effectiveness of these isolates, individually and in a consortium, at 2 and 15 days, to determine their capacity to reduce Salmonella typhimurium growth in soil. The pot experiment was conducted as a factorial in a completely randomized design with three replications. The factors included inoculation with antagonistic bacteria (control (without antagonistic isolates and test strain), test strain Salmonella typhimurium, Bacillus sp. strain SS4, Bacillus rugosus strain CS5, Priestia aryabhattai strain CL1, Bacillus vallismortis strain AS4, and a microbial consortium comprising a mixture of 4 bacterial isolates) and the period of incubation time (2 and 15 days). To confirm the bacteria grown in the Salmonella-specific medium, DNA extraction was performed, and electrophoresis of the PCR products was conducted.

Results The results of laboratory study indicated that out of the 9 antagonistic isolates tested, four bacteria—B. rugosus CS5, B. vallismortis AS4, P. aryabhattai CL1, and Bacillus sp. SS4—exhibited the highest inhibition of Salmonella growth in liquid culture. In solid culture, B. vallismortis AS4, B. rugosus CS5, P. aryabhattai CL1, and Bacillus sp. SS4, respectively, showed the greatest suppression of this pathogen. The four selected bacteria demonstrated the capability to produce siderophores. The highest protease production index, with values of 2.9 and 2.5, were observed in B. rugosus CS5 and P. aryabhattai CL1, respectively. B. vallismortis AS4 was noted for its exclusive production of lipase. Additionally, P. aryabhattai CL1 and B. rugosus CS5 were capable of producing hydrogen cyanide, with concentrations of 0.48 mg/mL and 0.2 mg/mL, respectively. The pot experiment results indicated that all four bacteria significantly reduced the pathogen population in soil 15 days post-inoculation. Notably, Bacillus sp. SS4 and the microbial consortium were more effective, decreasing the population by 40.30×103 and 221.8×103CFU/g of soil, respectively. The electrophoresis results of the PCR products confirmed the presence of the Salmonella typhimurium in the plates.

Conclusion

Given the substantial impact of plant growth-promoting bacteria in diminishing the Salmonella typhimurium population in soil after 15 days, it is advisable to inoculate these bacteria into the soil as a method of pathogen control, in accordance with the procedures outlined in this study.

Author Contributions

Motahare Abedinzadeh carried out the experiment and wrote the original draft of the manuscript. Naeimeh Enayatizamir and Ehsan Shokri verified the analytical methods and supervised the research. Ehsan Shokri handled the laboratory analysis. Shahla Kian Amiri investigated the findings. All authors discussed the results and contributed to the final manuscript, but Naeimeh Enayatizamir wrote the final version of the manuscript. All authors have read and agreed to the published version of the manuscript.

Data Availability Statement

Data underpinning the findings of this study can be found within the tables, figures, and detailed explanations provided in this article.

Acknowledgements

The authors would like to express their gratitude to Shahid Chamran University of Ahvaz (SCU.AS1402.248) and the Agricultural Biotechnology Research Institute of Iran (081-05-05-002-9901-01003).

Ethical considerations

The authors avoided data fabrication, falsification, plagiarism, and misconduct.

Conflict of interest

 The author declares no conflict of interest.

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Iranian Journal of Soil and Water Research
Volume 56, Issue 1
April 2025
Pages 265-280

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