BIODIVERSITAS ISSN: 1412-033X Volume 24, Number 8, August 2023 E-ISSN: 2085-4722 Pages: 4334-4341 DOI: 10.13057/biodiv/d240815 Genetic development of Stx phage TESSA SJAHRIANI 1, , MALA KURNIATI 2 , DEBI ARIVO 1 , DWI MARLINA SYUKRI 1 , WIWIEK TYASNINGSIH 3 1 Department of Microbiology, Faculty of Medicine, Universitas Malahayati. Jl. Pramuka 27, Bandar Lampung 35158, Lampung, Indonesia. Tel./fax.: +62-819-7971580,  email: tessasah@malahayati.ac.id 2 Department of Biology, Faculty of Medicine, Universitas Malahayati. Jl. Pramuka 27, Bandar Lampung 35158, Lampung, Indonesia 3 Department of Microbiology, Faculty of Veterinary Medicine, Universitas Airlangga. Jl. Mayjen. Prof. Dr. Moestopo 47, Surabaya 60131, East Java, Indonesia Manuscript received: 19 December 2022. Revision accepted: 14 July 2023. Abstract. Sjahriani T, Kurniati M, Arivo D, Syukri DM, Tyasningsih W. 2023. Genetic development of Stx phage. Biodiversitas 24: 4334-4341. Stx phages have been studied and described due to their involvement in shigatoxin. The variety, evolution, dispersion, and molecular mechanisms of STEC and Stx phages have all been widely studied, but further research is still required. The objective of this study was to investigate the unique sequences of bacteriophage DNA from the NCBI database and to identify the changes in the DNA sequence of both regular and Stx-carrying bacteriophages. It was first reported in 2001, but studies have not been carried out. The Stx- carrying bacteriophage DNA sequence, similarity of identity, e-value, and gaps were analyzed using PCR and DNA sequencing. The number of bacteriophage used in this study was eight. The results revealed the similarity of bacteriophage to Stx-carrying bacteriophages by 82.67% to 88.5%, and exhibited notable variations in comparison to each phage 1 to 8 (Eco157 phage WASJ/2022) (p<0.001). Moreover, results exhibited an e-value ranging from 7e-85 to 3e-44. In addition, results showed gaps between 1-2% in comparison to other Stx-carrying bacteriophages in NCBI with bacteriophage alterations on the bases of Cytosine and Thymine in 184 th and 263 rd . Therefore, this work provided the potency of bacteriophage as an alternative pharmaceutical in controlling foodborne diseases. Keywords: Foodborne disease, genetic, phage, STEC, Stx INTRODUCTION In 2019, the largest contribution of cattle in Indonesia was in East Java, there were approximately 4.705.067 (27.79%) of cattle (Central Statistics Agency of Indonesia 2019a). East Java produces around 21.95% of its meat from cattle slaughtered, and the majority of them are consumed by local people (Central Statistics Agency of Indonesia, 2019b). The disposal from abattoir causes a pollution in the Kalimas River due to the dysfunction of wastewater treatment. Fundamentally, the food industry in East Java has employed a range of sanitation procedures to improve the fresh produce's microbiological safety and quality (Fisher et al. 2016; Joshi et al. 2018). The government of East Java has been working to raise the Human Development Index and reduce the poverty rate. However, the percentage of people in poverty in East Java is higher by 10.86% compared with the national average of poverty rate, particularly in rural areas (Sjahriani et al. 2021a). In Indonesia, the prevalence of food poisoning cases has reached by 80.83% (National Agency of Drug and Food Control of Indonesia 2016). Foodborne diseases and food poisoning have gained attention of medical community, particularly in developing countries (Carbas et al. 2013). It is estimated that Escherichia coli O157:H7 causes 63,000 cases of foodborne infections, 2,100 hospitalizations, 20 fatalities, and high cost of $271 million (Hoffmann et al. 2015; Poxleitner et al. 2016). Escherichia coli O157:H7 can produce shigatoxin that is suspected to alter the type and extent the impairment of human tissue, leading to catastrophic renal failure, bloody diarrhea, and death (Melton-Celsa 2014; Luna-Gierke et al. 2014). Therefore, the food industries use a variety of sanitation techniques to improve the microbiological safety and quality of fresh food. Bacteriophages get attached to the cells surfaces of bacteria through certain surface receptors of host cell (Zhan et al. 2015). Phage is believed to extract and infect E. coli serotypes or other members of the Enterobacteriaceae family (Khalil et al. 2016). Furthermore, the genetic material of bacteriophage is injected into the cytoplasm of the bacterial cell, resulting an infection (Silva et al. 2016). Bacteriophages provide a safe and natural way to prevent the spread of pathogen in humans, due to their natural component (Bhardwaj et al. 2015). It is primarily composed a non-harmful nucleic acid and a member of the human microbiome (Huh et al. 2019). The infection of bacteriophage is initiated by perforation of bacterial cell wall, and viral DNA is transferred into the cytoplasm of host cells (Fokine and Rossmann 2014). The Rz/Rz1 protein involved in the outer membrane breakdown, resulting in bacterial lysis (Catalao et al. 2013). The analysis of DNA phage sequences offers an alternative treatment of foodborne infections due to the growing incidence of antibiotics resistance (Aslam et al. 2018). This work provided a profitable, prices and targeted approach since the variety, evolution, dissemination, and molecular mechanisms of STEC and Stx phages have been studied in