(IJIRSE) International Journal of Innovative Research in Science & Engineering ISSN (Online) 2347-3207 Characterization of Proteus mirabilis Lytic Phage from Situ Letik River, Bogor, Indonesia Rachmi Afriani 1 , Iman Rusmana 2 , Sri Budiarti 3* Graduate Student 1 , Lecturer 2 , Lecturer 3 Bogor Agricultural University – Biology Department Bogor – Indonesia *s_budiarti@yahoo.com Abstract-Proteus mirabilis isolated from the feces of diarrhea patients was resistant to ampicillin, amoxicillin and trimethoprim-sulfomethaxazole. Therefore, to reduce potential infection of antibiotic- resistant P. mirabilis, alternative solutions such as application of lytic phage were needed. The aims of this study were to isolate and characterize phage that can lyse P. mirabilis cells. Phages were isolated from poultry waste and river water in Situ Letik village. Phage isolates were determined by their ability to form plaques, specificity, characteristics of protein and morphology. Phage was specific to P. mirabilis as its host. Sodium Dodecyl Sulphate-Poly Acrilamide Gel Electrophoresis (SDS-PAGE) analysis showed that the phage virion contains 8 proteins with molecular weight between 20-91 kDa. Transmission electron microscope observation showed that the phage was the member of Myoviridae family with icosahedral hexagonal head, the diameter about 68.185 nm, the length of tail about 109.019 nm, and the diameter of the tail about 17.89 nm width. Keywords: diarrhea, lytic phage, Proteus mirabilis I. INTRODUCTION Diarrheal is one of the most common disease leading cause of death in developing countries, including Indonesia. Diarrhea can be caused by the variety of microorganisms such as viruses, protozoa and bacterial enteropathogens. Proteus mirabilis is a Gram-negative bacteria (enteropathogens) which can cause diarrhea. There are 8 bacterial enteropathogens including Proteus spp that were isolated from stool samples of 100 children with diarrhea aged under 5 years [14]. The most of treatment stage to control diarrheal disease caused by P. mirabilis is using antibiotics. However, several studies reported that the current use of antibiotics to control bacterial pathogens becomes ineffective due to the emergence antibiotics resistance of P. mirabilis. Some isolates of P. mirabilis multiresistant to antibiotics, namely ampicillin, gentamicin, ceftazidime, cefotaxime, cefuroxime, cefalothin, cefepime, piperacillin, trimethoprim-sulfamethoxazole and ciprofloxacin [15]. Some isolates of P. mirabilis have ESBL (Extended Spectrum -Lactamases) that play an important role to increase the resistancy of P. mirabilis to the -lactam group antibotic [18]. Antibiotic-resistant bacteria P. mirabilis can contaminate food and water. Along with the rapid development of science and technology, many researches were done to look for the biocontrol agents that are eco-friendly in purpose to control water and food pollution by P. mirabilis. One of the alternative is by using lytic phages. The use of lytic phages in controlling the growth of human pathogenic bacteria is a natural alternative methods that are environmentally friendly. The most common source of phage to reduce pathogenic bacterial contaminants of water and food can be isolated from the stool, a wide variety of food, water, wastewater, soil and infected tissue. Some phages have been applied as biocontrol of food contamination, including Enterobacter sakazaki phage lytic applied to infant formula and lytic phage of Salmonella and Campylobacter on chicken skin [11]. Lytic phages as biocontrol bacterial pathogens has been applied to the cheese address to control the listeriosis disease caused by Listeria monocytogenes [6]. Phage has ability to lysis indigenous Salmonella P38 from feces of diarrheal patient [23]. Lytic phage infectivity presents against enteropathogenic Escherichia coli (EPEC K.1.1) antibiotic-resistant isolates from patients with diarrhea in Indonesia [4]. Research characterization of lytic phages against antibiotic-resistant bacteria P.mirabilis pathogens that can contaminate water, food and the cause of diarrhea has not been done. Therefore, the objective of this research are to characterize lytic phage of P. mirabilis to be used as biocontrol water and food contamination caused by P. mirabilis. II. MATERIALS AND METHODS Identification and Pathogenicity Test of Proteus mirabilis Identification of P. mirabilis was determined using gram staining test and API 20E KIT. Pathogenicity test was conducted at blood agar plates. Observations of media discoloration blood was done on 3 hours and 5