Rom J Leg Med [27] 16-21 [2019] DOI: 10.4323/rjlm.2019.16 © 2019 Romanian Society of Legal Medicine 16 FORENSIC PATHOLOGY Determination the presence of amplifcation products of 16s rRNA microcystis aeruginosa as a biomarker of drowning Volodymyr M. Voloshynovych 1,* , Roman O. Kasala 2 , Uliana Ya. Stambulska 3 , Marian S. Voloshynovych 4 _________________________________________________________________________________________ Abstract: Forensic medical diagnostics of drowning now is a difcult issue to resolve. Determination of diatom plankton with light microscopy is one of the supplementary methods for diagnostics of drowning. Te disadvantage of this method is the use of concentrated acids to destroy the tissues of the organs, which greatly complicates, and sometimes precludes the detection of diatom plankton. In this case, the detection of other phytoplankton species in internal organs is treated as pseudoplankton, but does not have a diagnostic value. We have developed a sensitive and specifc method of drowning diagnostics using a pair of specifc oligonucleotide primers by the polymerase chain reaction (PCR) method to determine the presence of DNA of Cyanobacteria of the genus Microcystis, namely a fragment of the 16S rRNA gene in the tissues of mice and water samples in order to establish the fact and place of drowning. In order to evaluate the diagnostic value of this method, we conducted an experimental study to detect fragments of the 16S rRNA gene in mice tissues during drowning and post-mortem immersion. Te amplifcation products were found in the tissues of heart, kidneys, liver, spleen, bone tissue, brain tissue, and lungs in case of drowning. During post-mortem immersion products of amplifcation are detected only in the tissues of lungs. Te results indicate that the proposed PCR method is a potentially useful tool for diagnosing of mechanical asphyxia as a result of drowning. Key Words: Drowning, Forensic pathology, DNA, Polymerase chain reaction, Cyanobacteria. 1) Ivano-Frankivsk National Medical University, Department of Pathomorphology and Forensic Medicine, Ivano- Frankivsk, Ukraine * Corresponding author: Tel: +380509569868, E-mail: voloshynovych@yahoo.com 2) Kharkiv Sate Zoo Veterinary Academy, Mala Danylivka, Ukraine 3) Vasyl Stefanyk Precarpathian National University, Department of Biochemistry and Biotechnology, Ivano-Frankivsk, Ukraine 4) Ivano-Frankivsk National Medical University, Department of Dermatology and Venereology, Ivano-Frankivsk, Ukraine INTRODUCTION According to a global report by the World Health Organization, drowning occurs annually with 360,000 people worldwide dying as a result of drowning, accounting for more than 9% of global mortality [1]. More than 90% of deaths due to drowning are recorded in low and middle-income countries. Mechanical asphyxia as a result of drowning in Ivano-Frankivsk region amounted to 354 cases for the period from 2012 to 2016. Tis is 4,5% of all types of expertise conducted by the Ivano-Frankivsk Regional Bureau of Forensic Medical Examination [2]. Forensic medical diagnostics mechanical asphyxia as a result of drowning is currently a difcult issue for resolution [3-8]. Drowning is a leading cause of unintentional deaths [9]. A number of methods have been developed over the years to determine mechanical asphyxia as a result of drowning [10]. Determination of the presence of diatom algae is one of the supplementary methods of diagnosis, not only drowning but sometimes the place of drowning [11-13]. Te scientifc community is constantly discussing the authenticity and reliability of the diatom test at drowning [14]. Tere are even several cases where the expert's fndings obtained using diatoms have been challenged and sometimes canceled in court [15].