Many criminals, offenders often use biological material to cause crime and harm the victims in a group. But here Forensics come up with the new and growing techniques that are Microbial Forensics. Microbial forensics is the study of science for testing evidence from a bioterrorism act, bio crime, toxin release for attribution purposes by criminals in the form of microbes. Fungi, viruses, and bacteria are microorganism which is tested by microbial forensics. Bacteria, fungi, viruses all are eukaryotes and can cause disease in any living organism. In Bioterrorism criminal motives is to terrorize and make people worry about their health and in some cases murder them by using biological evidence. It has five phases that are preparedness, early warning, notification, response, and recovery. Many criminal organizations mostly terrorists and bio war scientists change little things in a virus or mutate it to make it toxic or lethal; insertions, deletions, and single nucleotide polymorphism are few techniques to cause changes. One such bio crime is white powder crime. Many microorganisms are altered by using techniques called genetic engineering. Scientists have discovered that modified microorganisms are resistant to many conditions like heat. Modified microorganisms also help in the production of food. They have been used in dairy, pharmaceutical, wine, beer etc.

In 2001, The United States was under Anthrax caused by the bacteria Bacillus anthracis which almost killed 5 people of 22 who got anthrax. This bacteria in a powdered form were put into the letters. Also, anthrax spores can be easily released into the air and within a couple of minutes, thousands of people can get affected by inhaling contaminated air. In the year 1990s United States formed a unit called Hazardous Material Response Unit (HMRU) to investigate bioterrorism with scientific evidence. The Microbial Forensics field includes computer science, microbiology, phylogenetic, microbial genomes etc. The duty of microbial forensics is to identify the sample; collect the sample from the victims or sometimes from the crime scene, preserve the sample in a well proper way without causing any contamination and handle the sample in a well a mannered way. After lab analysis and they must provide the interpretation of tests with quality assurance. The microbial attack occurs in a group of people usually are of two types: covert way and overt way. In an overt way, it is caused by terrorist like for example transferring a package which contains biological agents and can harm people are released into the environment. In this case, there is a huge involvement of law enforcement and their concerns are who caused it, isolate identification, production process, and Epidemiologic concerns. Whereas in covert way attacks are natural disease break in an environment, for example, COVID 19. Public health administrators play a major role. They are usually concerned about the treatment process, how many people are infected, what is the causative agent, and the origin of the initial case. The term Validation is mostly used to signify reliance on a test or process, it is also important for scientific disciplines, like in the progressing field of microbial forensics. There is a minimal set of criteria that are supposed to be considered in evolving plans of validation of microbial forensics methods. A few of them are Description of scope, intended application, the purpose of the method, end results to be assessed, and so on. These criteria are applied to global goals, in the collection process, in transportation and storage under a chain of custody, in laboratory procedures, and an interpretation as well. It is not practicable for it to be all-inclusive due to the wide range of diversity of samples and their types for which microbial forensic methods are needed to be applied. Some of the application of microbial forensics in biological warfare is to trace outbreaks of microbial diseases, also many surveillances store the DNA fingerprints for microbes and maintain a database. Usually, the database contains all the information on biological and genetic data on the microbial disease. This data takes information from many organisms and compare one genome with another genome. In a crime aspect, microbial forensics can identify and give a clear explanation about the cause of death by any microorganisms. It also helps in finding the location by analyzing soil microbiomes and with sexually transmitted microbial diseases.

Also, forensic scientists take help of microorganisms to identify the time of death by studying the bacteria cycle. During the decomposition of dead body, the microbes present on it have microbial succession and this succussion can help in finding the accurate time of death. During the unattended and suspicious criminal investigation ‘microbial clock’ is very helpful. Microbes are attached to body and biological conditions, so if in case the dead body is shifted to secondary crime scene, the microbes from primary crime scene can be found on the body. Its type of corroborative evidence leads crime investigator to primary crime scene. Microbes can also be found in lungs of victim, in case of drowning. Tools used by microbial forensics are electron microscopy, crystallography, molecular sequencing, microbial culture, chemistry, biochemistry, and mass spectrometry. Also, for finding causative agents lab should be sophisticated ventilated and scientists should wear Personal Protective Equipment (PPE) like gloves, masks, face shields, etc. Techniques used for finding infected agents as defined by the Sampling and Identification of Biological and Chemical Agents (SIBCA) handbook. It mainly has three techniques; firstly, it is Nucleic Acid Amplification Techniques, here microorganisms like Bacillus Anthrax, Brucella spp, Francisella tularensis, and Yersinia pestis re-identified by the Reverse transcription-polymerase chain reaction test (RT PCR). Due to the low number of bacteria samples, DNA dehydration, inappropriate DNA preparation lead to negative PCR results. Serology is the second technique used for the identification of microorganisms like Brucellosis, tularemia, and plague. This technique can give false-positive results due to the cross-reactions and can also give false negative due to extra or loss of antigenic structures. And lastly its Typing and Stain identification technique, here microbes are analyzed to see whether strains are from the same source or not. The result in this technique is obtained by Variable Number Tandem Repeat (VNTR), Multilocus Sequence Typing (MLST), Single Nucleotide Polymorphism (SNP), and other tools. Here the quality of the database is sometimes not reliable because it totally depended on the accuracy of submitted sequences.

Conclusion: – Microbial Forensics is still developing in India, it connects medicines with forensics. We need much research and awareness about bio crime and the future is also about bio war. Corona is one such example. With different tools and techniques, microbial forensics help in catching the perpetrators who cause bio crime and with advanced medical tools on microbial forensics help the medical to protect the people from biothreat.