Zoonotic diseases are described as diseases that are transmitted between animals and humans through direct or indirect contact or by any other route. Many microorganisms causing recently identified zoonotic infections in humans were initially detected in animals (specifically, wildlife) or products of animal origin. Dogs and cats are the most common pets which can disseminate zoonotic diseases. Rabies, ringworms, Campylobacteriosis, Salmonellosis, and Leptospirosis are the most common diseases that can spread through dogs. All domesticated animals, including poultry, have the potential to harbor germs that cause food-borne diseases. Around 90% of bacteria-related food-borne infections are caused by Salmonella spp. and Campylobacter spp. Molecular biology methods are increasingly being used in small animal veterinary care to diagnose infectious diseases. Understanding of infectious disease agents has improved because of techniques like polymerase chain reaction (PCR), Real-time PCR, Restriction fragment length polymorphism (RFLP), ELISA and CRISPR-CAS. Approximately 50% of bacterial and 87% of viral genomes have been identified so far by different molecular techniques. To subtype organisms beyond phenotypic categories, genotyping techniques such as PFGE, RAPD, REP-PCR, and AFLP are especially applied. As DNA is the starting material, these molecular biology techniques are often frequently referred to as "DNA fingerprinting." In addition to epidemiology, these techniques are also employed in forensic medicine and evolutionary biology research. Compared to phenotyping procedures, genotyping techniques frequently offer better discriminating power. Another way to identify genomic sequences from various microbial communities is through metagenomics. Over the last few decades, metagenomics-based methodologies have been developed to assess, analyze, and utilize biodiversity across a wide range of various environmental niches. Metagenomics has helped characterize the microbiomes in many samples, such as the gastrointestinal tract of various creatures (e.g., feline, canine, human, mouse, and chicken), in addition to the discoveries of viral genomes. As technology continues to evolve, the integration of molecular diagnostics into routine surveillance and healthcare systems holds promise for more effective prevention and control of zoonotic diseases, ultimately safeguarding both humans and animals.