CHARACTERIZING DRUG RESISTANT BETA-LACTAMASES PRODUCED BY ISOLATEDBACTERIA FROM FIELD SAMPLES, TOWARDS THE UNDERSTANDING OF AMR AND DEVELOPMENT OF NOVEL DIAGNOSTICS

Abstract

Antimicrobial resistance (AMR) poses a critical global health challenge, driven by microbial microevolution due to selection pressure imposed by higher-concentration antibiotics. The development of antimicrobial resistance is a natural phenomenon, but anthropogenic activity drastically enhanced this process. If we look at the anthropogenic side, the usage, and dosage of antibiotics in the medical and nonmedical sector is exponentially increasing. If we analyze the distribution of antibiotics in the human and animal sector, we find that food animal farms utilize more than 70% of the total antibiotics. Cattle, poultry, sheep, and swine are major food animal sectors in the modern world. Organized and unorganized farming sectors are majorly contributed to producing meat-based products, in which unorganized sectors are mainly involved. Unprescribed and indiscriminate use of antibiotics in food animals assists in the emergence of antimicrobial resistance as a global health hazard problem. The animal products in the food chain are the primary source of antimicrobial resistance and therapeutic failure in humans. Veterinarians and health workers mainly use Tetracycline and beta-lactam antibiotics on animal farms. Bacteria have evolved different novel mechanisms to combat these antibiotics. Excess antibiotics are not completely assimilated in the gut and are excreted in the feces and urine. Drug residues, discharge in the soil, wastewater, and compost can put selection pressure on the naturally present bacterial population and allow to grow of only drug-resistant bacteria in the vicinity. High concentration of antibiotic residues detected in freshwater bodies, oceans, and animal products ultimately goes to humans. These antibiotic residues directly disturb the environment's micro-ecology and push the growth of antimicrobial-resistant bacteria. Detection of XDR (extensively drug-resistant) from animal farm soil, compost, and vermicompost, defining the effect of the misuse of antibiotics in the animals. One of the challenging parts of drug resistance is detecting and diagnosing the resistant bacteria in the samples in a short period. Gold standard methods are prolonged and need experts with laboratory set-up to detect the drug-resistant bacteria in the samples. At the same time, the advanced molecular AMR detection methods are quick and reliable but costly and also need experts with laboratory setups to perform experiments on AMR genes or detect resistant bacteria. To minimize drug-resistant bacteria detection time, cost, and dependency on the laboratory set-up, we developed the “BL-Tester” device. This device was based on a dye that can change color in the presence of beta-lactamase-producing bacteria in the samples. At room temperature, Dye with β-lactamase producing bacteria gives a color change from yellow to red within 8 hrs. Dye optimal concentration provides the visible color signal, and the limit-of-detection (LOD) of final β-lactamase was 10 mU/mL. This system needs only 1ml or 1gm of samples and can work on wastewater, soil, urine, vermicompost, compost, and animal feces samples. By developing the “BL-Tester” device, we aimed to develop a culture-free, point-of-need, and point-of-care testing kit with a final product that can carry into the field to screen drug-resistant bacteria load and collect the samples. Fast, less expensive, and reliable results are the important points to developing the “BL-Tester” AMR diagnostic kit.