I always knew that reading about something does not equate to experiencing it - but I recently realised that I didn't truly understand the extent of this difference.
On June 15th, 2022, I stepped into the Tata Institute of Fundamental Research, a little lost as I tried to find my way to the Anand Research Group - Dr. Amitesh Anand’s microbiology lab in the Department of Biological Sciences.
In school, I had been learning about bacteria since the 8th grade. I started by learning that microscopic organisms also carry out the 7 life processes (movement, respiration, sensitivity, growth, reproduction, excretion, nutrition) and then began to understand how these very microorganisms can be deadly. Over the years, I had seen textbook diagrams and online videos - but never held a plate of bacteria in my hand (with gloves, of course!). That was until I entered the microbiology lab.
I was the first high school student to have entered their lab, and at first, it felt intimidating to be surrounded by post-doctorates, PhD and grad students. However, over the course of the month I spent there, I formed a close relationship with all the lab members. Despite there being age differences between us, they always involved me - whether it was in their experiments or in their conversations. I began with simply observing how the agarose gel for gel electrophoresis is made, why bacterial cultures are incubated in the shaker incubator, and how DNA is amplified by a Polymerase Chain Reaction (PCR) machine.
In school, I had learned the steps of a PCR - denaturation, annealing and extension. But in the lab, I experienced setting up a PCR reaction and determining the optimum conditions required for each step.
My first experiment in the lab was plotting the bacteria growth curve. I knew the basics of this sigmoidal curve - with the lag, log, stationary and death phase. With my sterile hands in the laminar flow hood, I inoculated a primary culture of two strains of Escherichia coli in a minimal glucose medium. After a day of incubation, I calculated the Optical Density (OD) values using a spectrophotometer - once again, a device I had only read about.
I conducted a few more experiments but what fascinated me the most was isolating a plasmid from a bacteria. A plasmid is a small, circular, double-stranded DNA molecule that is different from a cell's chromosomal DNA. Genes found in plasmid DNA are hence, different from those found in the chromosomal DNA. This plasmid DNA often contains antibiotic-resistant genes.
Antibiotic resistance can be developed in three ways: 1.) enzymatic degradation of drugs, 2.) alteration of bacterial proteins that are antimicrobial targets and 3.) changes in membrane permeability to antibiotics. When plasmid-harbouring bacteria grow and divide, the antimicrobial-resistant gene is passed on to daughter cells, usually in the form of a plasmid.
Antimicrobial resistance (AMR) can make the treatment of infectious diseases more difficult. The microbes stop responding to medicines, thereby increasing the risk of disease spread and mortality. Due to the negligence in the use of antibiotics during a viral infection, the issue of AMR has become worse in recent decades. Even though antibiotics do not have any effect during a viral infection, they are still prescribed to patients. This misuse increased a lot during the COVID-19 pandemic, thereby worsening the problem of bacterial AMR. Thus, AMR can be reduced by the prevention of misuse and overuse of antibiotics.
Again, these were things I had just read about in school and online. In the lab, I got to actually separate the plasmid DNA which, in certain cases, can be a culprit for AMR in bacteria. It was fascinating to understand how such a minuscule molecule can lead to drastic consequences in different contexts.