A sneak peek into biofilms in wound infections

Snehal Kadam’s interview with Bio Patrika hosting “Vigyaan Patrika”, a series of author interviews. Snehal is a first year PhD student with Dr. Angela Oates at the Hull York Medical School in the United Kingdom. Fascinated by all things bacterial, she is particularly interested in understanding biofilms, their role in infections and antibiotic resistance. She is also passionate about science communication and runs the Talk To A Scientist program with her co-founder Dr. Karishma Kaushik. When not in the lab, one can find her exploring the best food in the town! Here, Snehal talks about her work on biofilms carried out as a research assistant in Dr. Karishma Kaushik’s group at the Institute of Bioinformatics and Biotechnology, Savitribai Phule Pune University.

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How would you explain your paper’s key results to the non-scientific community?

We have all had a wound on our skin at some point in our life. It may have been a tiny paper cut, or a more serious injury from trauma. For wounds that are not as simple as a paper cut, we may have visited the doctor. They may prescribe us antibiotics, to prevent the wound from getting infected. When wounds do get infected with different micro-organisms (very tiny organisms), wound healing gets delayed. Another major problem with infected wounds is that infecting bacteria form ‘biofilms’. Biofilms are communities or aggregates of bacteria, living together in a self-secreted slimy 3D matrix. This matrix is like a covering around the bacteria which makes it difficult for antibiotics as well as our own immune system to attack and remove them. Since the bacteria are protected in this form, treating such infections becomes quite difficult. The bacteria in wounds interact constantly with various biological and chemical factors, such as our own immune and skin cells and the molecules they secrete. This means that the environment in the wound surrounding bacteria is very complex and has various components.

Schematic

In the laboratory, when scientists study wound infecting microorganisms, they make use of various nutrient broths. These broths are a mix of different chemicals and nutrients which facilitate bacterial growth in laboratory conditions. Although this approach works to an extent, these broths do not mimic the actual environment present in wounds and hence are not the most relevant way of studying wound infections in the lab. In our recent work, we developed an artificial wound fluid (called an in vitro wound milieu (IVWM)) that incorporates wound-relevant components such as serum, collagen, fibrinogen, fibronectin, lactoferrin, and lactic acid. This composition is quite close to clinical wound fluid, and hence closely represents the wound milieu. We found that two bacteria commonly found in wounds (Pseudomonas aeruginosa and Staphylococcus aureus), have different characteristics when grown in the IVWM compared to the nutrient broth (Luria-Bertani (LB) broth). For example, the bacteria grow at different rates, form biofilms of different thicknesses, and the effects of antibiotics on these biofilms is also different in the IVWM than laboratory nutrient broths.

By using an in vitro wound milieu, we can take a step towards understanding bacteria in wounds better!

What are the possible consequences of these findings for your research area?

Our findings show that incorporating human-relevant factors into laboratory studies of wound is important. By showing how characteristics like bacterial growth, biofilms and susceptibility to antibiotics differ when such factors are incorporated, we make a case for more studies to use this approach in biofilm and wound research. By using an in vitro wound milieu, we can take a step towards understanding bacteria in wounds better!

What was the exciting moment (eureka moment) during your research?

To visualize what biofilms looked like in the wound milieu we developed and in other media, we were using a ‘tile scan’ approach with a confocal microscope. The method involved taking images of individuals regions of the biofilm in 3D, and then stitching such adjoining regions to get a whole image. The approach we used allowed us to image the whole biofilm and not just a part of it. This was crucial to visualize the 3D structure of the biofilm. This experiment was something we had been working on to get more accurate images but got further delayed due to the pandemic. When I finally standardized the method to do this and obtained beautiful fluorescent images of the biofilms – that was the most exciting moment! It took us almost a year to be able to achieve this.

What do you hope to do next?

In terms of this piece of research, the wound milieu has great potential to explore further. Given its ease of formulation and compatibility with many standard biofilm assays, its recipe can be revised to incorporate even more components relevant to wound study. For example, when studying bacteria in diabetic wounds, adding glucose to the IVWM could help to mimic more relevant physiological conditions.

At present I am pursuing my PhD at the Hull York Medical School, United Kingdom where I am exploring antibiotic resistance in skin wound infections for my PhD project.

Where do you seek scientific inspiration?

Science is so much more than just ‘facts’. Through my journey in science, I have realized that the more I learn, the more inspired I am! My interests in microbiology and infections also stems from this – the more I learn about these tiny microorganisms causing infections in an organism as big as us humans, the more fascinated I become. I consider myself fortunate to have had great mentors through my scientific journey. Their support and encouragement have played a very important role in my journey so far. Dr. Karishma Kaushik, the corresponding author on this recent study, has been not only a great supervisor, but also an amazing mentor and friend!

How do you intend to help Indian science improve?

I think it is important to contribute to science not only through research, but also by contributing to the community itself. The Indian scientific community is constantly evolving and has so much potential. I, along with Dr. Karishma Kaushik, co-founded Talk To A Scientist, an online interactive platform for young minds (ages 6-16) to interact with real scientists live. In our weekly webinars we discuss various scientific topics and invite guest scientists to share their expertise. We are now funded by the IndiaBioscience Outreach Grant, and have completed one year of this platform. We are currently hosting Indian PhD students and postdoctoral researchers to share their work and research areas with our young participants.

Additionally, I have contributed to setting up the India Biofilms Society along with Dr. Karishma Kaushik and others. We are a group of researchers across India, exploring biofilms in health and environment research. We recently hosted ‘Biofilm Baithak’, our first India based biofilm two-day webinar to highlight biofilm research in India and identify ways to take it forward. We have also hosted international webinars with UK and Singapore based biofilm researchers.

During the initial months of the lockdown in India in 2020, Dr. Karishma and I hosted a series of webinars (along with guest speakers) for the academic community addressing different topics like academic writing, applying for internships in academia, social media for academics, return to India journeys for early career researchers and science careers outside academia.  I have also authored multiple articles for the academic community on IndiaBioscience and SciSoup.

Apart from this, I, along with a batchmate from IISER-Pune (Naven Narayanan), have started the Mentorship for Applications to PhDs (MAP) Program this year. Through this, we hope to create a network of mentors to guide Indian PhD aspirants through the PhD application process in India and around the world.

Reference

Kadam S, Madhusoodhanan V, Dhekane R, Bhide D, Ugale R, Tikhole U, Kaushik KS. Milieu matters: An in vitro wound milieu to recapitulate key features of, and probe new insights into, mixed-species bacterial biofilms. Biofilm. 2021 Apr 3;3:100047. doi: 10.1016/j.bioflm.2021.100047.

Dr. KS Kaushik Lab: https://www.karishmakaushiklab.com/

Edited by: Pratibha Siwach

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