While our time in Nepal is sprinkled with plenty of extracurriculars, the real reason for our travels is to finish lab work for the Living with Leopards Project. The Living with Leopards Project, Shashank Poudel’s Ph.D. dissertation, has been ongoing for a number of years prior to our arrival. Starting before COVID, camera trap data was collected from the hills region of Nepal, just north of Chitwan. We started analyzing some of this data during the spring semester, going through AI-scrubbed photos and tagging animal images with the corresponding species. In addition, leopard scat samples were collected from the camera-trap survey area. These scats were stored in a -20 degrees Celsius freezer at the NTNC BCC during COVID and were thawed when we arrived on site. While there, we worked with these scat samples to conduct a diet analysis through physical and molecular methods. I learned a lot about wildlife research techniques while working here, and I’ll hope to illustrate a little bit about what I learned through this post.

Our research started with the dirty work: washing out the leopard scats and drying them to identify leopard prey species. For two days, we sieved out all the hair, bones, and other prey species markers and sun dried them on the hot tin rooves at the NTNC. Luckily, this didn’t take very long in the heat of the day, and Puja—the NTNC’s resident orphaned rhino—was never far away, guarding our coveted specimen.

When all of our samples had dried, we bagged them and moved to the cool of the downstairs lab. In the lab, we randomly drew 40 hairs/feathers from each sample. Then, with the training of other researchers staying at the NTNC, we learned how to prep and mount these hairs on microscope slides.

Viewed under the microscope, the cuticular pattern of each hair is unique to its species. Through this method, we are able to document the diet of each leopard, and Shashank hopes to correlate this data to patterns of human conflict in his final dissertation.

While it is easy to forget the significance of our work on a day-to-day basis, I was shocked out of my routine when I found one scat that looked eerily familiar to me. The length of the hair was unmistakable. It looked like a clump of hair I might have pulled out of a shower drain back home.

With a pit in my stomach, I pulled out one of my own hairs and mounted it on a reference slide. I labeled the species “human.” Though the results of the hair analysis are still pending, I would not be surprised to find our worst suspicions confirmed. We’d personally witnessed a leopard during our time in Chitwan, and stories of leopards preying on children are all too common in nearby communities. Through this work I’m reminded of the reality of leopard-human conflict, but I’m hopeful the study will help mitigating this tragedy in the future.

After all of our hair slides had been mounted for each sample, we took to the air-conditioned molecular lab upstairs. There, the wildlife veterinarian demoed our DNA extraction protocol. Using the equipment provided by USAID, the WWF and others, we enjoyed the air-conditioned facilities which had been built with grant money just a few years prior. We spent three full days in the lab extracting all the DNA from our samples. The next week, our leopard-specific primers arrived from Kathmandu, and we were ready for the final test: PCR amplification.

DNA extraction and amplification is notoriously hard to perform on leopard scat. Furthermore, I had never attempted a scat extraction prior to my time in Nepal. While tissue and hair samples are more straightforward to isolate, our scats were collected directly in the field where they may have been degraded. This kind of DNA isolation is nonetheless necessary to collect minimally-invasive samples from elusive species such as leopard. The experts at the NTNC already confirmed the scats as leopard based on their appearance, however PCR amplification is still the gold standard to positively identify our samples. We were advised to remain cautious so as not to get our hopes up as we poured hours into the DNA extraction and PCR amplification. When it came time for the results of the first test batch to be read, I waited with bated breath. The results? Two positives out of two samples tested. Success!

Using the optimized protocol from the test batch, we electrophoresed the rest of our samples in mass during our last few days at the NTNC. The PCR results are still pending, though the test batch proved we’d at least successfully extracted the DNA from leopard scat. This is arguably the hardest part of the procedure. I felt accomplished knowing I’d added a new skill to my repertoire and was able to contribute a meaningful piece to the Living with Leopards Project.