The awarded project aims to investigate how ecogeographic barriers impact the population structure of the Critically Endangered yellow-tailed woolly monkey (Lagothrix flavicauda) in Peru. Our focus is on understanding genetic differentiation between northern and southern populations, which appear separated by extensive land uninhabited by the species. We also explore genetic adaptation to high-altitude environments, assessing whether low-altitude habitats may pose barriers to dispersal. Overall, we aim to quantify genetic diversity across sampled populations to evaluate how varied landscapes—both natural and human-induced—affect this species' migration.

With the financial support from the NSF DDRIG, Co-PI Zarate sampled from five L. flavicauda populations in Peru, increasing our sample size to a total of 125 individuals. The provided map illustrates where Co-PI Zarate searched for and sampled from during this collection period and during a pilot collection in 2019. We now have a sufficient sample coverage over the known range of this species, having sampled from the northern and southern populations, and an area that lies at the mid-point of these populations, which will allow us to characterize how evolutionarily separated the northern and southern populations are from one another.

At Boston University’s Sensory Morphology and Anthropological Genomics Lab (SMAGL), we developed protocols for sequencing L. flavicauda mitochondrial DNA using Oxford Nanopore technology and built a reference mitochondrial genome using samples previously collected by PI Schmitt. We used the remainder of the funds provided by DDRIG to acquire lab kits and reagents and to return to the Wildlife Conservation Lab (WCL) at Los Amigos Biological Station (LABS) in Peru. Here we extracted DNA from the collected fecal samples, amplified it using PCR with established long-range primers, and sequenced these products using portable Nanopore technology. Accomplishing this lab work in Peru is an important component of this work and our Broader Impacts, as a bulk of the project was completed in-country and involved collaboration with local researchers in the area.

We faced challenges with degraded samples that hindered the isolation of long mitochondrial fragments. In such cases, we successfully amplified smaller fragments targeting regions important for assessing population diversity (e.g., COX2, D-Loop). Currently, we have isolated long fragments of the mitochondrial genome from 79 individuals and smaller fragments from an additional 22. We are currently analyzing the data and using GIS to characterize landscape features and barriers within and between areas where we collected L. flavicauda samples. So far, we have aligned data from 24 samples to a reference genome with high coverage depth (mean = 1713). As we continue data alignment, we intend to investigate genetic differences across groups living in fragmented habitats and search for signs of selection related to high-altitude adaptation.

In addition to research progress, we have fulfilled our Broader Impact goals. Co-PI Zarate held workshops with her Colombian field assistant Isabella Monedero Rodríguez at three schools in communities living sympatrically with L. flavicauda. These workshops (photos provided) involved activities fueling discussions about the importance of protecting the local forest concession – including hand-made coloring books and arts & crafts projects that emphasized the resources provided by the primary forest habitat of the monkeys. They also handed out informational pamphlets and coloring books to families with the help of the concession association employees. This work was largely successful due to Isabella's prior experience with Peruvian communities.

Co-PI Zarate developed an internship program for high school students from New Mission High School in Boston, where 55% of students are Black or African American, and 45% are Latino or Hispanic. This initiative helps students meet graduation requirements while fostering a long-term relationship between SMAGL and the school. In May 2023, we hosted two interns who learned DNA extraction, PCR, and Nanopore sequencing techniques. The following year, Co-PI Zarate trained two Ph.D. students to mentor three more high school interns. She (with support from PI Schmitt) also provided wet lab training in the BU SMAGL to a postdoctoral research scholar, several Ph.D. students, and two undergraduate students while establishing the genetics protocol used in this project. One of the undergrads, Sofia Weaver, spent time at the WCL this summer where she worked on amplifying additional samples for the mitochondrial DNA fragment as well as beginning her own project with the samples investigating potential sex-biased dispersal in L. flavicauda through the amplification of Y chromosome fragments using the protocols established for this project.

Our preliminary bioinformatic analyses and related projects have been accepted or presented at four conferences, including the upcoming 2024 London Calling. We are also preparing our reference mitogenomes for L. flavicauda and closely related species for publication. As the project nears completion, we are observing increased interest in using conservation genetics to protect L. flavicauda, both within our lab and among Peruvian researchers. Our collaborations with two Peruvian NGOs will ensure that our findings contribute to conservation policy and decisions.

Last Modified: 09/26/2024
Modified by: Melissa Zarate