An expedition to uncover hidden biodiversity across South America and Antarctica

Methods

Summary

Within each region, sampling locations will be selected based on their relative exposure to human activities, such as tourism, deforestation, and infrastructure, as well as climate change impacts, including wildfire-prone zones and areas of glacial retreat. We will collect water samples following standardized eDNA protocols to minimize contamination and ensure replicability. Multiple replicate samples will be taken at each site to account for local variability. We will conduct temporal replication where feasible to capture seasonal shifts in biodiversity.  

Samples will be filtered on-site, preserved with buffer solutions, and transported to Dr. Susana Caballero’s lab at Nova Southeastern University for analysis. DNA will be extracted using DNeasy Blood and Tissue Kits (Qiagen), then amplified and sequenced on the Illumina MiSeq platform. Sequences will be matched against reference databases to generate site-specific species inventories.  

To analyze biodiversity patterns, we will calculate alpha diversity (species richness and Shannon index) to evaluate species variety within sites and beta diversity (Bray-Curtis and Jaccard) to compare community composition across sites. Ordination methods (NMDS, PCoA) will visualize these differences. Statistical tests, including PERMANOVA and generalized linear models (GLMs), will be used to assess how biodiversity varies between pristine and impacted areas, across regions, and over time. Where available, we will compare eDNA results with traditional survey data to assess overlap, detect cryptic species, and validate the monitoring potential of eDNA.  

A key part of this project involves engaging local communities along the expedition route. In areas, particularly throughout the Amazon and Patagonia, we will collaborate with schools and community members to collect samples as part of a citizen science initiative. A lecture will introduce community members to the project and train them in following standardized eDNA protocols. We will use pre- and post-participation surveys or interviews to generate a report on the changes in environmental awareness or conservation attitudes. All activities will adhere to ethical research standards, including obtaining informed consent and demonstrating cultural respect. Biodiversity sampling will comply with all relevant national and international regulations. 

Challenges

Some challenges we anticipate overcoming are finding and working with schools and communities. To overcome this, we are partnering with universities and foundations that work within each region. 

Pre Analysis Plan

By utilizing eDNA analysis, we will produce an updated census of vertebrate biodiversity across the Amazon, Patagonia, and Antarctica. Capturing genetic material shed by organisms into the environment, eDNA enables the detection of species that are rare, cryptic, endangered, or otherwise difficult to monitor using traditional survey methods. These findings will provide essential baseline data for evaluating biodiversity trends and ecological shifts in some of the planet’s most vulnerable ecosystems. 

The results will inform conservation priorities by identifying current species distributions, biodiversity hotspots, and areas under ecological stress from climate change and human impacts. This study could detect signals of environmental degradation or the emergence of invasive species. Understanding the current state of the world’s remote biodiversity is crucial for informing future generations, guiding conservation efforts, and shaping global policy decisions. Maintaining these records will be essential for future generations to track shifts in ecological baselines and to understand the consequences of the current state of affairs.  

We will involve local communities, educators, and students in a citizen science initiative. This involvement aims to enhance the relationship between the local cultures and their environment. The results, stories, and visuals from this study will be shared through Mike Horn’s social channels, as well as short documentaries and photo essays that highlight the sponsors who enabled this research. Your brand will be a part of scientific development and adventures, expanding public awareness of biodiversity science and the conservation challenges faced in remote, critical areas of the world.