Illuminating Painted Bunting breeding season diets using eDNA

University of North Texas
Corinth, Texas
DOI: 10.18258/54159
Raised of $5,000 Goal
Funded on 11/09/23
Successfully Funded
  • $5,250
  • 105%
  • Funded
    on 11/09/23

About This Project

Given rapid declines in songbird populations, we must understand their trophic relationships to aid in conservation efforts. By utilizing fecal eDNA, this project will provide comprehensive diet information for the breeding population of Painted Buntings (Passerina ciris, PABU) at Lewisville Lake Environmental Learning Area (LLELA). Robust PABU diet information will aid ongoing restoration efforts at LLELA and inform their broader conservation.

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What is the context of this research?

The diet for Painted Buntings was established in the early 1900s using stomach content analysis, a method that is fatal for the bird, can miss soft-bodied prey items due to digestion, and relies on taxonomic expertise. Given human impacts on songbird habitats since this assessment, this diet information can be updated and greatly expanded using modern genomics approaches. Collaborating with the UNT Painted Bunting Project, which has been researching PABU ecology at LLELA since 2017, I non-invasively collected 192 fecal samples across 3 critical periods of the PABU breeding season: pre-breeding, breeding, and post-breeding. I plan to use the novel method of fecal eDNA metabarcoding to determine the presence of diet items from DNA in PABU feces.

What is the significance of this project?

Anthropogenic pressures have resulted in a ~30% decline in adult breeding birds since 1970, equating to ~3 billion birds lost in 50 years. Given these rapid declines, it is critcally important that the information used to support the conservation and habitat maintenance of songbirds is robust. This research will serve to update and expand upon the existing diet information for PABUs in order to provide a more holistic view of their feeding ecology by considering demographics and life history events. This research will inform future restoration decisions by the Conservation Manager at LLELA, as well as provide baseline data for PABU diets during critical life history stages in order to better inform international conservation efforts.

What are the goals of the project?

Given that 1) protein is essential for maintenance of body conditions and egg formation, 2) female Painted Buntings are the sole incubators of eggs and parental caregivers, 3) arachnids have been observed to be a key subsidy for nestlings of another species, and 4) that migration is energtically expensive, the aim of this project is to use fecal eDNA metabarcoding to test the following hypotheses:

H1: a) There will be a shift to incorporating more arthropods in the diet of female Painted Buntings during pre-breeding, a pattern that will be less pronounced in males. b) Arthropods will be the dominant food item in diets of Painted Bunting nestlings.

H2: In the post-rearing period leading up to migration, the diets of Painted Buntings will converge across bird demographics.


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Sample collection has been completed with support from the Fort Worth Audubon Society. I am now seeking funding to send fecal DNA extracts to a third party for DNA amplification and sequencing. I will extract fecal DNA from the samples in my lab at UNT, which greatly reduces the processing costs. It will cost ~$100 per sample to amplify and sequence extracted fecal DNA. Given the large number of previously collected samples (n = 192), I have identified 3 levels of funding that will enable specific project goals to be achieved:

Goal 1: $5000 to sequence fecal DNA from 50 fecal samples

20 pre-breeding, 10 breeding/nesting, and 20 post-breeding samples

Goal 2: + $5000 to sequence diet item DNA from 50 more fecal samples

G1 plus 20 pre-breeding and 30 post-breeding samples

Goal 3: + $5000 to sequence diet item DNA from 50 more fecal samples

G2 plus 10 pre-breeding, 10 breeding/nesting, and 30 post-breeding samples

Endorsed by

This is a really exciting project and will be a breakthrough in our understanding of the diet ecology of passerine birds. Haley does outstanding work in both the field and lab and is a perfect fit for succeeding with this groundbreaking science.
Painted Buntings are a vibrant but declining part of the North American avifauna. Most prior research on diet in this species took place over 100 years ago, using methods that are considered outdated, and did not go into demographic detail or have any focus on potential seasonal differences. This project will not only update and refine our knowledge of diet in this species, the use of fecal DNA metabarcoding (which is a fairly new method for studying bird diets) will help create a valuable roadmap for future studies of avian diet.
Haley is currently my graduate student working on her M.Sc. in Environmental Science at UNT. The research she is proposing capitalizes on a novel form of diet assessment (fecal DNA metabarcoding) to illuminate the diet of a declining bird spcies, the Painted Bunting, at key life history stages. Detailed assessments of songbird diets are either non-existent, or based on rudimentary approaches that can be fatal (e.g., gut content analysis). This research will be critical for determining how best to conserve Painted Buntings in the Anthropocene.
Haley Daniels is an up-and-coming player on the environmental stage and my experiences working with her over the last several years has shown me that once she starts something, she never lets go. This research is the first of hopefully many species of songbirds that can be sampled to help us determine if our restoration efforts are being utilized as a food source. This information will also shed a light on how this species' diet has or has not adapted as the habitat has been degraded and thereby guide our continuing habitat restoration efforts.

Project Timeline

I have spent the 2023 PABU breeding season collecting fecal samples at LLELA. Next, I plan to extract diet DNA from these fecal samples. I anticipate completing this by the end of the year and shipping these samples to a third party for DNA amplification and sequencing at the beginning of 2024. The turnaround time is relatively quick, so I expect to begin analyzing the resulting data by February. From there I will complete my thesis with the goal of publishing my findings.

May 23, 2023

Pre-breeding period fecal collection complete

Jul 14, 2023

Breeding/Nesting period fecal collection complete

Aug 04, 2023

Post-breeding period fecal collection complete

Sep 25, 2023

Project Launched

Dec 31, 2023

Fecal DNA extractions complete

Meet the Team

Haley Daniels
Haley Daniels
M.Sc. Student


University of North Texas
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Haley Daniels

Haley joined Dr. Zacchaeus Compson's lab in 2021 as an undergraduate field technician and stayed on with the lab to pursue her MSc in Environmental Science. She has previous experience focusing on Blackland prairie restoration, bird banding and stream ecology. Haley is equally interested in avian ecology and habitat restoration.

Additional Information



UNT Painted Bunting Project

Fort Worth Audubon Society


Cover photo credit: Susan Supak

Project Backers

  • 23Backers
  • 105%Funded
  • $5,250Total Donations
  • $119.57Average Donation
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