1. Title: Aquatic microbial communities influence the larval amphibian gut microbiota, neurodevelopment, and behavior

2. Author Information
   A. Principal Investigator Contact Information
   Name: Dr. Sarah Woodley
   Institution: Duquesne University
   Address: Pittsburgh, PA USA
   Email: woodleys@duq.edu

   B. Associate or Co-investigator Contact Information
   Name: Kyle Emerson
   Institution: Duquesne University
   Address: Pittsburgh,PA USA
   Email: emersonk@duq.edu

3. Date of data collection (single date, range, approximate date): 2021-2022

4. Geographic location of data collection: Pittsburgh, PA USA

5. Information about funding sources that supported the collection of the data: NIH grant P51 OD011092 for operation of the Oregon National Primate Research Center.

SHARING/ACCESS INFORMATION

1. Links to publications that cite or use the data:

Emerson, Kyle; Woodley, Sarah (Forthcoming 2024). Something in the water: Aquatic microbial communities influence the larval amphibian gut microbiota, neurodevelopment, and behavior. Proceedings of the Royal Society B.

1. Recommended citation for this dataset:

Emerson, Kyle; Woodley, Sarah (Forthcoming 2024). Something in the water: Aquatic microbial communities influence the larval amphibian gut microbiota, neurodevelopment, and behavior [Dataset]. Dryad. https://doi.org/10.5061/dryad.wstqjq2t2

DATA & FILE OVERVIEW

1. File List:

A) Emerson_2023_data.xlsx
B) Emerson - Sheet Information.csv
C) Emerson - metadata.csv
D) Emerson - Baseline Behavior.csv
E) Emerson - Empty Vial Behavior.csv
F) Emerson - Vial With Food Behavior.csv
G) Emerson - Food Slurry Behavior.csv

1. Relationship between files, if important: File A is the entire data set. File B contains all of the abbreviations used in the data files. Files D - G are separate files for behavioral assays.

2. Additional related data collected that was not included in the current data package: None

3. Are there multiple versions of the dataset? No
   A. If yes, name of file(s) that was updated: NA
   i. Why was the file updated? NA
   ii. When was the file updated? NA

DATA-SPECIFIC INFORMATION FOR: Emerson - metadata.csv

1. Number of variables: 40

2. Number of cases/rows: 72

3. Variable List:

* Microbial_Trtmt: Whether tadpoles were raised in Natural pond water (1) or autoclaved pond water (2)

* Replicate: What bin they were housed in

* Date_Euthanized: Date of euthanasia after dependent variable collection

* Date_Died: Date a tadpole naturally died during the experiment

* Corticosterone: Plasma Corticosterone levels (ng/mL)

* TW: Telencephalon Width (mm)

* TL: Telencephalon Length (mm)

* OTW: Optic Tectum Width (mm)

* OTL: Optic Tectum Length (mm)

* DW: Diencephalon Width (mm)

* DL: Diencephalon Length (mm)

* MW: Medulla Width (mm)

* RC1 - RC3: Principal Components 1-3 (PCs) calculated for Relative Brain Shape using Principal Component Analysis

• ASV: Amplicon Sequence Variants calculated for the gut microbiota

  • Shannon: Shannon Diversity Index calculated for the gut microbiota
  • Faiths: Faith's Phylogenetic Diversity calculated for the gut microbiota
  • VisEmpty_RC1 - RC3: Principal Components 1-3 (PCs) calculated for tadpole behavioral responses during the Empty Vial test using Principal Component Analysis

• VisFood_RC1 - RC3: Principal Components 1-3 (PCs) calculated for tadpole behavioral responses during the Vial filled with Frog Brittle test using Principal Component Analysis.

• Olfactory_RC1 - RC3: Principal Components 1-3 (PCs) calculated for tadpole behavioral responses during the Food Slurry test using Principal Component Analysis.

1. Missing data codes: None

2. Specialized formats or other abbreviations used: None

DATA-SPECIFIC INFORMATION FOR: Emerson - Baseline Behavior.csv, Emerson - Empty Vial Behavior.csv, Emerson - Vial with Food Behavior. csv, and Emerson - Food Slurry Behavior.csv

1. Number of variables: 40

2. Number of cases/rows: 20

3. Variable List:

* Microbial_Trtmt: Whether tadpoles were raised in Natural pond water (1) or autoclaved pond water (2)

* Av. Speed: Average Speed of tadpoles during behavioral test

* Mob. Av. Speed: Average speed of tadpoles when mobile during behavioral test

* Average Accel: Average acceleration of tadpoles during behavioral test

* Mobility Rate: Percentage of time mobile of tadpoles during behavioral test

* Visible Frames: Number of video frames in which the tadpole was visible during the behavioral test.

* Invisible Frames: Number of video frames in which the tadpole was invisible during the behavioral test.

* Explored areas: Number of different segmented areas tadpoles entered during the behavioral test.

* Exploration Rate: Percentage of total areas tadpoles entered during the behavioral test.

* Total Distance: Total distance a tadpole traveled during a behavioral test.

* Frozen Events: Number of instances that a tadpole was not moving for longer than one cumulative second during a behavioral test.

* Total Time Frozen: Total time a tadpole spent stationary during a behavioral test.

* Average time frozen: Average amount of cumulative time a tadpole spent stationary during a behavioral test.

• Time in Center: Total time a tadpole spent in an area of the arena that was not adjacent to the walls of the arena during a behavioral test.

  • Time near object/cue: Time a tadpole spent in the same area as an added visual or olfactory cue during a behavioral test.

1. Missing data codes: None

2. Specialized formats or other abbreviations used: None

Abstract and Further Experimental Information

We tested the hypothesis that larval amphibian neurodevelopment is affected by the aquatic microbial community present in their housing water. Newly hatched Northern Leopard Frog (Lithobates pipiens) tadpoles were raised in pond water that was unmanipulated (natural) or autoclaved. Tadpoles raised in autoclaved pond water had a gut microbiota with reduced bacterial diversity and altered community composition, had decreased behavioral responses to sensory stimuli, were larger in overall body mass, had relatively heavier brains, and had altered brain shape when compared with tadpoles raised in natural pond water. Further, the diversity and composition of the gut microbiota were associated with tadpole behavioral responses and brain measurements. Our results suggest that aquatic microbial communities shape tadpole behavior and brain development, providing strong support for the occurrence of the MGB axis in amphibians. Lastly, the dramatic role played by aquatic microbial communities on vertebrate neurodevelopment and behavior should be considered in future wildlife conservation efforts.