Abstract

We used the glass frog E. prosoblepon as a study model to explore how oviposition substrate influences embryo survival. We examined how oviposition site selection affects reproductive success. We combined field monitoring of egg clutches and oviposition site choice experiments to explore how microclimatic conditions, hatching success, and mortality causes affect the selected substrates for oviposition in this species. We found that microclimatic conditions did not differ between substrates, but hatching success was significantly higher on the substrate S. diffusa, which also experienced less predation. In the two-choice experiment, all females laid their eggs on S. diffusa, and those clutches had higher hatching success and faster embryonic development rates than those on leaves. These results indicate that non-random oviposition site selection in E. prosoblepon is driven by the maximization of embryo survival.

Contact

For questions regarding code and data analysis, please contact:

J. Sebastián Curaca-Fierro
University of Missouri
juansecurak97@gmail.com

Project Overview

Structure of the Data Folder

Save files to a folder named Data and create an additional folder named Figures. Below is a detailed description of the datasets used for the analysis, organised by year and processing stage.

Directory Tree

Data/ ├── Data Fieldwork 2024/ │ ├── Processed/ │ │ ├── all_substrates_temperatures.xlsx │ │ ├── diffusa_temperature.xlsx │ │ ├── hatching_success_and_mortality_2024.xlsx │ │ ├── leaf_temperature.xlsx │ │ ├── mortality_data_2021.csv │ │ └── rainfall_2024.xlsx │ └── Raw/ │ └── oviposition_substrates_2024.xlsx └── Data Fieldwork 2025/ └── Processed/ ├── devrate_2025_analysis.xlsx ├── egg_survival_matrix.xlsx ├── hatch_success_2025.xlsx └── Two_choice_matrix.xlsx Figures/ R_code/ └── code_Ovi_site_E.prosoblepon.qmd

Folder Contents

Data Fieldwork 2024 — Processed
1.1 all_substrates_temperatures.xlsx
Temperature and humidity data for all substrate types of the monitored clutches in 2024.
  • date Date (mm/dd/yyyy) and hour of the log.
  • temp Micro-environmental temperature of the substrate where the clutch was found.
  • humid Micro-environmental humidity of the substrate where the clutch was found.
  • datalog Identifier of the data logger used for the measurement.
  • ClutchID Unique identifier for the clutch.
  • susbstrate Type of substrate (following Goyes Vallejos et al. 2024).
1.2 diffusa_temperature.xlsx
Humidity and temperature specifically for monitored clutches on S. diffusa in 2024.
  • Hour_date_SellDiff Date (mm/dd/yyyy) and hour of the log.
  • SellDif_Micro_temp Micro-environmental temperature on the substrate.
  • SellDif_Micro_hum Micro-environmental humidity on the substrate.
  • Datalogger Identifier of the data logger used.
  • Clutch Unique identifier for the clutch.
1.3 hatching_success_and_mortality_2024.xlsx
Combined data for hatching success and mortality in egg clutches (2024).
  • Clutch_size Number of eggs per clutch.
  • Substrate Categorisation of the substrates used for oviposition.
  • Predation, Dry, Fungal, Failed, Rain, Unknown Counts of eggs dead per mortality source. Predation = embryos predated; Dry = desiccated; Fungal = fungal infections; Rain = rain stripped; Unknown = uncertain cause.
  • Num_hatched Total number of tadpoles successfully hatched per clutch.
  • Num_died Total number of eggs that died during development.
  • Prop.hatched Proportion of eggs hatched (Num_hatched / Clutch_size).
  • Prop.mortality Proportion of eggs dead (Num_died / Clutch_size).
1.4 leaf_temperature.xlsx
Humidity and temperature specifically for monitored clutches on leaves in 2024.
  • Date Date (mm/dd/yyyy) and hour of the log.
  • Temperature Micro-environmental temperature on the substrate.
  • Humidity Micro-environmental humidity on the substrate.
  • Datalog Identifier of the data logger used.
  • ClutchID Unique identifier for the clutch.
  • Susbstrate Type of substrate (following Goyes Vallejos et al. 2024). In this case, leaves only.
1.5 mortality_data_2021.csv
Substrate types and mortality sources for egg clutches from Goyes Vallejos et al. (2024) collected in 2021 along the same two transects of the Botanical Garden.
  • clutch_size Number of eggs per clutch.
  • substrate_type Categorisation of the substrates used for oviposition.
  • predation, dry, fungal, failed, rain, unknown Counts of eggs dead per mortality source (same definitions as 1.3, lowercase).
  • num_hatched Total number of tadpoles successfully hatched per clutch.
  • num_died Total number of eggs that died during development.
  • prop.hatched Proportion of eggs hatched (num_hatched / clutch_size).
  • prop.mortality Proportion of eggs dead (num_died / clutch_size).
1.6 rainfall_2024.xlsx
Rainfall data and surface temperature for the monitored clutches in 2024.
  • transect Transect where the clutch was found.
  • substrate Categorisation of the substrates used for oviposition (following Goyes Vallejos et al. 2024).
  • day_rain Daily rainfall received on the clutch (mm in the last 24 hours).
Data Fieldwork 2024 — Raw

This directory (…/Data Fieldwork 2024/Raw) contains raw, unprocessed observations from the 2024 field season.

2.1 oviposition_substrates_2024.xlsx
All clutches found in 2024, including oviposition sites and physical/spatial variables measured during searches.
  • habitat Category indicating disturbed, secondary, or primary forest. (Note: Only clutches in the disturbed area, Botanical Garden OTS, were used for analyses.)
  • transect Name of the surveyed transect.
  • location Approximate spatial location where the clutch was found.
  • substrate General specification of the physical substrate used for oviposition.
  • substrate_type Categorisation of the substrates used for oviposition (following Goyes Vallejos et al. 2024).
  • stage_found Developmental stage of the clutches at time of encounter, following Gosner (1960).
  • clutch_size Total number of eggs per clutch.
  • neighbor_clutches Count of other clutches found nearby within a 20 cm radius.
  • distance_water_cm Horizontal distance (cm) from the clutch to the border of the creek.
  • height_from_soil_cm Vertical distance (cm) of the clutch location relative to the soil.
  • monitored Indicates ('Y') if clutches were found at Gosner stage prior to 18 and subsequently monitored daily.
Data Fieldwork 2025 — Processed

This directory (…/Data Fieldwork 2025/Processed) contains processed data from the 2025 two-choice experiments.

3.1 devrate_2025_analysis.xlsx
Developmental data per clutch for the 2025 choice experiments.
  • pair_clutchID Unique identifier for the clutch.
  • substrate Substrate type used (either leaf or S. diffusa).
  • treatment Type of experiment (either no-choice or two-choice).
  • gosnerStage Gosner developmental stage for the monitored day. Dead clutches coded as 999.
  • daysfromOvip Number of days elapsed since oviposition.
  • earlydead Indicates if the clutch died before reaching hatching competency (Y/N).
3.2 egg_survival_matrix.xlsx
Wide-format matrix for the Kaplan–Meier analyses of embryos in 2025.
  • clutchsize Total number of eggs in the clutch.
  • substrate Substrate used in the experiment (either S. diffusa or leaf).
  • egg Identifies an individual egg within a clutch (development tracked per embryo).
  • stageday0, stageday1, stageday2, … Embryo developmental stage (Gosner 1960) on each day since oviposition. Coded 25 at hatching competency (and until hatched, then 1). Coded 0 on the day of death.
3.3 hatch_success_2025.xlsx
Hatching success and mortality sources for both experiments in 2025 (two-choice and no-choice).
  • clutch_ID Unique identifier for the clutch.
  • substrate Substrate type (either S. diffusa or leaf).
  • treatment Type of experiment (either two-choice or no-choice).
  • clutchSize Total number of eggs per clutch.
  • predated, fungi, non_dev, rainStrip, dehydrated, unknown Counts of dead embryos per mortality category, following Warkentin (2000) and Goyes Vallejos & Ramirez-Soto (2020).
  • hatched Total number of eggs successfully hatched.
  • hatching_onset Hatching start day (days since oviposition).
  • hatching_offset Hatching completion day (days since oviposition).
  • totaldead Total number of dead embryos in the clutch.
  • hatchSuccess Proportion of successfully hatched eggs (hatched / clutchSize).
3.4 Two_choice_matrix.xlsx
Log of two-choice experiments in 2025, indicating the substrate preferred by females for oviposition.
  • hourStart Sampling start time.
  • hourEnd Sampling end time.
  • hourFound Time when the pair in amplexus was spotted.
  • transect Transect in which the pair was found.
  • pair_clutchID Unique ID for the pair and resulting clutch.
  • clutchSize Number of eggs per clutch.
  • substrateChosen Substrate selected by the female for oviposition.
  • cage Identifier of the cage used.

Software and Dependencies