DOI: 10.5061/dryad.xsj3tx9s3
title: Costs of delayed mating opportunities, overripening, and mate choice in Killifish

description: Females of highly iteroparous, oviparous species must mate multiply and
 potentially choose mates hundreds to thousands of times throughout their
 lifespan. Theory indicates that costly female mating preferences may favor
 low levels of preference in species that have to mate repeatedly. One
 potential cost of mating preference is overripening, which occurs in some
 external fertilizers when females ovulate but do not spawn their eggs.
 Overripening may create a trade-off between the benefits of mate choice
 (i.e., rejecting males of poor quality) and egg viability. This study
 tested for overripening as a cost of delayed mating and determined its
 effects on female mating preferences in the bluefin killifish, Lucania
 goodei. Female L. goodei often mate multiple times each day across
 multiple days when gravid. Females were divided into three treatments:
 mate every day, mate every two days, and mate every four days. In the
 first experiment, females were paired with either a red or a yellow
 conspecific male on spawning days.  In the second experiment,
 females were paired with either a conspecific male (L. goodei) or a
 heterospecific male (L. parva) on spawning days. In both experiments, the
 eggs were retrieved following spawning, and their viability (i.e.,
 survival) was monitored for five days. Females that were forced to ‘hold
 onto’ their eggs for extended periods had smaller clutch sizes and lower
 egg viability, demonstrating that costs to rejecting males potentially
 exist. In the first experiment, there was no evidence of female choice for
 red versus yellow males. In the second experiment, female preference for
 conspecifics versus heterospecifics declined when females could only mate
 once every four days, but the differences among treatments did not reach
 statistical significance. The results of this study illustrate a potential
 cost to female mate choice and suggest that the benefits of mate choice
 may not outweigh the costs due to egg overripening.
description: We collected bluefin and rainwater killifish. We conducted two
 experiments. In the first, females were allowed to mate daily, every other
 day, or once every four days. The goal was to determine whether egg
 viability decreased when females were precluded from spawning daily. We
 also sought to determine whether decreases in egg viability due to
 overripening caused a decrease in female mating preference. In the first
 experiment, female <em>L. goodei </em>could spawn with either
 a red or yellow color morph of their own species. Females were paired
 either every day, every other day, or once every four days with either a
 red or yellow male and given the opportunity to spawn for four hours.
 After this time, the eggs were collected, counted, and their survival to
 hatching was recorded. In the second experiment, female <em>L.
 goodei </em>were allowed to spawn with either a
 conspecific <em>L. goodei </em>male or a
 heterospecific <em>L. parva </em>male. Again, on spawning
 days, females could spawn for four hours. The eggs were then collected,
 counted, and monitored for survival until hatching.  We
 used these data to examine the effects of holding females on egg survival.
 We also compared the number of eggs laid with either red or yellow males
 (experiment 1) or conspecific or heterospecific males (experiment
 2).  The data sets have information on the replicate
 female, the treatment, the identity of the male, the number of eggs
 obtained, and the number of eggs that eventually hatch. 
description: # Costs of delayed mating opportunities, overripening, and mate choice in
 Killifish
 [https://doi.org/10.5061/dryad.xsj3tx9s3](https://doi.org/10.5061/dryad.xsj3tx9s3) ## Description of the data and file structure **Data Files** **The first experiment uses the 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)Egg_Hold_Exp1_data.csv data file.** There are 8 columns. They are the following: **Day** - This experiment took place across sequential days. Day 1 is '1', the first day of experimentation. The last day is 16. **Rep** - There were 6 replicate females in each treatment. These are numerical, but the R-code creates a unique ID for each female **treatment**   - This is a number code for the treatments. This helps with graphing. The numbers are 1, 2,3. These refer to three different treatments. Treatment 1 = spawn every day, Treatment 2 = spawn every other day, Treatment 3 = spawn every 4 days; The R code turns these into factors. **malecolor**   - refers to whether the female was paired with male having 'red' or 'yellow' anal fins **female_size_start**    - female standard length at the start of the experiment. It is not used in the analyses   **total**   - total number of eggs spawned on that day **alive**    - number of eggs that lived to hatching **dead** - number of eggs that died before hatching **The second experiment uses file 'EggHold_heterospecific_data.csv'. It is very similar to the first file. It has the following 8 columns:** **Day** - This experiment took place across sequential days. The first day is 17 because this is a continuation from the previous experiment. The last day is 32. (numerical) **Rep** - There were 6 replicate females in each treatment. These are numerical, but the R-code creates a unique ID for each female **treatment**   - This is a number code for the treatments. This helps with graphing. The numbers are 1, 2,3. These refer to three different treatments. Treatment 1 = spawn every day, Treatment 2 = spawn every other day, Treatment 3 = spawn every 4 days; The R code turns these into factors. **male_species** - refers to whether the female was paired with a conspecific *L. goodei* male or a heterospecific *L. parva *male **female_size_start**    - female standard length at the start of the experiment. It is not used in the analyses   **total**   - total number of eggs spawned on that day **alive**    - number of eggs that survived to hatching **dead** - number of eggs that died before hatching ## Code/software Chanetal.R is the R script file that has the data analysis and the commands used to make the figures. It uses the following packages and libraries: dplyr, ggplot2, lme4, lmerTest, car, RVAideMemoire, emmeans, tidyr, ggpubr, and grid. The scripts were run on R version 4.4.2 (2024-10-31) -- "Pile of Leaves".