DOI: 10.5061/dryad.jh9w0vtjt
title: Data from: Colony discrimination and competition in the eusocial trematode, Himasthla rhigedana

description: The California horn snail (Cerithideopsis californica) hosts a diverse
 community of trematode parasite species, yet these species rarely co-occur
 in the same host. Some trematodes in this community competitively exclude
 conspecifics and heterospecifics using a soldier caste. How these
 trematodes can distinguish colonymates from competitors is unknown. Here
 we examine patterns of colony discrimination in Himasthla rhigedana, a
 marsh-dwelling species of parasitic trematode that possesses a soldier
 caste in their intermediate snail hosts. Aggression assays pairing
 colonies against multiple opponents demonstrate that H. rhigedana
 distinguish between conspecific colonies, consistently directing more
 attacks towards colonies collected from a distant marsh. We demonstrate
 that conspecific interactions between colonies are predominantly
 symmetrical (both colonies attack during encounters), and that the
 likelihood of aggression is the same whether the attacker soldier is
 “sterile” (soldier redia with no germinal balls) or an “intermediate”
 (soldier redia with developing germinal balls). Recognizing heterospecific
 or conspecific threats is a necessary function for the evolution of
 soldier castes, which almost exclusively occur within eusocial insects. By
 finding parallels in disparate phyla, our results in H. rhigedana provide
 a foundation for understanding the evolution of colony discrimination
 generally, as well as in trematode species and other parasite taxa with
 similar competitive interactions.
description: Methodology can be found in the corresponding pre-printed
 manuscript.
description: # Colony discrimination and competition in the eusocial trematode,
 *Himasthla rhigedana* 
 ONLINE RESOURCE 1 (.zip)
 ################################################## *Decription:* A zip
 file of videos (.wmv) of cercariae collected from each colony used in our
 manuscript. Cercariae are how morphological species claims are made for
 our trematode species, so we include these videos as reference for how we
 identified each species used in our experiments. These videos comply with
 the CC0 license waiver required by Dryad for publication, and we allow any
 readers to make their own morphospecies claims as desired from these
 videos. Within the zip file is two folders, one for each assay (ABC, ABX).
 Each file name indicates the colony the cercariae were collected from. For
 example, "A1_Cercs_1.wmv" in the ABC folder shows the cercariae
 from colony A1 in the ABC assays. ONLINE RESOURCE 2 (.txt)
 ################################################## *Description:* A text
 file showing the final alignments of each COI sequence from the trematode
 colonies used in the ABC assays. This file is used in the accompanying R
 code to generate our haplotypes for Figure 4. More information on how
 these alignments were chosen can be found in the manuscript methods.
 ONLINE RESOURCE 3 (.csv)
 ################################################## *Description:* A csv
 dataset recording the aggressive interactions between trematodes in our
 ABC and ABX assays. This file was used in the accompanying R code to
 generate our boxplots and bar graphs for Figures 2 and 3, as well as the
 statistical tests using the binary attack outcomes as dependent variables
 in our pairwise t-tests (Table 1). All of these figures as well as the
 information found in our tables can be replicated by inputting this
 dataset into our R code and following the embedded instructions.
 *Explanation of data by column:* ASSAY = Categorical, binary. Indicates
 whether the sample in a row was used in the "ABC" or
 "ABX" assays described in the main manuscript. TRIAL =
 Numerical, discrete. A trial is one set of an ABC or ABX assay, labelled
 in chronological order. COLONY = Categorical, binary. Only "A"
 or "B" are possible. Indicates which colony, within an ABC or
 ABX assay, a soldier trematode was sampled from. ENEMY = Categorical.
 Indicates which colony, within an ABC or ABX assay, a recipient trematode
 was sampled from. TYPE = Categorical. An abbreviation that indicates the
 location and species sampled from the field. For example,
 "BL-Hirh" means a trematode samples from a Bolinas Lagoon snail
 host, identified as the morphospecies Himasthla rhigedana. More
 information about these abbreviations can be found in the main manuscript.
 BITES = Numerical, discrete. The number of soldier worms that engaged in
 biting their paired recipient. This can be from witnessing a
 "bite" on camera, or by witnessing the devoured remains that
 suggest aggression/biting occured. BITES.C = Numerical, discrete. The same
 data as the BITES column, but manually corrected, removing cases where the
 soldier trematode appeared to be biting or had previously consumed an
 enemy cercariae, not the enemy redia they were paired against. This is the
 column we call upon in our measurements of aggression. ONLINE RESOURCE 4
 (.csv) ################################################## Description: A
 csv dataset used specifically for the logistic regression models (Table
 2). All the information in table 2 can be replicated by inputting this
 dataset into our R code and following the embedded instructions. However,
 the table output provided by the stargazer package will be slightly
 different than what is observed in the manuscript (Table 2), as slight
 visual changes were made in latex before adding the table to the
 manuscript. *Explanation of data by column:* ASSAY = Categorical, binary.
 Indicates whether the sample in a row was used in the "ABC" or
 "ABX" assays described in the main manuscript. TRIAL =
 Numerical, discrete. A trial is one set of an ABC or ABX assay, labelled
 in chronological order. TREATMENT = Categorical, binary. Indicates what
 type of pairing the trematodes in the corresponding row were in, as well
 as the directionality of aggression. For example, "AB" means a
 soldier from colony A paired against a recipient from colony B. COLONY =
 Categorical. Indicates the colony ID and what trial the colony was in. For
 example, "A1" is the "A" colony from Trial 1 of either
 the ABC or ABX assay. A1 can exist twice, one for ABC and one for ABX, but
 this does not confound the data, as only one assay type is called when
 each logistic regression model was generated in R. ENEMY = Categorical,
 binary. Indicates which geographic region the recipient worm in an assay
 came from. N.ENEMY = Numerical, binary. A dummy variable for the ENEMY
 column. WELL ID = Numerical, discrete. Indicates which arena the trematode
 was assayed in. Every pairing had its own unique arena. BITES = Numerical,
 discrete. The number of soldier worms that engaged in biting their paired
 recipient. This can be from witnessing a "bite" on camera, or by
 witnessing the devoured remains that suggest aggression/biting occured.
 BITES.C = Numerical, discrete. The same data as the BITES column, but
 manually corrected, removing cases where the soldier trematode appeared to
 be biting or had previously consumed an enemy cercariae, not the enemy
 redia they were paired against. This is the column we call upon in our
 measurements of aggression. GERMS = Numerical, discrete. The maximum
 number of germinal balls observed within soldiers on video. COI-N =
 Numerical, discrete. The number of nucleotide base pair differences found
 between the two colonies paired in assay. Essentially the hamming distance
 between two sequences pulled, unaltered, from our "ONLINE RESOURCE
 2" supplemental file. COI-K80 = Numerical, continuous. The Kimura
 parameter distance between the COI sequences of the paired colonies. More
 information on this model of genetic distance measurement can be found in
 the main manuscript, in the documentation for the dist.dna function in R.
 COI-TN93 = Numerical, continuous. Another version of the Kimura parameter
 distance between the COI sequences of paired colonies. More information on
 this model of genetic distance measurement can be found in the main
 manuscript, in the documentation for the dist.dna function in R. ONLINE
 RESOURCE 5 (.R) ##################################################
 *Description* = An R code file, allowing any reader to replicate the
 figures and tables and statistical results shown in the corresponding
 pre-printed manuscript. Instructions for how to upload data and output the
 figures and tables are included inside, but these instructions assume the
 reader already has a basic knowledge of using R. All relevant outputs for
 this code file are included in the supplemental materials of this Dryad
 submission.