INTERSEXUAL CONFLICT IN ANDRODIOECIOUS CLAM SHRIMP SPECIES:

DO ANDRODIOECIOUS HERMAPHRODITES EVOLVE

TO AVOID MATING WITH MALES?

A Thesis Presented to

The Graduate Faculty of The University of Akron

In Partial Fulfillment

of the Requirements for the Degree

Master of Science

Rebecah Ford

May, 2017 INTERSEXUAL CONFLICT IN ANDRODIOECIOUS CLAM SHRIMP SPECIES:

DO ANDRODIOECIOUS HERMAPHRODITES EVOLVE

TO AVOID MATING WITH MALES?

Rebecah E Ford

Thesis

Approved: Accepted:

______Advisor Interim Dean of the college Dr. Stephen Weeks Mr. John Green

______Committee Member Dean of the Graduate School Dr. Randall Mitchell Dr. Chand Midha

______Committee Member Date Dr. Todd Blackledge

______Department Chair Dr. Stephen Weeks

ii ABSTRACT

How species pass on their genes to the next generation varies considerably. Through the evolutionary transition of reproductive types, it is hypothesized that a species can go from having males and females (dioecy) to hermaphrodites, with a few possible intermediate stages (Pannell, 2002; Weeks 2006b & 2012, Eppley & Jesson, 2008): hermaphrodites and females (gynodioecy) as well as hermaphrodites and males (androdioecy). Clam shrimp crustaceans have mating systems including dioecy (males and females) and androdioecy which makes them good systems for studying this transition. Chasnov

(2010) hypothesized that in androdioecious populations, hermaphrodites may evolve to avoid or evade males if the cost of outcrossing with males no longer outweighs the benefit. To test this hypothesis in clam shrimp, two questions were asked: 1) Have hermaphrodites evolved so that males cannot effectively recognize them?; and 2) Do androdiecious hermaphrodites prefer to be left alone? To answer the first question, mate guarding was observed in two dioecious and two androdioecious clam shrimp species. I found that dioecious males and androdioecious males guarded significantly differently

(p< 0.0001) in that androdioecious males made more mistakes when guarding potential mates. To test the second question, a three chambered tank was constructed wherein a focal male or hermaphrodite/female was given a choice between the same sex or the opposite sex. There was a significant sex effect (p< 0.0001) in that males of both reproductive types stayed closer to the opposite sex and females/hermaphrodites stayed iii closer to the same sex. There was also a marginally significant reproductive type effect

(p=0.0567) where androdioecious species were more likely to stay near the same sex and dioecious species were likely to stay near the opposite sex. Together, these two project support Chasnov’s 2010 theory and suggest that hermaphrodites are indeed evolving to avoid and evade males.

iv ACKNOWLEDGEMENTS

I would like to thank my advisor, Dr. Stephen Weeks, for guidance throughout the whole development of this project. I would also like to thank Dr. Randall Mitchell and Dr. Todd

Blackledge for their helpful comments during the process. Additional thanks to undergraduates Shara Ross, Paul Toth, and Nick Kasmar for rearing and care of the clam shrimp, Brent Ford for help in making the chamber, and to all friends and family for their support along the way.

v TABLE OF CONTENTS

Page

LIST OF TABLES…………………………………………………………………….. viii

LIST OF FIGURES……………………………………………..………………………. ix

CHAPTER

I. INTRODUCTION………………………………………………………..…………… 1

II. MATERIALS AND METHODS……………………………………………………... 8

General Methods……………………………………………………….………… 8

Mate Guarding Project………………………………………………………….. 11

Statistical Analysis………………………………………………...……. 12

Chamber Project………………………………………………………...………. 12

Statistical Analysis…………………………………………..………….. 14

III. RESULTS……………………………………………………………………..……. 15

Mate Guarding Project………………………………………………………….. 15

Chamber Project………………………………………………..……………….. 17

vi IV. DISCUSSION…………………………………………………………...………….. 19

REFERENCES…………………………………………………………………………. 26

APPENDICES…………………………………………….……………………………. 29

APPENDIX A. Data Collected for Mate Guarding Project…………………….. 30

APPENDIX B. Data Collected for Three Chamber Project……………...…….. 48

vii LIST OF TABLES Table Page 1.1 Location of clam shrimp soil samples……………………………………..…… 10 3.1 Two-way ANOVA of time spent in the chamber for each reproductive type and focal sex………………………………………………….……………. 18

viii LIST OF FIGURES Figure Page 3.1 Comparison of androdioecious and dioecious guarding…………………...…… 16 3.2 Time (in minutes) spent for each sex in the two reproductive types…………… 18

ix CHAPTER I

INTRODUCTION

In order to pass genes to the next generation, every living thing must reproduce; but how species do so varies considerably. It would seem that having a mixture of males and females (dioecy) is most advantageous in the animal kingdom, considering dioecy is the most common reproductive system among animals (Leonard, 2010; Weeks et al.

2006b). This makes sense in view of the wide range of benefits gained in a dioecious population. In dioecy, males and females can put all their resources into specific male or female functions compared to being a hermaphrodite that has to “choose” which gamete to produce, by when, and how many resources to put towards which sex (Charnov 1979).

Additionally, the avoidance of inbreeding is one of the most important benefits to dioecious species (Charlesworth & Charlesworth 1987; Baker, 1959). Inbreeding reduces the survival and decreases fertility (among other problems) of inbred offspring

(Charlesworth & Charlesworth 1987). Furthermore, in both the plant and animal kingdoms, mechanisms to avoid inbreeding have repeatedly evolved. For example, members of the animal kingdom often have one sex disperse away from the home location to avoid mating with their relatives (Pusey, 1987). In some plants, a mechanism

1 called heterostyly, where species have differing lengths of anthers and stigmas, is just one mechanism by which these plant species avoid creating inbred offspring (Ganders, 1979).

However, there are instances where self-fertilization would be advantageous for an individual’s fitness. For example, in low density situations, self-fertilization in hermaphrodites may be optimal considering it is better to run the risk of inbreeding than to not pass on your genes at all because one cannot find a mate (termed “reproductive assurance”; Baker, 1955). Thus, the most likely candidates to evolve selfing hermaphroditism are organisms living in environments where it is hard to find a mate or where your population is small and gene dispersal is rare (Ghiselin, 1969).

Through this evolutionary transition from having males and females (dioecy) to hermaphrodites, there are a few possible intermediate stages (Pannell, 2002; Weeks

2006b & 2012, Eppley & Jesson, 2008): hermaphrodites and females (gynodioecy) and hermaphrodites and males (androdioecy). Androdioecy is found scattered throughout the animal kingdom while gynodioecy is quite rare in animals (Weeks, 2012). Having a dioecious, intermediately hermaphroditic (androdioecious or gynodiecious) or purely hermaphroditic mating system has different costs and benefits (Ghiselin 1969; Weeks &

Bernhardt, 2004; Weeks et al., 2006b, 2009; Chasnov, 2010) and therefore which mating system evolves should be situational to the species.

If the evolutionary transition from dioecy to a different mating system occurs in the animal kingdom, it is has been proposed that the species would first evolve to be androdioecious and then potentially evolve to being purely hermaphroditic (Pannell,

2 2002; Weeks et al, 2006b, 2012). However, if it is beneficial to stay androdioecious and gain the benefits of being able to outcross or self-fertilize (Pannell, 2002), then the evolutionary transition from androdioecy to hermaphroditism may not occur. But if reproducing with males via outcrossing no longer has a benefit, then full selfing hermaphroditism should evolve (Ghiselin, 1969; Charnov, 1979; Pannell, 2002; Weeks et al., 2006b; Eppley & Jesson 2008). Therefore, androdioecious populations may either completely evolve into a population consisting of only hermaphrodites, or be

“constrained” to coexist with males in androdioecy. Those that are “constrained” usually occurs in plants and animals that have quick life cycles and pioneer new environments

(Pannell 1997, 2002; Weeks et al., 2006b; Weeks, 2012).

Androdioecy is one of the rarest mating systems only being found in 115 animal species including two cnidarians, 27 nematodes, one nemertean, 72 crustaceans, three insects, seven annelids, and three vertebrates (Weeks, 2012). Because of the unique reproductive dynamic androdioecy presents, several experiments have focused on the intersexual conflict created between the males and hermaphrodites of these species

(Chasnov, 2010; Benvenuto & Weeks, 2011). Having both selfing and outcrossing in a single species sets up an asymmetrical sexual conflict wherein males need to outcross with hermaphrodites to pass on their genes but hermaphrodites do not need males to pass on their genes (Chasnov, 2010). In other words, the males need the hermaphrodites more than the hermaphrodites need the males for fitness, creating the asymmetrical sexual conflict (Benvenuto and Weeks, 2012). This sexual conflict can create an evolutionary arms race where males and females try to outcompete one another to “win” the contest

3 (Chapman & Partridge, 1996). The winner often depends on factors such as the fitness pay off, the conflict cost, the power of the opponent, and the quality of the adaptations, such as size or secondary sexual characteristics used for controlling the opponent (Parker,

1979; Yamamura & Jormalainen, 1996).

A model was developed by Chasnov (2010) as a potential explanation for why androdioecy may exist and persist, taking into consideration the potential costs and benefits of being a self-fertilizing hermaphrodite. Chasnov’s model states that first a mutation in a female must occur, allowing her to self-fertilize (i.e., produce sperm). This would lead to a population with males, females, and newly evolved hermaphrodites.

Next, the hermaphrodites would need to outcompete the females to evolve a population consisting of males and hermaphrodites (i.e., androdioecy). If selfed offspring have a fitness of one half (or more) that of the outcrossed offspring, selfing would be advantageous because selfed offspring have double the relatedness to their parents relative to outcrossed offspring (Chasnov, 2010). This situation is more likely to evolve if there is reduced inbreeding depression and an increased need for reproductive assurance (Ghiselin 1969). Over time, the hermaphrodites may then evolve to avoid males, thereby becoming “reluctant” mates (Chasnov, 2010).

There are a variety of factors to consider when deciphering whether androdioecious hermaphrodites should evolve to avoid males (Chasnov, 2010; Weeks,

2012). If the benefits of selfing outweigh the costs of outcrossing, perhaps natural selection would favor hermaphrodites that evade males. Perhaps the hermaphrodites do not need the males at all? If this were the case, then when hermaphrodites are given a 4 choice, they should choose to self more often than outcross with a male. “If natural selection favors hermaphrodites that self-fertilize, then even persistent males may have difficulty finding and mating with hermaphrodites” (Chasnov 2010, page 545).

Of the androdioecious animals, nematodes and branchiopod crustaceans have been the most extensively studied (Chasnov 2010). In these species, the hermaphrodites can only reproduce by outcrossing with a male or self-fertilizing; they cannot outcross with other hermaphrodites. On the other hand, males need hermaphrodites to reproduce and pass on their genes. If the hermaphrodites do not need the males as strongly as the males need them, then why are males maintained in these androdiecious systems? The two simplest options are: 1) the males are indeed beneficial to the hermaphrodites and are maintained through forces such as avoidance of inbreeding depression, or 2) the males do not benefit the hermaphrodites and thus the males are maintained by rare undesirable copulations initiated by males as hypothesized by Chasnov (2010).

In the nematodes, most research on androdioecy has been done on Caenorhabditis elegans and Caenorhabditis briggsae which have been compared to the closely related dioecious species, Caenorhabditis remanei and Caenorhabditis brenneri. In these androdioecious nematode species, hermaphrodites lose many “female traits” when compared with the dioecious species, making the hermaphrodites hard to recognize by males. For example, the androdioecious C. elegans and C. briggsae do not have the same sex pheromone expressed by dioecious females of their family (Chasnov et al., 2007).

Additionally, males of C. elegans and C. briggsae seem to have trouble copulating with hermaphrodites, whereas males of the dioecious species easily mate with their respective 5 females. For example, C. elegans and C. briggsae hermaphroditic movement often impedes males from gaining contact with them, thus hindering copulation (Garcia et al.,

2007). Additionally, these hermaphrodites no longer respond to a factor males produce to nullify female resistance, compared to dioecious females who do respond (Garcia et al.,

2007). Some hermaphrodites also appear to be expel some or all of the semen after mating occurs (Barker, 1994).

These androdioecious Caenorhabditis hermaphrodites evolved behaviors that reduce receptivity to males; however there does not appear to have been any corresponding selection on males, such as adaptations to better recognize the hermaphrodites. Androdioecious Caenorhabditis males are still attracted to sex pheromones produced by closely related dioecious females, even though they are no longer created by hermaphrodites of their own species (Chasnov et al., 2007).

Additionally, males in androdioecious populations cannot recognize hermaphrodites from a distance (Chasnov et al., 2007). It would seem that in Caenorhabditis, androdioecious hermaphrodites are “reluctant” mates compared to dioecious females, and seem to have evolved adaptations to help them avoid males (Chasnov, 2010).

Similar studies have yet to be conducted in the branchiopod crustaceans. Clam shrimp are branchiopod crustaceans that are an ideal model system for studying sexual conflict (Benvenuto & Weeks, 2011; Weeks & Benvenuto 2008). Like the Rhabititdae nematodes, these crustaceans are differentiated based on their mating systems with both dioecy and androdioecy found throughout the family Limnadiidae (Weeks 2012). In particular, Eulimnadia species have an androdiecious mating system where 6 hermaphrodites can either outcross with males or self-fertilize. If the costs of outcrossing are greater than the costs of selfing, then selfing should be favored (Chasnov, 2010). For androdioecious clam shrimp, when the cost of outcrossing is weighed with the benefits of self-fertilization, it may be possible that hermaphrodites are evolving to avoid and possibly even evade males.

The following study focused on two questions to get at these ideas: 1) Have hermaphrodites evolved so that males cannot effectively recognize them? and 2) Do androdiecious hermaphrodites prefer to be left alone? If (1) is true, then dioecious males should more effectively recognize females for mate guarding while androdiecious males should be more prone to make mistakes. If so, dioecious males should be more frequently found attached to their opposite sex compared to androdioecious males, and the latter should be more likely to mistakenly “guard” other things. If (2) is true, then androdioecious hermaphrodites should avoid males and instead spend time near other hermaphrodites. These two questions will be addressed in two separate experiments delineated below.

7 CHAPTER II

METHODS AND MATERIALS

General Methods

Encysted eggs of two dioecious species (Eocyzicus argillaquus, Paralimnadia stanleyana) and two androdioecious species (Eulimnadia texana, and Eulimnadia dahli) were hatched from previously collected field soil samples (Table 1.1). Soil was put into the bottom of a tank until approximately 5 millimeters depth of the bottom was covered with the soil containing the eggs. Each individual soil sample containing the species’ eggs was put in separate tanks in the lab and then hydrated with deionized water. Each tank was kept under 24-hour Durotest sunlight-simulating fluorescent lights and had an air-stone to provide aeration. Once per day the tanks were fed a portion of food mixture made with a 0.5 grams brewer’s yeast, 0.5 grams of ground algae fish food flakes, and

100mL water. Any hatched tadpole shrimp were removed from the tanks, as these species will eat the clam shrimp and their eggs. The clam shrimp were then allowed to sexually mature before being used in the experiments. Shrimp were considered mature once the presence of claspers on males and eggs on hermaphrodites/females were seen.

Once maturity was reached, males and females were marked so they could be differentiated. To mark the clam shrimp, they were placed under a dissecting microscope

8 and the water was removed from around the clam shrimp using a pipette and then the carapace was further dried off using kimwipes. A thin paintbrush was then dipped into nail polish and a series of dots were placed on the carapace to differentiate individuals within males and females. After the marking was done, a small amount of water was put around the shrimp until the mark completely dried. Once the mark dried, the shrimp was returned to a temporary holding container momentarily until it was used for the experiment.

9 Table 1.1 Location of clam shrimp soil samples Species Reproductive Location name Coordinates mode Eocyzicus argillaquus Dioecious Cullimbin pools 1 & 5, S30°50.972 Australia E117°14.751

Paralimnadia Dioecious Kanangra Walls Pool 1, S33°59.933’ stanleyana Australia E150° 5.133’

Eulimnadia texana Androdioecious Wallace (previously 31°57.387′N; WAL, Sassaman & 109°08.998′W Weeks, 1993), Arizona, USA

Eulimnadia dahli Androdioecious The humps pool 3 & 6, S32°19.034 Australia E118°57.525

Elachbutting pool 5 & S30°35.984; 6, Australia E118°36.447

10 Mate Guarding Project

To observe the mate guarding behavior of clam shrimp species, a 1000 mL beaker was filled with 900mL of water from the species rearing tank. To this environment, ten previously molted carapaces and 10 pieces of debris from their tank (leaf litter and/or small sticks that were approximately the size of a carapace) were added. Five males and five female/hermaphrodites of the same species were then randomly collected from the home tank using a large-bore pipette and placed into the observation environment. Each of the males was marked with a different pattern of paint dots on their carapace using nail polish (as noted above) so the focal and other males could be differentiated from each other during the observation. Lastly, the focal male was added to the environment and allowed to acclimate for approximately two minutes. Afterwards, the focal male was observed for 20 minutes and scored using the JWatcher computer application (Blumstein

& Daniel, 2007) based on the following behaviors: guard same sex, guard opposite sex, guard debris, guard molted carapace, and swimming. “Guarding” was defined as a clasping of one of the above categories during the observational period. JWatcher recorded the number of times a behavior occurred as well as the length of time it occurred. Once a trial was completed, the focal male was removed and returned to a separate tank so as to not be used again, and a new focal male was used in the next trial.

11 Statistical Analysis

Behavioral observation trials were conducted for the two dioecious species

(Eocyzicus argillaquus, n=55; Paralimnadia stanleyana, n=35) and the two androdioecious species (Eulimnadia texana, n=50; Eulimnadia dahli, n=46). Counts of the number of times and what an individual guarded were used to perform the analyses.

First, a chi-square was run to determine whether there was a difference between the two species within each reproductive type. There were no significant differences in

2 guarding behavior between either of the two androdioecious species ( (3) = 1.76; P =

2 0.62) or the dioecious species ( (3) = 6.97; P = 0.07). Thus, the results were combined for the two species within each reproductive type (androdioecious and dioecious) and a second a chi-square was run to determine whether there was an overall difference between reproductive types.

Chamber Project

The following methods were adapted from Medland et al (2000) for quantifying clam shrimp encounter behavior in the lab. To test whether hermaphrodites/females and/or males have a preference for being near the same or opposite sex, shrimp were placed in a three chambered (larger center chamber + two smaller peripheral chambers)

Plexiglas container with the following dimensions: Overall - 35cm long, 15cm wide; center chamber - 25cm x 15cm; each peripheral chamber - 5cm x 15 cm. All chambers were 3 cm deep. The perimeter of the whole container was opaque to reduce any

12 distraction from outside the chamber and the dividers between sub-chambers were clear with holes smaller than the clam shrimp (1 millimeter diameter) drilled through them to allow for both visual and chemical cues between the shrimp in the different chambers.

The center chamber was marked with two lines drawn on the bottom of the tank allowing the identification of three equal portions of that central chamber.

The Plexiglas container was then filled with water from the species’ rearing tank.

Five randomly selected hermaphrodites or females (based on species) were then added to one of the peripheral chambers and five randomly selected males were added to the opposite peripheral chamber. A focal hermaphrodite/female or male was then added to the center chamber and allowed to acclimate for 2 minutes. After the acclimation period,

JWatcher (Blumstein & Daniel, 2007) was used to score which third of the center chamber the focal clam shrimp was in for a total of 10 minutes. JWatcher totaled the time a focal shrimp entered each third of the central chamber. This set up allowed the focal individual to make a choice to be near the same sex, the opposite sex, or to have no preference. After being used as a focal individual, the clam shrimp was returned to a separate tank, to eliminate the possibility of being used again. After each trial, the chamber was physically reoriented to remove any preference for a certain side of the experimental setup.

13 Statistical Analysis

Observations were collected for two dioecious species (Eocyzicus argillaquus, n=55; Paralimnadia stanleyana, n=56) and two androdioecious species (Eulimnadia texana, n=63; Eulimnadia dahli, n=53).

To analyze these data, the time a focal individual spent in the third of the chamber closest to the same sex was subtracted from the amount of time an individual spent in the third of the chamber nearest the opposite sex. Hence, if the value is positive, the focal individual spent more time near the opposite sex and if the value is negative, the focal individual spent more time near the same sex. A value near zero suggests no preference of either sex.

ANOVA’s were then computed to assess the difference in male and females/hermaphrodites in time spent near either sex. First, the two species within reproductive type were compared to each other to note any species-specific behavioral differences. There was no significant difference between species either for the androdioecious (F1,112 = 0.715; P = 0.3996) or the dioecious species (F1,107 = 1.640; P =

0.2030). Thus, the results were combined for each species within reproductive type. Next, a two-way ANOVA was run to assess behavioral differences in where the shrimp spent their time in the focal chambers between reproductive types (androdioecious compared to dioecious) and sexes (male compared to females/hermaphrodite).

14 CHAPTER III

RESULTS

Mate Guarding project

2 The reproductive types were significantly different in how they guarded (χ (3) =

92.38; P < 0.0001; Fig. 3.1). The androdioecious shrimp made many more guarding mistakes than the dioecious clam shrimp. The androdioecious males mistakenly guarded debris, empty carapaces, and the same sex in about two-thirds of the guarding attempts, and only correctly guarded hermaphrodites about one-third of the time (Fig. 3.1). The opposite was true for the dioecious males: they guarded females almost two thirds of the time (64%) while making guarding mistakes slightly over a third of the time (Fig. 3.1). In fact, the androdioecious species guarded empty carapaces, the opposite sex, and the same sex almost equally whereas the dioecious males clearly were the most effective at finding their opposite sex (Fig. 3.1).

15 Figure 3.1 Comparison of androdioecious and dioecious guarding

16 Not only did androdioecious and dioecious species guard differently (Figure 3.1), they also differed in the total number of guarding attempts. Androdioecious shrimp guarded a total of 633 individual times while dioecious only guarded 247 times throughout all the trials in approximately the same amount of total time, meaning androdioecious clam shrimp guarded over double the amount of times that dioecious shrimp did (Fig. 3.1).

Chamber Project

The time spent either near to or away from the opposite sex, differed in the two reproductive types (Table 3.1; Fig. 3.2). In both reproductive types, the males tended to spend more time near the opposite sex whereas the females/hermaphrodites tended to spend more time near their own sex (Focal Sex effect, Table 3.1; Fig. 3.2). There was a marginally significant reproductive type effect (Table 3.1) with androdioecious individuals tending to spend time nearer the same sex whereas dioecious individuals tending to spend time nearer the opposite sex (Fig. 3.2). However, this result was mainly driven by androdioecious hermaphrodites having a greater affinity for other androdioecious hermaphrodites than females being attracted to other females while dioecious males were more strongly attracted to females than androdioecious males were attracted to hermaphrodites (Fig. 3.2). In fact, the only sex/reproductive type combinations that showed a significant tendency towards one sex or the other (i.e., that the Opposite – Same ≠ 0) were androdioecious hermaphrodites being more attracted to other hermaphrodites and dioecious males being more attracted to dioecious females (Fig.

3.2). 17 Table 3.1 Two-way ANOVA of time spent in the chamber for each reproductive type and focal sex

Main Effect DF SS F-ratio Prob>F Reproductive Type 1 97.45 3.67 0.0567 Focal Sex 1 827.38 31.16 <.0001 Reprod* Focal Sex 1 0.09 0.003 0.9549

Figure 3.2 Time (in minutes) spent for each sex in the two reproductive types. Error bars depict 95% confidence intervals

18 CHAPTER IV

DISCUSSION

Androdioecy in clam shrimp presents a unique situation where hermaphrodites can reproduce via outcrossing with a male or by self-fertilization but cannot outcross with other hermaphrodites (Weeks et al., 2006a, 2006b). This situation presents a conflict between males and hermaphrodites because males benefit from having hermaphrodites more than hermaphrodites benefit from males (Benvenuto & Weeks 2011). Therefore, the costs vs. benefits of outcrossing compared to “selfing” may select for hermaphrodites that only self-fertilize (Chasnov, 2010). If so, hermaphrodites may evolve to avoid or evade males (Chasnov, 2010). To determine whether androdioecious hermaphrodites are evolving to avoid males, two questions were asked herein: 1) Have hermaphrodites evolved so that males cannot effectively recognize them? and 2) Do androdiecious hermaphrodites avoid areas with males?

Herein, we found evidence consistent with the notion that clam shrimp hermaphrodites are evolving to evade males. To answer the first of our two questions, we observed mate guarding behavior and found androdiecious clam shrimp guarded

19 significantly differently than dioecious clam shrimp (Fig. 3.1). The androdioecious males guarded empty carapaces, other males, and hermaphrodites at almost the same rate, suggesting that androdioecious males cannot easily recognize the opposite sex. On the other hand, dioecious males guarded females much more often than anything else (Fig.

1), showing that dioecious males can effectively find their opposite sex, although occasional mistakes are still made. Additionally, androdioecious males attempted guarding over two times more often than dioecious males, again suggesting that androdioecious males cannot easily find their intended target: the opposite sex. This evidence points to androdioecious hermaphrodites evolving to evade males, perhaps by losing the characteristics that make hermaphrodites easily identifiable as the opposite sex.

The observation that androdioecious males guard empty carapaces and other males similarly, but not debris, could suggest that males are attracted to texture or shape of the carapace because of a loss of attraction to the opposite sex via pheromones or other female cues. Additional observations showed that males would sometimes guard an empty carapace for an extended time but would grab other males and then let go quickly.

If androdioecious males cannot recognize hermaphrodites by some sort of female cue due to hermaphrodites evolving to avoid them, perhaps males are finding other ways to recognize the opposite sex. Perhaps after males find the appropriate texture or shape, they take additional cues from the amount of resistance, such as kicking, that they get from what they are guarding. This could explain why an empty carapace, which gives no

20 resistance and has the appropriate texture and/or shape, is guarded only 2% less than the intended opposite sex. Androdioecious males also attempt to guard other males often which could also be due to them having the correct texture or shape; however, these males show resistance which may be why they are guarded 5% less than the opposite sex.

To answer the second question, we observed clam shrimp in a divided chamber.

Androdioecious hermaphrodites were strongly attracted to other hermaphrodites whereas males were weakly (but not significantly) attracted to the hermaphrodites (Fig. 3.2). The dioecious females were weakly (marginally significantly) attracted to other females and the dioecious males were strongly attracted to females. Although androdioecious males were more attracted to hermaphrodites than conspecific males, the relationship was not as strong as dioecious males being attracted to conspecific females. This evidence shows that dioecious males are able to find the opposite sex more easily than androdiecious males, possibly because hermaphrodites are no longer producing chemical attractants for androdioecious males to respond to. Similar evidence suggesting hermaphrodites are evolving to avoid males was noted by the tendency of hermaphrodites to stay closer to other hermaphrodites and away from males. Females also stay closer to their own sex but not nearly to the same extent as seen in the hermaphrodites (Fig. 3.2). These observations are most easily explained by hermaphrodites actively avoiding males (question 2 above), supporting the prediction (Chasnov 2010) that androdioecious hermaphrodites should evolve to avoid males to increase their rate of self-fertilization.

Other research has touched on the question as to whether androdioecious clam shrimp are evolving to avoid males or not in clam shrimp. Contrary to Chasnov’s 2010 21 theory (and our current research), there seems to be some evidence that androdioecious

Eulimnadia hermaphrodites do prefer to outcross. Females in dioecious clam shrimp populations will not move their eggs into their brood chamber until a male is present

(Weeks et al., 2008). However, Eulimnadia hermaphrodites that are isolated will move eggs into their brood chamber more slowly than hermaphrodites in the presence of a male

(Zucker et al., 2002). It has also been shown that Eulimnadia males swim faster while hermaphrodites swim slower in the presence of males, which could be a hermaphroditic strategy to increase the likelihood of encountering a male (Medland et al., 2000). In this same study, Medland et al. (2000) found that hermaphrodites were attracted to being near males in an observation chamber. Hermaphrodites of Eulimnadia texana also kick and resist mate guarding less when they are closer to being receptive (Benvenuto & Weeks

2011). At face value, the above examples point towards hermaphrodites “wanting” to outcross.

However, the above studies do not necessarily imply that they are still attracted to males. Although isolated hermaphrodites do wait longer before they move their eggs into their brood chamber than paired hermaphrodites, they nonetheless will move the eggs in the absence of males. Dioecious females will never move their eggs into their brood chamber without a male present. Therefore, the fact that hermaphrodites move their eggs into their brood chamber even without a male present suggests that they are evolving away from the need to wait for males. The fact that they still wait for some time may be a legacy of the previous behavior of waiting indefinitely. In terms of hermaphrodites swimming slower than males (Medland et al. 2000), hermaphrodites have been seen

22 laying still on the substrate in their tank while males swim around quickly in the water above (personal observation). Hermaphrodites swimming more slowly or stopping in the presence of males could be interpreted as hermaphrodites separating themselves from males, either in the water column or via swimming speed, to avoid males rather than a behavior to obtain copulations. Additionally, Medland et al. (2000) finding that hermaphrodites were attracted to males could be due to a flaw in their design. In their experiment, the chamber was only divided into two parts: a focal individual portion

(containing a hermaphrodite or male) and a smaller portion containing only the opposite sex. Thus, the hermaphrodites were not given a choice between the same sex or opposite sex, and therefore the attraction of the hermaphrodite to the opposite sex may not have been completely accurate; hermaphrodites could have been attracted to movement or other clam shrimp rather than the opposite sex. In the current experiment, when both opposite and same sex options were presented, the hermaphrodites clearly preferred the same sex (Fig. 3.2), suggesting they are not attracted to males when given a proper choice. Thus, even though previous studies have reported evidence that hermaphrodites

“want” to outcross, these previous observations, combined with the current evidence, is supportive of Chasnov’s 2010 theory that hermaphrodites are evolving to avoid mating with males.

Additional evidence also points towards androdioecious clam shrimp evolving to primarily self-fertilize. Weeks et al (2000), showed that even when mating occurs, less than half of the eggs are actually outcrossed by the male. However, it is not known if this is from inviable male sperm or hermaphrodite choice. There are also some clam shrimp

23 populations which are purely hermaphroditic (Weeks et al., 2005 & 2008), which could be because these populations have evolved from an androdioecious ancestor by successfully eliminating outcrossing and thus losing males. Eulimnadia agassizii is one such species in which no males have been found throughout their populations (Weeks et al., 2005). Additionally, there are several populations within various Eulimnadia species

(Weeks et al., 2006a) that have no males. In all these cases, it is possible that hermaphrodites have been selected to avoid males, although it is also possible that males may have been lost due to their poorer dispersal abilities (Pannell, 1997, 2002). Future studies of hermaphroditic mating behavior that compares hermaphrodites from male+hermaphrodite relative to hermaphrodite-only populations may shed light on these two options.

Chasnov (2010) theorized that in androdioecious systems, if the benefits of selfing are greater than the benefits of outcrossing, then androdioecious hermaphrodites should evolve away from outcrossing. One such way a hermaphrodite could evolve away from outcrossing is by becoming unrecognizable to males by losing female characteristics to evade males. Another way to evolve away from outcrossing would be to avoid males all together. Together, the two experiments in this study show androdioecious hermaphrodites evading and avoiding males at a higher rate than their dioecious close relatives. Additionally, previous clam shrimp research is also consistent with androdioecious hermaphrodites evolving away from characteristics that promote outcrossing (Zucker et al., 2002; Weeks et al. 2000; Benvenuto & Weeks, 2011). This combined evidence supports Chasnov’s (2010) theory.

24 Researchers have suggested that androdioecy is an intermediate step in the evolution of hermaphroditism from dioecy (Pannell, 2002; Weeks 2006b & 2012, Eppley

& Jesson, 2008). If the pattern shown in the current projects of evolving to avoid males continues in clam shrimp, this could result in androdioecious species eventually becoming purely hermaphroditic, which may indeed explain why some clam shrimp populations and species are currently purely hermaphroditic (Weeks et al., 2005 & 2008).

Alternatively, the fact that males are still found in the majority of Eulimnadia populations could mean that clam shrimp species that are androdioecious are “stuck” in that reproductive type because males force copulations with hermaphrodites, even if hermaphrodites are evolving to avoid males (Chasnov, 2010). Future research in both clam shrimp and nematodes may shed light on the importance of this form of sexual conflict for the maintenance of androdioecy in these two lineages.

25 REFERENCES

Baker, H.G. 1955. Self-compatibility and establishment after 'long-distance' dispersal. Evolution, 9(3): 347-349 Baker, H.G. 1959. Reproductive methods as factors in speciation in flowering plants. Cold Spring Harbor Symposia on Quantitative Biology, 24: 177-191. Barker, D.M. 1994. Copulatory plugs and paternity assurance in the nematode Caenorhabditis elegans. Anim. Behav. 48: 147–156. Benvenuto, C. & Weeks, S.C. 2011. Mate guarding behavior in clam shrimp: the influence of mating system on intersexual conflict. Behavioral Ecology and Sociobiology, 65: 1899-1907 Blumstein, D. T., & Daniel, J. C. (2007). Quantifying behavior the JWatcher way. Sinauer Associates Incorporated. Chapman T, Partridge L. 1996. Sexual conflict as fuel for evolution. Nature 381:189-190. Charlesworth, C. & Charlesworth, D. 1987. Inbreeding depression and its evolutionary consequences. Annual Review of Ecology and Systematics, 18: 237-268 Charnov, E.L. 1979. Simultaneous hermaphroditism and sexual selection. Population Biology, 76(5): 2480-2484. Chasnov, J.R. 2010. The evolution from females to hermaphrodites results in a sexual conflict over mating in androdioecious nematode worms and clam shrimp. Journal of Evolutionary Biology, 23: 539-556. Chasnov, J.R., So, W.K., Chan, C.M., & Chow, K.L. 2007. The species, sex, and stage specificity of a Caenorhabditis sex pheromone. PNAS, 104(16):6730-6735. Eppley, S. M., and L. K. Jesson. 2008. Moving to mate: the evolution of separate and combined sexes in multicellular organisms. J. Evol. Biol. 21:727–736. Ganders, F. 1979. The biology of heterostyly. New Zealand Journal of Botany, 17: 607- 635. Garcia, L.R., LeBoeuf, B., Koo, P. 2007. Diversity in mating behavior of hermaphroditic and male–female caenorhabditis nematodes. Genetics, 175: 1761-1771.

26 Ghiselin, M.T. 1969. The Evolution of hermaphroditism among animals. The Quarterly Review of Biology, 44(2): 189-208. Leonard, J. L. 2010. The evolution of sexes, anisogamy, and sexual systems. Natural versus sexual selection. Pp. 15–39 in J. L. Leonard and A. Cordoba-Aguilar, ed. The Evolution of primary sexual characters in animals. Oxford Univ. Press, New York.

Medland, V.L., Zucker, N., & Weeks, S.C. 2000. Implications for the maintenance of androdioecy in the freshwater shrimp, Eulimnadia texana: Encounters between males and hermaphrodites are not random. Ethology, 106: 839-848. Pannell, J. 1997. The maintenance of gynodioecy and androdioecy in a metapopulation. Evolution, 10-20. Pannell, J.R. 2002. The evolution and maintenance of androdioecy. Annual Review of Ecology and Systematics, 33: 397-425. Parker, G. A. 1979. Sexual selection and sexual conflict. Sexual selection and reproductive competition in insects, 123-166. Pusey, A.E. 1987. Sex-based dispersal and inbreeding avoidance in birds and mammals. Tree, 2(10): 295-299. Sassaman, C. & Weeks, S.C. 1993. The genetic mechanism of sex determination in the conchostracan shrimp Eulimnadia texana. Am. Nat. 141: 314–328. Weeks, S.C. 2012. The role of androdioecy and gynodioecy in mediating evolutionary transitions between dioecy and hermaphroditism in the Animalia. Evolution, 66(12): 3670-3686. Weeks, S.C. & Benvenuto, C. 2008. Mate guarding in the androdioecious clam shrimp Eulimnadia texana: Male assessment of hermaphrodite receptivity. Ethology, 114: 64-74. Weeks, S. C., C. Benvenuto, and S. K. Reed. 2006b. When males and hermaphrodites coexist: a review of androdioecy in animals. Integrative and Comparitive Biology 46:449–464.

27 Weeks, S.C. & Bernhardt, R.L. 2004. Maintenance of androdioecy in the freshwater shrimp, Eulimnadia texana: field estimates of inbreeding depression and relative male survival. Evolutionary Ecology Research, 6: 227-242. Weeks, S.C., Crosser, B.R., Gray, M.M., Matweyou, J.A., & Zucker,N. 2000. Is there sperm storage in the clam shrimp Eulimnadia texana? Invertebrate Biology, 119(2): 215-221. Weeks, S. C., Posgai, R. T., Cesari, M., & Scanabissi, F. 2005. Androdioecy inferred in the clam shrimp Eulimnadia agassizii (Spinicaudata: Limnadiidae). Journal of Crustacean Biology, 25(3), 323-328. Weeks, S.C., Reed, S.K., Ott, D.W. & Scanabissi, F. 2009. Inbreeding effects on sperm production in clam shrimp (Eulimnadia texana). Evol. Ecol. Res. 11: 125–134. Weeks, S.C., Sanderson, T.F., Reed, S.K., Zofkiova, M., Knott, B., Balaraman, U., Pereira,G., Senyo, D.M., & Hoeh, W.R. 2006a. Ancient androdioecy in the freshwater crustacean Eulimnadia. Proceedings of the Royal Society, 273: 725- 734. Weeks, S. C., Sanderson, T. F., Zofkova, M., & Knott, B. 2008. Breeding systems in the clam shrimp family Limnadiidae (Branchiopoda, Spinicaudata). Invertebrate Biology, 127(3), 336-349. Yamamura N. & Jormalainen V. 1996. Compromised strategy resolves intersexual conflict over pre copulatory guarding duration. Evolutionary Ecology 10:661-680. Zucker, N., Aguilar, G.A., Weeks, S.C., & McCandless,L.G. 2002. Impact of males on variation in the reproductive cycle in an androdioecious desert shrimp. Invertebrate Biology, 121(1):66-72.

28 APPENDICES

29 APPENDIX A. Data Collected for Mate Guarding Project

# of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 1 empty carapace 0 0 dioeious Eocyzicus argillaquus 1 debris 0 0 dioeious Eocyzicus argillaquus 1 same sex 0 0 dioeious Eocyzicus argillaquus 1 opposite sex 1 1080167 dioeious Eocyzicus argillaquus 2 empty carapace 0 0 dioeious Eocyzicus argillaquus 2 debris 0 0 dioeious Eocyzicus argillaquus 2 same sex 1 2553 dioeious Eocyzicus argillaquus 2 opposite sex 2 12178 dioeious Eocyzicus argillaquus 3 empty carapace 0 0 dioeious Eocyzicus argillaquus 3 debris 0 0 dioeious Eocyzicus argillaquus 3 same sex 0 0 dioeious Eocyzicus argillaquus 3 opposite sex 2 1179815 dioeious Eocyzicus argillaquus 4 empty carapace 0 0 dioeious Eocyzicus argillaquus 4 debris 0 0 dioeious Eocyzicus argillaquus 4 same sex 0 0 dioeious Eocyzicus argillaquus 4 opposite sex 2 3361 dioeious Eocyzicus argillaquus 5 empty carapace 0 0 dioeious Eocyzicus argillaquus 5 debris 0 0 dioeious Eocyzicus argillaquus 5 same sex 0 0 dioeious Eocyzicus argillaquus 5 opposite sex 1 245767 dioeious Eocyzicus argillaquus 6 empty carapace 0 0 dioeious Eocyzicus argillaquus 6 debris 0 0 dioeious Eocyzicus argillaquus 6 same sex 1 6467 dioeious Eocyzicus argillaquus 6 opposite sex 4 357582 dioeious Eocyzicus argillaquus 7 empty carapace 0 0 dioeious Eocyzicus argillaquus 7 debris 0 0 dioeious Eocyzicus argillaquus 7 same sex 0 0 dioeious Eocyzicus argillaquus 7 opposite sex 7 52409 dioeious Eocyzicus argillaquus 8 empty carapace 0 0 dioeious Eocyzicus argillaquus 8 debris 3 9928 dioeious Eocyzicus argillaquus 8 same sex 1 1974 dioeious Eocyzicus argillaquus 8 opposite sex 2 680008

30 # of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 9 empty carapace 0 0 dioeious Eocyzicus argillaquus 9 debris 0 0 dioeious Eocyzicus argillaquus 9 same sex 1 1380 dioeious Eocyzicus argillaquus 9 opposite sex 1 20385 dioeious Eocyzicus argillaquus 10 empty carapace 2 3323 dioeious Eocyzicus argillaquus 10 debris 0 0 dioeious Eocyzicus argillaquus 10 same sex 2 3965 dioeious Eocyzicus argillaquus 10 opposite sex 3 28514 dioeious Eocyzicus argillaquus 11 empty carapace 1 6424 dioeious Eocyzicus argillaquus 11 debris 0 0 dioeious Eocyzicus argillaquus 11 same sex 0 0 dioeious Eocyzicus argillaquus 11 opposite sex 2 6859 dioeious Eocyzicus argillaquus 12 empty carapace 0 0 dioeious Eocyzicus argillaquus 12 debris 0 0 dioeious Eocyzicus argillaquus 12 same sex 0 0 dioeious Eocyzicus argillaquus 12 opposite sex 2 264519 dioeious Eocyzicus argillaquus 13 empty carapace 0 0 dioeious Eocyzicus argillaquus 13 debris 0 0 dioeious Eocyzicus argillaquus 13 same sex 1 722 dioeious Eocyzicus argillaquus 13 opposite sex 1 178199 dioeious Eocyzicus argillaquus 14 empty carapace 0 0 dioeious Eocyzicus argillaquus 14 debris 2 73907 dioeious Eocyzicus argillaquus 14 same sex 1 2552 dioeious Eocyzicus argillaquus 14 opposite sex 3 113502 dioeious Eocyzicus argillaquus 15 empty carapace 0 0 dioeious Eocyzicus argillaquus 15 debris 0 0 dioeious Eocyzicus argillaquus 15 same sex 0 0 dioeious Eocyzicus argillaquus 15 opposite sex 1 1196677 dioeious Eocyzicus argillaquus 16 empty carapace 0 0 dioeious Eocyzicus argillaquus 16 debris 0 0 dioeious Eocyzicus argillaquus 16 same sex 4 5634 dioeious Eocyzicus argillaquus 16 opposite sex 5 379935 dioeious Eocyzicus argillaquus 17 empty carapace 1 1039 dioeious Eocyzicus argillaquus 17 debris 0 0 dioeious Eocyzicus argillaquus 17 same sex 0 0 dioeious Eocyzicus argillaquus 17 opposite sex 1 173872 dioeious Eocyzicus argillaquus 18 empty carapace 0 0 dioeious Eocyzicus argillaquus 18 debris 0 0 dioeious Eocyzicus argillaquus 18 same sex 0 0 dioeious Eocyzicus argillaquus 18 opposite sex 3 28561 dioeious Eocyzicus argillaquus 19 empty carapace 0 0 31 # of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 19 debris 0 0 dioeious Eocyzicus argillaquus 19 same sex 0 0 dioeious Eocyzicus argillaquus 19 opposite sex 3 754172 dioeious Eocyzicus argillaquus 20 empty carapace 0 0 dioeious Eocyzicus argillaquus 20 debris 0 0 dioeious Eocyzicus argillaquus 20 same sex 2 3323 dioeious Eocyzicus argillaquus 20 opposite sex 0 0 dioeious Eocyzicus argillaquus 21 empty carapace 0 0 dioeious Eocyzicus argillaquus 21 debris 0 0 dioeious Eocyzicus argillaquus 21 same sex 0 0 dioeious Eocyzicus argillaquus 21 opposite sex 1 1177521 dioeious Eocyzicus argillaquus 22 empty carapace 0 0 dioeious Eocyzicus argillaquus 22 debris 0 0 dioeious Eocyzicus argillaquus 22 same sex 1 4844 dioeious Eocyzicus argillaquus 22 opposite sex 2 110922 dioeious Eocyzicus argillaquus 23 empty carapace 0 0 dioeious Eocyzicus argillaquus 23 debris 0 0 dioeious Eocyzicus argillaquus 23 same sex 2 2392 dioeious Eocyzicus argillaquus 23 opposite sex 1 88909 dioeious Eocyzicus argillaquus 24 empty carapace 0 0 dioeious Eocyzicus argillaquus 24 debris 0 0 dioeious Eocyzicus argillaquus 24 same sex 0 0 dioeious Eocyzicus argillaquus 24 opposite sex 1 438824 dioeious Eocyzicus argillaquus 25 empty carapace 0 0 dioeious Eocyzicus argillaquus 25 debris 0 0 dioeious Eocyzicus argillaquus 25 same sex 0 0 dioeious Eocyzicus argillaquus 25 opposite sex 2 49630 dioeious Eocyzicus argillaquus 26 empty carapace 0 0 dioeious Eocyzicus argillaquus 26 debris 0 0 dioeious Eocyzicus argillaquus 26 same sex 1 99824 dioeious Eocyzicus argillaquus 26 opposite sex 1 201469 dioeious Eocyzicus argillaquus 27 empty carapace 0 0 dioeious Eocyzicus argillaquus 27 debris 0 0 dioeious Eocyzicus argillaquus 27 same sex 1 1432 dioeious Eocyzicus argillaquus 27 opposite sex 2 960624 dioeious Eocyzicus argillaquus 28 empty carapace 0 0 dioeious Eocyzicus argillaquus 28 debris 0 0 dioeious Eocyzicus argillaquus 28 same sex 0 0 dioeious Eocyzicus argillaquus 28 opposite sex 1 3630 dioeious Eocyzicus argillaquus 29 empty carapace 0 0 dioeious Eocyzicus argillaquus 29 debris 0 0 32

# of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 29 same sex 0 0 dioeious Eocyzicus argillaquus 29 opposite sex 2 101439 dioeious Eocyzicus argillaquus 30 empty carapace 0 0 dioeious Eocyzicus argillaquus 30 debris 0 0 dioeious Eocyzicus argillaquus 30 same sex 4 9878 dioeious Eocyzicus argillaquus 30 opposite sex 4 465585 dioeious Eocyzicus argillaquus 31 empty carapace 0 0 dioeious Eocyzicus argillaquus 31 debris 0 0 dioeious Eocyzicus argillaquus 31 same sex 2 3996 dioeious Eocyzicus argillaquus 31 opposite sex 2 7088 dioeious Eocyzicus argillaquus 32 empty carapace 0 0 dioeious Eocyzicus argillaquus 32 debris 0 0 dioeious Eocyzicus argillaquus 32 same sex 1 1891 dioeious Eocyzicus argillaquus 32 opposite sex 3 53972 dioeious Eocyzicus argillaquus 33 empty carapace 1 2274 dioeious Eocyzicus argillaquus 33 debris 0 0 dioeious Eocyzicus argillaquus 33 same sex 1 1896 dioeious Eocyzicus argillaquus 33 opposite sex 1 94882 dioeious Eocyzicus argillaquus 34 empty carapace 1 1542 dioeious Eocyzicus argillaquus 34 debris 0 0 dioeious Eocyzicus argillaquus 34 same sex 0 0 dioeious Eocyzicus argillaquus 34 opposite sex 1 87218 dioeious Eocyzicus argillaquus 35 empty carapace 0 0 dioeious Eocyzicus argillaquus 35 debris 0 0 dioeious Eocyzicus argillaquus 35 same sex 0 0 dioeious Eocyzicus argillaquus 35 opposite sex 1 1069 dioeious Eocyzicus argillaquus 36 empty carapace 1 1388 dioeious Eocyzicus argillaquus 36 debris 0 0 dioeious Eocyzicus argillaquus 36 same sex 0 0 dioeious Eocyzicus argillaquus 36 opposite sex 3 6027 dioeious Eocyzicus argillaquus 37 empty carapace 0 0 dioeious Eocyzicus argillaquus 37 debris 0 0 dioeious Eocyzicus argillaquus 37 same sex 1 2563 dioeious Eocyzicus argillaquus 37 opposite sex 2 175799 dioeious Eocyzicus argillaquus 38 empty carapace 0 0 dioeious Eocyzicus argillaquus 38 debris 0 0 dioeious Eocyzicus argillaquus 38 same sex 1 4440 dioeious Eocyzicus argillaquus 38 opposite sex 1 1629 dioeious Eocyzicus argillaquus 39 empty carapace 0 0 dioeious Eocyzicus argillaquus 39 debris 0 0 dioeious Eocyzicus argillaquus 39 same sex 0 0 33

# of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 39 opposite sex 2 671417 dioeious Eocyzicus argillaquus 40 empty carapace 0 0 dioeious Eocyzicus argillaquus 40 debris 0 0 dioeious Eocyzicus argillaquus 40 same sex 0 0 dioeious Eocyzicus argillaquus 40 opposite sex 1 1181638 dioeious Eocyzicus argillaquus 41 empty carapace 1 1297 dioeious Eocyzicus argillaquus 41 debris 1 2290 dioeious Eocyzicus argillaquus 41 same sex 1 2311 dioeious Eocyzicus argillaquus 41 opposite sex 1 2067 dioeious Eocyzicus argillaquus 42 empty carapace 0 0 dioeious Eocyzicus argillaquus 42 debris 0 0 dioeious Eocyzicus argillaquus 42 same sex 4 10400 dioeious Eocyzicus argillaquus 42 opposite sex 5 76472 dioeious Eocyzicus argillaquus 43 empty carapace 0 0 dioeious Eocyzicus argillaquus 43 debris 1 2614 dioeious Eocyzicus argillaquus 43 same sex 0 0 dioeious Eocyzicus argillaquus 43 opposite sex 4 401492 dioeious Eocyzicus argillaquus 44 empty carapace 1 2009 dioeious Eocyzicus argillaquus 44 debris 0 0 dioeious Eocyzicus argillaquus 44 same sex 2 4560 dioeious Eocyzicus argillaquus 44 opposite sex 4 41961 dioeious Eocyzicus argillaquus 45 empty carapace 0 0 dioeious Eocyzicus argillaquus 45 debris 0 0 dioeious Eocyzicus argillaquus 45 same sex 0 0 dioeious Eocyzicus argillaquus 45 opposite sex 3 677858 dioeious Eocyzicus argillaquus 46 empty carapace 0 0 dioeious Eocyzicus argillaquus 46 debris 0 0 dioeious Eocyzicus argillaquus 46 same sex 0 0 dioeious Eocyzicus argillaquus 46 opposite sex 3 87451 dioeious Eocyzicus argillaquus 47 empty carapace 0 0 dioeious Eocyzicus argillaquus 47 debris 0 0 dioeious Eocyzicus argillaquus 47 same sex 2 3628 dioeious Eocyzicus argillaquus 47 opposite sex 1 15804 dioeious Eocyzicus argillaquus 48 empty carapace 0 0 dioeious Eocyzicus argillaquus 48 debris 0 0 dioeious Eocyzicus argillaquus 48 same sex 1 1653 dioeious Eocyzicus argillaquus 48 opposite sex 0 0 dioeious Eocyzicus argillaquus 49 empty carapace 0 0 dioeious Eocyzicus argillaquus 49 debris 0 0 dioeious Eocyzicus argillaquus 49 same sex 1 999 dioeious Eocyzicus argillaquus 49 opposite sex 1 957 34 # of length of grab reproduction species trial grab type grabs (ms) dioeious Eocyzicus argillaquus 50 empty carapace 0 0 dioeious Eocyzicus argillaquus 50 debris 0 0 dioeious Eocyzicus argillaquus 50 same sex 0 0 dioeious Eocyzicus argillaquus 50 opposite sex 1 1081203 dioeious Eocyzicus argillaquus 51 empty carapace 1 2980 dioeious Eocyzicus argillaquus 51 debris 0 0 dioeious Eocyzicus argillaquus 51 same sex 1 1509 dioeious Eocyzicus argillaquus 51 opposite sex 1 1114908 dioeious Eocyzicus argillaquus 52 empty carapace 1 918 dioeious Eocyzicus argillaquus 52 debris 0 0 dioeious Eocyzicus argillaquus 52 same sex 2 4163 dioeious Eocyzicus argillaquus 52 opposite sex 1 110591 dioeious Eocyzicus argillaquus 53 empty carapace 1 1395 dioeious Eocyzicus argillaquus 53 debris 0 0 dioeious Eocyzicus argillaquus 53 same sex 1 3487 dioeious Eocyzicus argillaquus 53 opposite sex 1 5129 dioeious Eocyzicus argillaquus 54 empty carapace 0 0 dioeious Eocyzicus argillaquus 54 debris 0 0 dioeious Eocyzicus argillaquus 54 same sex 2 7479 dioeious Eocyzicus argillaquus 54 opposite sex 3 287271 dioeious Eocyzicus argillaquus 55 empty carapace 0 0 dioeious Eocyzicus argillaquus 55 debris 0 0 dioeious Eocyzicus argillaquus 55 same sex 1 785 dioeious Eocyzicus argillaquus 55 opposite sex 1 691303 dioeious Paralimnadia stanleyana 1 empty carapace 2 2361 dioeious Paralimnadia stanleyana 1 debris 0 0 dioeious Paralimnadia stanleyana 1 same sex 1 2422 dioeious Paralimnadia stanleyana 1 opposite sex 1 702 dioeious Paralimnadia stanleyana 2 empty carapace 0 0 dioeious Paralimnadia stanleyana 2 debris 1 638 dioeious Paralimnadia stanleyana 2 same sex 0 0 dioeious Paralimnadia stanleyana 2 opposite sex 1 752 dioeious Paralimnadia stanleyana 3 empty carapace 0 0 dioeious Paralimnadia stanleyana 3 debris 0 0 dioeious Paralimnadia stanleyana 3 same sex 0 0 dioeious Paralimnadia stanleyana 3 opposite sex 1 21868 dioeious Paralimnadia stanleyana 4 empty carapace 0 0 dioeious Paralimnadia stanleyana 4 debris 1 2009 dioeious Paralimnadia stanleyana 4 same sex 0 0 dioeious Paralimnadia stanleyana 4 opposite sex 2 28172 dioeious Paralimnadia stanleyana 5 empty carapace 0 0 35 # of length of grab reproduction species trial grab type grabs (ms) dioeious Paralimnadia stanleyana 5 debris 0 0 dioeious Paralimnadia stanleyana 5 same sex 0 0 dioeious Paralimnadia stanleyana 5 opposite sex 1 2657 dioeious Paralimnadia stanleyana 6 empty carapace 1 2205 dioeious Paralimnadia stanleyana 6 debris 0 0 dioeious Paralimnadia stanleyana 6 same sex 0 0 dioeious Paralimnadia stanleyana 6 opposite sex 1 20239 dioeious Paralimnadia stanleyana 7 empty carapace 1 695 dioeious Paralimnadia stanleyana 7 debris 0 0 dioeious Paralimnadia stanleyana 7 same sex 2 3346 dioeious Paralimnadia stanleyana 7 opposite sex 0 0 dioeious Paralimnadia stanleyana 8 empty carapace 0 0 dioeious Paralimnadia stanleyana 8 debris 0 0 dioeious Paralimnadia stanleyana 8 same sex 1 13618 dioeious Paralimnadia stanleyana 8 opposite sex 2 692068 dioeious Paralimnadia stanleyana 9 empty carapace 0 0 dioeious Paralimnadia stanleyana 9 debris 0 0 dioeious Paralimnadia stanleyana 9 same sex 0 0 dioeious Paralimnadia stanleyana 9 opposite sex 2 26021 dioeious Paralimnadia stanleyana 10 empty carapace 0 0 dioeious Paralimnadia stanleyana 10 debris 0 0 dioeious Paralimnadia stanleyana 10 same sex 1 6144 dioeious Paralimnadia stanleyana 10 opposite sex 0 0 dioeious Paralimnadia stanleyana 11 empty carapace 0 0 dioeious Paralimnadia stanleyana 11 debris 0 0 dioeious Paralimnadia stanleyana 11 same sex 0 0 dioeious Paralimnadia stanleyana 11 opposite sex 2 78853 dioeious Paralimnadia stanleyana 12 empty carapace 0 0 dioeious Paralimnadia stanleyana 12 debris 0 0 dioeious Paralimnadia stanleyana 12 same sex 0 0 dioeious Paralimnadia stanleyana 12 opposite sex 1 833791 dioeious Paralimnadia stanleyana 14 empty carapace 0 0 dioeious Paralimnadia stanleyana 14 debris 0 0 dioeious Paralimnadia stanleyana 14 same sex 0 0 dioeious Paralimnadia stanleyana 14 opposite sex 6 32962 dioeious Paralimnadia stanleyana 15 empty carapace 1 1524 dioeious Paralimnadia stanleyana 15 debris 0 0 dioeious Paralimnadia stanleyana 15 same sex 0 0 dioeious Paralimnadia stanleyana 15 opposite sex 1 1353 dioeious Paralimnadia stanleyana 16 empty carapace 1 1952 dioeious Paralimnadia stanleyana 16 debris 0 0 36 # of length of grab reproduction species trial grab type grabs (ms) dioeious Paralimnadia stanleyana 16 same sex 1 1811 dioeious Paralimnadia stanleyana 16 opposite sex 0 0 dioeious Paralimnadia stanleyana 17 empty carapace 0 0 dioeious Paralimnadia stanleyana 17 debris 0 0 dioeious Paralimnadia stanleyana 17 same sex 1 1842 dioeious Paralimnadia stanleyana 17 opposite sex 1 1360 dioeious Paralimnadia stanleyana 18 empty carapace 1 1010 dioeious Paralimnadia stanleyana 18 debris 0 0 dioeious Paralimnadia stanleyana 18 same sex 0 0 dioeious Paralimnadia stanleyana 18 opposite sex 3 7390 dioeious Paralimnadia stanleyana 19 empty carapace 0 0 dioeious Paralimnadia stanleyana 19 debris 0 0 dioeious Paralimnadia stanleyana 19 same sex 1 1339 dioeious Paralimnadia stanleyana 19 opposite sex 1 21813 dioeious Paralimnadia stanleyana 21 empty carapace 0 0 dioeious Paralimnadia stanleyana 21 debris 0 0 dioeious Paralimnadia stanleyana 21 same sex 0 0 dioeious Paralimnadia stanleyana 21 opposite sex 1 2663 dioeious Paralimnadia stanleyana 22 empty carapace 0 0 dioeious Paralimnadia stanleyana 22 debris 0 0 dioeious Paralimnadia stanleyana 22 same sex 0 0 dioeious Paralimnadia stanleyana 22 opposite sex 1 3094 dioeious Paralimnadia stanleyana 23 empty carapace 0 0 dioeious Paralimnadia stanleyana 23 debris 0 0 dioeious Paralimnadia stanleyana 23 same sex 0 0 dioeious Paralimnadia stanleyana 23 opposite sex 1 6359 dioeious Paralimnadia stanleyana 24 empty carapace 0 0 dioeious Paralimnadia stanleyana 24 debris 0 0 dioeious Paralimnadia stanleyana 24 same sex 2 8849 dioeious Paralimnadia stanleyana 24 opposite sex 2 544810 dioeious Paralimnadia stanleyana 25 empty carapace 0 0 dioeious Paralimnadia stanleyana 25 debris 0 0 dioeious Paralimnadia stanleyana 25 same sex 0 0 dioeious Paralimnadia stanleyana 25 opposite sex 1 7040 dioeious Paralimnadia stanleyana 27 empty carapace 0 0 dioeious Paralimnadia stanleyana 27 debris 0 0 dioeious Paralimnadia stanleyana 27 same sex 0 0 dioeious Paralimnadia stanleyana 27 opposite sex 1 22933 dioeious Paralimnadia stanleyana 29 empty carapace 0 0 dioeious Paralimnadia stanleyana 29 debris 0 0 dioeious Paralimnadia stanleyana 29 same sex 0 0 37 # of length of grab reproduction species trial grab type grabs (ms) dioeious Paralimnadia stanleyana 29 opposite sex 1 2826 dioeious Paralimnadia stanleyana 30 empty carapace 0 0 dioeious Paralimnadia stanleyana 30 debris 0 0 dioeious Paralimnadia stanleyana 30 same sex 0 0 dioeious Paralimnadia stanleyana 30 opposite sex 1 138738 dioeious Paralimnadia stanleyana 31 empty carapace 0 0 dioeious Paralimnadia stanleyana 31 debris 0 0 dioeious Paralimnadia stanleyana 31 same sex 0 0 dioeious Paralimnadia stanleyana 31 opposite sex 1 279527 dioeious Paralimnadia stanleyana 34 empty carapace 1 19346 dioeious Paralimnadia stanleyana 34 debris 0 0 dioeious Paralimnadia stanleyana 34 same sex 0 0 dioeious Paralimnadia stanleyana 34 opposite sex 2 101708 dioeious Paralimnadia stanleyana 35 empty carapace 0 0 dioeious Paralimnadia stanleyana 35 debris 0 0 dioeious Paralimnadia stanleyana 35 same sex 0 0 dioeious Paralimnadia stanleyana 37 empty carapace 1 1284 dioeious Paralimnadia stanleyana 37 debris 0 0 dioeious Paralimnadia stanleyana 37 same sex 0 0 dioeious Paralimnadia stanleyana 37 opposite sex 1 27073 dioeious Paralimnadia stanleyana 38 empty carapace 0 0 dioeious Paralimnadia stanleyana 38 debris 0 0 dioeious Paralimnadia stanleyana 38 same sex 0 0 dioeious Paralimnadia stanleyana 38 opposite sex 1 1502 dioeious Paralimnadia stanleyana 39 empty carapace 1 893 dioeious Paralimnadia stanleyana 39 debris 0 0 dioeious Paralimnadia stanleyana 39 same sex 1 1784 dioeious Paralimnadia stanleyana 39 opposite sex 1 31487 dioeious Paralimnadia stanleyana 41 empty carapace 0 0 dioeious Paralimnadia stanleyana 41 debris 0 0 dioeious Paralimnadia stanleyana 41 same sex 0 0 dioeious Paralimnadia stanleyana 41 opposite sex 2 10282 dioeious Paralimnadia stanleyana 44 empty carapace 1 824 dioeious Paralimnadia stanleyana 44 debris 0 0 dioeious Paralimnadia stanleyana 44 same sex 0 0 dioeious Paralimnadia stanleyana 44 opposite sex 1 4216 dioeious Paralimnadia stanleyana 45 empty carapace 0 0 dioeious Paralimnadia stanleyana 45 debris 0 0 dioeious Paralimnadia stanleyana 45 same sex 0 0 dioeious Paralimnadia stanleyana 45 opposite sex 2 531012 androdiecious Eulimnadia texana 1 empty carapace 1 458290 38

# of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia texana 1 debris 1 1455 androdiecious Eulimnadia texana 1 same sex 0 0 androdiecious Eulimnadia texana 1 opposite sex 3 181673 androdiecious Eulimnadia texana 2 empty carapace 3 10149 androdiecious Eulimnadia texana 2 debris 0 0 androdiecious Eulimnadia texana 2 same sex 1 6322 androdiecious Eulimnadia texana 2 opposite sex 4 68989 androdiecious Eulimnadia texana 3 empty carapace 5 186746 androdiecious Eulimnadia texana 3 debris 3 6317 androdiecious Eulimnadia texana 3 same sex 5 21396 androdiecious Eulimnadia texana 3 opposite sex 0 0 androdiecious Eulimnadia texana 4 empty carapace 3 45375 androdiecious Eulimnadia texana 4 debris 1 3464 androdiecious Eulimnadia texana 4 same sex 3 38909 androdiecious Eulimnadia texana 4 opposite sex 1 7047 androdiecious Eulimnadia texana 5 empty carapace 10 60214 androdiecious Eulimnadia texana 5 debris 1 3214 androdiecious Eulimnadia texana 5 same sex 1 17295 androdiecious Eulimnadia texana 5 opposite sex 5 6881 androdiecious Eulimnadia texana 6 empty carapace 5 182040 androdiecious Eulimnadia texana 6 debris 1 795 androdiecious Eulimnadia texana 6 same sex 3 12066 androdiecious Eulimnadia texana 6 opposite sex 3 60198 androdiecious Eulimnadia texana 7 empty carapace 2 29999 androdiecious Eulimnadia texana 7 debris 0 0 androdiecious Eulimnadia texana 7 same sex 0 0 androdiecious Eulimnadia texana 7 opposite sex 0 0 androdiecious Eulimnadia texana 8 empty carapace 2 29999 androdiecious Eulimnadia texana 8 debris 0 0 androdiecious Eulimnadia texana 8 same sex 0 0 androdiecious Eulimnadia texana 8 opposite sex 0 0 androdiecious Eulimnadia texana 9 empty carapace 2 16530 androdiecious Eulimnadia texana 9 debris 1 593 androdiecious Eulimnadia texana 9 same sex 0 0 androdiecious Eulimnadia texana 9 opposite sex 6 52944 androdiecious Eulimnadia texana 10 empty carapace 0 0 androdiecious Eulimnadia texana 10 debris 2 1832 androdiecious Eulimnadia texana 10 same sex 2 14746 androdiecious Eulimnadia texana 10 opposite sex 2 21202 androdiecious Eulimnadia texana 11 empty carapace 1 6989 androdiecious Eulimnadia texana 11 debris 1 749 39

# of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia texana 11 same sex 4 17734 androdiecious Eulimnadia texana 11 opposite sex 2 17504 androdiecious Eulimnadia texana 12 empty carapace 2 9086 androdiecious Eulimnadia texana 12 debris 0 0 androdiecious Eulimnadia texana 12 same sex 2 30415 androdiecious Eulimnadia texana 12 opposite sex 4 137542 androdiecious Eulimnadia texana 13 empty carapace 2 28286 androdiecious Eulimnadia texana 13 debris 3 14147 androdiecious Eulimnadia texana 13 same sex 2 13653 androdiecious Eulimnadia texana 13 opposite sex 7 32173 androdiecious Eulimnadia texana 14 empty carapace 0 0 androdiecious Eulimnadia texana 14 debris 0 0 androdiecious Eulimnadia texana 14 same sex 0 0 androdiecious Eulimnadia texana 14 opposite sex 4 937094 androdiecious Eulimnadia texana 15 empty carapace 1 15869 androdiecious Eulimnadia texana 15 debris 0 0 androdiecious Eulimnadia texana 15 same sex 0 0 androdiecious Eulimnadia texana 15 opposite sex 2 8168 androdiecious Eulimnadia texana 16 empty carapace 3 10346 androdiecious Eulimnadia texana 16 debris 5 21413 androdiecious Eulimnadia texana 16 same sex 0 0 androdiecious Eulimnadia texana 16 opposite sex 1 11855 androdiecious Eulimnadia texana 17 empty carapace 4 48786 androdiecious Eulimnadia texana 17 debris 3 10753 androdiecious Eulimnadia texana 17 same sex 9 153456 androdiecious Eulimnadia texana 17 opposite sex 6 14124 androdiecious Eulimnadia texana 18 empty carapace 3 55702 androdiecious Eulimnadia texana 18 debris 1 5150 androdiecious Eulimnadia texana 18 same sex 1 16227 androdiecious Eulimnadia texana 18 opposite sex 0 0 androdiecious Eulimnadia texana 19 empty carapace 8 32115 androdiecious Eulimnadia texana 19 debris 1 894 androdiecious Eulimnadia texana 19 same sex 1 3543 androdiecious Eulimnadia texana 19 opposite sex 2 120746 androdiecious Eulimnadia texana 20 empty carapace 3 11467 androdiecious Eulimnadia texana 20 debris 1 618 androdiecious Eulimnadia texana 20 same sex 5 29763 androdiecious Eulimnadia texana 20 opposite sex 1 21549 androdiecious Eulimnadia texana 21 empty carapace 6 37010 androdiecious Eulimnadia texana 21 debris 2 2055 androdiecious Eulimnadia texana 21 same sex 1 3631 40 # of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia texana 21 opposite sex 3 13255 androdiecious Eulimnadia texana 22 empty carapace 1 25547 androdiecious Eulimnadia texana 22 debris 0 0 androdiecious Eulimnadia texana 22 same sex 1 4898 androdiecious Eulimnadia texana 22 opposite sex 3 45288 androdiecious Eulimnadia texana 23 empty carapace 1 970 androdiecious Eulimnadia texana 23 debris 0 0 androdiecious Eulimnadia texana 23 same sex 2 56654 androdiecious Eulimnadia texana 23 opposite sex 0 0 androdiecious Eulimnadia texana 24 empty carapace 1 503 androdiecious Eulimnadia texana 24 debris 0 0 androdiecious Eulimnadia texana 24 same sex 0 0 androdiecious Eulimnadia texana 24 opposite sex 0 0 androdiecious Eulimnadia texana 25 empty carapace 0 0 androdiecious Eulimnadia texana 25 debris 3 21752 androdiecious Eulimnadia texana 25 same sex 3 125013 androdiecious Eulimnadia texana 25 opposite sex 1 80995 androdiecious Eulimnadia texana 26 empty carapace 0 0 androdiecious Eulimnadia texana 26 debris 0 0 androdiecious Eulimnadia texana 26 same sex 1 6116 androdiecious Eulimnadia texana 26 opposite sex 1 658050 androdiecious Eulimnadia texana 27 empty carapace 0 0 androdiecious Eulimnadia texana 27 debris 0 0 androdiecious Eulimnadia texana 27 same sex 1 2280 androdiecious Eulimnadia texana 27 opposite sex 0 0 androdiecious Eulimnadia texana 28 empty carapace 0 0 androdiecious Eulimnadia texana 28 debris 0 0 androdiecious Eulimnadia texana 28 same sex 0 0 androdiecious Eulimnadia texana 28 opposite sex 1 1046222 androdiecious Eulimnadia texana 29 empty carapace 5 57690 androdiecious Eulimnadia texana 29 debris 0 0 androdiecious Eulimnadia texana 29 same sex 5 25751 androdiecious Eulimnadia texana 29 opposite sex 9 81206 androdiecious Eulimnadia texana 30 empty carapace 2 361898 androdiecious Eulimnadia texana 30 debris 0 0 androdiecious Eulimnadia texana 30 same sex 3 17528 androdiecious Eulimnadia texana 30 opposite sex 3 58981 androdiecious Eulimnadia texana 31 empty carapace 5 140823 androdiecious Eulimnadia texana 31 debris 0 0 androdiecious Eulimnadia texana 31 same sex 3 85297 androdiecious Eulimnadia texana 31 opposite sex 5 42145 41 # of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia texana 32 empty carapace 3 26682 androdiecious Eulimnadia texana 32 debris 1 833 androdiecious Eulimnadia texana 32 same sex 7 36493 androdiecious Eulimnadia texana 32 opposite sex 13 117220 androdiecious Eulimnadia texana 33 empty carapace 2 37454 androdiecious Eulimnadia texana 33 debris 0 0 androdiecious Eulimnadia texana 33 same sex 2 11976 androdiecious Eulimnadia texana 33 opposite sex 2 6709 androdiecious Eulimnadia texana 34 empty carapace 2 86692 androdiecious Eulimnadia texana 34 debris 1 839 androdiecious Eulimnadia texana 34 same sex 2 23997 androdiecious Eulimnadia texana 34 opposite sex 0 0 androdiecious Eulimnadia texana 35 empty carapace 7 11241 androdiecious Eulimnadia texana 35 debris 1 1425 androdiecious Eulimnadia texana 35 same sex 9 44451 androdiecious Eulimnadia texana 35 opposite sex 12 394722 androdiecious Eulimnadia texana 36 empty carapace 4 7203 androdiecious Eulimnadia texana 36 debris 3 5852 androdiecious Eulimnadia texana 36 same sex 3 8394 androdiecious Eulimnadia texana 36 opposite sex 4 11329 androdiecious Eulimnadia texana 37 empty carapace 1 690 androdiecious Eulimnadia texana 37 debris 0 0 androdiecious Eulimnadia texana 37 same sex 0 0 androdiecious Eulimnadia texana 37 opposite sex 0 0 androdiecious Eulimnadia texana 38 empty carapace 5 29116 androdiecious Eulimnadia texana 38 debris 3 3368 androdiecious Eulimnadia texana 38 same sex 10 62634 androdiecious Eulimnadia texana 38 opposite sex 4 238771 androdiecious Eulimnadia texana 39 empty carapace 0 0 androdiecious Eulimnadia texana 39 debris 3 4512 androdiecious Eulimnadia texana 39 same sex 0 0 androdiecious Eulimnadia texana 39 opposite sex 1 1207 androdiecious Eulimnadia texana 40 empty carapace 0 0 androdiecious Eulimnadia texana 40 debris 0 0 androdiecious Eulimnadia texana 40 same sex 0 0 androdiecious Eulimnadia texana 40 opposite sex 1 1183862 androdiecious Eulimnadia texana 41 empty carapace 3 6800 androdiecious Eulimnadia texana 41 debris 1 905 androdiecious Eulimnadia texana 41 same sex 1 5107 androdiecious Eulimnadia texana 41 opposite sex 4 11485 androdiecious Eulimnadia texana 42 empty carapace 0 0 42 # of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia texana 42 debris 0 0 androdiecious Eulimnadia texana 42 same sex 0 0 androdiecious Eulimnadia texana 42 opposite sex 1 1182328 androdiecious Eulimnadia texana 43 empty carapace 1 2515 androdiecious Eulimnadia texana 43 debris 1 1325 androdiecious Eulimnadia texana 43 same sex 1 1017 androdiecious Eulimnadia texana 43 opposite sex 3 10427 androdiecious Eulimnadia texana 44 empty carapace 2 12405 androdiecious Eulimnadia texana 44 debris 1 12377 androdiecious Eulimnadia texana 44 same sex 3 19690 androdiecious Eulimnadia texana 44 opposite sex 4 448194 androdiecious Eulimnadia texana 45 empty carapace 1 8371 androdiecious Eulimnadia texana 45 debris 0 0 androdiecious Eulimnadia texana 45 same sex 0 0 androdiecious Eulimnadia texana 45 opposite sex 1 3612 androdiecious Eulimnadia texana 46 empty carapace 6 25701 androdiecious Eulimnadia texana 46 debris 4 7265 androdiecious Eulimnadia texana 46 same sex 8 43266 androdiecious Eulimnadia texana 46 opposite sex 3 28718 androdiecious Eulimnadia texana 47 empty carapace 4 5114 androdiecious Eulimnadia texana 47 debris 1 1242 androdiecious Eulimnadia texana 47 same sex 2 6432 androdiecious Eulimnadia texana 47 opposite sex 6 9327 androdiecious Eulimnadia texana 48 empty carapace 1 2451 androdiecious Eulimnadia texana 48 debris 0 0 androdiecious Eulimnadia texana 48 same sex 0 0 androdiecious Eulimnadia texana 48 opposite sex 1 1377 androdiecious Eulimnadia texana 49 empty carapace 2 2624 androdiecious Eulimnadia texana 49 debris 2 4317 androdiecious Eulimnadia texana 49 same sex 1 3409 androdiecious Eulimnadia texana 49 opposite sex 1 17677 androdiecious Eulimnadia texana 50 empty carapace 1 6226 androdiecious Eulimnadia texana 50 debris 0 0 androdiecious Eulimnadia texana 50 same sex 3 8991 androdiecious Eulimnadia texana 50 opposite sex 2 28701 androdiecious Eulimnadia dahli 3 empty carapace 0 0 androdiecious Eulimnadia dahli 3 debris 0 0 androdiecious Eulimnadia dahli 3 same sex 1 27155 androdiecious Eulimnadia dahli 3 opposite sex 0 0 androdiecious Eulimnadia dahli 4 empty carapace 0 0 androdiecious Eulimnadia dahli 4 debris 1 716 43

# of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia dahli 4 same sex 2 4160 androdiecious Eulimnadia dahli 4 opposite sex 0 0 androdiecious Eulimnadia dahli 5 empty carapace 2 93610 androdiecious Eulimnadia dahli 5 debris 0 0 androdiecious Eulimnadia dahli 5 same sex 0 0 androdiecious Eulimnadia dahli 5 opposite sex 2 175211 androdiecious Eulimnadia dahli 6 empty carapace 3 4103 androdiecious Eulimnadia dahli 6 debris 1 1531 androdiecious Eulimnadia dahli 6 same sex 1 16944 androdiecious Eulimnadia dahli 6 opposite sex 3 17849 androdiecious Eulimnadia dahli 7 empty carapace 1 1554 androdiecious Eulimnadia dahli 7 debris 1 2074 androdiecious Eulimnadia dahli 7 same sex 1 78997 androdiecious Eulimnadia dahli 7 opposite sex 0 0 androdiecious Eulimnadia dahli 9 empty carapace 1 391633 androdiecious Eulimnadia dahli 9 debris 0 0 androdiecious Eulimnadia dahli 9 same sex 1 25087 androdiecious Eulimnadia dahli 9 opposite sex 0 0 androdiecious Eulimnadia dahli 10 empty carapace 1 16249 androdiecious Eulimnadia dahli 10 debris 1 2667 androdiecious Eulimnadia dahli 10 same sex 1 8774 androdiecious Eulimnadia dahli 10 opposite sex 0 0 androdiecious Eulimnadia dahli 12 empty carapace 2 98288 androdiecious Eulimnadia dahli 12 debris 2 3663 androdiecious Eulimnadia dahli 12 same sex 1 1006 androdiecious Eulimnadia dahli 12 opposite sex 1 6436 androdiecious Eulimnadia dahli 13 empty carapace 1 497690 androdiecious Eulimnadia dahli 13 debris 0 0 androdiecious Eulimnadia dahli 13 same sex 1 1450 androdiecious Eulimnadia dahli 13 opposite sex 0 0 androdiecious Eulimnadia dahli 14 empty carapace 2 5131 androdiecious Eulimnadia dahli 14 debris 0 0 androdiecious Eulimnadia dahli 14 same sex 5 19862 androdiecious Eulimnadia dahli 14 opposite sex 2 47159 androdiecious Eulimnadia dahli 15 empty carapace 2 329909 androdiecious Eulimnadia dahli 15 debris 0 0 androdiecious Eulimnadia dahli 15 same sex 0 0 androdiecious Eulimnadia dahli 15 opposite sex 1 199488 androdiecious Eulimnadia dahli 16 empty carapace 1 4760 androdiecious Eulimnadia dahli 16 debris 1 714 androdiecious Eulimnadia dahli 16 same sex 3 10408 44

# of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia dahli 16 opposite sex 5 130309 androdiecious Eulimnadia dahli 17 empty carapace 2 51667 androdiecious Eulimnadia dahli 17 debris 0 0 androdiecious Eulimnadia dahli 17 same sex 1 5451 androdiecious Eulimnadia dahli 17 opposite sex 0 0 androdiecious Eulimnadia dahli 18 empty carapace 0 0 androdiecious Eulimnadia dahli 18 debris 0 0 androdiecious Eulimnadia dahli 18 same sex 1 2264 androdiecious Eulimnadia dahli 18 opposite sex 1 419004 androdiecious Eulimnadia dahli 19 empty carapace 6 112443 androdiecious Eulimnadia dahli 19 debris 1 752 androdiecious Eulimnadia dahli 19 same sex 3 32447 androdiecious Eulimnadia dahli 19 opposite sex 5 26942 androdiecious Eulimnadia dahli 20 empty carapace 1 162616 androdiecious Eulimnadia dahli 20 debris 1 1422 androdiecious Eulimnadia dahli 20 same sex 0 0 androdiecious Eulimnadia dahli 20 opposite sex 0 0 androdiecious Eulimnadia dahli 21 empty carapace 0 0 androdiecious Eulimnadia dahli 21 debris 0 0 androdiecious Eulimnadia dahli 21 same sex 0 0 androdiecious Eulimnadia dahli 21 opposite sex 1 969 androdiecious Eulimnadia dahli 22 empty carapace 4 15820 androdiecious Eulimnadia dahli 22 debris 0 0 androdiecious Eulimnadia dahli 22 same sex 2 45005 androdiecious Eulimnadia dahli 22 opposite sex 2 4674 androdiecious Eulimnadia dahli 23 empty carapace 0 0 androdiecious Eulimnadia dahli 23 debris 0 0 androdiecious Eulimnadia dahli 23 same sex 0 0 androdiecious Eulimnadia dahli 23 opposite sex 3 28884 androdiecious Eulimnadia dahli 24 empty carapace 4 20677 androdiecious Eulimnadia dahli 24 debris 1 4545 androdiecious Eulimnadia dahli 24 same sex 3 37834 androdiecious Eulimnadia dahli 24 opposite sex 0 0 androdiecious Eulimnadia dahli 25 empty carapace 1 3261 androdiecious Eulimnadia dahli 25 debris 0 0 androdiecious Eulimnadia dahli 25 same sex 0 0 androdiecious Eulimnadia dahli 25 opposite sex 1 77863 androdiecious Eulimnadia dahli 26 empty carapace 2 35167 androdiecious Eulimnadia dahli 26 debris 0 0 androdiecious Eulimnadia dahli 26 same sex 2 10305 androdiecious Eulimnadia dahli 26 opposite sex 0 0 45

# of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia dahli 27 empty carapace 0 0 androdiecious Eulimnadia dahli 27 debris 0 0 androdiecious Eulimnadia dahli 27 same sex 0 0 androdiecious Eulimnadia dahli 27 opposite sex 2 14384 androdiecious Eulimnadia dahli 28 empty carapace 2 12942 androdiecious Eulimnadia dahli 28 debris 0 0 androdiecious Eulimnadia dahli 28 same sex 0 0 androdiecious Eulimnadia dahli 28 opposite sex 1 2445 androdiecious Eulimnadia dahli 29 empty carapace 2 9837 androdiecious Eulimnadia dahli 29 debris 3 2661 androdiecious Eulimnadia dahli 29 same sex 2 27822 androdiecious Eulimnadia dahli 29 opposite sex 2 33074 androdiecious Eulimnadia dahli 30 empty carapace 0 0 androdiecious Eulimnadia dahli 30 debris 0 0 androdiecious Eulimnadia dahli 30 same sex 1 1197039 androdiecious Eulimnadia dahli 30 opposite sex 0 0 androdiecious Eulimnadia dahli 31 empty carapace 4 35472 androdiecious Eulimnadia dahli 31 debris 0 0 androdiecious Eulimnadia dahli 31 same sex 0 0 androdiecious Eulimnadia dahli 31 opposite sex 4 163615 androdiecious Eulimnadia dahli 32 empty carapace 1 23873 androdiecious Eulimnadia dahli 32 debris 0 0 androdiecious Eulimnadia dahli 32 same sex 0 0 androdiecious Eulimnadia dahli 32 opposite sex 1 4811 androdiecious Eulimnadia dahli 33 empty carapace 3 4675 androdiecious Eulimnadia dahli 33 debris 0 0 androdiecious Eulimnadia dahli 33 same sex 1 10249 androdiecious Eulimnadia dahli 33 opposite sex 2 28036 androdiecious Eulimnadia dahli 34 empty carapace 1 4578 androdiecious Eulimnadia dahli 34 debris 1 1582 androdiecious Eulimnadia dahli 34 same sex 4 33954 androdiecious Eulimnadia dahli 34 opposite sex 2 4417 androdiecious Eulimnadia dahli 35 empty carapace 1 1950 androdiecious Eulimnadia dahli 35 debris 0 0 androdiecious Eulimnadia dahli 35 same sex 0 0 androdiecious Eulimnadia dahli 35 opposite sex 3 8489 Eulimnadia dahli androdiecious 36 empty carapace 1 1787 androdiecious Eulimnadia dahli 36 debris 1 1004 androdiecious Eulimnadia dahli 36 same sex 0 0 androdiecious Eulimnadia dahli 36 opposite sex 4 8871

46 # of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia dahli 37 empty carapace 1 2087 androdiecious Eulimnadia dahli 37 debris 0 0 androdiecious Eulimnadia dahli 37 same sex 2 4434 androdiecious Eulimnadia dahli 37 opposite sex 1 7650 androdiecious Eulimnadia dahli 38 empty carapace 0 0 androdiecious Eulimnadia dahli 38 debris 1 4352 androdiecious Eulimnadia dahli 38 same sex 3 24653 androdiecious Eulimnadia dahli 38 opposite sex 0 0 androdiecious Eulimnadia dahli 39 empty carapace 2 17819 androdiecious Eulimnadia dahli 39 debris 2 2181 androdiecious Eulimnadia dahli 39 same sex 2 23001 androdiecious Eulimnadia dahli 39 opposite sex 2 3601 androdiecious Eulimnadia dahli 40 empty carapace 2 43135 androdiecious Eulimnadia dahli 40 debris 0 0 androdiecious Eulimnadia dahli 40 same sex 2 30708 androdiecious Eulimnadia dahli 40 opposite sex 1 15818 androdiecious Eulimnadia dahli 41 empty carapace 2 17831 androdiecious Eulimnadia dahli 41 debris 1 1480 androdiecious Eulimnadia dahli 41 same sex 0 0 androdiecious Eulimnadia dahli 41 opposite sex 0 0 androdiecious Eulimnadia dahli 42 empty carapace 1 1951 androdiecious Eulimnadia dahli 42 debris 0 0 androdiecious Eulimnadia dahli 42 same sex 1 14523 androdiecious Eulimnadia dahli 42 opposite sex 1 37167 androdiecious Eulimnadia dahli 43 empty carapace 2 5881 androdiecious Eulimnadia dahli 43 debris 0 0 androdiecious Eulimnadia dahli 43 same sex 5 71371 androdiecious Eulimnadia dahli 43 opposite sex 0 0 androdiecious Eulimnadia dahli 44 empty carapace 0 0 androdiecious Eulimnadia dahli 44 debris 0 0 androdiecious Eulimnadia dahli 44 same sex 0 0 androdiecious Eulimnadia dahli 44 opposite sex 2 7889 androdiecious Eulimnadia dahli 45 empty carapace 0 0 androdiecious Eulimnadia dahli 45 debris 1 9560 androdiecious Eulimnadia dahli 45 same sex 1 3254 androdiecious Eulimnadia dahli 45 opposite sex 0 0 androdiecious Eulimnadia dahli 46 empty carapace 0 0 androdiecious Eulimnadia dahli 46 debris 0 0 androdiecious Eulimnadia dahli 46 same sex 0 0 androdiecious Eulimnadia dahli 46 opposite sex 0 0 androdiecious Eulimnadia dahli 47 empty carapace 2 4897 47 # of length of grab reproduction species trial grab type grabs (ms) androdiecious Eulimnadia dahli 47 debris 0 0 androdiecious Eulimnadia dahli 47 same sex 3 67419 androdiecious Eulimnadia dahli 47 opposite sex 3 11372 androdiecious Eulimnadia dahli 48 empty carapace 0 0 androdiecious Eulimnadia dahli 48 debris 0 0 androdiecious Eulimnadia dahli 48 same sex 1 31154 androdiecious Eulimnadia dahli 48 opposite sex 0 0 androdiecious Eulimnadia dahli 49 empty carapace 0 0 androdiecious Eulimnadia dahli 49 debris 0 0 androdiecious Eulimnadia dahli 49 same sex 0 0 androdiecious Eulimnadia dahli 49 opposite sex 1 4627 androdiecious Eulimnadia dahli 50 empty carapace 1 17627 androdiecious Eulimnadia dahli 50 debris 0 0 androdiecious Eulimnadia dahli 50 same sex 1 13139 androdiecious Eulimnadia dahli 50 opposite sex 1 61135

48 APPENDIX B. Data Collected for Chamber Project

Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Eocyzicus argillaquus 1 male opposite sex 23 383842 dioecious Eocyzicus argillaquus 1 male middle 29 186874 dioecious Eocyzicus argillaquus 1 male same sex 7 26924 dioecious Eocyzicus argillaquus 2 male opposite sex 15 472835 dioecious Eocyzicus argillaquus 2 male middle 19 84617 dioecious Eocyzicus argillaquus 2 male same sex 4 41095 dioecious Eocyzicus argillaquus 3 male opposite sex 12 117711 dioecious Eocyzicus argillaquus 3 male middle 47 160287 dioecious Eocyzicus argillaquus 3 male same sex 36 321314 dioecious Eocyzicus argillaquus 4 male opposite sex 36 214346 dioecious Eocyzicus argillaquus 4 male middle 53 209339 dioecious Eocyzicus argillaquus 4 male same sex 19 175502 dioecious Eocyzicus argillaquus 5 female opposite sex 36 214346 dioecious Eocyzicus argillaquus 5 female middle 53 209339 dioecious Eocyzicus argillaquus 5 female same sex 19 175502 dioecious Eocyzicus argillaquus 6 female opposite sex 14 409575 dioecious Eocyzicus argillaquus 6 female middle 21 134096 dioecious Eocyzicus argillaquus 6 female same sex 8 55517 dioecious Eocyzicus argillaquus 7 female opposite sex 11 283364 dioecious Eocyzicus argillaquus 7 female middle 16 71806 dioecious Eocyzicus argillaquus 7 female same sex 7 243471 dioecious Eocyzicus argillaquus 8 female opposite sex 20 424929 dioecious Eocyzicus argillaquus 8 female middle 26 149023 dioecious Eocyzicus argillaquus 8 female same sex 6 25173 dioecious Eocyzicus argillaquus 9 female opposite sex 14 382685 dioecious Eocyzicus argillaquus 9 female middle 16 146283 dioecious Eocyzicus argillaquus 9 female same sex 3 69470 dioecious Eocyzicus argillaquus 10 female opposite sex 9 56273 dioecious Eocyzicus argillaquus 10 female middle 15 120158 dioecious Eocyzicus argillaquus 10 female same sex 8 421928 dioecious Eocyzicus argillaquus 11 male opposite sex 9 397716 dioecious Eocyzicus argillaquus 11 male middle 19 133096 dioecious Eocyzicus argillaquus 11 male same sex 11 67641 dioecious Eocyzicus argillaquus 12 male opposite sex 8 572624 dioecious Eocyzicus argillaquus 12 male middle 8 23648

49 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Eocyzicus argillaquus 12 male same sex 1 2494 dioecious Eocyzicus argillaquus 13 male opposite sex 18 336113 dioecious Eocyzicus argillaquus 13 male middle 28 103769 dioecious Eocyzicus argillaquus 13 male same sex 12 157975 dioecious Eocyzicus argillaquus 14 male opposite sex 3 163603 dioecious Eocyzicus argillaquus 14 male middle 18 81824 dioecious Eocyzicus argillaquus 14 male same sex 17 353972 dioecious Eocyzicus argillaquus 15 male opposite sex 22 276025 dioecious Eocyzicus argillaquus 15 male middle 36 174185 dioecious Eocyzicus argillaquus 15 male same sex 14 149326 dioecious Eocyzicus argillaquus 16 male opposite sex 13 326669 dioecious Eocyzicus argillaquus 16 male middle 20 115565 dioecious Eocyzicus argillaquus 16 male same sex 7 159480 dioecious Eocyzicus argillaquus 17 male opposite sex 15 302075 dioecious Eocyzicus argillaquus 17 male middle 29 151005 dioecious Eocyzicus argillaquus 17 male same sex 14 146423 dioecious Eocyzicus argillaquus 18 female opposite sex 9 453878 dioecious Eocyzicus argillaquus 18 female middle 9 95209 dioecious Eocyzicus argillaquus 18 female same sex 2 48081 dioecious Eocyzicus argillaquus 19 female opposite sex 0 0 dioecious Eocyzicus argillaquus 19 female middle 1 599098 dioecious Eocyzicus argillaquus 19 female same sex 0 0 dioecious Eocyzicus argillaquus 20 female opposite sex 1 189914 dioecious Eocyzicus argillaquus 20 female middle 3 399886 dioecious Eocyzicus argillaquus 20 female same sex 2 8277 dioecious Eocyzicus argillaquus 21 female opposite sex 7 217956 dioecious Eocyzicus argillaquus 21 female middle 14 77279 dioecious Eocyzicus argillaquus 21 female same sex 7 302588 dioecious Eocyzicus argillaquus 22 female opposite sex 15 232372 dioecious Eocyzicus argillaquus 22 female middle 22 174914 dioecious Eocyzicus argillaquus 22 female same sex 7 191692 dioecious Eocyzicus argillaquus 23 female opposite sex 8 240352 dioecious Eocyzicus argillaquus 23 female middle 17 93554 dioecious Eocyzicus argillaquus 23 female same sex 9 264049 dioecious Eocyzicus argillaquus 24 male opposite sex 5 47123 dioecious Eocyzicus argillaquus 24 male middle 17 165547 dioecious Eocyzicus argillaquus 24 male same sex 12 386053

50 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Eocyzicus argillaquus 25 male opposite sex 8 156782 dioecious Eocyzicus argillaquus 25 male middle 17 166539 dioecious Eocyzicus argillaquus 25 male same sex 9 275302 dioecious Eocyzicus argillaquus 26 male opposite sex 1 19447 dioecious Eocyzicus argillaquus 26 male middle 3 21533 dioecious Eocyzicus argillaquus 26 male same sex 2 558407 dioecious Eocyzicus argillaquus 27 male opposite sex 4 573237 dioecious Eocyzicus argillaquus 27 male middle 5 20864 dioecious Eocyzicus argillaquus 27 male same sex 2 5317 dioecious Eocyzicus argillaquus 28 female opposite sex 1 5805 dioecious Eocyzicus argillaquus 28 female middle 6 40811 dioecious Eocyzicus argillaquus 28 female same sex 6 552025 dioecious Eocyzicus argillaquus 29 male opposite sex 15 444853 dioecious Eocyzicus argillaquus 29 male middle 20 122397 dioecious Eocyzicus argillaquus 29 male same sex 6 32075 dioecious Eocyzicus argillaquus 30 male opposite sex 3 381235 dioecious Eocyzicus argillaquus 30 male middle 5 183190 dioecious Eocyzicus argillaquus 30 male same sex 2 34705 dioecious Eocyzicus argillaquus 31 male opposite sex 16 452094 dioecious Eocyzicus argillaquus 31 male middle 17 133349 dioecious Eocyzicus argillaquus 31 male same sex 3 13658 dioecious Eocyzicus argillaquus 32 male opposite sex 10 524157 dioecious Eocyzicus argillaquus 32 male middle 13 43365 dioecious Eocyzicus argillaquus 32 male same sex 4 32025 dioecious Eocyzicus argillaquus 33 male opposite sex 2 33658 dioecious Eocyzicus argillaquus 33 male middle 9 311451 dioecious Eocyzicus argillaquus 33 male same sex 7 254102 dioecious Eocyzicus argillaquus 34 female opposite sex 3 519251 dioecious Eocyzicus argillaquus 34 female middle 5 56993 dioecious Eocyzicus argillaquus 34 female same sex 2 23740 dioecious Eocyzicus argillaquus 35 female opposite sex 1 57068 dioecious Eocyzicus argillaquus 35 female middle 6 71230 dioecious Eocyzicus argillaquus 35 female same sex 5 471586 dioecious Eocyzicus argillaquus 36 female opposite sex 0 0 dioecious Eocyzicus argillaquus 36 female middle 3 25129 dioecious Eocyzicus argillaquus 36 female same sex 3 574854 dioecious Eocyzicus argillaquus 37 female opposite sex 3 299591 dioecious Eocyzicus argillaquus 37 female middle 4 287201 dioecious Eocyzicus argillaquus 37 female same sex 1 12471 dioecious Eocyzicus argillaquus 38 female opposite sex 5 361447 51 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Eocyzicus argillaquus 38 female middle 7 207703 dioecious Eocyzicus argillaquus 38 female same sex 1 30832 dioecious Eocyzicus argillaquus 40 female opposite sex 0 0 dioecious Eocyzicus argillaquus 40 female middle 4 49350 dioecious Eocyzicus argillaquus 40 female same sex 4 550561 dioecious Eocyzicus argillaquus 41 female opposite sex 2 45000 dioecious Eocyzicus argillaquus 41 female middle 7 113080 dioecious Eocyzicus argillaquus 41 female same sex 4 441855 dioecious Eocyzicus argillaquus 42 female opposite sex 10 213963 dioecious Eocyzicus argillaquus 42 female middle 20 153832 dioecious Eocyzicus argillaquus 42 female same sex 10 232140 dioecious Eocyzicus argillaquus 43 male opposite sex 3 370327 dioecious Eocyzicus argillaquus 43 male middle 5 79115 dioecious Eocyzicus argillaquus 43 male same sex 4 150488 dioecious Eocyzicus argillaquus 44 female opposite sex 0 0 dioecious Eocyzicus argillaquus 44 female middle 2 8904 dioecious Eocyzicus argillaquus 44 female same sex 2 591038 dioecious Eocyzicus argillaquus 45 male opposite sex 3 208399 dioecious Eocyzicus argillaquus 45 male middle 11 87583 dioecious Eocyzicus argillaquus 45 male same sex 8 303890 dioecious Eocyzicus argillaquus 46 female opposite sex 12 342301 dioecious Eocyzicus argillaquus 46 female middle 21 115981 dioecious Eocyzicus argillaquus 46 female same sex 9 141695 dioecious Eocyzicus argillaquus 47 male opposite sex 5 455424 dioecious Eocyzicus argillaquus 47 male middle 6 127421 dioecious Eocyzicus argillaquus 47 male same sex 1 16938 dioecious Eocyzicus argillaquus 48 female opposite sex 4 322178 dioecious Eocyzicus argillaquus 48 female middle 7 199581 dioecious Eocyzicus argillaquus 48 female same sex 1 78153 dioecious Eocyzicus argillaquus 49 male opposite sex 4 116187 dioecious Eocyzicus argillaquus 49 male middle 18 134248 dioecious Eocyzicus argillaquus 49 male same sex 14 349440 dioecious Eocyzicus argillaquus 50 female opposite sex 5 51671 dioecious Eocyzicus argillaquus 50 female middle 13 200692 dioecious Eocyzicus argillaquus 50 female same sex 10 347425 dioecious Eocyzicus argillaquus 51 male opposite sex 3 595390 dioecious Eocyzicus argillaquus 51 male middle 2 4514 dioecious Eocyzicus argillaquus 51 male same sex 0 0 dioecious Eocyzicus argillaquus 52 female opposite sex 1 214443 dioecious Eocyzicus argillaquus 52 female middle 8 88113 52 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Eocyzicus argillaquus 52 female same sex 7 297377 dioecious Eocyzicus argillaquus 53 male opposite sex 3 502824 dioecious Eocyzicus argillaquus 53 male middle 3 84416 dioecious Eocyzicus argillaquus 53 male same sex 1 12606 dioecious Eocyzicus argillaquus 54 female opposite sex 2 130832 dioecious Eocyzicus argillaquus 54 female middle 11 183327 dioecious Eocyzicus argillaquus 54 female same sex 9 285730 dioecious Eocyzicus argillaquus 55 male opposite sex 7 561528 dioecious Eocyzicus argillaquus 55 male middle 6 38406 dioecious Eocyzicus argillaquus 55 male same sex 0 0 dioecious Eocyzicus argillaquus 56 female opposite sex 0 0 dioecious Eocyzicus argillaquus 56 female middle 15 123270 dioecious Eocyzicus argillaquus 56 female same sex 16 474934 dioecious Paralimnadia stanleyana 1 male opposite sex 11 176582 dioecious Paralimnadia stanleyana 1 male middle 20 189379 dioecious Paralimnadia stanleyana 1 male same sex 10 233716 dioecious Paralimnadia stanleyana 2 female opposite sex 1 291424 dioecious Paralimnadia stanleyana 2 female middle 5 86223 dioecious Paralimnadia stanleyana 2 female same sex 4 222115 dioecious Paralimnadia stanleyana 3 male opposite sex 13 224194 dioecious Paralimnadia stanleyana 3 male middle 25 278336 dioecious Paralimnadia stanleyana 3 male same sex 12 97308 dioecious Paralimnadia stanleyana 4 female opposite sex 4 77421 dioecious Paralimnadia stanleyana 4 female middle 21 214773 dioecious Paralimnadia stanleyana 4 female same sex 16 307563 dioecious Paralimnadia stanleyana 5 male opposite sex 6 88589 dioecious Paralimnadia stanleyana 5 male middle 18 159321 dioecious Paralimnadia stanleyana 5 male same sex 11 351950 dioecious Paralimnadia stanleyana 6 female opposite sex 7 122624 dioecious Paralimnadia stanleyana 6 female middle 22 148414 dioecious Paralimnadia stanleyana 6 female same sex 15 328698 dioecious Paralimnadia stanleyana 7 male opposite sex 8 139377 dioecious Paralimnadia stanleyana 7 male middle 17 111493 dioecious Paralimnadia stanleyana 7 male same sex 10 349114 dioecious Paralimnadia stanleyana 8 female opposite sex 6 159061 dioecious Paralimnadia stanleyana 8 female middle 18 134443 dioecious Paralimnadia stanleyana 8 female same sex 10 306267 dioecious Paralimnadia stanleyana 9 male opposite sex 10 105807 dioecious Paralimnadia stanleyana 9 male middle 17 373117 dioecious Paralimnadia stanleyana 9 male same sex 6 120469 53 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Paralimnadia stanleyana 10 female opposite sex 8 433957 dioecious Paralimnadia stanleyana 10 female middle 9 117674 dioecious Paralimnadia stanleyana 10 female same sex 3 48368 dioecious Paralimnadia stanleyana 11 male opposite sex 3 550511 dioecious Paralimnadia stanleyana 11 male middle 3 49234 dioecious Paralimnadia stanleyana 11 male same sex 0 0 dioecious Paralimnadia stanleyana 12 female opposite sex 4 267546 dioecious Paralimnadia stanleyana 12 female middle 13 126200 dioecious Paralimnadia stanleyana 12 female same sex 9 206137 dioecious Paralimnadia stanleyana 13 male opposite sex 13 176647 dioecious Paralimnadia stanleyana 13 male middle 28 214288 dioecious Paralimnadia stanleyana 13 male same sex 15 208921 dioecious Paralimnadia stanleyana 14 female opposite sex 1 7220 dioecious Paralimnadia stanleyana 14 female middle 3 590629 dioecious Paralimnadia stanleyana 14 female same sex 1 2045 dioecious Paralimnadia stanleyana 15 male opposite sex 9 242094 dioecious Paralimnadia stanleyana 15 male middle 14 186163 dioecious Paralimnadia stanleyana 15 male same sex 5 171566 dioecious Paralimnadia stanleyana 16 female opposite sex 15 220647 dioecious Paralimnadia stanleyana 16 female middle 24 288786 dioecious Paralimnadia stanleyana 16 female same sex 9 88948 dioecious Paralimnadia stanleyana 17 male opposite sex 16 359031 dioecious Paralimnadia stanleyana 17 male middle 19 208028 dioecious Paralimnadia stanleyana 17 male same sex 4 32118 dioecious Paralimnadia stanleyana 18 female opposite sex 6 205480 dioecious Paralimnadia stanleyana 18 female middle 9 141512 dioecious Paralimnadia stanleyana 18 female same sex 4 252949 dioecious Paralimnadia stanleyana 19 male opposite sex 20 188380 dioecious Paralimnadia stanleyana 19 male middle 30 257046 dioecious Paralimnadia stanleyana 19 male same sex 10 154244 dioecious Paralimnadia stanleyana 20 female opposite sex 15 271995 dioecious Paralimnadia stanleyana 20 female middle 24 167436 dioecious Paralimnadia stanleyana 20 female same sex 10 160198 dioecious Paralimnadia stanleyana 21 female opposite sex 21 249078 dioecious Paralimnadia stanleyana 21 female middle 31 288108 dioecious Paralimnadia stanleyana 21 female same sex 9 61377 dioecious Paralimnadia stanleyana 22 male opposite sex 0 0 dioecious Paralimnadia stanleyana 22 male middle 15 123270 dioecious Paralimnadia stanleyana 22 male same sex 16 474934 dioecious Paralimnadia stanleyana 23 female opposite sex 0 0 54 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Paralimnadia stanleyana 23 female middle 15 123270 dioecious Paralimnadia stanleyana 23 female same sex 16 474934 dioecious Paralimnadia stanleyana 24 female opposite sex 10 220128 dioecious Paralimnadia stanleyana 24 female middle 17 158878 dioecious Paralimnadia stanleyana 24 female same sex 7 220941 dioecious Paralimnadia stanleyana 25 male opposite sex 10 220128 dioecious Paralimnadia stanleyana 25 male middle 17 158878 dioecious Paralimnadia stanleyana 25 male same sex 7 220941 dioecious Paralimnadia stanleyana 26 female opposite sex 0 0 dioecious Paralimnadia stanleyana 26 female middle 1 442748 dioecious Paralimnadia stanleyana 26 female same sex 1 157217 dioecious Paralimnadia stanleyana 27 male opposite sex 3 297185 dioecious Paralimnadia stanleyana 27 male middle 4 301315 dioecious Paralimnadia stanleyana 27 male same sex 1 1468 dioecious Paralimnadia stanleyana 28 female opposite sex 1 18509 dioecious Paralimnadia stanleyana 28 female middle 4 28971 dioecious Paralimnadia stanleyana 28 female same sex 4 552315 dioecious Paralimnadia stanleyana 29 male opposite sex 1 268052 dioecious Paralimnadia stanleyana 29 male middle 3 330241 dioecious Paralimnadia stanleyana 29 male same sex 1 1603 dioecious Paralimnadia stanleyana 30 female opposite sex 7 74474 dioecious Paralimnadia stanleyana 30 female middle 13 166351 dioecious Paralimnadia stanleyana 30 female same sex 8 359162 dioecious Paralimnadia stanleyana 31 male opposite sex 2 165628 dioecious Paralimnadia stanleyana 31 male middle 4 400334 dioecious Paralimnadia stanleyana 31 male same sex 1 33662 dioecious Paralimnadia stanleyana 32 female opposite sex 0 0 dioecious Paralimnadia stanleyana 32 female middle 1 599931 dioecious Paralimnadia stanleyana 32 female same sex 0 0 dioecious Paralimnadia stanleyana 33 male opposite sex 1 276802 dioecious Paralimnadia stanleyana 33 male middle 1 323131 dioecious Paralimnadia stanleyana 33 male same sex 0 0 dioecious Paralimnadia stanleyana 34 female opposite sex 0 0 dioecious Paralimnadia stanleyana 34 female middle 2 407789 dioecious Paralimnadia stanleyana 34 female same sex 1 192080 dioecious Paralimnadia stanleyana 35 male opposite sex 3 75390 dioecious Paralimnadia stanleyana 35 male middle 16 160757 dioecious Paralimnadia stanleyana 35 male same sex 12 363813 dioecious Paralimnadia stanleyana 36 female opposite sex 13 186252 dioecious Paralimnadia stanleyana 36 female middle 19 217535 55 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Paralimnadia stanleyana 36 female same sex 7 196099 dioecious Paralimnadia stanleyana 37 male opposite sex 1 599862 dioecious Paralimnadia stanleyana 37 male middle 0 0 dioecious Paralimnadia stanleyana 37 male same sex 0 0 dioecious Paralimnadia stanleyana 38 female opposite sex 0 0 dioecious Paralimnadia stanleyana 38 female middle 1 599954 dioecious Paralimnadia stanleyana 38 female same sex 0 0 dioecious Paralimnadia stanleyana 39 male opposite sex 1 599978 dioecious Paralimnadia stanleyana 39 male middle 0 0 dioecious Paralimnadia stanleyana 39 male same sex 0 0 dioecious Paralimnadia stanleyana 40 female opposite sex 0 0 dioecious Paralimnadia stanleyana 40 female middle 0 0 dioecious Paralimnadia stanleyana 40 female same sex 1 598674 dioecious Paralimnadia stanleyana 41 male opposite sex 1 3878 dioecious Paralimnadia stanleyana 41 male middle 4 476801 dioecious Paralimnadia stanleyana 41 male same sex 2 119239 dioecious Paralimnadia stanleyana 42 female opposite sex 2 35726 dioecious Paralimnadia stanleyana 42 female middle 7 45591 dioecious Paralimnadia stanleyana 42 female same sex 5 518629 dioecious Paralimnadia stanleyana 43 male opposite sex 2 589643 dioecious Paralimnadia stanleyana 43 male middle 2 6765 dioecious Paralimnadia stanleyana 43 male same sex 1 3584 dioecious Paralimnadia stanleyana 44 female opposite sex 0 0 dioecious Paralimnadia stanleyana 44 female middle 1 12144 dioecious Paralimnadia stanleyana 44 female same sex 1 587840 dioecious Paralimnadia stanleyana 45 male opposite sex 3 154830 dioecious Paralimnadia stanleyana 45 male middle 4 396621 dioecious Paralimnadia stanleyana 45 male same sex 1 48502 dioecious Paralimnadia stanleyana 46 female opposite sex 0 0 dioecious Paralimnadia stanleyana 46 female middle 1 599907 dioecious Paralimnadia stanleyana 46 female same sex 0 0 dioecious Paralimnadia stanleyana 47 male opposite sex 1 198545 dioecious Paralimnadia stanleyana 47 male middle 1 399054 dioecious Paralimnadia stanleyana 47 male same sex 0 0 dioecious Paralimnadia stanleyana 48 female opposite sex 0 0 dioecious Paralimnadia stanleyana 48 female middle 1 598397 dioecious Paralimnadia stanleyana 48 female same sex 0 0 dioecious Paralimnadia stanleyana 49 male opposite sex 0 0 dioecious Paralimnadia stanleyana 49 male middle 1 599611 dioecious Paralimnadia stanleyana 49 male same sex 0 0 56 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) dioecious Paralimnadia stanleyana 50 female opposite sex 0 0 dioecious Paralimnadia stanleyana 50 female middle 1 599448 dioecious Paralimnadia stanleyana 50 female same sex 0 0 dioecious Paralimnadia stanleyana 51 male opposite sex 2 161152 dioecious Paralimnadia stanleyana 51 male middle 8 286499 dioecious Paralimnadia stanleyana 51 male same sex 4 151941 dioecious Paralimnadia stanleyana 52 female opposite sex 2 283482 dioecious Paralimnadia stanleyana 52 female middle 2 316149 dioecious Paralimnadia stanleyana 52 female same sex 0 0 dioecious Paralimnadia stanleyana 53 female opposite sex 5 170592 dioecious Paralimnadia stanleyana 53 female middle 7 132566 dioecious Paralimnadia stanleyana 53 female same sex 2 295974 dioecious Paralimnadia stanleyana 54 male opposite sex 1 204608 dioecious Paralimnadia stanleyana 54 male middle 1 394751 dioecious Paralimnadia stanleyana 54 male same sex 0 0 dioecious Paralimnadia stanleyana 55 male opposite sex 0 0 dioecious Paralimnadia stanleyana 55 male middle 1 596547 dioecious Paralimnadia stanleyana 55 male same sex 0 0 dioecious Paralimnadia stanleyana 56 female opposite sex 0 0 dioecious Paralimnadia stanleyana 56 female middle 3 98502 dioecious Paralimnadia stanleyana 56 female same sex 3 500592 androdiecious Eulimnadia texana 1 female opposite sex 3 92788 androdiecious Eulimnadia texana 1 female middle 10 117034 androdiecious Eulimnadia texana 1 female same sex 8 390043 androdiecious Eulimnadia texana 2 male opposite sex 2 43324 androdiecious Eulimnadia texana 2 male middle 10 83687 androdiecious Eulimnadia texana 2 male same sex 10 472173 androdiecious Eulimnadia texana 3 female opposite sex 0 0 androdiecious Eulimnadia texana 3 female middle 0 0 androdiecious Eulimnadia texana 3 female same sex 1 599837 androdiecious Eulimnadia texana 4 male opposite sex 25 346140 androdiecious Eulimnadia texana 4 male middle 39 123664 androdiecious Eulimnadia texana 4 male same sex 15 129826 androdiecious Eulimnadia texana 5 female opposite sex 7 563700 androdiecious Eulimnadia texana 5 female middle 7 35710 androdiecious Eulimnadia texana 5 female same sex 0 0 androdiecious Eulimnadia texana 6 male opposite sex 18 309846 androdiecious Eulimnadia texana 6 male middle 30 159961 androdiecious Eulimnadia texana 6 male same sex 11 130120 androdiecious Eulimnadia texana 7 female opposite sex 10 155377 57 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia texana 7 female middle 26 122874 androdiecious Eulimnadia texana 7 female same sex 13 321306 androdiecious Eulimnadia texana 8 male opposite sex 3 33774 androdiecious Eulimnadia texana 8 male middle 6 226650 androdiecious Eulimnadia texana 8 male same sex 4 339413 androdiecious Eulimnadia texana 9 male opposite sex 8 486999 androdiecious Eulimnadia texana 9 male middle 10 78329 androdiecious Eulimnadia texana 9 male same sex 4 33437 androdiecious Eulimnadia texana 10 female opposite sex 5 97688 androdiecious Eulimnadia texana 10 female middle 13 158745 androdiecious Eulimnadia texana 10 female same sex 8 343129 androdiecious Eulimnadia texana 11 male opposite sex 3 15465 androdiecious Eulimnadia texana 11 male middle 8 179178 androdiecious Eulimnadia texana 11 male same sex 6 405304 androdiecious Eulimnadia texana 12 female opposite sex 7 345378 androdiecious Eulimnadia texana 12 female middle 13 171345 androdiecious Eulimnadia texana 12 female same sex 7 82992 androdiecious Eulimnadia texana 13 female opposite sex 6 234954 androdiecious Eulimnadia texana 13 female middle 9 133843 androdiecious Eulimnadia texana 13 female same sex 4 230921 androdiecious Eulimnadia texana 14 female opposite sex 14 230377 androdiecious Eulimnadia texana 14 female middle 28 144780 androdiecious Eulimnadia texana 14 female same sex 13 224599 androdiecious Eulimnadia texana 15 male opposite sex 12 123472 androdiecious Eulimnadia texana 15 male middle 32 116025 androdiecious Eulimnadia texana 15 male same sex 21 360421 androdiecious Eulimnadia texana 16 female opposite sex 0 0 androdiecious Eulimnadia texana 16 female middle 1 47808 androdiecious Eulimnadia texana 16 female same sex 2 551925 androdiecious Eulimnadia texana 17 male opposite sex 26 394141 androdiecious Eulimnadia texana 17 male middle 37 115292 androdiecious Eulimnadia texana 17 male same sex 12 90292 androdiecious Eulimnadia texana 18 female opposite sex 0 0 androdiecious Eulimnadia texana 18 female middle 2 6143 androdiecious Eulimnadia texana 18 female same sex 3 593744 androdiecious Eulimnadia texana 19 male opposite sex 12 458073 androdiecious Eulimnadia texana 19 male middle 15 97013 androdiecious Eulimnadia texana 19 male same sex 4 43995 androdiecious Eulimnadia texana 20 female opposite sex 3 52472 androdiecious Eulimnadia texana 20 female middle 7 69884 58 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia texana 20 female same sex 5 477495 androdiecious Eulimnadia texana 21 male opposite sex 9 206627 androdiecious Eulimnadia texana 21 male middle 17 177713 androdiecious Eulimnadia texana 21 male same sex 8 215374 androdiecious Eulimnadia texana 22 male opposite sex 8 209419 androdiecious Eulimnadia texana 22 male middle 18 236301 androdiecious Eulimnadia texana 22 male same sex 10 154269 androdiecious Eulimnadia texana 23 female opposite sex 1 2838 androdiecious Eulimnadia texana 23 female middle 2 11244 androdiecious Eulimnadia texana 23 female same sex 3 585212 androdiecious Eulimnadia texana 24 male opposite sex 13 211825 androdiecious Eulimnadia texana 24 male middle 25 129152 androdiecious Eulimnadia texana 24 male same sex 13 258454 androdiecious Eulimnadia texana 25 female opposite sex 7 346727 androdiecious Eulimnadia texana 25 female middle 9 249364 androdiecious Eulimnadia texana 25 female same sex 1 3440 androdiecious Eulimnadia texana 26 male opposite sex 6 330226 androdiecious Eulimnadia texana 26 male middle 9 205646 androdiecious Eulimnadia texana 26 male same sex 3 64034 androdiecious Eulimnadia texana 27 female opposite sex 0 0 androdiecious Eulimnadia texana 27 female middle 1 599874 androdiecious Eulimnadia texana 27 female same sex 0 0 androdiecious Eulimnadia texana 28 male opposite sex 18 257410 androdiecious Eulimnadia texana 28 male middle 29 215818 androdiecious Eulimnadia texana 28 male same sex 12 126688 androdiecious Eulimnadia texana 29 female opposite sex 7 87672 androdiecious Eulimnadia texana 29 female middle 20 98533 androdiecious Eulimnadia texana 29 female same sex 12 413628 androdiecious Eulimnadia texana 30 male opposite sex 2 25853 androdiecious Eulimnadia texana 30 male middle 9 79653 androdiecious Eulimnadia texana 30 male same sex 8 494359 androdiecious Eulimnadia texana 31 female opposite sex 1 104256 androdiecious Eulimnadia texana 31 female middle 3 247003 androdiecious Eulimnadia texana 31 female same sex 1 248664 androdiecious Eulimnadia texana 32 male opposite sex 3 432680 androdiecious Eulimnadia texana 32 male middle 4 167258 androdiecious Eulimnadia texana 32 male same sex 0 0 androdiecious Eulimnadia texana 33 female opposite sex 3 531067 androdiecious Eulimnadia texana 33 female middle 5 37167 androdiecious Eulimnadia texana 33 female same sex 2 31651 59

Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia texana 34 male opposite sex 0 0 androdiecious Eulimnadia texana 34 male middle 2 153753 androdiecious Eulimnadia texana 34 male same sex 1 446195 androdiecious Eulimnadia texana 35 female opposite sex 1 3500 androdiecious Eulimnadia texana 35 female middle 2 3505 androdiecious Eulimnadia texana 35 female same sex 2 592945 androdiecious Eulimnadia texana 36 male opposite sex 1 158288 androdiecious Eulimnadia texana 36 male middle 3 100454 androdiecious Eulimnadia texana 36 male same sex 2 339960 androdiecious Eulimnadia texana 37 female opposite sex 0 0 androdiecious Eulimnadia texana 37 female middle 3 139833 androdiecious Eulimnadia texana 37 female same sex 3 460138 androdiecious Eulimnadia texana 38 male opposite sex 4 482657 androdiecious Eulimnadia texana 38 male middle 5 18205 androdiecious Eulimnadia texana 38 male same sex 2 99078 androdiecious Eulimnadia texana 39 female opposite sex 1 9710 androdiecious Eulimnadia texana 39 female middle 5 136367 androdiecious Eulimnadia texana 39 female same sex 3 453883 androdiecious Eulimnadia texana 40 male opposite sex 1 395392 androdiecious Eulimnadia texana 40 male middle 2 119613 androdiecious Eulimnadia texana 40 male same sex 1 84952 androdiecious Eulimnadia texana 41 male opposite sex 16 298053 androdiecious Eulimnadia texana 41 male middle 27 102333 androdiecious Eulimnadia texana 41 male same sex 10 195679 androdiecious Eulimnadia texana 42 male opposite sex 0 0 androdiecious Eulimnadia texana 42 male middle 15 61618 androdiecious Eulimnadia texana 42 male same sex 14 537800 androdiecious Eulimnadia texana 43 male opposite sex 11 530635 androdiecious Eulimnadia texana 43 male middle 13 41130 androdiecious Eulimnadia texana 43 male same sex 3 27550 androdiecious Eulimnadia texana 44 male opposite sex 14 492509 androdiecious Eulimnadia texana 44 male middle 16 77495 androdiecious Eulimnadia texana 44 male same sex 3 29407 androdiecious Eulimnadia texana 45 male opposite sex 26 298981 androdiecious Eulimnadia texana 45 male middle 36 216216 androdiecious Eulimnadia texana 45 male same sex 12 83891 androdiecious Eulimnadia texana 46 male opposite sex 11 201808 androdiecious Eulimnadia texana 46 male middle 18 132496 androdiecious Eulimnadia texana 46 male same sex 8 265127 androdiecious Eulimnadia texana 47 female opposite sex 10 292956 60

Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia texana 47 female middle 18 212849 androdiecious Eulimnadia texana 47 female same sex 8 93512 androdiecious Eulimnadia texana 48 female opposite sex 0 0 androdiecious Eulimnadia texana 48 female middle 4 368337 androdiecious Eulimnadia texana 48 female same sex 3 230993 androdiecious Eulimnadia texana 49 female opposite sex 1 5736 androdiecious Eulimnadia texana 49 female middle 1 2236 androdiecious Eulimnadia texana 49 female same sex 1 591482 androdiecious Eulimnadia texana 50 female opposite sex 1 115283 androdiecious Eulimnadia texana 50 female middle 8 59513 androdiecious Eulimnadia texana 50 female same sex 7 424437 androdiecious Eulimnadia texana 51 female opposite sex 10 129929 androdiecious Eulimnadia texana 51 female middle 26 107685 androdiecious Eulimnadia texana 51 female same sex 17 361718 androdiecious Eulimnadia texana 52 female opposite sex 3 40250 androdiecious Eulimnadia texana 52 female middle 10 71926 androdiecious Eulimnadia texana 52 female same sex 8 485781 androdiecious Eulimnadia texana 53 female opposite sex 9 286081 androdiecious Eulimnadia texana 53 female middle 19 58328 androdiecious Eulimnadia texana 53 female same sex 11 254903 androdiecious Eulimnadia texana 54 female opposite sex 5 562593 androdiecious Eulimnadia texana 54 female middle 4 36502 androdiecious Eulimnadia texana 54 female same sex 0 0 androdiecious Eulimnadia texana 55 female opposite sex 4 28553 androdiecious Eulimnadia texana 55 female middle 12 76108 androdiecious Eulimnadia texana 55 female same sex 7 494325 androdiecious Eulimnadia texana 56 female opposite sex 4 566434 androdiecious Eulimnadia texana 56 female middle 4 30360 androdiecious Eulimnadia texana 56 female same sex 1 2340 androdiecious Eulimnadia texana 57 male opposite sex 6 264752 androdiecious Eulimnadia texana 57 male middle 12 60811 androdiecious Eulimnadia texana 57 male same sex 6 273798 androdiecious Eulimnadia texana 58 female opposite sex 4 383859 androdiecious Eulimnadia texana 58 female middle 6 95951 androdiecious Eulimnadia texana 58 female same sex 2 119538 androdiecious Eulimnadia texana 59 female opposite sex 7 512446 androdiecious Eulimnadia texana 59 female middle 6 86370 androdiecious Eulimnadia texana 59 female same sex 0 0 androdiecious Eulimnadia texana 60 male opposite sex 3 527105 androdiecious Eulimnadia texana 60 male middle 6 38710 61

Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia texana 60 male same sex 3 33161 androdiecious Eulimnadia texana 61 male opposite sex 16 206833 androdiecious Eulimnadia texana 61 male middle 29 124328 androdiecious Eulimnadia texana 61 male same sex 14 266542 androdiecious Eulimnadia texana 62 male opposite sex 6 544600 androdiecious Eulimnadia texana 62 male middle 7 40403 androdiecious Eulimnadia texana 62 male same sex 2 13866 androdiecious Eulimnadia texana 63 male opposite sex 9 286081 androdiecious Eulimnadia texana 63 male middle 19 58328 androdiecious Eulimnadia texana 63 male same sex 11 254903 androdiecious Eulimnadia dahli 1 female opposite sex 0 0 androdiecious Eulimnadia dahli 1 female middle 9 160098 androdiecious Eulimnadia dahli 1 female same sex 9 439480 androdiecious Eulimnadia dahli 2 male opposite sex 4 141860 androdiecious Eulimnadia dahli 2 male middle 11 73276 androdiecious Eulimnadia dahli 2 male same sex 7 384741 androdiecious Eulimnadia dahli 3 female opposite sex 4 141860 androdiecious Eulimnadia dahli 3 female middle 11 73276 androdiecious Eulimnadia dahli 3 female same sex 7 384741 androdiecious Eulimnadia dahli 4 male opposite sex 7 258469 androdiecious Eulimnadia dahli 4 male middle 13 127026 androdiecious Eulimnadia dahli 4 male same sex 7 214104 androdiecious Eulimnadia dahli 5 female opposite sex 0 0 androdiecious Eulimnadia dahli 5 female middle 0 0 androdiecious Eulimnadia dahli 5 female same sex 2 599909 androdiecious Eulimnadia dahli 6 male opposite sex 10 419952 androdiecious Eulimnadia dahli 6 male middle 13 82994 androdiecious Eulimnadia dahli 6 male same sex 3 95526 androdiecious Eulimnadia dahli 7 female opposite sex 0 0 androdiecious Eulimnadia dahli 7 female middle 1 598625 androdiecious Eulimnadia dahli 7 female same sex 0 0 androdiecious Eulimnadia dahli 8 female opposite sex 0 0 androdiecious Eulimnadia dahli 8 female middle 14 172463 androdiecious Eulimnadia dahli 8 female same sex 12 426771 androdiecious Eulimnadia dahli 9 male opposite sex 6 116143 androdiecious Eulimnadia dahli 9 male middle 19 173978 androdiecious Eulimnadia dahli 9 male same sex 12 306941 androdiecious Eulimnadia dahli 10 female opposite sex 0 0 androdiecious Eulimnadia dahli 10 female middle 1 599547 androdiecious Eulimnadia dahli 10 female same sex 0 0 62 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia dahli 11 female opposite sex 0 0 androdiecious Eulimnadia dahli 11 female middle 1 599515 androdiecious Eulimnadia dahli 11 female same sex 0 0 androdiecious Eulimnadia dahli 12 male opposite sex 12 281229 androdiecious Eulimnadia dahli 12 male middle 26 106753 androdiecious Eulimnadia dahli 12 male same sex 14 210143 androdiecious Eulimnadia dahli 13 male opposite sex 6 466969 androdiecious Eulimnadia dahli 13 male middle 9 103155 androdiecious Eulimnadia dahli 13 male same sex 4 26798 androdiecious Eulimnadia dahli 14 male opposite sex 10 256035 androdiecious Eulimnadia dahli 14 male middle 18 162721 androdiecious Eulimnadia dahli 14 male same sex 8 179806 androdiecious Eulimnadia dahli 15 male opposite sex 4 333435 androdiecious Eulimnadia dahli 15 male middle 7 60327 androdiecious Eulimnadia dahli 15 male same sex 3 198528 androdiecious Eulimnadia dahli 16 male opposite sex 8 167283 androdiecious Eulimnadia dahli 16 male middle 11 69935 androdiecious Eulimnadia dahli 16 male same sex 3 362378 androdiecious Eulimnadia dahli 17 male opposite sex 5 228883 androdiecious Eulimnadia dahli 17 male middle 12 171363 androdiecious Eulimnadia dahli 17 male same sex 7 193712 androdiecious Eulimnadia dahli 18 male opposite sex 13 308097 androdiecious Eulimnadia dahli 18 male middle 22 98610 androdiecious Eulimnadia dahli 18 male same sex 9 191618 androdiecious Eulimnadia dahli 19 female opposite sex 3 60482 androdiecious Eulimnadia dahli 19 female middle 6 382183 androdiecious Eulimnadia dahli 19 female same sex 3 155969 androdiecious Eulimnadia dahli 20 female opposite sex 4 29588 androdiecious Eulimnadia dahli 20 female middle 17 47711 androdiecious Eulimnadia dahli 20 female same sex 14 522662 androdiecious Eulimnadia dahli 21 male opposite sex 15 223825 androdiecious Eulimnadia dahli 21 male middle 23 172613 androdiecious Eulimnadia dahli 21 male same sex 10 203497 androdiecious Eulimnadia dahli 22 female opposite sex 7 273696 androdiecious Eulimnadia dahli 22 female middle 16 143230 androdiecious Eulimnadia dahli 22 female same sex 9 182952 androdiecious Eulimnadia dahli 23 male opposite sex 5 73279 androdiecious Eulimnadia dahli 23 male middle 28 174160 androdiecious Eulimnadia dahli 23 male same sex 24 352320 androdiecious Eulimnadia dahli 24 female opposite sex 0 0 63 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia dahli 24 female middle 0 0 androdiecious Eulimnadia dahli 24 female same sex 1 599882 androdiecious Eulimnadia dahli 25 male opposite sex 2 595856 androdiecious Eulimnadia dahli 25 male middle 1 4029 androdiecious Eulimnadia dahli 25 male same sex 0 0 androdiecious Eulimnadia dahli 26 female opposite sex 3 184889 androdiecious Eulimnadia dahli 26 female middle 7 50842 androdiecious Eulimnadia dahli 26 female same sex 5 364172 androdiecious Eulimnadia dahli 27 male opposite sex 9 368241 androdiecious Eulimnadia dahli 27 male middle 18 102263 androdiecious Eulimnadia dahli 27 male same sex 10 129452 androdiecious Eulimnadia dahli 28 female opposite sex 5 167715 androdiecious Eulimnadia dahli 28 female middle 13 103492 androdiecious Eulimnadia dahli 28 female same sex 8 328637 androdiecious Eulimnadia dahli 29 male opposite sex 2 9132 androdiecious Eulimnadia dahli 29 male middle 8 174950 androdiecious Eulimnadia dahli 29 male same sex 5 415850 androdiecious Eulimnadia dahli 30 female opposite sex 0 0 androdiecious Eulimnadia dahli 30 female middle 1 98950 androdiecious Eulimnadia dahli 30 female same sex 1 501044 androdiecious Eulimnadia dahli 31 male opposite sex 1 11080 androdiecious Eulimnadia dahli 31 male middle 3 24595 androdiecious Eulimnadia dahli 31 male same sex 4 564296 androdiecious Eulimnadia dahli 32 female opposite sex 0 0 androdiecious Eulimnadia dahli 32 female middle 0 0 androdiecious Eulimnadia dahli 32 female same sex 1 599977 androdiecious Eulimnadia dahli 33 male opposite sex 5 336109 androdiecious Eulimnadia dahli 33 male middle 10 164697 androdiecious Eulimnadia dahli 33 male same sex 6 99117 androdiecious Eulimnadia dahli 34 female opposite sex 0 0 androdiecious Eulimnadia dahli 34 female middle 4 126798 androdiecious Eulimnadia dahli 34 female same sex 5 473184 androdiecious Eulimnadia dahli 35 male opposite sex 4 117808 androdiecious Eulimnadia dahli 35 male middle 7 162519 androdiecious Eulimnadia dahli 35 male same sex 4 319537 androdiecious Eulimnadia dahli 36 female opposite sex 1 57960 androdiecious Eulimnadia dahli 36 female middle 2 10873 androdiecious Eulimnadia dahli 36 female same sex 2 530713 androdiecious Eulimnadia dahli 37 male opposite sex 11 451724 androdiecious Eulimnadia dahli 37 male middle 13 64276 64 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia dahli 37 male same sex 4 83946 androdiecious Eulimnadia dahli 38 female opposite sex 2 57150 androdiecious Eulimnadia dahli 38 female middle 4 540133 androdiecious Eulimnadia dahli 38 female same sex 1 2664 androdiecious Eulimnadia dahli 39 male opposite sex 3 322738 androdiecious Eulimnadia dahli 39 male middle 7 223876 androdiecious Eulimnadia dahli 39 male same sex 3 53353 androdiecious Eulimnadia dahli 40 female opposite sex 0 0 androdiecious Eulimnadia dahli 40 female middle 1 599897 androdiecious Eulimnadia dahli 40 female same sex 0 0 androdiecious Eulimnadia dahli 41 male opposite sex 5 481603 androdiecious Eulimnadia dahli 41 male middle 9 39725 androdiecious Eulimnadia dahli 41 male same sex 5 78662 androdiecious Eulimnadia dahli 42 female opposite sex 1 29317 androdiecious Eulimnadia dahli 42 female middle 1 570545 androdiecious Eulimnadia dahli 42 female same sex 0 0 androdiecious Eulimnadia dahli 43 male opposite sex 9 341702 androdiecious Eulimnadia dahli 43 male middle 13 45104 androdiecious Eulimnadia dahli 43 male same sex 5 213146 androdiecious Eulimnadia dahli 44 female opposite sex 10 189769 androdiecious Eulimnadia dahli 44 female middle 15 70393 androdiecious Eulimnadia dahli 44 female same sex 6 339554 androdiecious Eulimnadia dahli 45 male opposite sex 5 383507 androdiecious Eulimnadia dahli 45 male middle 10 85519 androdiecious Eulimnadia dahli 45 male same sex 5 130881 androdiecious Eulimnadia dahli 46 female opposite sex 2 170718 androdiecious Eulimnadia dahli 46 female middle 3 175846 androdiecious Eulimnadia dahli 46 female same sex 1 253368 androdiecious Eulimnadia dahli 47 male opposite sex 6 200443 androdiecious Eulimnadia dahli 47 male middle 15 112752 androdiecious Eulimnadia dahli 47 male same sex 10 286555 androdiecious Eulimnadia dahli 48 female opposite sex 0 0 androdiecious Eulimnadia dahli 48 female middle 1 599950 androdiecious Eulimnadia dahli 48 female same sex 0 0 androdiecious Eulimnadia dahli 49 male opposite sex 11 361475 androdiecious Eulimnadia dahli 49 male middle 15 117621 androdiecious Eulimnadia dahli 49 male same sex 6 120836 androdiecious Eulimnadia dahli 50 female opposite sex 4 239870 androdiecious Eulimnadia dahli 50 female middle 6 47341 androdiecious Eulimnadia dahli 50 female same sex 3 312774 65 Crosses time in focal into chamber reproduction species trial sex chamber chamber (s) androdiecious Eulimnadia dahli 51 male opposite sex 8 229010 androdiecious Eulimnadia dahli 51 male middle 16 89771 androdiecious Eulimnadia dahli 51 male same sex 9 281147 androdiecious Eulimnadia dahli 52 female opposite sex 0 0 androdiecious Eulimnadia dahli 52 female middle 1 100317 androdiecious Eulimnadia dahli 52 female same sex 3 499677 androdiecious Eulimnadia dahli 53 male opposite sex 14 245403 androdiecious Eulimnadia dahli 53 male middle 28 156187 androdiecious Eulimnadia dahli 53 male same sex 14 198369

66