Simultaneous Hermaphrodites Research Articles

Control of social monogamy through aggression in a hermaphroditic shrimp.

IntroductionSex allocation theory predicts that in small mating groups simultaneous hermaphroditism is the optimal form of gender expression. Under these conditions, male allocation is predicted to be very low and overall per-capita reproductive output maximal. This is particularly true for individuals that live in pairs, but monogamy is highly susceptible to cheating by both partners. However, certain conditions favour social monogamy in hermaphrodites. This study addresses the influence of group size on group stability and moulting cycles in singles, pairs, triplets and quartets of the socially monogamous shrimp Lysmata amboinensis, a protandric simultaneous hermaphrodite.ResultsThe effect of group size was very strong: Exactly one individual in each triplet and exactly two individuals in each quartet were killed in aggressive interactions, resulting in group sizes of two individuals. All killed individuals had just moulted. No mortality occurred in single and pair treatments. The number of moults in the surviving shrimp increased significantly after changing from triplets and quartets to pairs.ConclusionSocial monogamy in L. amboinensis is reinforced by aggressive expulsion of supernumerous individuals. We suggest that the high risk of mortality in triplets and quartets results in suppression of moulting in groups larger than two individuals and that the feeding ecology of L. amboinensis favours social monogamy.

Larval settlement preference maximizes genetic mixing in an inbreeding population of a simultaneous hermaphrodite (Bugula stolonifera, Bryozoa)

Conspecific aggregations in terrestrial and aquatic organisms can have a significant effect on an individual's survival, growth and reproductive fitness, particularly if these aggregations are composed of closely related individuals. Such aggregations can form passively, as a consequence of dispersal, or actively, as a consequence of kin recognition. In this study, we investigated the genetic composition of individuals in conspecific aggregations in the simultaneous hermaphroditic marine bryozoan Bugula stolonifera. Conspecific larvae routinely metamorphose on adult colonies; the possibility that larvae select or avoid their maternal colony was investigated utilizing 10 newly developed polymorphic microsatellite loci. Adult colonies were collected from Eel Pond, Woods Hole, Massachusetts and inspected for the presence of attached individuals. Adult colonies and their attached individuals were genotyped and compared to assess genetic relatedness within and among these groups relative to the overall genetic variability of the sampling site. Overall, the population of B. stolonifera at this site was found to be outside Hardy-Weinberg equilibrium because of significant levels of inbreeding. No significant genetic differentiation, however, was found between any groups, documenting that a group containing an adult colony and its attached individuals had as much genetic variability as was found for the entire sampling site. Parentage-exclusion analyses showed that the vast majority of attached individuals (>93%) could not have derived from the colony on which they were attached. Kinship analyses showed that the majority of attached individuals (≈63%) shared less than a half-sibling relationship. These results suggest that a colony's nearest neighbours are not composed of siblings, and thus, larval settlement preference can maximize outcrossing in this inbreeding population.

Sperm as a paternal investment: a model of sex allocation in sperm-digesting hermaphrodites

Some simultaneously hermaphroditic animals are known to digest received sperm. To investigate the effect of sperm digestion on the sex allocation of simultaneous hermaphrodites, we constructed models about evolutionarily stable resource allocation between male and female functions. We assumed that resource obtained from sperm digestion is used for gametogenesis (sperm and/or egg production). As a result, we found that sperm digestion increases the evolutionarily stable allocation to male function under finite number of matings. This is because excess sperm function as nuptial gift or paternal investment when at least a fraction of digested sperm is translated into eggs. Therefore, some factors which affect the assurance of paternity for sperm donors, such as cryptic female choice and/or sperm displacement, may change the result. In addition, this result implies that sperm digestion does not necessarily make male role less preferred. Further studies on the usage of donated sperm are required to test the validity of our models.

The gametogenic cycle of Leguminaia whaetleyi (Lea, 1862) in lake Gölbaşı, Turkey (Bivalvia: Unionidae)

The gonadal structure and cycle of the Leguminaia whaetleyi is described for the first time indicating that 5% of this species is simultaneous hermaphrodite. A total of 420 individuals were collected in monthly samples from October 2008 to September 2009 in Gölbaşı Lake, Turkey. Calculation of the condition index and histological examination of the gonads showed that gametogenesis began in January. Spawning occurred between May and August with one maximum peak in July. Annual maximum oocytes size peaked in July. The population consisted of simultaneous hermaphrodite individuals. Sex ratio of L. whaetleyi was significantly different from the expected 1:1 ratio (P< 0.05), and also female biased sex ratio was recorded. In hermaphrodite specimens, male and female follicles were mixed in the visceral mass but can be clearly distinguished by light microscopy study. L. whaetleyi appears to be a dioecious species in which 25 specimens were simultaneous hermaphrodite and under certain environmental conditions may be capable of self-fertilization. This suggests that detailed studies on sex ratio of L. whaetleyi may be helpful in understanding unionid phylogeny.

Estimating divergence times of lizardfishes and their allies (Euteleostei: Aulopiformes) and the timing of deep-sea adaptations

The divergence times of lizardfishes (Euteleostei: Aulopiformes) are estimated utilizing a Bayesian approach in combination with knowledge of the fossil record of teleosts and a taxonomic review of fossil aulopiform taxa. These results are integrated with a study of character evolution regarding deep-sea evolutionary adaptations in the clade, including simultaneous hermaphroditism and tubular eyes. Divergence time estimations recover that the stem species of the lizardfishes arose during the Early Cretaceous/Late Jurassic in a marine environment with separate sexes, and laterally directed, round eyes. Tubular eyes have arisen independently at different times in three deep-sea pelagic predatory aulopiform lineages. Simultaneous hermaphroditism evolved a single time in the stem species of the suborder Alepisauroidei, the clade of deep-sea aulopiforms during the Early Cretaceous. This result indicates the oldest known evolutionary event of simultaneous hermaphroditism in vertebrates, with the Alepisauroidei being the largest vertebrate clade with this reproductive strategy.

Influence of heterotrophic feeding on the sexual reproduction of Pocillopora verrucosa in aquaria

Recent research has shown that heterotrophic feeding has a strong effect on both maintenance and growth of corals by providing mainly a nutritional source of nitrogen and phosphorus. Nevertheless, no study has yet been undertaken on the interactions between feeding and sexual reproduction in corals. This study examines the effects of heterotrophic feeding on the sexual reproduction of Pocillopora verrucosa in aquaria. Rotifers were used as a live food source at two concentrations ((LFC = low feed colonies) = 5 × 10 2 organisms l − 1 ; (HFC = high feed colonies) = 15 × 10 2 organisms l − 1 ) with an unfed control (UC = unfed colonies). Three replicates of five colonies were used for each treatment. Rotifers were distributed among the nine aquaria four times per week for 3 h. Histological sections of coral polyps were prepared to monitor the development of gametogenic stages and the fecundity of the colonies. The number and size of oocytes, sperm sacs and gonad stages were determined in each polyp. Both fed and starved colonies of P. verrucosa proved to be simultaneous hermaphrodites and broadcast spawners. The gametogenesis period was short (3 months) and occurred from October to December 2007. No spawning event was observed in the aquaria. However, the disappearance of mature oocytes in samples collected in January 2008 suggested that spawning took place between December 2007 and January 2008. Heterotrophic feeding appeared to have a negative effect on reproduction in P. verrucosa. The results showed that both the proportion of polyps with gametes and the reproductive effort were lower in the fed than in starved colonies. It is likely that an energetic trade-off occurred between reproduction and other metabolic functions. However, oocytes were bigger in fed corals compared with the unfed controls. Several hypotheses are proposed to explain these metabolic/energy distribution patterns.

Circannual Gonad Activity in Two Species of the GenusVestiaP. Hesse (Gastropoda: Pulmonata: Clausiliidae)

Vestia gulo and V. turgida are iteroparous simultaneous hermaphrodites. In the wild they are active from April till October; they reproduce in the spring and summer (egg retention from May to August), and hibernate from November till March. Their gonads show the greatest activity in the spring and summer (maturation of oocytes, intensive vitellogenesis: March–May, numerous mature oocytes: May–July; production and maturation of spermatozoa: March–May; numerous packets of mature spermatozoa: May–October), which coincides with the reproductive season. The onset of reproduction is determined by the size of the pool of vitellogenic and mature oocytes; oocyte production starts in the summer of the previous vegetation season and lasts till next spring. Mature spermatozoa are present in the gonad from spring till autumn which indicates an ability to mate during the whole active period. During hibernation the gonads contain no spermatids, mature spermatozoa or advanced vitellogenic oocytes.

Bateman Gradients in Hermaphrodites: An Extended Approach to Quantify Sexual Selection

Sexual selection is often quantified using Bateman gradients, which represent sex-specific regression slopes of reproductive success on mating success and thus describe the expected fitness returns from mating more often. Although the analytical framework for Bateman gradients aimed at covering all sexual systems, empirical studies are biased toward separate-sex organisms, probably because important characteristics of other systems remain incompletely treated. Our synthesis complements the existing Bateman gradient approach with three essential reproductive features of simultaneous hermaphrodites. First, mating in one sex may affect fitness via the opposite sex, for example, through energetic trade-offs. We integrate cross-sex selection effects and show how they help characterizing sexually mutualistic versus antagonistic selection. Second, male and female mating successes may be correlated, complicating the interpretation of Bateman gradients. We show how to quantify the impact of this correlation on sexual selection and propose a principal component analysis on male and female mating success to facilitate interpretation. Third, self-fertilization is accounted for by adding selfed progeny as a separate category of reproductive success to analyses of Bateman gradients. Finally, using a worked example from the snail Biomphalaria glabrata, we illustrate how the extended analytical framework can enhance our understanding of sexual selection in hermaphroditic animals and plants.

Intra-locus sexual conflict and sexually antagonistic genetic variation in hermaphroditic animals

Intra-locus sexual conflict results when sex-specific selection pressures for a given trait act against the intra-sexual genetic correlation for that trait. It has been found in a wide variety of taxa in both laboratory and natural populations, but the importance of intra-locus sexual conflict and sexually antagonistic genetic variation in hermaphroditic organisms has rarely been considered. This is not so surprising given the conceptual and theoretical association of intra-locus sexual conflict with sexual dimorphism, but there is no a priori reason why intra-locus sexual conflict cannot occur in hermaphroditic organisms as well. Here, I discuss the potential for intra-locus sexual conflict in hermaphroditic animals and review the available evidence for such conflict, and for the existence of sexually antagonistic genetic variation in hermaphrodites. I argue that mutations with asymmetric effects are particularly likely to be important in mediating sexual antagonism in hermaphroditic organisms. Moreover, sexually antagonistic genetic variation is likely to play an important role in inter-individual variation in sex allocation and in transitions to and from gonochorism (separate sexes) in simultaneous hermaphrodites. I also describe how sequential hermaphrodites may experience a unique form of intra-locus sexual conflict via antagonistic pleiotropy. Finally, I conclude with some suggestions for further research.

Population dynamics, sex ratio and size at sex change in a protandric simultaneous hermaphrodite, the spiny shrimp Exhippolysmata oplophoroides

One of the main goals of sex allocation theory is understanding sex ratio evolution. However, theoretical studies predicting sex ratios in species with unusual sexual systems, such as protandric simultaneous (PS) hermaphroditism, are rare. In PS hermaphrodites, juveniles first develop into functional males that mature into simultaneous hermaphrodites later in life. Here, we report on the sex ratio (males/males + hermaphrodites) in the PS hermaphroditic shrimp Exhippolysmata oplophoroides. A 2-year study demonstrated that hermaphrodites dominated the population in two different bays. This skewed sex ratio may be explained by limited encounter rates among conspecifics. In agreement with this idea, the density of shrimps was extremely low (≤1 shrimp km−2) at the two study sites. Size at sex phase change and sex ratios remained relatively stable through time at the two bays. The stability of these parameters might be explained by the rather steady population structure of this species during the study period. A review of sex ratios in PS hermaphroditic shrimps (Lysmata and Exhippolysmata) revealed considerable variation; some species have male- and others hermaphrodite-skewed sex ratios. The conditions explaining inter- and intra-specific sex ratio variation in protandric simultaneous hermaphroditic species remain to be addressed.

Effects of Selfing on Offspring Survival and Reproduction in a Colonial Simultaneous Hermaphrodite (Bugula stolonifera, Bryozoa)

Understanding the consequences of selfing in simultaneous hermaphrodites requires investigating potential deleterious effects on fitness at all stages of life. In this study, I examined the effects of selfing throughout the life cycle of the marine bryozoan Bugula stolonifera, a colonial simultaneous hermaphrodite. In 2008, larvae from field-collected colonies were cultured through metamorphosis to reproductively mature colonies either in the presence of one other colony, the paired treatment, or alone, the solitary treatment. Results demonstrated that selfing in this species is possible, in that colonies in the solitary treatment produced viable larvae that successfully completed metamorphosis. On average, however, these colonies released significantly fewer larvae, which experienced reduced rates of metamorphic initiation and completion compared to the paired treatment. These experiments were extended in 2009, when metamorphs from colonies reared in the solitary (n = 58) and paired (n = 61) treatments were transferred to the field for growth to reproductive maturity and then brought back to the laboratory for larval collection. Results revealed additional deleterious effects associated with selfing, as no viable larvae were recovered from colonies deriving from the solitary treatment. In contrast, offspring from the paired treatment released 1030 larvae and 99% initiated metamorphosis, 97% of which completed metamorphosis. Overall, selfed larvae not only had significantly decreased chances of survival, but those that did survive did not successfully reproduce.

Life history of a simultaneously hermaphroditic fish, Diplectrum formosum

Sand perch Diplectrum formosum were collected in the Atlantic Ocean waters off the south-eastern U.S. for life history analyses between April 2001 and July 2004 and ranged in size from 63 to 236 mm standard length (L(S)) and 0-8 years of age. Diplectrum formosum are simultaneous hermaphrodites that reach 50% sexual maturity at 12.2 months and 122.8 mm L(S) for testicular tissue and 13.6 months and 129.3 mm L(S) for ovarian tissue. The gonad contains ovarian and testicular tissue separated by a thin basement membrane, with no means of internal self-fertilization. Spawning females were found between March and January with a peak of spawning activity in May and a realized spawning periodicity of 2 days, equivalent to a maximum of 168 spawning events per year. Fish with testicular tissue in spawning condition were obtained throughout the year in every month sampled with a maximum frequency between June and September. There were also trends indicating that testicular tissue was more likely to be in spawning condition in the presence of hydrated oocytes within the accessory structure. This may indicate the use of the accessory structure as a storage site for hydrated oocytes until a mate can be located.

Reproductive system of the genus Crasiella (Gastrotricha, Macrodasyida)

Crasiella diplura from Sweden and Crasiella sp. from Italy were studied alive and with transmission electron microscopy (TEM). The two species are simultaneous hermaphrodites and share the same reproductive system lay-out: paired ovaries extend along the posterior part of the intestine and join mid-dorsally, while bilateral, club-shaped testes lie at the sides of the anterior gut, extending as deferentia that fuse on the mid-ventral plane and open into a single pore; gametes mature in a caudocephalic and centripetal direction. The bulky, sac-like, frontal organ is lined by a simple epithelium and lies dorsolaterally to the intestine, on the left side of the body. The spindle-shaped caudal organ is musculo-glandular and is located ventrolaterally to the gut on the right side. It is characterized by the presence of a roughly Y-shaped internal channel that opens into two pores close to each other, which function for the intake and outlet of the (auto)sperm, respectively. The spermatozoa, which are peculiar and similar in the two species, are characterized by a long and complex acrosome consisting of four ultrastructurally distinct regions, three of which find equivalence in other gastrotrich species. The flagellum lacks a striated cylinder. Anatomy and ultrastructure enable us to hypothesize a modality of sperm transfer in Crasiella that is similar to that observed in Macrodasys.

Determinants of female fecundity in a simultaneous hermaphrodite: the role of polyandry and food availability

Classical sexual selection theory assumes that the reproductive success of females is primarily limited by the resources available for egg production rather than by the number of mating partners. However, there is now accumulating evidence that multiple mating can entail fitness costs or benefits for females. In this study we investigated the effect of polyandry (i.e., the mating with different mating partners) and food availability on the reproductive output of the female sex function in an outcrossing simultaneous hermaphrodite, the free-living flatworm Macrostomum lignano. We exposed virgin worms to different group sizes, a treatment that has previously been shown to affect the level of polyandry in this species. Moreover, we manipulated the food availability throughout the subsequent egg laying period, during which the worms were kept in isolation. The number of offspring produced was used as an estimate of female fecundity. We found that food availability, but not group size, had a significant effect on female fecundity. Additionally, female fecundity was positively correlated with the number of stored sperm in the female sperm-storage organ at the time of isolation, but it was not correlated with body or ovary size of the worms. Our results suggest that female fecundity in M. lignano is primarily determined by the resources available for egg production, and not by the level of polyandry, confirming classic sexual selection theory for simultaneous hermaphrodites.

The symbiotic lifestyle and its evolutionary consequences: social monogamy and sex allocation in the hermaphroditic shrimp Lysmata pederseni

Sex allocation theory predicts female-biased sex allocation for simultaneous hermaphrodites with a monogamous mating system. Mating systems theory predicts that monogamy is advantageous in environments where refuges are discrete, scarce, relatively small, and when predation risk is high outside of these refuges. These predictions were tested with the Caribbean shrimp Lysmata pederseni, a simultaneous hermaphrodite which has an early male phase and lives inside tubes of the sponge Callyspongia vaginalis. This host sponge is a scarce resource that, together with the high predation risk typical of tropical environments, should favor monogamy in the shrimp. Field observations demonstrated that shrimps were frequently encountered as pairs within these tube sponges. Pairs were equally likely to comprise two hermaphrodites or one hermaphrodite and one male. Several of these pairs were observed for long periods of time in the field. Experiments demonstrated that hermaphrodites tolerated other hermaphrodites but not males in their host sponge. These results suggest that pairs of hermaphroditic L. pederseni are socially monogamous; they share the same host individual and might reproduce exclusively with their host partners for long periods of time. Nevertheless, males appeared less likely to establish long-term associations with hermaphrodites as indicated by the rate of their disappearance from their hosts (greater than that of hermaphrodites). Sex allocation was female biased in monogamous hermaphrodites. On average, hermaphrodites invested 34 times more to female than to male reproductive structures. Monogamy and female-biased sex allocation seem to be evolutionary consequences of adopting a symbiotic lifestyle in simultaneous hermaphrodites.

Molecular systematics of peppermint and cleaner shrimps: phylogeny and taxonomy of the genera Lysmata and Exhippolysmata (Crustacea: Caridea: Hippolytidae)

Shrimps from the ecologically diverse genera Lysmata and Exhippolysmata are rare among marine invertebrates because they are protandric simultaneous hermaphrodites: shrimps initially mature and reproduce solely as males, and later in life become functional simultaneous hermaphrodites. Considerable progress on the reproductive ecology of members from these two genera has been achieved during the last decade. However, several outstanding issues of systematic nature remain to be addressed. Here, a molecular phylogeny of these two genera was used to examine the overall evolutionary relationship within and between species and genera, and to answer various questions related to the systematic status of several species. The present phylogenetic analysis, including 53 sequences and 26 species of Lysmata and Exhippolysmata, indicates that semiterrestrial shrimps from the genus Merguia represent the sister group to a second natural clade composed by shrimps from the genera Lysmata and Exhippolysmata. Also, the phylogenetic analysis confirmed that the genus Lysmata is paraphyletic, and includes the genus Exhippolysmata, as noted in a preliminary study. The tree partially supports the separation of species with or without a developed accessory branch into two different genera or subgenera (i.e. Lysmata and Hippolysmata having a well-developed accessory branch, or not, respectively). The genetic distance between the cleaner shrimps Lysmata amboinensis and Lysmata grabhami was smaller than has been observed between other sister species. On the other hand, the topology of the tree indicates that these two entities are reciprocally monophyletic. Thus, this latter result, together with minor but constant differences in the colour pattern reported for these two entities, indicates that there is no reason to stop treating them as different valid species. This study enabled the long overdue resolution of standing taxonomic questions in shrimps from the genera Lysmata and Exhippolysmata. In the future, this phylogeny will help to reveal the conditions favouring the origins of several behavioural and morphological novelties in these unique shrimps. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 160, 254–265.

The evolution of mating systems in barnacles

Thoracican barnacles exhibit tremendous diversity among species in reproductive life histories, including hermaphroditism, dioecy and androdioecy. Androdioecy (where populations contain a mix of males and hermaphrodites) is rare among animals, but has been described in more than 30 species of barnacles and appears to have evolved independently at least four times in this clade. Several models have been suggested to explain the evolution and stability of different mating systems in barnacles, including the relatively high frequency of androdioecy. We review this body of theory and compare predictions to existing empirical data on interspecific patterns of sex allocation. Allocation to male and female reproduction may also vary within barnacle species, and these patterns may provide additional insight into the evolutionary forces that select for different mating systems. Theory predicts that for simultaneous hermaphrodites, the optimal allocation to male function should increase with the size of mating groups, due to sperm competition. This prediction is supported by the only previously published study to address this issue. We present new data from another intertidal barnacle ( Tetraclita rubescens) that do not support the predicted association between investment in male reproduction and local density of conspecifics. We suggest that this departure from the expected relationship may arise from a reduced importance of sperm competition in this species. Despite over 150 years of interest in barnacle mating systems, many questions remain unanswered. We argue that barnacles provide a superb opportunity to test predictions of sex allocation theory and we offer suggestions for promising areas of future research.

Cytodifferentiation during the spermatogenesis of the hermaphrodite caridea Exhippolysmata oplophoroides

Among the decapods, the caridean Exhippolysmata oplophoroides has been described as a simultaneous protandric hermaphrodite, seeing that it presents a male initial stage followed by a hermaphrodite one in which it can function as male and as female. This work had the aims of characterizing the microscopical morphology of the male portion of the ovotestes gonads from E. oplophoroides, at the different development stages, identifying each cell from the germ lines during spermatogenesis, as well as describing the ultramorphology of spermatozoans in the terminal region of the vasa deferentia. Shrimps were collected in Ubatuba, north coast of São Paulo, and their male gonads and the ampoule were removed, fixed and processed according to histological routine and for scanning electron microscopy. The testicular portion is divided in lobes, inside which cells at the same stage of the spermatogenic cycle are observed, with prevalence of spermatogonia and spermatocytes at stages I, II and V of gonad development, whereas spermatids and spermatozoans are found at stages III and IV, respectively. Ultramorphology of the terminal portion of the vasa deferentia exhibits mature aflagellated spike-shaped spermatozoans, encased in secretion and between membrane foldings that will constitute the spermatophores. Despite presenting reproductive characteristics common to other decapods, E. oplophoroides shows spermatozoans as well as spermatophore with typical morphology, which is important for its identification and taxonomy. Further, this species showed polysaccharide secretions where the spermatozoa are immerse as far as the testicular portion, which could have a role in their transport and nutrition as well as spermatophore constitution and/or fixation; differently, other caridean species begin spermatophore formation during the passage of the gametes through the vasa deferentia.

Sperm competition affects sex allocation but not sperm morphology in a flatworm

Sperm competition has been shown to be an important evolutionary agent affecting the behaviour, physiology, and morphology of both males and females. One morphological trait that is particularly likely to be affected by sperm competition is sperm size because it is thought to influence the competitiveness of sperm by determining sperm longevity, motility, and/or their ability to displace competing sperm. Most comparative studies across taxa have found a positive relationship between the level of sperm competition and sperm length, but very few studies have tested for a phenotypically plastic adjustment of sperm morphology in response to sperm competition. In this study, we experimentally tested for an effect of sperm competition on phenotypic plasticity in sperm morphology in an obligately outcrossing simultaneous hermaphrodite, the free-living flatworm Macrostomum lignano, by either raising worms in monogamous pairs (no sperm competition) or in promiscuous groups (intense sperm competition). Worms in groups produced larger testes and smaller ovaries as predicted by sex allocation theory and as previously documented in this species. However, we found no evidence for an effect of group size on sperm morphology, measured as total sperm length, sperm body length, and the length of two different sperm appendages. We conclude that M. lignano may either be incapable of adjusting the sperm morphology in a phenotypically plastic way and/or that there might be no benefit of phenotypic plasticity in sperm traits in this species.

Male Accessory Gland Protein Reduces Egg Laying in a Simultaneous Hermaphrodite

Seminal fluid is an important part of the ejaculate of internally fertilizing animals. This fluid contains substances that nourish and activate sperm for successful fertilization. Additionally, it contains components that influence female physiology to further enhance fertilization success of the sperm donor, possibly beyond the recipient's optimum. Although evidence for such substances abounds, few studies have unraveled their identities, and focus has been exclusively on separate-sex species. We present the first detailed study into the seminal fluid composition of a hermaphrodite (Lymnaea stagnalis). Eight novel peptides and proteins were identified from the seminal-fluid-producing prostate gland and tested for effects on oviposition, hatching and consumption. The gene for the protein found to suppress egg mass production, Ovipostatin, was sequenced, thereby providing the first fully-characterized seminal fluid substance in a simultaneous hermaphrodite. Thus, seminal fluid peptides and proteins have evolved and can play a crucial role in sexual selection even when the sexes are combined.