Male reproductive system of the arrow crab Stenorhynchus seticornis (Inachoididae)

Our aim was to describe the reproductive system of males and the formation of sperm packages in the seminal receptacle (SR) of recently mated females of the arrow crab Stenorhynchus seticornis. The male reproductive system was analyzed, and was described using light microscopy and histological and histochemical methods. The first pair of gonopods was described by means of scanning electron microscopy. Additionally, the dehiscence of spermatophores was tested using samples obtained from the vas deferens of males and from the seminal receptacle of recently mated females. Testes were tubular type, and each vas deferens consisted of three regions: the anterior vas deferens (AVD), including a proximal portion that was filled with free spermatozoa and a distal portion contained developing spermatophores; the median vas deferens (MVD) that contained completely formed spermatophores; and the posterior vas deferens (PVD), which contained only granular secretions. The accessory gland, which was filled with secretions, was located in the transition region between the MVD and the PVD. The spermatophores from the MVD were of different sizes, and none of them showed dehiscence in seawater, whereas those spermatophores in contact with the seminal receptacle were immediately broken. The ultrastructure of the gonopods revealed the presence of denticles at the distal portion, which contribute to the mechanical rupture of the spermatophore wall during the transfer of sperm. The contents of the PVD and accessory gland of males are transferred together with the spermatophores, and are responsible for the secretions observed among the sperm packets in the SR of the female. We suggest that these secretions formed the layers found in the SR of recently mated females, and may play a role in sperm competition in arrow crabs.     

Functional morphology of the female reproductive system of a crab with highly extensible seminal receptacles and extreme sperm storage capacity

We studied the functional morphology of the female reproductive system of the purple stone crab Danielethus crenulatus . The most remarkable feature is the relative storage capacity and extensibility of the seminal receptacles. These receptacles are a pair of simple sacs that lack internal structures dividing the internal lumen. Differences in seminal receptacle size and contents are accompanied by conspicuous changes in receptacle lining at a tissue level. Full seminal receptacles contain discrete sperm masses formed by hardened fluid and densely packed spermatophores. Different sperm masses are likely from different mates and their stratified disposition within the seminal receptacles is compatible with rival sperm displacement and last sperm precedence. Additionally, the anatomical structure of the vulva and vagina suggest active female control over copula. We discuss our results in the general context of sperm storage in brachyurans and the implications for the mating system of this species.     

Sperm competition and reproductive success in the decapod Inachus phalangium (Majidae): a male ghost spider crab that seals off rivals' sperm

Parker's (1970a) hypothesis that the overlap of multiple mating sperm in the female's storage organs promotes sperm competition is tested here for the first time in Crustacea: specifically, the mechanisms and consequences of sperm competition are detailed for the spider crab Inachus phalangium . Females of this species store ejaculates from successive copulations with different males discretely and consecutively in sac-like twin seminal receptacles. During copulation males transfer a large quantity of a sperm-free seminal plasma, followed by the sperm which is stored in small spermatophores and forms a densely-packed sperm packet. It was shown, using ³H-thymidine-labelled ejaculate, that the last male to mate displaces the ejaculate of his predecessors dorsally into the apex of the receptacle. Sperm of previous matings are sealed in with the hardening seminal plasma (sperm gel) and are thus prevented from being used to fertilize eggs, while the last male to mate places his sperm closest to the oviduct and vaginal openings. In experiments using the 'sterile-male' method, sperm from the last male to mate gained all fertilizations in subsequent broods. The seminal plasma forms the sperm gel in ghost spider crabs which is used for displacement of previously stored sperm, whereas various other brachyuran taxa use seminal plasma to produce the sperm plug, which prevents a male's sperm from being displaced.     

Fishery Induces Sperm Depletion and Reduction in Male Reproductive Potential for Crab Species under Male-Biased Harvest Strategy

Sperm depletion in males can occur when polygynous species are intensively exploited under a male-biased management strategy. In fisheries involving crabs species, the effects of this type of management on the reproductive potential is far from being understood. This study tests whether male-biased management of the principal Chilean crab fishery is able to affect the potential capacity of Metacarcinus edwardsii males to transfer sperm to females. Five localities in southern Chile, recording contrasting crab fishery landing, were selected to assess the potential of sperm depletion triggered by fishery. Seasonally, male crabs from each locality were obtained. Dry weight and histological condition of vasa deferentia and the Vaso-Somatic Index (VSI) were determined in order to use them as proxies for sperm depletion and male reproductive condition. A manipulative experiment was performed in the laboratory to estimate vasa deferentia weight and VSI from just-mated males in order to obtain a reference point for the potential effects of the fishery on sperm reserves. Sperm storage capacity is significantly affected by fisheries; during the mating season vasa deferentia from localities with low fishery intensity were heavier than those from high intensity fisheries, and these differences were even more evident in large males. Histological section showed that this disparity in vasa deferentia weight was explained principally by differences in the quantity of spermatophores rather than other seminal material. VSI was always higher in males from localities with low fishery intensity. Males from localities with high fishery intensity showed little capacity to recover sperm reserves and the VSI of these males remained below the values of the just-mated males. Detriment in the capacity of males to transfer sperm is the first step to sperm limitation in an exploited population, thus detection of sperm depletion can be an alert to introduce changes in the current management of crabs.     

The ontogeny of the female reproductive system in the parasitic castrator pea crab Calyptraeotheres garthi: Implications for its mating system

The knowledge of the mating system of pea crabs is still fragmentary as it remains dubious whether females copulate in the juvenile and free‐living ‘hard’ or in the obligatory symbiotic stages (adult stage ‘V’ or intermediate stages II to IV). To discriminate between these two possibilities, we analysed the female seminal receptacles, vagina and opercula, and the sperm content in different stages of the pea crab Calyptraeotheres garthi. Our histology and scanning electron microscopy results revealed that in the hard stage the seminal receptacle is simple without secretory epithelia, and vagina and opercula are not controlled by musculature. In stages II to IV, the seminal receptacles, vagina, and opercula are under development and these structures reach maturity in stage V. These results suggest that females become receptive in stage V and not during predating stages. We found no spermatozoa in SR of ‘hard’ and stage II to IV females while these structures were loaded of sperm in most stage V, indicating that females start to mate in stage V. Our results support the notion that males of C. garthi roam among hosts in search for sedentary stage V females, as predicted by Baeza and Thiel's ( 2007 ) model of mating systems for symbiotic crustaceans. Nevertheless, we failed to reveal whether females mate repeatedly: the accumulation of sperm in larger females might indicate occurrence of multiple copula or a high variability in male sperm transfer.     

Female reproductive morphology of coral‐inhabiting gall crabs (Crustacea: Decapoda: Brachyura: Cryptochiridae)

Gall crabs are obligate associates of stony corals in which they induce skeletal modifications. In some cryptochirid species, females live in open depressions accessible to males; while in others, females are rather isolated in semiclosed galls, which necessitates elaborate sperm storage capabilities by the female. In this study we investigate the female gross morphology and reproductive systems of Fungicola syzygia lodged in semiclosed flattened pits, Opecarcinus cathyae with semiopen pits and Pseudocryptochirus viridis from shallow open depressions using line drawings and histological methods. The general morphology of the cryptochirids' reproductive systems is uniform and conforms to other thoracotreme brachyurans: paired muscular vaginae of the concave pattern lead from the sternal gonopores into paired seminal receptacles where sperm is stored. The seminal receptacle is internally lined by distinct types of epithelia: a cuticle underlined by a columnar epithelium ventrally, a monolayered secretory epithelium dorsally and a multilayered transfer tissue where the oviducts enter the seminal receptacle. In all studied specimens, the seminal receptacle contained free spermatozoa; however, in specimens of Pseudocryptochirus viridis it also contained spermatophores, indicating a recent insemination. In contrast to most other brachyurans ovaries of the investigated cryptochirids extend into the pleon. The specific degree of ovary extension differs between the studied species and is closely related to female body shape.     

Biochemistry of seminal secretions of the crab Scylla serrata with reference to sperm metabolism and storage in the female.

Biochemical studies on the male reproductive tissues and seminal secretions have been made with reference to sperm metabolism and different stages of maturity in the crab Scylla serrata. The results reveal that the seminal plasma and spermatophores are rich in protein, carbohydrate, and lipid. In general, organic components of spermatophores are considerably higher than those of seminal plasma. Enzyme studies show that the succinate dehydrogenase (SDH) activity is very low, whereas fumarate reductase (FR) and lactate dehydrogenase (LDH) exhibit high activity. Electrophoretic studies on LDH show that, in addition to the occurrence of a sperm-specific fraction, LDHx, the M-type subunits are predominant in the mature spermatophores. These results from enzyme studies suggest that sperm metabolism is mainly anaerobic, utilizing the carbohydrates as substrates. The results for maturational changes reveal that the male reproductive tissues and their secretions contain lesser quantity of organic components in the immature crabs; as the maturity proceeds, there is not only concentration of organic substances but also an increase in the size of spermatophores. The concentration of biochemical constituents is highest in the proximal vas deferens (PVD), suggesting that the granular seminal plasma as well as the sperm-agglutinating substance and spermatophoric wall are secreted in this region. The spermatheca of the unmated female crabs are poor in organic constituents. After mating, their contents are enriched by organic substances derived from contributions of the seminal substances. During sperm storage in the spermatheca, only the carbohydrates decline steeply. A low activity of SDH, but a moderate level of LDH and a high level of FR activity, is recorded in the spermathecal content of mated crabs, providing further evidence for anaerobic metabolism of sperm during storage in female. A sharp fall in the stored carbohydrates constitutes further evidence in this regard.     

A hypothesis about the origin of sperm storage in the Eubrachyura, the effects of seminal receptacle structure on mating strategies and the evolution of crab diversity: How did a race to be first become a race to be last?

The origins and evolution of sperm storage in Brachyura are enigmatic: sperm is either stored in seminal receptacles, accessible via the vulvae on the sixth thoracic sternite, or in spermathecae at the border between the seventh and eighth sternites. Crabs with spermathecae are collectively referred to as “podotremes” while crabs with seminal receptacles belong to the Eubrachyura. The position of gonopores is the primary basis for subdividing the Eurachyura into the Heterotremata (female vulvae + males with coxal gonopores) and Thoracotremata (female vulvae + males with sternal gonopores). We present a hypothesis about the evolution of seminal receptacles in eubrachyuran female crabs and argue that the sternal gonopore has been internalized into chitin-lined seminal receptacles and the vulva is in fact a secondary aperture. The loss of some or all of the ancestral chitinous seminal receptacle lining was linked to ventral migration of the oviduct connection. Male and female strategies are to maximize gamete fertilization. The most important variable for females is sperm supply, enhanced by long-term storage made possible by the seminal receptacle. To maximize their fertilization rates males must adapt to the structure of the seminal receptacle to ensure that their sperm are close to the oviduct entrance. The major evolutionary impetus for female mating strategies was derived from the consequences of better sperm conservation and the structure of the seminal receptacle. The advantages were all to the females because their promiscuity and sperm storage allowed them to produce more genetically variable offspring, thereby enhancing variation upon which natural selection could act. We extend our arguments to Brachyura as a whole and offer a unifying explanation of the evolution of seminal receptacles, comparing them with the spermathecae found in “Podotremata”: they were independent solutions to the same problem: maintaining sperm supply during evolutionary carcinization.Explanation of eubrachyuran mating strategies requires analysis of the mating–moulting link, indeterminate vs. determinate growth format and seminal receptacle structure. Two alternatives for each of these characters means that there are eight possible outcomes. Six of these outcomes have been realized, which we term Portunoid, Majoid, Eriphoid, Xanthoid, Cancroid, and Grapsoid–Ocypodoid strategies, respectively. Mapping these characters on to a workable phylogeny (wherein some changes to the seminal receptacle + moulting–mating links are assumed to have occurred more than once) produces the following relationships: Portunoids + Majoids are a sister group to the rest of the Eubrachyura, which fall into two sister groups, Eriphoids + Xanthoids and Cancroids + Grapsoid–Ocypodoids and the “Podotremata” is sister group to all the Eubrachyura. We conclude that what began as a race to be the first to mate was turned on its head to become a race to be last, by the evolutionary changes to the seminal receptacle. Eubrachyuran females were advantaged by greater reproductive autonomy, more opportunity to mate with other males, resulting in more genetically variable progeny and leading to the evolution of much greater taxonomic diversity compared to “podotremes”.     

Morphological analysis of the female reproductive system of Stenorhynchus seticornis (Brachyura: Inachoididae) and comparisons with other Majoidea

We examined the female reproductive system of the yellowline arrow crab Stenorhynchus seticornis by means of histological and histochemical techniques. Mature specimens obtained in the field were kept in the laboratory for mating experiments. After 24 h, newly mated females were dissected, and their reproductive trait routinely processed for embedding in historesin. The specimens examined each possessed a pair of kidney‐shaped seminal receptacles (SR), and these we classified as ventral type, based on the location of the oviduct opening. The mesodermal dorsal region (DR) of SR consisted of a stratified epithelium with scaly cells, while the ectodermal ventral region (VR) was composed of a simple epithelium covered by a cuticle. The oviduct opened at the transition region (TR) between DR and VR and had no velum. The simple epithelium of TR had more folds on the face of the oviduct opening. The vagina exhibited the same features as the TR epithelium and was contiguous to VR, anchored by muscles. In the lumen, from one to three strata of sperm packets were observed, the dorsal one containing free sperm, and the most ventral stratum, spermatophores. An acidophilic glycoprotein layer enclosed the strata. Spermatophores in the ventral stratum were enclosed in a voluminous secretion, composed by acid polysaccharides most likely from the last male mated. The ventral‐type receptacle, stratified sperm packets, and polyandry, usually observed in females of Majoidea, suggest the occurrence of sperm competition in S. seticornis, favoring the sperm of the last male mated, as its sperm mass is located near the opening of the female oviduct.     

The functional morphology and role of the thelycum in insemination,and its relation to the mating system in the seagrass shrimp Ambidexter symmetricus (Decapoda: Processidae)

There is considerable variation in structures known to function in the transfer and storage of sperm in female decapod crustaceans. The thelycum is a secondary sexual character that forms from the posterior thoracic sterna of female shrimps (especially penaeoids and sergestoids). Females in the caridean shrimp family Processidae have a thelycum‐like structure which rarely occurs in other caridean females. We tested the hypothesis that the processid thelycum serves as a spermatheca for either short‐term attachment or long‐term sperm storage. When inseminated females of the processid Ambidexter symmetricus were isolated after mating, newly spawned and then incubated embryos hatched, but in the continued absence of a male, females were unable to fertilize a subsequent spawn. Our observations on A. symmetricus show that sperm were not retained after female spawning, and thus the thelycum is not used for long‐term sperm storage as in many penaeoids. In A. symmetricus, the thelycum may serve as an external median spermatheca (seminal receptacle) for temporary attachment and storage of a sperm mass during the 2–3 h interval between mating and spawning. Observations on mating behavior support the hypothesis of a pure‐search (promiscuous) mating system in A. symmetricus, with males showing little interest before, and copulating with females only after, the female parturial molt. Mating encounters were short (<2 min). This mating system is like that of other caridean shrimps with populations structured similarly to those in A. symmetricus: a relatively high density of mobile individuals and sexual dimorphism in body size (reproductive females larger than males) but not in cheliped weaponry (similar in males and females).     

Morphology and function of the reproductive tract of the spider crab Libinia spinosa (Crustacea, Brachyura, Majoidea): pattern of sperm storage

Morphology and function of the male reproductive tract, female spermatheca and patterns of sperm storage were assessed in the crab Libinia spinosa using histological methods. Testes are characterized by the presence of peripheral spermatogonia and different sequences of sperm maturity. Spermatophores begin to be packed in the last portion. The vas deferens consists of three sections: anterior, with undeveloped spermatophores and free sperm; median, with well-developed spermatophores; and posterior with granular secretions. Female spermathecae are of the ventral type, with a velum separating dorsal and ventral chambers. Live individuals were kept in the laboratory and arranged in pairs. An experiment was conducted toward the end of the reproductive season, in which males with the right gonopod excised were placed with receptive females. After mating, females were killed and the spermathecae dissected for histological study and observation of the pattern of sperm storage. Spermatozoa were found forming discrete sperm packages. New ejaculates can fill the entire spermatheca or be restricted to the ventral chamber; sperm are rounded, with a distinguishable acrosomal core. Old ejaculates are restricted to the dorsal chamber and are of irregular shape and larger size; an acrosomal core was not distinguishable. The secretions produced by the glandular epithelium of the dorsal chamber of the spermathecae are likely to have a role in the removal of dead sperm.     

Pattern of sperm storage and migration in the reproductive tract of the swallowtail butterfly Papilio xuthus: cryptic female choice after second mating

Females of the swallowtail butterfly Papilio xuthus L. (Lepidoptera: Papilionidae) mate multiply during their life span and use the spermatophores transferred to increase their longevity as well as fecundity. Sperm from different males may be stored in the sperm storage organs (bursa copulatrix and spermatheca). To clarify the pattern of sperm storage and migration in the reproductive tract, mated females are dissected after various intervals subsequent to the first mating, and the type and activity of sperm in the spermatheca are observed. When virgin females are mated with virgin males, the females store sperm in the spermatheca for more than 10 days. Sperm displacement is found in females that are remated 7 days after the first mating. Immediately after remating, these females flush out the sperm of the first male from the spermatheca before sperm migration of the second male has started. However, females receiving a small spermatophore at the second mating show little sperm displacement, and the sperm derived from the small spermatophore might not be able to enter the spermatheca. Females appear to use spermatophore size to monitor male quality.     

Density affects mating mode and large male mating advantage in a fiddler crab

Fiddler crabs show two different mating modes: either females search and crabs mate underground in male burrows, or males search and crabs mate on the surface near female burrows. We explored the relationship between crab density, body size, the searching behavior of both sexes, and the occurrence of both mating modes in the fiddler crab Uca uruguayensis. We found that crabs change their mating mode depending on their size and crab density. Crabs mated mostly on the surface at low densities, and underground at high densities. The proportion of wandering receptive females but not courting males accounted for the variation in mating modes. This suggests that whether crabs mate underground (or on the surface) is determined by the presence (or absence) of searching females. We found that the change in the mating mode affected the level of assortative mating; males mating underground were bigger than those mating on the surface, suggesting active female choice. Given that fiddler crabs experience multiple reproductive cycles, they are prone to showing behavioral plasticity in their mating strategy whenever the payoffs of using different mating modes differ between reproductive events. Our results suggest that the incorporation of different levels of environmental variability may be important in theoretical models aimed at improving our understanding of the evolution of alternative mating tactics and strategies.     

Male age, mating status and nuptial gift quality in a bushcricket

Female mate choice occurs in many animals, and in some species females prefer older males. Because older males have demonstrated their survival ability, they may be of higher genetic quality, providing genetic benefits to the offspring of their mates. However, in species where females receive direct benefits of matings, younger males may be more likely to provide more fertile or more nutritious ejaculates, so females may discriminate against older males. Males of the bushcricket Ephippiger ephippiger (Orthoptera: Tettigoniidae) produce large spermatophores at mating (>30% of body weight, circa 10% protein content). Female E. ephippiger discriminate against the song of older males. We examined the effects of male age and mating history on male reproductive investment (spermatophore size, sperm number, nitrogen content). Males produced spermatophores with significantly fewer sperm and of lower nitrogen content on their fourth mating, despite free access to food and a 1-week interval between matings, indicating that there is a cost of mating to males. There was no indication that older virgin males produced lower-quality spermatophores. Rather, older males produced bigger spermatophores of higher nutritional value and containing more sperm. Male age and mating history seem likely to be strongly correlated in the field. We conclude that female E. ephippiger probably prefer the songs of younger males, because in the field, this preference correlates with male mating history and therefore resources provided at mating. Thus, female preference for younger males could reflect discrimination against low-quality nuptial gifts.     

Parental care and reproductive behavior of the clown goby, <Emphasis Type="Italic">Microgobius gulosus</Emphasis>, with observations on predator interactions

Describe reproductive behavior and mating system of the clown goby from field observations. Clown gobies exhibit a loosely haremic mating system. Pairs construct burrows at the base of cattails, the roots of which provide structural support and a spawning substrate. Larger males monopolize multiple burrows, each with an individual female. After spawning, males camouflage burrow entrances with sand and females brood developing young for 4 days. Males continue to guard the covered nests in 50% of observed brooding periods. Burrows are also used as shelter from predators. Both sexes confront intruders but only males exhibit a distinct color response to juvenile blue crabs, Callinectes sapidus, the most significant predator. The male color response appeared to mimic the color of adult blue crabs, a known predator of juvenile crabs, perhaps acting as a deterrent. The presence of the predatory blue crab may require one parent to perform deterrent displays, promoting female care in this mating system.     

A Noninvasive Method For In situ Determination of Mating Success in Female American Lobsters (Homarus americanus)

Despite being one of the most productive fisheries in the Northwest Atlantic, much remains unknown about the natural reproductive dynamics of American lobsters. Recent work in exploited crustacean populations (crabs and lobsters) suggests that there are circumstances where mature females are unable to achieve their full reproductive potential due to sperm limitation. To examine this possibility in different regions of the American lobster fishery, a reliable and noninvasive method was developed for sampling large numbers of female lobsters at sea. This method involves inserting a blunt-tipped needle into the female''s seminal receptacle to determine the presence or absence of a sperm plug and to withdraw a sample that can be examined for the presence of sperm. A series of control studies were conducted at the dock and in the laboratory to test the reliability of this technique. These efforts entailed sampling 294 female lobsters to confirm that the presence of a sperm plug was a reliable indicator of sperm within the receptacle and thus, mating. This paper details the methodology and the results obtained from a subset of the total females sampled. Of the 230 female lobsters sampled from George''s Bank and Cape Ann, MA (size range = 71-145 mm in carapace length), 90.3% were positive for sperm. Potential explanations for the absence of sperm in some females include: immaturity (lack of physiological maturity), breakdown of the sperm plug after being used to fertilize a clutch of eggs, and lack of mating activity. The surveys indicate that this technique for examining the mating success of female lobsters is a reliable proxy that can be used in the field to document reproductive activity in natural populations.     

Pairing and insemination patterns in a giant weta (Deinacrida rugosa: Orthoptera; Anostostomatidae)

Positive size assortative mating can arise if either one or both sexes prefer bigger mates or if the success of larger males in contests for larger females leaves smaller males to mate with smaller females. Moreover, body size could not only influence pairing patterns before copulation but also the covariance between female size and size of ejaculate (number of spermatophores) transferred to a mate. In this field study, we examine the pre-copulatory mate choice, as well as insemination, patterns in the Cook Strait giant weta (Deinacrida rugosa). D. rugosa is a large orthopteran insect that exhibits strong female-biased sexual dimorphism, with females being nearly twice as heavy as males. Contrary to the general expectation of male preference for large females in insects with female-biased size dimorphism, we found only weak support for positive size assortative mating based on size (tibia length). Interestingly, although there was no correlation between male body size and the number of spermatophores transferred, we did find that males pass more spermatophores to lighter females. This pattern of sperm transfer does not appear to be a consequence of those males that mate heavier females being sperm depleted. Instead, males may provide lighter females with more spermatophores perhaps because these females pose less of a sperm competition risk to mates.     

Functional morphology of the sperm-containing chambers of the sea slug Okenia polycerelloides in the context of sexual selection

Sea slugs are interesting models to study post-copulatory sexual selection in simultaneous hermaphrodites due to the enormous variation of their reproductive systems. However, the knowledge of the functional morphology of their reproductive system is limited to few species, and it is rarely discussed in the context of sexual selection theory. In this study, we investigated the functional morphology of the sperm-containing chambers (i.e., ampulla, seminal receptacle, and bursa copulatrix) of the reproductive system of Okenia polycerelloides (Ortea & Bouchet, 1983), based on light, confocal, and electron microscopy. Although the morphology of the ampulla is similar to other species, indicating that it is a site for autosperm storage, we found some sperm facing the ampullar epithelium, a feature commonly regarded as characteristic of the seminal receptacle of sea slugs. The seminal receptacle of O. polycerelloides showed secretory activity and contained sperm with distribution and orientation suggestive of stratification of allosperm from distinct mating events, a feature that would affect sperm competition. The bursa copulatrix had epithelial cells with secretory and absorptive characteristics, and contained degraded sperm and yolk granules within its lumen. Comparative analyses of the contents of each organ demonstrated that sperm digestion occurs in the bursa copulatrix and affects sperm heads first, changing their morphology from slender and curved to shorter and ellipsoid before complete lysis. Although digestion and absorption of surplus sperm are currently the main hypothesized functions for the bursa copulatrix, its role in cryptic female choice should not be ruled out. The close structural connection between the seminal receptacle and bursa copulatrix, as well as their muscular walls, would enable control over the fate of the sperm received in each mating event, that is, storage or digestion.