Determinants of paternity in the garden snail Helix aspersa

Despite the likely importance of post-copulatory sexual selection in simultaneous hermaphrodites, the factors influencing sperm competition in these organisms are generally unknown. We have investigated the effects of dart-shooting, mating order, and several other predictors on the proportion of offspring fathered by penultimate (P_n-1) and ultimate (P_n) sperm donors in multiply mated garden snails, Helix aspersa. While paternity ratios were biased towards the penultimate donor (mean P_n-1=0.61), the magnitude of this advantage was dependent upon which of the two donors successfully darted the recipient. Mean P_n-values increased from 0.17 when the recipient was hit by the penultimate donor to 0.39 when it was hit by the ultimate donor. Furthermore, the effect of the dart was more pronounced in the clutches of smaller recipients. From these results, and observations of live sperm in the storage organs, we propose a novel mechanism to explain the detected pattern of sperm utilization in helicid snails. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00265-002-0519-6.     

Among- and within-population variation in sperm quality in the simultaneously hermaphroditic land snail Arianta arbustorum

Sperm competition models on the evolution of sperm size assume associations with another sperm quality trait, sperm longevity. Sperm length can also provide an indication of possible mechanisms affecting motility and thus fertilization success. Despite their importance, however, detailed mechanisms of sperm competition at the gamete level are poorly understood. In simultaneously hermaphroditic land snails, sperm traits and cryptic female choice are assumed to be crucial in determining fertilization success. We examined the variation in sperm length and number among individuals from four natural populations of the land snail Arianta arbustorum, a species with multiple mating and long-term sperm storage. We also assessed variation in velocity, motility and longevity of sperm in snails from two of the four populations. Independent of shell size, sperm length differed among populations and, to a minor extent, even among individuals within populations. Mean sperm length of a snail was not correlated with the number of sperm delivered in a spermatophore. The mean sperm velocity (=VCL) did not differ between snails from two populations. However, VCL varied among snails. Percentage motility and longevity of sperm differed between snails from the two populations. No correlations were found between length, velocity, percentage motility and longevity of sperm. To conclude, individual snails differed in sperm quality, and this variation may partly explain the differential fertilization success between A. arbustorum snails. Moreover, our findings did not support the positive association between sperm length and longevity assumed by sperm competition models for internally fertilizing species.     

Multiple paternity and individual variation in sperm precedence in the simultaneously hermaphroditic land snail Arianta arbustorum

Intraspecific variation in the proportion of offspring sired by the second mate with a female (P₂) is an aspect of sperm competition that has received little attention. I examined the effects of delay between copulations (range 9–380 days) and size of sperm donor on sperm precedence in double-mated individuals of the simultaneously hermaphroditic land snail Arianta arbustorum. Using shell colour as a genetic marker, paternity was analysed in 132 broods produced by 35 snails that had mated with two partners of different genotype. Sperm precedence (P₂) was influenced by the time between the two matings when the mating delay exceeded 70 days (one reproductive season). P2 averaged 0.34 in the first brood of snails that mated twice within 70 days indicating first mate sperm precedence. In contrast, P₂ averaged 0.76 in broods of snails that remated in the following season, indicating a decreased viability of sperm from the first mate. The size of sperm-donating individuals had no effect on the fertilization success of their sperm in the first brood produced after the second copulation. Analysis of long-term sperm utilization in 23 snails that laid three to nine egg batches over 2 years revealed striking differences among individuals. Five snails (21.7%) exhibited first-mate sperm precedence throughout, eight snails (34.8%) showed second-mate sperm precedence throughout, whereas ten snails (43.5%) exhibited sperm mixing in successive batches. It is suggested that the individual variation in sperm precedence in A. arbustorum may partly be due to differences in the amount of sperm transferred. Paternity analysis in 34 batches laid by 19 wild-caught individuals of A. arbustorum indicated that at least 12 snails (63.2%) used sperm from two or more mates for the fertilization of their eggs. This suggests a high incidence of multiple paternity in broods of A. arbustorum under field conditions.     

Independence, not conflict, characterizes dart-shooting and sperm exchange in a hermaphroditic snail

Although the sexes are united in hermaphrodites, conflict can still occur because the male and female functions have separate interests. We examined the evidence for conflict in the mating system of the terrestrial snail Cantareus aspersus (formerly Helix aspersa) where sharp, calcareous darts are ‘shot’ during courtship. We predicted that the use of the dart would either reflect or create conflict and this would be evident in either the courtship behavior or the transference of sperm. Previous studies demonstrated that the dart functions after sperm transfer to increase sperm survival. Using detailed observations of mating snails, we examined the factors that determine dart shooting order, the behavioral responses after being hit by a dart, the accuracy of dart shooting, and the allocation of sperm resources. We found that each dart was shot independently, and each animal appeared to be interested only in getting off the best possible shot, probably one that penetrates deeply near the genital pore. There is no evidence of mating conflict. Every snail transfers sperm to its partner, and the size of the donation does not depend on the success or failure of either snail’s dart shot. Although the receipt of a dart does not appear to cause harm, it may produce indirect costs due to the partial loss of control over fertilization. We conclude that mating in C. aspersus is a partnership in which independent actors demonstrate unconditional reciprocity during courtship and sperm transfer.     

Sperm precedence and sperm storage in multiply mated red flour beetles

In insects, the last male to mate with a female often gains access to a disproportionate number of subsequent fertilizations. This study examined last-male sperm precedence patterns in doubly and triply mated Tribolium castaneum females. Sperm storage processes were investigated by measuring the quantity of sperm stored within the female spermatheca following single, double, and triple matings. Both doubly mated and triply mated females exhibited high last-male sperm precedence for progeny produced during the first 48 h following the last mating, with females in both groups exhibiting parallel declines in sperm precedence 1 and 2 weeks later. The number of sperm stored by females increased by 33% between singly mated and doubly mated females, indicating that the spermatheca is filled to only two-thirds capacity following insemination by the first male. Based on the proportion of stored sperm from first and second matings, we tested predictions about sperm precedence values based on models of random sperm mixing. High initial last-male sperm precedence strongly supports stratification of last-male sperm. By 1–2 weeks after double matings, sperm precedence declined to levels indistinguishable from values expected under random mixing. These results provide insight into mechanisms of sperm storage and utilization in this species. Received: 20 August 1997 / Accepted after revision: 24 May 1998     

Sperm transfer and storage in relation to sperm competition in Callosobruchus maculatus

This paper examines the underlying mechanisms of sperm competition in the beetle Callosobruchus maculatus (F.) (Coleoptera: Bruchidae). Recently developed mathematical models of sperm competition are combined with an empirical investigation of the processes of sperm transfer and storage. During a single insemination virgin males transfer approximately 46000 sperm, 85% more sperm than females can effectively store in their spermathecae. Many of these sperm remain in the bursa copulatrix where they are apparently rapidly degraded and can therefore play no role in fertilization. The spermatheca (primary site of sperm storage) is filled by a single insemination and sperm are lost from this organ at a constant rate. This rate of sperm loss from the spermatheca is insufficient for sperm mixing (without displacement) or sperm stratification to account for the degree of last male sperm precedence measured as P ₂; the proportion of offspring fathered by the second male to mate reported for this species (P ₂ = 0.83, when two inseminations are separated by 24 h). Models of sperm displacement correctly predict high levels of sperm precedence although the precision of these predictions is limited because the proportion of sperm entering the spermatheca cannot be accurately determined. The results suggested that last male sperm precedence in C. maculatus the result of sperm displacement, although the exact mechanism of displacement (sperm-for-sperm or fluid displacement) remains unknown. Possible constraints imposed by female genital anatomy on sperm displacement are discussed.     

Females influence sperm storage and use in the yellow dung fly Scathophaga stercoraria (L.)

Summary The influence of the female on the process of sperm storage and use was examined. Copula duration, the condition of the female and whether or not a copula terminated naturally influenced the number of spermathecae (of three) in which once-mated females stored sperm. Females stored more sperm the larger their mate and the sperm from larger males were stored more unevenly amongst the spermathecae than were those from smaller males. Double-mated females had sperm in fewer spermathecae the larger the second of their mates and these spermathecae tended to be the ones which lay together within the female. The P2 values over three successive clutches were constant and sperm precedence was complete when the larger male was second to mate but began low and increased over subsequent clutches when the smaller male mated second. These results suggest females prefer, and are able, to use the sperm of larger males to fertilise their eggs. It is proposed that multiple spermathecae in Diptera have evolved to give females better control over offspring paternity.     

Spermatogenesis,sperm storage and comparative sperm morphology in nine species of Capitella,Capitomastus and Capitellides (Polychaeta: Capitellidae)

Light microscopy and transmission electron microscopy (TEM) were used to describe spermatogenesis and the morphology of mature sperm and sperm storage organs in five sibling species of Capitella, three species in the related genus Capitomastus, and one species in the genus Capitellides. These capitellids lack a well-developed testis, but young males have a few specialized regions of the peritoneum in the eighth setiger, where germ cells proliferate and spermatogonia are released into the coelom, and spermiogenesis is completed. Mature sperm are stored in the central regions of paired genital ducts (coelomoducts), which lie between the seventh and eighth setigers. The cells forming the walls of the coelomostome and central region of the duct are ciliated and have large glycogen deposits. The lumenal borders have extensive microvilli and there is evidence that they secrete glycogen-containing materials into the duct. All species have modified primitive sperm with a conical acrosome, elongated nucleus, and long middle piece extending along the proximal portion of the flagellum. A single ring-shaped mitochondrion encircles the centriolar region of the middle piece and the cytoplasm is filled with glycogen. The sperm of all nine species differ significantly in the lengths of their middle pieces, acrosomes and especially in their nuclear lengths. The nuclear lengths have a twofold range among the sibling species of Capitella and Capitomastus. Subtle differences in the shape and volume of the acrosomal vesicle and acrosomal space characteristic of the Capitella sibling species seem to correlate with a basic division of these species into those with diploid chromosome numbers of 20 or 26. Spermiogenesis, the number of sperm produced, and the method of sperm storage are appropriate for efficient sperm utilization in fertilization. No evidence indicates that spermatophores are formed and transferred between individuals and the method of sperm transfer is not understood. The differences in the dimensions and acrosome morphology of mature sperm, and the previously demonstrated specializations in the egg envelopes in the Capitella sibling species, are characteristic features of the reproductive isolation that exists among these capitellid species.     

A comparative analysis of sperm storage and mating strategies in five species of brachyuran crabs

Variation in female sperm storage is explained, in part, by the amount of sperm transferred at mating. Laboratory mating experiments were conducted on Eurypanopeus depressus and Rhithropanopeus harrisii from the Chesapeake Bay and Pachygrapsus transversus from Florida, while mated pairs of Uca beebei and U. terpsichores were collected from mudflats in Panama. All experiments and collections were conducted during the summer of 2006 and 2007. More sperm was transferred to larger than smaller females, and by species with long copulation durations (R. harrisii and E. depressus). These two species live in cryptic habitats, have high sperm/egg ratios, and likely store sperm across multiple broods. In contrast, P. transversus and U. beebei mate conspicuously, have short copulations, transfer fewer sperm, and have low sperm/egg ratios. Comparisons of sperm transfer across different mating strategies and habitats provide a better understanding of female sperm storage in the Brachyura.     

Autoradiographic investigation of uptake and storage of exogenous sperm by the ovary of the compound ascidian Diplosoma listerianum

The movement of radiolabelled non-self sperm within the female reproductive tract of cultured specimens of the hermaphroditic ascidian Diplosoma listerianum Milne Edwards was investigated by light-microscope autoradiography. Passage of male gametes up the oviduct (fertilization canal) to the ovary was demonstrated for sperm known unequivocally to be of external (non-self) origin. The lumen of the ovary was directly confirmed as a site of long-term storage of sperm, as had previously been suggested from circumstantial evidence. The ovary had a branching structure. Sperm entered blind diverticula leading from the main lumen of the ovary, and persisted there adjacent to the oocyte at the end of each of these side-branches. This arrangement included the association of stored sperm with very small, immature oocytes. An investigation of the time-course of sperm uptake showed that exogenous sperm did not enter the oviducts of recipient ramets for some time (>3 h) after they were initially available, with first presence of sperm noted in the distal half of the duct at 9 h and greatest recorded uptake at 27 h. Sperm had reached the ovary 6.6 h after the first observation of their occurrence in the distal part of the oviduct. Uptake declined steeply between 27 and 46.8 h of continual exposure to sperm released by a single genetic source, and remained low at 81 h. The significance of these observations for the mating pattern of D. listerianum is discussed.     

Habitat choice in the intertidal snail Tegula funebralis

Intraspecific variation in habitat preference was studied in the black turban snail Tegula funebralis (Adams, 1854), at two locations on the northern U.S. Pacific Coast. Studies in 1977 using a mass-marking technique showed that most snails found either above or in permanent tidepools at low tide return to their original habitats within a few days after experimental habitat reversal. This return is not due to homing behavior, but is apparently based on the recognition of ecological characteristics of the two habitats. Experiments in 1978 with individually-marked snails suggest that they prefer specific intertidal levels, and not merely above-pool or in-pool habitats. Theoretical models predict that this behavior could play a major role in the maintenance of genetic polymorphism in a species like T. funebralis, whose intertidal environment is characterized by extreme spatial heterogeneity.     

Factors influencing clustering in the intertidal snail Cerithium moniliferum

The snail Cerithium moniliferum Kiener at Heron Island (southern end of the Great Barrier Reef) clusters at outgoing tide and disperses at incoming tide. In containers with a constant water level, but exposed to the normal day and night rhythm, the snails cluster and disperse rhythmically for up to several days. Graphical simulation and spectral analysis, preceded by first difference filtering, showed two components, one corresponding to the tidal, and the other to the diurnal rhythm. Snails kept under continuous light also exhibit a two-component rhythm of clustering and dispersal. In containers with high (40 cm) water level and under continuous light or normal day and night rhythm, the two-component rhythm may—for some unknown reason—be replaced by long fluctuations of dispersal, with clustering occurring in the afternoon only in snails kept under normal alternation of day and night. Snails kept for 24 h in dryness prior to putting them into containers with water, remain clustered (sometimes with an initial dispersal “shock”) for a long time, with subsequent slow fluctuations of clustering and dispersal, probably induced by the light conditions. Factors contributing to clustering and dispersal are: (1) increased rate of dropping from container walls at outgoing tide; (2) lack of locomotory activity at low tide; (3) negative geotaxis at incoming tide; (4) positive geotaxis at outgoing tide; (5) attraction of snails by other snails (positive chemotaxis) throughout the rhythm. Sudden exposure to light increases the rate of dropping from the container walls and may be a contributing factor to stronger clustering during the day. Factors 1–4 show endogenous periodicities.     

Sexual reproduction in the compound ascidian Diplosoma listerianum (Tunicata). I. Metamorphosis,storage and phagocytosis of sperm in female duct

In the colonial ascidian Diplosoma listerianum (collected in the Lagoon of Venice in 1986 and 1991), ovulating eggs detach themselves from the ovary wall and segregate in the tunic without exposure to seawater. With the aim of finding evidence of the pathway followed by spermatozoa to approach the oocyte, histological and ultrastructural observations were made. The results showed that the hollow ovary elongates in a fertilization canal which flanks the sperm duct and opens externally, next to the anus. Intercellular extended tight junctions isolate the lumen of this canal from the blood. Sperm were found in the fertilization canal, both free and in intracellular vacuoles of wandering cells (phagocytes). These sperm showed differences in comparison to those from the sperm duct, which are possible signs of a sperm reaction. The main modifications were shown by the dense groove, a narrow invagination of the plasmalemma bound to the nuclear envelope by dense material. In non-modified sperm, the groove runs spirally all along the head, while in those in the fertilization canal, it coils in the anterior half of the head, pressing back the long mitochondrion and endoplasmic tubules. These modifications were interpreted to be result of the release of the dense groove, considered to be a sort of stretched, contractile spring. The presence of sperm in the fertilization canal and particularly in phagocytes is discussed in relation to the capacity for exogenous sperm storage by zooids and to the necessity of waste clearance in order for successive waves of fresh sperm to be able to move towards fully grown oocytes. The origin of the female fertilization canal, the role of its tight junctions, and the complex sperm morphology are discussed as adaptations to internal fertilization in D. listerianum.     

Meiofauna and the thiobios in the east flower garden brine seep

Special hydrodynamic-chemical conditions at the East Flower Garden brine seep have provided the opportunity to examine the community structure of the thiobios and the oxybiotic-thiobiotic boundary. The boundary between the thiobios, whose population maxima occur in sulfidedependent chemoclines and which presumably have an ecologic requirement for sulfide, and the oxybios, which occur in oxidized zones above the chemocline, is controlled by sulfide, not oxygen. The boundary, which may not be at zero sulfide, is determined by a time-concentration phenomenon based on a dynamic interplay of sulfide and oxygen supply rates and the biota's sulfide detoxification capabilities. In Gollum's Canyon, where oxygen is plentiful, the boundary is at 10–40 μg-atoms·l^-1 sulfide. Total abundances of organisms at thiobiotic stations were comparable to total abundances at oxybiotic stations. Highest thiobiotic abundance was 202 051 organisms per m²; highest oxybiotic abundance was 240 572 organisms per m². The thiobios is dominated by representatives of the lower Bilateria (viz. Gnathostomulida, Platyhelminthes and Aschelminthes). These groups accounted for 50–80% of all the organisms present in the thiobiotic stations but less than 20% of all organisms in the oxybiotic stations. At two thiobiotic stations, over 50% of all organisms were gnathostomulids. Thiobios included macrofaunal as well as meiofaunal components. Peak abundances of amphipods were associated with the thiobiotic environment.     

Multiple paternity, sperm storage, and reproductive success of female and male painted turtles (Chrysemys picta) in nature

When females receive no direct benefits from multiple matings, concurrent multiple paternity is often explained by indirect genetic benefits to offspring. To examine such possibilities, we analyzed genetic paternity for 1,272 hatchlings, representing 227 clutches, from a nesting population of painted turtles (Chrysemys picta) on the Mississippi River. Goals were to quantify the incidence and distribution of concurrent multiple paternity across clutches, examine temporal patterns of sperm storage by females, and deduce the extent to which indirect benefits result from polyandrous female behaviors. Blood samples from adult males also allowed us to genetically identify the sires of surveyed clutches and to assess phenotypic variation associated with male fitness. From the genetic data, female and male reproductive success were deduced and then interpreted together with field data to evaluate possible effects of female mating behaviors and sire identity on offspring fitness. We document that more than 30% of the clutches were likely fathered by multiple males, and that presence of multiple paternity was positively correlated with clutch size. Furthermore, the data indicate that the second male to mate typically had high paternity precedence over the first.     

Induction and reversion of multixenobiotic resistance in the marine snail Monodonta turbinata

Many marine organisms inherently express the presence and function of a new defence mechanism, termed multixenobiotic resistance mechanism (MXRM) because of its similarity to multidrug resistance (MDR) found in tumor cell lines resistant to chemotherapeutic drugs. However, previously no information was available on a possible induction of the activity of MXRM in organisms living at polluted sites. The purpose of the present study was to demonstrate the inducibility of this defence mechanism in the marine snail Monodonta turbinata, an organism known to inhabit a wide range of environmental conditions existing along a pollution gradient. Specimens of M. turbinata used in the present study were collected from March 1993 through March 1994 near Rovinj, Croatia. The accumulation of generally labelled ³H-vincristine (³H-VCR) in the gills of snails living at an unpolluted site was 67% higher and very sensitive to verapamil, an inhibitor of P-glycoprotein activity in comparison to snails living at a polluted site. The accumulation of vincristine (VCR) in snails from a polluted site was reduced and insensitive to verapamil due to the induced state of P-glycoprotein, as was demonstrated immunochemically by the enhanced concentration of a 140 kDa protein cross-reacting with the polyclonal antibodies raised against mammalian P-glycoprotein. MXRM could be induced in snails from the unpolluted site to the level found in snails living at a polluted site either by transplantation for 3 d to a polluted site, or by exposure for 3 d to sea water polluted with Diesel-2 oil (hydrocarbon concentration equivalent to 50 ppb of Kuwait oil). Discovery of the inducibility of this new defence mechanism in aquatic organisms may have important implications in ecotoxicology, as was demonstrated by a 104% enhanced accumulation of VCR in the presence of the MXRM-inhibitor, the so-called chemosensitizer. Besides, a striking difference in the levels of MXRM-elements (the sensitivity of the accumulation rate of xenobiotics to verapamil, immunochemical expression of P-glycoprotein, and the sensitivity of the binding of xenobiotics on membrane vesicles to verapamil) found between specimens living at the polluted and unpolluted sites offers a new molecular biomarker for exposure to pollutants.     

The effect of postcopulatory male behaviour on ejaculate distribution within the female sperm storage organs of the fly,Dryomyza anilis (Diptera : Dryomyzidae)

Summary After sperm transfer, male Dryomyza anilis increase their fertilization success by tapping the female external genitalia with their claspers. In order to clarify the mechanism, we examined the effect of tapping movements on the distribution and fate of the male ejaculate within the female's sperm storage organs. The structure of the female internal genitalia is described and the volume of sperm found within the female is estimated. Experiments show that males place their sperm near the exit of the female's bursa copulatrix: most of the last male's sperm are expelled before oviposition and only 10–30% remain in the female. The results suggest that the mechanism by which male D. anilis gain last male sperm precedence is complex, and two possibilities are suggested. Offprint requests to: M. Otronen     

Daily cycling of the crystalline style in the omnivorous,Deposit-feeding estuarine snail Ilyanassa obsoleta

Crystalline styles are present in nearly all bivalve mollusks, and are known to undergo cyclic changes in size and/or occurrence in many species. A few gastropod species are known to possess styles, but such cycles as may exist are unknown or not well documented. Results reported here show that Ilyanassa obsoleta (Say) in a sandflat population lose and regain their crystalline styles each day. Styles were consistently low in frequency just after dawn. The presence of this kind of rhythm in a deposit-feeding gastropod has not previously been noted. It is suggested that this cycle may have played a key role in the evolution of this widespread and numerically abundant species.