Spermatozoan ultrastructure and spermatogenesis in aplousobranch ascidians,with some phylogenetic considerations

Spermatozoa and some stages of spermatogenesis were studied for four species of aplousobranch ascidians. Spermatozoa of Clavelina lepadiformis (Müller) (family Clavelinidae) are plesiomorphous in that they have apical acrosomal vesicles and a moderately elongated, cylindrical nucleus. The elongated mitochondrion is twisted ea. 11/2 times around the nucleus. In their ultrastructural morphology, C. lepadiformis sperm conform in some respects to the least-derived ascidian sperm, those of the phlebobranch ascidian Ciona intestinalis L.; however, the sperm of Clavelina lepadiformis have two apomorphies not shared with those of Ciona sp.: (1) the mitochondrion of Clavelina lepadiformis is long and spiralled along the entire nucleus rather than being comparatively compact and not at all helical; (2) the mitochondrial cristae are elongated parallel to the long axis of the nucleus, whereas in Ciona sp. sperm the cristae are unmodified. In Distaplia sp., Aplidium sp. and Synoicum pulmonaria (Ellis and Solander) the spermatozoa are more derived and consist of a proximal cylindrical and a distal corkscrew-like part. The mitochondrion in Distaplia sp. and Aplidium sp. contains electron-dense material and extends in a long thread around the nucleus. In S. pulmonaria the mitochondrion surrounds the anterior part of the nucleus in mature spermatozoa, and an elongated, dense structure displaying fine striation is enclosed in the mitochondrion in late spermatids. The sperm ultrastructural morphology observed in this study is consistent with the majority view that clavelinids are closest to the ancestral ascidian but is also consistent with other conclusions, particularly that the Cionidae are closest to the stem ascidian.     

Differential use of stored sperm during oviposition in the damselfly Calopteryx splendens xanthostoma (Charpentier)

Female Calopteryx splendens xanthostoma (Charpentier) (Odonata), have two discrete sperm storage organs, the bursa copulatrix and the paired spermathecae. We used the Random Amplified Polymorphic DNA(RAPD) profiling technique to determine from which sperm storage organ spermatozoa were used to fertilise eggs during behaviourally distinct oviposition bouts. During oviposition bouts following remating sperm from the bursa copulatrix are largely used to fertilise eggs, but when females avoid remating before an oviposition bout the sperm used to fertilise eggs are derived mainly from the paired spermathecae. Our observations suggest that female C. s. xanthostoma can avoid male paternity assurance mechanisms by a combination of behavioural and anatomical adaptation: the reason(s) they do so is unclear. Received: 28 November 1995/Accepted after revision: 5 August 1996     

Multiple insemination increases reproductive success of female Montandon’s newt (<Emphasis Type="Italic">Triturus montandoni</Emphasis>, Caudata,Salamandridae)

The adaptive significance of multiple matings for females is a matter of much controversy. In insects, supplying the female’s sperm reserves with portions of fresh spermatozoa may be the main function of multiple matings. This simple explanation may also be applied to other animals which produce large numbers of eggs over prolonged periods of time. We tested the fertility insurance hypothesis in Montandon’s newt (Triturus montandoni, Amphibia, Salamandridae). T. montandoni females are inseminated internally by spermatophores they have picked up, and subsequently lay eggs fertilized by spermatozoa released from the spermatheca. We compared the reproductive success of singly and multiply inseminated females of Montandon’s newt in the laboratory. Multiply inseminated females laid more eggs and had a lower percentage of non-developing eggs than females who mated only once. Our data suggest that remating increases the reproductive success of multiply inseminated females by replenishing sperm reserves in the spermatheca or by supplying females with fresh portions of spermatozoa with high fertilizing capacity. Received: 7 January 2000 / Revised: 13 September 2000 / Accepted: 7 October 2000     

Variations in sperm motility of the Atlantic herring Clupea harengus

Sperm characteristics and fertilization success were measured in two groups of Manx autumn-spawning herring, Clupea harengus L., captured 1 wk apart. Samples contained motile sperm for up to 45 min after activation (average 7 min). Individual spermatozoa were motile for up to 5 min. The activation of individual spermatozoa is probably phased, although the mechanism for the delay is not clear. This can account for the overall low levels of active sperm at any one time, the periodic fluctuations observed in the proportion of motile sperm, and for successful fertilization rates achieved using samples of apparently immotile sperm. The proportion of motile sperm was not affected by the presence of eggs, but the duration of motility was longer when sperm was activated in the presence of eggs rather than in seawater alone. Individual males differed significantly in the duration of motility in their sperm samples and in the fertilization rates achieved, but not in the proportion of motile sperm in each sample. There were no consistent changes in the characteristics of the spawners or the sperm between the two sample dates. Sperm motility and duration of motility were significantly correlated, but none of the sperm characteristics measured was significantly related to fertilization success. Received: 13 March 1999 / Accepted: 20 May 1999     

Storage of exogenous sperm by the compound ascidianDiplosoma listerianum

Experimental evidence is presented for the storage of exogenous sperm by zooids of the simultaneously hermaphroditic didemnid ascidianDiplosoma listerianum. Cross-fertilized zygotes can be produced up to 1 mo after a ramet is placed in reproductive isolation. Previously published observations, considered in the light of the present data, suggest that the lumen of the ovary is the site of sperm storage. This is believed to be the first report of post-transmission storage of sperm by an ascidian. Existing records of storage of exogenous sperm in other taxa rarely involve aquatic animals in which spermatozoa are transmitted between mates singly through the surrounding water, as inD. listerianum. The potential significance of sperm storage in the mating systems of sessile aquatic invertebrates is discussed.     

In vitro effects of lead on fatty acid composition,oxidative stress biomarkers and quality of ram spermatozoa

The sperm quality and several parameters associated with oxidative stress were evaluated in ram (Ovis aries) spermatozoa suspensions incubated with 0, 50 or 500 ng/mL Pb during 0, 1, 3 or 6 h. The presence of Pb during incubation, reduced the integrity of the acrosome, % sperm motility and integrity, and the functionality of membrane. On the contrary, % polyunsaturated fatty acids and the activities of glutathione peroxidase and superoxide dismutase in the spermatozoa suspensions were not affected by Pb. Moreover, glutathione peroxidase activity was decreased and the fatty acid composition changed due to the relative increase in % stearic acid during the incubation time independently on Pb presence. Data showed that several effects of Pb on sperm quality usually observed in vivo also occurred in vitro, but without any relationship with oxidative stress biomarkers.     

Degrees of vacuolation of the absorptive intestinal cells of five Sagitta (Chaetognatha) species: possible ecophysiological implications

The cell organization and ultrastructure of the intestine have been studied in five species of Sagitta, one epiplanktonic (S. bipunctata Quoy and Gaimard, 1827), four mesoplanktonic (S. megalophthalma Dallot and Ducret, 1969, S. decipiens Fowler, 1905, S. minima Grassi, 1881, S. zetesios Fowler, 1905). The intestinal epithelium is composed of two ciliated cell types. The first (S-cell) principally occurs in the anterior intestine and represents typical secretory cells. The second (A-cell) occurs in the median and posterior intestine and displays ultrastructural features involved in absorption and intracellular digestion, that is, coated pits, endocytotic vesicles, cytoplasmic tubules, and two distinct types of digestive vacuoles. Whereas S-cells exhibit few ultrastructural differences among the five species, the vacuolar volume of the A-cells located in the mid portion of the intestine is higher in the mesoplanktonic species. In S. zetesios and S. megalophthalma, each side of the median intestine comprises several hypervacuolated A-cells visible in transverse section; their degree of vacuolation is inversely proportional to their number. The increased volume of the intestinal vacuoles is most marked in S. decipiens and S. minima; in both these species, each lateral side of the median intestine displays a single ultravacuolated A-cell. The possible ecophysiological implications of intestinal vacuoles are discussed in relation to patterns of the vertical distribution of several Sagitta species. The vacuoles are presumed to regulate buoyancy, enabling the mesoplanktonic species to make vertical diel migrations. Received: 22 January 2000 / Accepted: 28 July 2000     

Predicting the mating system from phenotypic correlations between life-history and sperm quality traits in the Alpine whitefish <Emphasis Type="Italic">Coregonus zugensis</Emphasis>

The mating behavior and reproductive strategies of Alpine whitefish like Coregonus zugensis (Nüsslin) are poorly understood, probably because they spawn in deep water where direct observations are difficult. In this study, we interpret life-history and sperm quality traits of fish that we caught from their spawning place. We found that males invest heavily into gonadal tissue (up to 5.6% of their body weight), which is, in comparison to other fish, consistent with external fertilization, distinct pairing and moderate to high communal spawning, or no pairing and low to moderate communal spawning. Sperm competition theory and recent experimental studies on other salmonids predict that males optimize ejaculate characteristics in relation to the costs of sperm and the level of competition they have to expect: dominant males are predicted to invest less into ejaculate quality and to have slower spermatozoa than subdominant males. We found that spermatozoa of older males are slower than those of younger males. Moreover, older males have larger breeding tubercles, a secondary sexual trait that has, in some previous studies, been found to be linked to good condition and to good genetic quality. Our results suggest that C. zugensis has age-linked reproductive strategies, that multimale spawning is common, i.e., that sperm competition plays a significant role, and that older males are on average dominant over younger males at the spawning place.     

Male characteristics, sperm traits, and reproductive success in winter-spawning Celtic Sea Atlantic herring, Clupea harengus

The relationship between sperm characteristics and reproductive success was examined in male herring, Clupea harengus L. Males were categorised as being first-time or repeat spawners on the basis of their age; they were also grouped according to whether their sperm were immediately active and exhibited forward motion on contact with seawater (FM) or had little or only vibratory motion (VM). Unlike the Pacific herring C. pallasii Valencienes, Atlantic herring sperm is usually motile on contact with seawater. The age, weight and gonadosomatic index (testes mass as a percentage of somatic mass = GSI) were measured and used as characteristics for individual fish. Sperm traits measured were (1) adenosine triphosphate (ATP) concentration, (2) sperm count, (3) duration of sperm motility. Reproductive success for each male was estimated from the fertilisation rate and from the length of larvae at hatching. Fertilisation rates for all fish were generally >80%. The ATP concentration of non-activated spermatozoa was negatively correlated with fertilisation rate. Among repeat spawners, fish with higher GSIs produced larvae that were larger at hatching. Although VM sperm fertilised eggs at rates equivalent to fertilisation by FM sperm, the larvae produced by VM sperm were significantly smaller at hatching. Larval length tended to increase in parallel with the duration of sperm motility, but the relationship was not significant in these tests. The results did not indicate any age or size pattern to spawning readiness in male herring. Sperm that are not yet ready to be shed are not fully motile on contact with seawater, but are still capable of fertilising eggs that hatch successfully. There is likely to be a progression of males which come into spawning readiness within a spawning shoal; therefore it is possible that paternal influences would result in a progressive decrease in larval size over the spawning period in winter-spawning Celtic Sea herring. Received: 22 November 1997 / Accepted: 8 June 1998     

Spermatogenesis inPyrosoma atlanticum (Tunicata: Thaliacea: Pyrosomatida): Implications for tunicate phylogeny

Colonies ofPyrosoma atlanticum were collected by submersible in October 1988 in the Caribbean Sea, and testes were studied by electron microscopy. Spermatogonia, spermatocytes and early spermatids have two centrioles. The proximal centriole subsequently disappears, its remains apparently persisting in the spermatozoon as dense material adjacent to the distal centriole, which gives rise to the axoneme. At the tip of early spermatids are several 50 nm proacrosomal vesicles, which disappear leaving no trace in early elongating spermatids. The spermatozoon lacks an acrosome and has a head 35µm long. The head is differentiated into a bulbous posterior portion 5µm long × 1µm wide, a thinner anterior portion 25µm long tapering from a width of 0.7µm to a width of 0.4µm, and a very thin anterior extension 5µm long × 0.5µm wide. At the start of elongation, the anterior extension begins to form just lateral to the proacrosomal vesicles as a spiral projection comprising part of the nucleus, covered by a thin sheath of cytoplasm. This sheath of cytoplasm undergoes a complex differentiation. Ultimately, the nucleus in the anterior extension is overlain by two membrane-bound sheaths of cytoplasm connected by a spiral flange of cytoplasm. Between these two sheaths is a spiral space, open to the exterior through a subterminal pore near the sperm tip. In early spermatids the mitochondria fuse into a single mitochondrion, which remains lateral to the nucleus. The cristae become modified late in spermatogenesis. Throughout elongation of the spermatid there are patches of dense material between the nucleus and mitochondrion. A manchette of microtubules transiently encircles the thin anterior portion of the nucleus during the last phase of elongation. A manchette is not present during most of elongation. In the spermatozoon the mitochondrion, which has reticulate cristae, spirals a few times about the nucleus and extends from the junction between the bulbous portion and the thinner anterior portion of the nucleus to the junction between the thinner anterior portion and the nuclear extension. Spermatogenesis inP. atlanticum, compared to that in other tunicates, most closely resembles that in colonial ascidians, and supports the majority view that pyrosomes arose from aplousobranch ascidians that lost their attachment to the substratum. Pyrosome sperm are more highly derived than doliolid sperm, which have an acrosome that is probably capable of exocytosis. When salp and pyrosome sperm are compared, both are highly derived, but neither shares any apomorphies with the other that it does not share with at least one other tunicate order. Thus, sperm morphology does not support the majority view that pyrosomes gave rise to doliolids and neither confirms nor denies the idea that pyrosomes are intermediate between aplousobranch ascidians and salps. Therefore, it is likely that the class Thaliacea is polyphyletic, with doliolids arising very early from the ascidian lineage and with salps and pyrosomes arising somewhat later.     

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.     

Spermatozoon structure of some North American prosobranchs from the families Lottiidae (Patellogastropoda) and Fissurellidae (Archaeogastropoda)

The spermatozoa of four species of the patellogastropod family Lottiidae (Lottia pelta, L. digitalis, L. strigatella, Tectura scutum) and one species of the archaeogastropod family Fissurellidae (Diodora aspera) were examined in 1990 using transmission electron microscopy. All have primitive or ect-aquasperm, typical of invertebrates using external fertilization. Sperm of the lottiid limpets are characterized by a 5 to 9 m-long head composed of a conical acrosome which constitutes >50% of the head length, and a cylindrical nucleus. The acrosome of all species of lottiids is differentiated internally, and has a posterior invagination 0.9 to 1 m in depth, into which an elongate acrosomal lobe protrudes. Between the posterior acrosomal lobe and the nucleus, the subacrosomal material is aggregated as a fibrous column. The midpiece of the sperm has a ring of 4 to 5 spherical mitochondria of 0.6 m diam, posterior to which is a collar of cytoplasm 1 m long, which sheaths the anterior portion of the axoneme. The size and morphology of the acrosome and large cytoplasmic collar clearly distinguish the spermatozoa of the Lottiidae from other families of Patellogastropoda. The sperm of D. aspera (Fissurellidae) is typical of the family of archaeogastropod; the head has a length to breadth ratio of 4:1, and the cylindrical nucleus is capped by a small acrosome, <25% of the total head length, which is deeply invaginated.     

Acrosome differentiation and the acrosome reaction in ascidian spermatozoa:<Emphasis Type="Italic"> Ascidiella aspersa</Emphasis> and<Emphasis Type="Italic"> Ascidia mentula</Emphasis> with some implications for tunicate phylogeny

The spermatozoa of both Ascidiella aspersa and Ascidia mentula have architectural features characteristic of ascidian spermatozoa that have previously been described. They have an elongated head (7 µm long for A. aspersa and 4 µm long for A. mentula), a single mitochondrion that is applied laterally to the nucleus and lacks a midpiece. The acrosome of A. aspersa spermatozoa is a flattened vesicle, about 200 nm×100 nm×40 nm (length, width and height). The acrosome of A. mentula spermatozoa consists of multiple vesicles; they are about 50 nm×50 nm×40 nm (length, width and height). During spermiogenesis in both species, several proacrosomal vesicles (50–70 nm in diameter) appear in a blister at the future apex of the spermatid. In A. aspersa, these vesicles fuse with each other to form a single acrosomal vesicle, while in A. mentula these vesicles do not fuse with each other, and form multiple acrosomal vesicles. In A. aspersa spermatozoa, calcium ionophore A23187 induces the acrosome reaction in which membrane fusion between the acrosomal apical membrane and the overlying sperm plasma membrane occurs along the peripheral margin of the acrosome, resulting in the release of a hybrid, membrane-bound, small vesicle. In A. mentula, multiple acrosomal vesicles disappear by releasing small vesicles after treatment with the calcium ionophore A23187; this also appears to be an acrosome reaction. This paper discusses the way in which acrosome structure and function may have changed during the evolution of the Tunicata.Communicated by T. Ikeda, Hakodate     

Bisphenol S induces oxidative stress and DNA damage in rat spermatozoa in vitro and disrupts daily sperm production in vivo

Bisphenol S has been introduced into some industrial applications and it may act as a xeno-estrogen that can alter endocrine functions and reproduction. The present study was carried out to examine the effect of bisphenol S exposure on oxidative stress, generation of reactive oxygen species, and DNA integrity in rat spermatozoa in vitro and daily sperm production and sperm DNA damage in vivo. Sperm were incubated with bisphenol S at concentrations of 0.5, 1, 10, and 100 µg/L. At the highest concentration, bisphenol S induced formation of reactive oxygen species, caused lipid peroxidation, affected superoxide dismutase levels, and increased DNA fragmentation. Adult rats were exposed to doses of 0.5, 5, 25, and 50 µg/kg/d for 28 days. Decrease in daily sperm production and an increase in sperm DNA damage was observed at the highest dose in the 50 µg/kg/d treated group, but sperm motility was not reduced.     

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.     

Sperm exchange in a simultaneous hermaphrodite

Many simultaneously hermaphroditic animals cross-inseminate when copulating, even though unilateral sperm exchange is functionally possible. The question is why reciprocity is the rule. This study addresses sperm exchange in a hermaphroditic planarian flatworm, Dugesia polychroa, in which self-fertilisation does not occur. We investigated the availability of self sperm for inseminating a partner, how self sperm affects the likelihood of sperm donation, and whether sperm donation depends on reciprocation by the partner. The amount of self sperm depended on body size, duration of isolation and experimental date, and animals that had more self sperm were more likely to mate. Depletion of allosperm (sperm received from partners) played only an indirect role, suggesting that animals copulate more to donate sperm rather than to replenish allosperm reserves. Among 60 copulating pairs, reciprocal insemination was more common (63.3%) than unilateral transfer (18.3%). A surprising 18.3% did not exchange sperm in either direction. Individuals were more likely to behave like their partners, resulting in more symmetrical matings (either reciprocity or no sperm exchange) than asymmetrical matings (unilateral sperm donation). The amount of self sperm donated during a copulation depended mainly on the amount available before the copulation and not on the amount received from the partner. The results suggest that conditional sperm exchange, or sperm trading, takes place in D. polychroa. Received: 31 May 1997 / Accepted after revision: 1 December 1997     

Tissular localization of the digestive proteases in the sedentary polychaetous annelid Sabellaria alveolata

The digestive tract of Sabellaria alveolata (L.) is partitioned into 4 main regions: oesophagus, gizzard, intestine and rectum. Histoenzymologic study of the repartition of the protease activities along the digestive tract has shown that the proteases are secreted in the anterior intestine, where numerous serous gland cells are present. These cells display many histochemical and cytological characteristics similar to those of the gland cells previously described in digestive organs of other polychaetes. The role of the other regions of the digestive tract is discussed.     

Spermiogenesis and spermatozoa in the relict bivalve genus Neotrigonia: relevance to trigonioid relationships,particularly Unionoidea

Spermiogenesis and spermatozoa in the relict trigonioid genus Neotrigonia are examined ultrastructurally. Mature Neotrigonia spp. spermatozoa exhibit the following features: (1) a blunt-conical nucleus; (2) an acrosomal complex composed of discoidal vesicles (9 to 15) arranged as a thin layer over the nuclear apex; (3) five (rarely four) spherical mitochondria positioned in depressions at the base of the nucleus; (4) proximal and distal centrioles (surrounded by the mitochondria); (5) a satellite complex anchoring the distal centriole and flagellum to the plasma membrane; and (6) a single flagellum. Spermatozoa of unionoids examined to date (Unionidae and Hyriidae only) exhibit similar features, including a nucleus with a blunt apex capped by a thin acrosomal complex (showing up to three vesicles). Among the Bivalvia, only Neotrigonia spp. and unionoid sperm possess an acrosomal complex composed of multiple vesicles. These data suggest that the Trigonioidea and Unionoidea are phylogenetically linked, but further work particularly on primitive unionoids is required to determine if this relationship is one of common ancestry or whether the Unionoidea have in fact been derived from the Trigonioidea.     

Levels of polyandry and intracolonial genetic relationships in Apis florea

DNA was extracted from worker and drone pupae of each of five colonies of the dwarf honey bee Apis florea. Polymerase chain reactions (PCR) were conducted on DNA extracts using five sets of primers known to amplify microsatellite loci in A. mellifera. Based on microsatellite allele distributions, queens of the five colonies mated with at least 5–14 drones. This is up to 3 times previous maximum estimates obtained from sperm counts. The discrepancy between sperm count and microsatellite estimates of the number of matings in A. florea suggests that despite direct injection of semen into the spermatheacal duct, either A. florea drones inject only a small proportion of their semen, or queens are able to rapidly expel excess semen after mating. A model of sexual selection (first proposed by Koeniger and Koeniger) is discussed in which males attempt to gain reproductive dominance by increasing ejaculate volume and direct injection of spermatozoa into the spermatheca, while queens attempt to maintain polyandry by retaining only a small fraction of each male's ejaculate. It is shown, at least in this limited sample, that the effective number of matings is lower in A. florea than in A. mellifera.     

Function of the mating plug in Drosophila hibisci Bock

The mating plug in Drosophila hibisci Bock is a firm, gelatinous structure that forms within the female’s uterus during copulation. Two non-mutually exclusive hypotheses for the function of the plug were evaluated. The plug may serve as a nutritional gift that females digest, using the constituents for somatic maintenance or to provision eggs as they mature within the ovaries. Alternatively, the plug may act as a chastity enforcement device by preventing subsequent copulations, and thereby reducing sperm competition. Plug size did not decrease within females over a period of 2 days, and dietary treatment in females did not affect plug size. The extent of ovarian provisioning was also not related to plug size. These results weaken the nutritional gift hypothesis. In contrast, the probability of a second copulation increased sharply with an experimental decrease in plug size. Moreover, females with plugs experimentally reduced in size were courted significantly more and mated significantly faster than females with larger plugs. These results support the chastity enforcement hypothesis. The plug retains the ejaculate and concentrates sperm at the anterior end of the uterus near the apertures of the sperm storage organs. The presence of the plug thus probably facilitates the movement of sperm into storage by retaining sperm at the anterior end of the uterus near the apertures of the sperm storage organs, which may be especially important for D. hibisci, in which sperm length is nearly twofold greater than ventral receptacle length. Matings with newly eclosed virgin females were significantly shorter than with older virgins, and copulations with the younger virgins ended more often without any sperm having yet entered into storage. The effectiveness of the plug in safeguarding a male’s ejaculate may have favoured the evolution of shortened copula durations with young virgins. One fitness advantage of shortened copula duration could be time liberated for the pursuit of further mating opportunities. Received: 12 May 2000 / Revised: 22 September 2000 / Accepted: 15 October 2000