Sperm transfer,displacement and precedence in Ischnura graellsii (Odonata : Coenagrionidae)

Summary Copulation in Ischnura graellsii may be divided into three stages, according to the movements and position of the male's abdomen. We measured sperm volumes in males and females interrupted at different phases of copulation in laboratory-reared and field specimens. The results showed that males remove sperm from the female during stage I, and do not transfer sperm until stage Il of the copulation. In the field females interrupted during stage I of copulation had less sperm than postcopula females, and the volume of sperm in laboratory females mated once or twice was similar. These results suggest that males can remove most of the sperm during stage I of copulation. Preparations of in-copula specimens showed the horns of the penis (used to remove sperm) inside the bursa copulatoox and the spermatheca. Therefore males can remove sperm from both organs, in contrast to the other Ischnura species so far studied, where males can empty only the bursa. The length of these horns is positively correlated with male body length, and there are significant differences in length between the left and right horns of individual males. This suggests great variability in the male's ability to remove sperm. On the other hand, ejaculate volume is positively related to male and female size, and negatively to male age. Males are likely to be able to detect the presence of sperm in females: if the effect of population density and time of start of copulation are taken into account, copulations are longer with mated than with virgin females. Using genetic markers, sperm precedence was studied by rearing the female offspring of 6 females mated with two males of different genotype. In 5 out of 6 crosses, the second male fertilized all the eggs laid by the female in her first clutch. On average, the following clutches were progressively more fertilized by the first male, but there were striking differences between crosses. These differences are probably due to the variability in the amount of sperm transferred and/or removed. Offprint requests to: A. Cordero     

Copulation duration and sperm competition in white-faced dragonflies (Leucorrhinia intacta; Odonata: Libellulidae)

Summary In many odonates, females mate with more than one male while laying a single clutch of eggs. We studied paternity of eggs laid by remated females of Leucorrhinia intacta, a small libellulid dragonfly, at a pond near Syracuse, NY, USA. The probability of a female remating is a function of male density on the pond. The length of copulations differs considerably among males active on the study pond at the same time. Much of this variation was correlated with differences in mating tactics of the males; copulations by males that stayed on their territories during copulation were shorter than those by other males (Fig. 2). Eggs collected from females mated to irradiated, sterile males and to free-living, fertile males indicated that the average paternity expectation was higher for long than for short copulations, and that the variance in paternity expectation was lower for long than for short copulations. Some possible causes of the high variation in paternity at low copulation durations and possible reasons for differences in copulation duration between male mating tactics are discussed.     

Variation in copulation duration and the resultant degree of sperm removal inOrthetrum cancellatum (L.) (Libellulidae: Odonata)

Summary Orthetrum cancellatum showed two distinct copulation durations at a study site in the south of France. These different durations are correlated with different degrees of sperm displacement from the female sperm stores; long copulations (894±142 s) resulting in almost 100% sperm removal, and short copulations (21.0±13.5 s) resulting in 10–15% removal. The difference in copulation duration is also related to the site of copulation and to the relative age of the copulating males. At oviposition sites males who gained copulations were relatively young and copulated for long periods. The difference in the duration of copulation and the degree of sperm removal is discussed with reference to the female habit of ovipositing before remating during oviposition episodes, and the possibility of differences in the rate at which males encounter receptive females at the two sites.     

Dominions and site-fixed aggressive behavior in breeding male Leucorrhinia intacta (Odonata: Libellulidae)

Summary We report aggressive spacing behavior in male dragonflies, Leucorrhinia intacta, that is characterized by variations in the probability of chasing conspecific male intruders within a defined area around a male's perch. The chase probability depends on the total intruder pressure and the behavior and distance of the intruder from the perched male (Fig. 2). This nonexclusive, site-fixed area has been called a dominion.We also examined the distribution of intruders among the various behavior-distance categories. Chase rates (per 15 min) were correlated mostly with number of intruders that hovered (rather than flew) close to the territorial male. Finally, we examined the impact of the changing chase probabilities on the variation in aggressive interactions as a function of intruder pressure. We conclude that, to some extent, territorial males stabilized defense costs across a variety of intruder pressures by maintaining dominions.     

Copulation duration and sperm precedence in the stalk-eyed fly Cyrtodiopsis whitei (Diptera : Diopsidae)

Summary By means of field observations and laboratory experiments on the Malaysian stalk-eyed fly Cyrtodiopsis whitei we examined the consequences of variation in copulation duration for sperm competition. In this sexually dimorphic species over 90% of all copulations occur in nocturnal aggregations with from one to four males and up to 24 females. Copulation duration observed in both the field and the laboratory exhibited a bimodal distribution with peaks at 10 and 50 s. In the field short copulations less than 30 s long occurred frequently when more than one male was present in an aggregation but most were not the direct result of male interference. Sperm counts from female spermathecae after artificial interruptions indicated sperm are not transferred during the first 40 s of a copulation. When solitary males mated up to five times in succession to virgin females, short copulations did not occur, nor was the number of sperm transferred reduced. However, short copulations did occur when we mated isolated females within 6 min of a previous copulation. By mating irradiated and non-irradiated males in reciprocal pairs we discovered that C. whitei exhibits both first-male sperm precedence and sperm mixing. More than half of the females mated first to sterile and then to fertile males failed to produce offspring. Such variation in copulation duration and sperm precedence is consistent with male placement and detection of a spermatophore that acts as a temporary mating plug. Our data suggest that those male C. whitei which successfully defend large aggregations of females reduce sperm waste and competition by preferentially transferring sperm to females that have not mated recently. Correspondence to: G.S. Wilkinson     

Sperm storage and mating in the deep-sea squid Taningia danae Joubin, 1931 (Oegopsida: Octopoteuthidae)

Spermatangium implantation is reported in the large oceanic squid Taningia danae, based on ten mated females from the stomachs of sperm whales. Implanted spermatangia were located in the mantle, head and neck (on both sides) or above the nuchal cartilage, under the neck collar and were often associated with incisions. These cuts ranged from 30 to 65 mm in length and were probably made by males, using the beak or arm hooks. This is the first time wounds facilitating spermatangium storage have been observed in the internal muscle layers (rather than external, as observed in some other species of squid). The implications of these observations for the mating behavior of the rarely encountered squid T. danae are discussed.     

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.     

Heavy metal contamination and physiological variability in the Brazilian mangrove crabs Ucides cordatus and Callinectes danae (Crustacea: Decapoda)

 Physiological studies were made on the crabs Ucides cordatus (L.) and Callinectes danae sampled from populations living in “polluted” mangroves on the southeast littoral of Brazil. Analysis of Cu, Cd, Zn, and Fe of sediments and crab tissues showed interspecific differences in tissue concentrations, and significantly higher levels of Cu, Cd, and Zn in “polluted” populations compared to “unpolluted” crabs living in uncontaminated mangrove in the same geographical area. Individuals of both species from the polluted site showed significantly greater capacities for regulating blood osmotic concentrations at low salinity (9‰). However, U. cordatus showed a reduced hypo-regulatory ability in 34‰S. Differences in ionoregulation were also seen. “Polluted”C. danae showed significantly higher Na/ K-ATPase levels in posterior gills compared to “unpolluted” crabs. Oxygen consumption rates (M˙ _O2) were elevated in U. cordatus, but depressed in C. danae from the “polluted” population. Individuals of both species from this site showed significantly lower O:N ratios, mainly because of an increased net efflux of ammonia. Adenylate energy charge (AEC) values of muscle and hepatopancreas in “unpolluted” and “polluted” populations of both species were not significantly different. These physiological differences are discussed in relation to the known acute physiological and metabolic effects of heavy metals in crustaceans, and interpretated in the light of possible adaptive changes following long-term exposure to contamination. Received: 6 August 1999 / Accepted: 22 June 2000     

Variation in copulation duration in Mnais pruinosa pruinosa Selys (Odonata: Calopterygidae)

Summary Males of the damselfly Mnais pruinosa pruinosa were observed to use three different tactics to secure mates. The mean duration of copulation differed between the three observed tatics and resulted in varying degrees of sperm removal and insemination. It is shown that the last male to mate had almost 100% sperm precedence immediately after copulation regardless of the duration of copulation and therefore the quantity of sperm removed. In situations where less than 100% of rivals' sperm was removed the sperm from different males mixed within the female sperm storage organs over a period of about 6 days: sperm mixing produced variation in last male sperm precedence. The significance of sperm mixing in M. p. pruinosa is discussed in the context of the observed matesecuring tactics and the frequent female habit (37% of observations) of ovipositing without remating during an oviposition bout.     

Variation in copulation duration in Mnais pruinosa pruinosa Selys (Odonata: Calopterygidae)

Summary The Japanese calopterygid damselfly Mnais pruinosa pruinosa shows three distinct copulation durations (Siva-Jothy and Tsubaki 1989). A variety of factors which might influence copulation duration were investigated. Results indicate that the temperature in the vicinity of copulating pairs, the rate at which males encountered receptive females, territory quality, the rate of agonistic interactions between males, female gravidity and female willingness to oviposit have no effect on copulation duration. The most significant factor was the location of the site at which males captured their mates. We suggest that under natural conditions the location of the capture site provides males with reliable, indirect information about a female's intention to oviposit: it is important for males of this species to acquire such information since sperm precedence, and therefore reproductive success, is in part dependant on the interval experienced by females between copulation and subsequent oviposition. The results are discussed and it is suggested that the three observed mate-securing tactics of M.p. pruinosa are facultative and information-dependant.     

Lifetime mating success in a natural population of the damselfly,Enallagma hageni (Walsh) (Odonata: Coenagrionidae)

Summary Variance in lifetime mating success was measured for individuals of a population of Enallagma hageni, a non-territorial damselfly in northern Michigan. E. hageni is an explosive breeder with scramble competition for mates. Highly skewed operational sex ratios resulted in intense male-male competition which took the form of interference with tandem pairs. 41% of the males failed to mate in their lifetime as opposed to only 3.6% mating failure in females. The effect on mating success of size, age, longevity, and time spent at the breeding site were investigated. Intermediate sized males obtained the most matings, and male lifetime mating success was highly correlated with longevity.     

Choice and utilization of oviposition sites by female Calopteryx maculata (Odonata: Calopterygidae)

Summary Females of the damselfly Calopteryx maculata (de Beauvois) initially choose the larger of a pair of adjacent oviposition sites, about 70% of the time (Table 1), or whichever of two equal sized sites had other ovipositing females on it (about 88% of the time-Table 2). These criteria for initial choice between a pair of sites also interact. Incoming females generally (57 to 74% of the time-Table 3) joined others on the small site rather than ovipositing alone at the adjacent, bigger site. When pairs of large and small sites were replicated across eight locations, there were nonsignificant trends towards greater utilization (eggs laid) of the larger of a pair of oviposition sites within locations (Table 4). The lack of agreement between initial choice and utilization shows that other factors besides size are important in the choice and use of oviposition sites. These include disturbance by males, the presence of other females and choice criteria that can only be assessed during oviposition. When all sites at the eight locations were equal in size, there was considerable day to day and location to location variation in eggs laid (Fig. 1). Viewed over periods of several days, some sites are obviously less attractive than others in terms of cumulative numbers of eggs laid at them. When the amount of vegetation was varied among locations, those with the bigger oviposition sites were used more often, somtimes significantly so, but there were also significant reversals (small sites used more often) (Table 5, Fig. 1). Thus, there is no simple effect of size on the utilization of oviposition sites by Calopteryx maculata females, despite a clear tendency for females to make initial choices based on this criterion. The considerable among and within location variation in number of eggs laid may reflect additional choice criteria or the interaction of size, the presence of other females, disturbance, and location.     

Sperm competition in the damselfly Enallagma hageni Walsh (Odonata: Coenagrionidae): benefits of multiple mating to males and females

Summary Sperm competition was investigated in the non-territorial damselfly, Enallagma hageni. Using irradiated (sterile) male techniques, the last male to mate was found to fertilize up to 95% of the eggs of the first clutch laid after mating. Dissection of females collected before, during, and after copula showed that a male removes a maximum of 87% of the sperm by volume of a previous mate. These data verify an earlier estimate of lifetime reproductive success in this species which was based on mating success, and suggest that indirect dissection methods offer minimum estimates of sperm precedence. Male E. hageni have ample opportunity to benefit from sperm precedence, since at least 10% of the receptive females encountered had already mated once that day, but still contained complete or partial clutches of eggs. Female E. hageni benefit directly from high sperm precedence because it allows them to exchange matings for guarding service by males during oviposition bouts under water.     

Underwater oviposition in a damselfly (Odonata: Coenagrionidae) favors male vigilance,and multiple mating by females

Summary Female Enallagma hageni oviposit underwater where they are inaccessible to males. I demonstrate that males guard submerged females rather than perch sites, and are behaviorally distinct from lone males at the water. In contrast to lone males, which always attempt to copulate with females presented to them, guarding males exhibit a conditional latency to remating which corresponds closely to the time required by females to oviposit a complete clutch of eggs. By ovipositing underwater, females decrease the risk that their eggs become exposed. Risks associated with submerged oviposition favor both mate guarding, and multiple, within-clutch matings by females. Both guarding mates and lone males rescue females that float on the water surface as the result of improper resurfacing. Such behavior reduces the mortality risk to females from 0.06 to 0.02 per oviposition bout. By remating between bouts, females benefit from the additional vigilance of lone males, who rescue floating females 1.4 times as often as original mates. A second consequence of multiple mating is an increase in the selective advantage of vigilance by mates. Because receptive females become scarce by early afternoon, whereas male density remains high, a male has little (3%) chance of encountering a second receptive female that day. However, he incurs a 42% risk of losing fertilizations if he abandons a mate. For male E. hageni mate guarding functions in the context of both natural and sexual selection. It insures that a mate lives to lay a complete egg clutch in addition to protecting a male's sperm investment.     

Variation in sperm morphometry and sperm competition among barn swallow (Hirundo rustica) populations

Spermatozoa vary greatly in size and shape among species across the animal kingdom. Postcopulatory sexual selection is thought to be the major evolutionary force driving this diversity. In contrast, less is known about how sperm size varies among populations of the same species. Here, we investigate geographic variation in sperm size in barn swallows Hirundo rustica, a socially monogamous passerine with a wide Holarctic breeding distribution. We included samples from seven populations and three subspecies: five populations of ssp. rustica in Europe (Czech, Italy, Norway, Spain, and Ukraine), one population of ssp. transitiva in Israel, and one population of ssp. erythrogaster in Canada. All sperm traits (head length, midpiece length, tail length, and total length) varied significantly among populations. The variation among the European rustica populations was much lower than the differences among subspecies, indicating that sperm traits reflect phylogenetic distance. We also performed a test of the relationship between the coefficient of between-male variation in total sperm length and extrapair paternity levels across different populations within a species. Recent studies have found a strong negative relationship between sperm size variation and extrapair paternity among species. Here, we show a similar negative relationship among six barn swallow populations, which suggests that the variance in male sperm length in a population is shaped by the strength of stabilizing postcopulatory sexual selection.     

Ultrastructure of sperm and spermathecae in Micromaldane spp. (Polychaeta: Capitellida: Maldanidae)

Assessing the possibility that external fertilization has re-evolved requires the study of monophyletic groups that exhibit various reproductive methods. Maldanid polychaetes show a range of reproductive mechanisms, though previous studies of reproduction have hitherto been restricted to larger species with external fertilization. Micromaldane pamelae Rouse and M. nutricula Rouse are small, gonochoristic maldanids that brood directly developing larvae. Both species have sperm with elongate nuclei and an acrosome extending down each side of the anterior end of the nucleus. A true midpiece is absent; two mitochondria extend along the posterior region of the nucleus. Spermatids develop synchronously in large clusters connected by a cytophore. In M. pamelae sperm are released into the water as spermatozeugmata. These are comprised of clusters of sperm with their tails oriented to the centre and the sperm heads facing outwards. Females of M. pamelae and M. nutricula bear pairs of spermathecae ventrally (M. pamelae three pairs, between setigers 10 and 11, 11 and 12 and 12 and 13 and M. nutricula two pairs, between setigers 10 and 11 and setigers 11 and 12). The blind sacs are epidermal invaginations bound closely together. The entrance to each spermatheca may only be 1 to 2 m across with each spermatheca holding several hundred sperm. This represents the first detailed study of spermathecae in the Capitellida. The occurrence and structure of spermathecae and spermatozeugmata in other groups are discussed and compared with Micromaldane spp. Comparisons are made with non-polychaetes with the purpose of discussing functional aspects of reproductive mechanisms in marine metazoans in general. Elongate sperm nuclei are associated with sperm storage and/or large egg size. The lack of an elongate sperm midpiece may be an indicator of having to swim in water but does not contraindicate sperm storage. Spermatozeugmata may serve as an indication of sperm storage and brooding of larvae. Speculations on the phylogenetic significance of these reproductive features are limited by the fact that supposedly modified (i.e., derived) states may reflect functional/structura, constraints of small body size.     

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.     

Female mediation of sperm competition in the millipede Alloporus uncinatus (Diplopoda: Spirostreptidae)

The tropical spirostreptid millipede Alloporus uncinatus has a polygynandrous mating pattern that is apparently shaped by sperm competition (Telford and Dangerfield 1990, 1993a). In the present study radioisotopic labelling of ejaculates was used to quantify the temporal effects of double mating sequences on sperm precedence patterns. Ejaculates of successive males mix completely within the sperm storage organs and are concentrated close to the site of fertilisation, the spermathecal-oviduct junction. When matings follow each other immediately (experiment 1), equal volumes of the ejaculates of each male are stored within the spermathecae, and both males have equal probability of paternity. Where matings are separated by a 24-h delay (experiment 2), the volumetric contribution of the first male is reduced by 54.8 % and last male precedence operates. Although the distal ends of the gonopods play a primary role in the redistribution of ejaculates (Barnett and Telford 1994), by using a single mating sequence we show that they are not responsible for the reduction in ejaculate volume. The most likely explanation is absorption and/or ejection of the first male's ejaculate by the female. Genital functional morphology is used to reconstruct the mechanism of ejaculate distribution.     

Timing of sperm shedding and release of aggregates in the salp Thalia democratica (Urochordata: Thaliacea)

Many holoplanktonic forms, such as medusae, spawn in response to environmental cues such as light or temperature. Few observations of the time of reproduction have been made in salps. Small groups of mature aggregates of the salp Thalia democratica (Forskal, 1775), collected off the Great Barrier Reef of Australia, were held under constant light in early to mid-December in the years 1990, 1993 and 1994. Sperm were shed between 0300 and 0600 hrs each day with a mean time of about 0450 hrs for all individuals for all years combined. Solitary stages released chains of fertilizable female individuals between 0500 and 0700 hrs each day, with a mean time of 0540 hrs. Field observations suggested that similar timing occurred in nature. Advancing the time of dusk from 1900 to 1700 or 1800 hrs. did not significantly alter the mean time of sperm shedding or chain release. T. democratica aggregates, collected in the Mediterranean Sea in 1995 and maintained in constant light, had a broader distribution of individual spawning times, with a mean time of 0625 hrs, which differed significantly from that of the Australian population. Coordination of the events of sexual reproduction in T. democratica may improve fertilization success and be responsible for recognizable age cohorts within blooms of this species. Received: 27 May 1997 / Accepted: 20 June 1997