Sperm Number,Spermatophore Weight and Remating in the Bushcricket Poecilimon veluchianus

The influence of the intermating interval on sperm number and spermatophore mass was examined in the bushcricket Poecilimon veluchianus. Males that remated after 1 d transferred about 50% of the sperm, but significantly more than 50% of the weight of spermatophores than males that remated after 2 d. Assuming a constant rate of replenishment of sperm and spermatophore material we concluded that available spermatophore material but not available sperm number influences remating interval in P. veluchianus males.     

Field mating rate of female meadow grasshoppers, Chorthippus parallelus, estimated from sperm counts

Abstract.  1. Female mating frequency is an important life-history component in many insect species. It has been suggested that its laboratory measurement may frequently represent an overestimation of the natural situation because laboratory females have little opportunity to avoid matings. Here, numbers of sperm stored by field-collected females are investigated in order to address whether this method is suitable for estimating field mating frequencies.
2. Data on sperm numbers stored by females and their estimated age based on mandible abrasion were fitted to a simple mechanistic model to obtain estimates of: (i) remating interval; (ii) ejaculate size, and (iii) the rate of sperm loss from the female sperm store. Females were calculated to remate every 5.9–7.0 days. By contrast, sperm numbers of field-collected females did not exceed ejaculate size, which would have resulted in the false conclusion that females mate only once.
3. In the laboratory, females first mated when aged 4–11 days and remated approximately every 2.6 days. Because females are not restricted by mate encounters under these conditions, this confirms the notion that female mating rates are likely to be lower in the field than in the laboratory. Such reduced field mating rates should be taken into account when predicting ecological or evolutionary parameters based on laboratory observed mating rates.     

Effects of male mating interval on spermatophore formation,transfer, and subsequent female receptivity and fecundity in the sorghum plant bug,Stenotus rubrovittatus

Males of the sorghum plant bug, Stenotus rubrovittatus (Matsumura) (Heteroptera: Miridae), transfer a spermatophore to females during copulation. After a 1‐day interval between the first and second copulation, males transferred both sperm and a spermatophore to females during the second copulation. However, when male mating interval was <1 h, they transferred sperm but no spermatophores to females during the second copulation. Therefore, the male mating interval probably produces two types of mated females, those with and those without a spermatophore. Mated females of S. rubrovittatus do not remate for at least 3 days after mating, even when courted, and lay more eggs than virgin females at the beginning of the oviposition period. The effects of spermatophores on female sexual receptivity and fecundity were examined using mated females with or without a spermatophore. Only one of the 40 (2.5%) mated females with a spermatophore remated, whereas 10 of the 26 (38.5%) without a spermatophore remated. Furthermore, mated females with a spermatophore laid more eggs than those without a spermatophore. These results suggest that spermatophores participate in reducing female sexual receptivity and enhancing female fecundity in S. rubrovittatus.     

Remating and male-derived nutrients in Drosophila melanogaster

Laboratory natural selection and environmental manipulations were used to investigate the importance of male-derived nutrients to female Drosophila melanogaster. No evidence for the importance of such nutrients was found. Females from the same wild type base stock exposed as adults to low quality food did not show elevated fecundity or survival when they remated more frequently, and on high quality food the females showed a ‘cost of mating’ in reduced survival. Laboratory evolution on low quality food did not lead to elevated rates of remating by females; females from each selection regimen remated more frequently than one another when kept on the food type to which they had been exposed for the previous 5 years, on which they also showed higher fecundity than one another. Even under conditions of extreme nutritional stress, when females were exposed to short term (4-day) cycle of exposure to very low and high quality food, they remated more frequently immediately after exposure to high quality food. The results of this last experiment suggested that, under these circumstances, current nutrition, fecundity or rate of sperm usage was more important than number of sperm in store or cumulative fecundity in determining the probability that a female would remate.     

Effect of gamma irradiation on sperm utilization in twice-mated female Phthorimaea operculella Zeller (Lep., Gelechiidae)

The effects of the second mating on fecundity and fertility of potato tuber moth (PTM) Phthorimaea operculella (Zeller), when females were mated with 450 Gy-irradiated and normal males or vice versa, were studied. The percentage of eggs fertilized by sperm of the second mating (P₂ value) was 0.99, indicating that sperm transferred during the last mating were predominantly utilized in egg fertilization. Females, mated first with irradiated males, remated after 2 days, whereas those mated with normal males, remated after 3.3 days. Fecundity of twice-mated females was higher than those mated only once. Females started to lay their eggs 1.9 days after the first mating, regardless of the type of male. However, virgin females did not lay eggs at all. Duration of copulation varied from 102 to 117 min for normal and irradiated males, respectively. The present study elucidated important aspects of mating behaviour of PTM which could improve the efficiency of its control by the sterile insect technique.     

Copulation, the dynamics of sperm transfer and female refractoriness in the leafhopper Balclutha incisa (Hemiptera: Cicadellidae: Deltocephalinae)

Abstract.  Mature sperm of the leafhopper Balclutha incisa (Matsumara) (Cicadellidae: Auchenorrhyncha: Hemiptera) are stored as a series of sperm bundles within seminal vesicles prior to ejaculation. During transfer, sperm are pumped from the vesicles into the ejaculatory duct to the complex aedeagus. Sperm transfer is marked by a c . 30-fold expansion of the spermatheca to accommodate both sperm and seminal fluid. Sperm number increases exponentially with male age, reaching a maximum of 700 000 after 14 days, while the number of sperm available on days 2–5 is between 70 000 and 100 000. During mating, maximum sperm transfer occurs after 7 min and mating is complete after about 10 min. Ejaculate size, defined by both sperm and associated accessory gland fluid, is influenced by male mating status and the interval since the previous mating. There is a positive correlation between duration of copulation and both ejaculate and the time to subsequent mating. Sperm are more likely to be retained in the testes during mating by males of 2–5 days post-emergence than older males. The number of sperm received by the female can be manipulated experimentally by mating males once (medium ejaculate) or twice (small ejaculate) immediately after their first mating. Females that receive small ejaculates from sperm-depleted males have a far shorter refractory period than females receiving medium to large ejaculates. Both ejaculate size and the time after males have mated influence the female post-mating refractory period as measured by the female's responsiveness to male sexual signalling.     

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.     

Effect of male body size on sperm precedence in the polyandrous butterfly Pieris napi L. (Lepidoptera: Pieridae)

Male Lepidoptera produce an ejaculate during copulation thatcontains both sperm and accessory gland nutrients and may functionas paternal investment and/or male mating effort Several studieshave examined how ejaculates function as paternal investment,but few have determined the influence of sperm competition onmale investment This study examines the effect of male bodysize on sperm precedence in the polyandrous butterfly Pierisnapi L. We used male body mass as an indicator of the size ofejaculate transferred and found that relative male size hada significant effect on paternity. The offspring of twice-matedfemales showed a low incidence of mixed paternity. Larger malesobtained the majority of fertilizations, and the degree of second-malesperm precedence was influenced by relative body size of matingmales. In general, second mates obtained fewer fertilizationsthe larger the size of the first mate. The interval betweenthe first and second mating was influenced by the size of thefirst male mate Females first mated to small males remated soonerthan females first mated to larger males Our results suggestthat large males may have a selective advantage over small maleswhen both a male's fertilization success and a female's refractoryperiod are influenced by the size of ejaculate transferred.Furthermore, the effect of male body size on the proportionof offspring sired lends support to the hypothesis that spermcompetition has played a major role in the evolution of ejaculatesize.     

Coevolution of male and female reproductive traits in a simultaneously hermaphroditic land snail

Inter- and intraspecific studies in gonochoristic animals reveal a covariation between sperm characteristics and the size of the female reproductive tract, indicating a rapid evolutionary divergence, which is consistent with the theory of post-copulatory sexual selection. Simultaneous hermaphrodites differ from species with separate sexes (gonochorists) in that they possess both functional male and female reproductive organs at the same time. We investigated whether in hermaphroditic animals intraspecific variation in reproductive traits results from divergent coevolution, by quantifying the variation in male and female traits among six natural populations of the snail Arianta arbustorum and examining the covariation in interacting traits. There was a significant among-population variation in spermatophore volume, number of sperm transferred and sperm length, as well as in volume of the sperm storage organ (spermatheca) and number of tubules, but not in spermatheca length. We found a positive association between sperm number transferred and spermatheca volume. This result suggests that the same post-copulatory mechanisms as in gonochorists drive the correlated evolution of reproductive characters in hermaphrodites.     

Multiple mating and lifetime reproductive output in female dobsonflies that receive nuptial gifts

In mating of the dobsonfly, Protohermes grandis (Thunberg), the male attaches the spermatophore externally to the female genitalia. The spermatophore includes a large gelatinous mass which the female detaches and feeds on after mating. While the female consumes this nuptial food gift, sperm is evacuated from the remaining portion of the spermatophore (sperm package) into her reproductive tract. Under laboratory conditions, mated females maintained receptivity throughout their lifetime, and they remated even on the day following copulation. A single insemination may supply enough sperm, as females mated only once deposited fertile eggs throughout life and, when dissected after death, all females had sperm in the spermatheca. There was a positive correlation between longevity and the number of matings. Lifetime fecundity also increased as mating multiplied. However, the size of eggs and hatchlings was not influenced by the number of matings. It seems that large spermatophore consumption by female P. grandis provides nutrients that increase fitness not in offspring quality, but in their quantity.     

SPERM DISPLACEMENT WITHOUT SPERM TRANSFER IN DROSOPHILA MELANOGASTER

In this paper we show that when Drosophila melanogaster females are mated twice, the semen of the second male causes a reduction of the effective number of resident sperm from the previous mating. This is demonstrated by two different kinds of experiments. In one set of experiments, mated females were remated to two different kinds of sterile males, one with normal semen and the other with deficient semen. The effect on the resident sperm was determined from the number of remaining progeny after mating to the sterile male, with the result that the normal semen reduced the amount of resident sperm in comparison with matings to the males with deficient semen. The second set of experiments employed interrupted matings. These experiments were based on the observation that semen is delivered before sperm during the first 5 min of copulation. The second matings were interrupted instantly, 2 min, and 4 min after the initiation of copulation. Compared to the instant interruptions, the two later interruptions had the effect of reducing the amount of resident sperm. The results of these two experiments clearly indicate that a sperm-incapacitation process plays a role in the well-documented phenomenon of sperm displacement (last-male advantage) in this species. Such a process could play a role in sperm displacement in the many cases where the mechanism is unknown.     

No reduction of female sexual receptivity following mating in a stalk‐eyed fly,Cyrtodiopsis dalmanni (Diptera: Diopsidae)

The level of female sexual receptivity is an important component of male and female reproductive success. In many insects, mating itself causes a sharp decline in female receptivity. This can be a direct result of the physical act of mating, or because of actions of sperm or seminal fluid proteins. The degree to which males can decrease female receptivity will directly affect their reproductive success, by altering the chance that their sperm will be used in fertilizations in the interval before the female mates again. In this study, we investigated the effect of mating on female receptivity in the sexually dimorphic stalk‐eyed fly, Cyrtodiopsis dalmanni. Our results showed no evidence for mating‐induced reductions in female receptivity. In addition, we found that matings with males that differed in eyespan did not cause differences in the level of female receptivity. There was also no evidence that females remated sooner when presented with large eyespan males. These results are surprising, given the indirect benefits that females gain from matings with large eyespan males. Finally we demonstrate that males do not appear to discriminate between females on the basis of female mating status.     

Seminal fluid causes temporarily reduced egg hatch in previously mated females

In Drosophila, male accessory gland fluid (seminal fluid) has multiple effects on the female's reproductive efficiency. Here, we show the effect of seminal fluid on rate of egg hatch immediately following mating. Singly mated females were remated to two classes of sterile males, one with seminal fluid and one without seminal fluid. Transfer of seminal fluid results in a strong reduction in egg hatch shortly after the mating. Also, it is shown that remating with normal males causes an immediate reduction of egg hatch followed by recovery to normal egg hatch. In all cases, unhatched eggs contained no sperm. These results are consistent with a role for seminal fluid in sperm competition, mediated by incapacitation or inefficient use of resident sperm.     

Female remating and the intensity of female choice in black-horned tree crickets, Oecanthus nigricornis

Repeated mating by females of many species occurs at frequenciesin excess of those needed to acquire additional sperm for fertilizingova. I tested three alternative hypotheses for the rate of rematingby females of the courtship-feeding tree cricket, Oecanthusnigricornis Walker, by manipulating diet quality and courtshipfeeding and measuring the time to remating by the female inrelation to four aspects of male phenotype (age, condition,fluctuating asymmetry, and size). First, in courtship-feedingspecies, remating may be due to selection to increase the amountof nutritional resources provided by males, with nutrient-deprivedfemales remating more quickly. Second, remating may functionas a mechanism of postcopulatory mate choice, with females rematingquickly when the quality of a previous mate is low. Third, quicknessof remating may be the consequence of precopulatory mate choiceprior to future matings, with females remating more quicklywith high-quality males, regardless of the quality of priormates. Females on a low-quality diet remated quickly, did notvary remating speed with the phenotype of their first mate,and did not differentially reject prospective second mates withdifferent phenotypes. In contrast, both the degree of coyness(measured as the frequency of mate rejection) and the intensityof female choice (measured as the size differential betweenaccepted and rejected mates) increased with diet quality. Theseresults support both the material-benefits and the precopulatorymate-choice hypotheses for remating speed of female tree crickets.There was mixed support for the postcopulatory choice hypothesis:females on the high-quality diet remated more slowly after firstmating with relatively large males, in support of the postcopulatorychoice hypothesis; however, the remating interval of femaleson the high-quality diet decreased with the condition of thefirst mate, opposite to the prediction of the postcopulatorychoice hypothesis     

Effect of sperm numbers and time of insemination relative to ovulation on sow fertility

We examined the effect of inseminating mixed parity sows (n = 231) once with fewer sperm at different times relative to ovulation. Lactation length was 19 days and sows received an IM injection of 600 IU equine chorionic gonadotrophin (eCG) 12 h before weaning. At 80 h after eCG injection, sows received an IM injection of 5 mg porcine luteinizing hormone (pLH). Predicted time of ovulation (PTO) was 38 h after pLH injection. Sows were assigned by parity to receive a single transcervical artificial insemination (AI) at either 6 or 24 h before PTO with semen doses containing either 2.5 or 1.25 × 10⁹ sperm. A positive control group of sows (n = 49) was subject to conventional AI 24 and 6 h before PTO. Detection of estrus was performed in the presence of a boar and only sows exhibiting estrous behavior at the assigned time of AI were included in the study. Farrowing rate for sows receiving 2.5 × 10⁹ sperm at 6 h before PTO was greater than that for sows receiving 1.25 × 10⁹ sperm at 24 h before PTO (85% versus 61%, P < 0.05). All other groups were intermediate. There was no effect of time of AI or sperm numbers on subsequent litter size. These data indicate that single insemination of fewer sperm may compromise sow fertility, even when performed transcervically, if not appropriately timed relative to ovulation.     

Sperm transfer, sperm storage, and sperm digestion in the hermaphroditic land snail Succinea putris (Gastropoda, Pulmonata)

Abstract. Many hermaphroditic species are promiscuous, have a sperm digesting organ and an allosperm storage organ (i.e., spermatheca) with multiple compartments (i.e., spermathecal tubules) providing opportunities for sperm competition. The relative paternity of a sperm donor drives the evolution of mating behaviors that allow manipulation of the sperm receiver's reproductive behavior or physiology. We studied the relationship between sperm transfer, sperm storage, sperm digestion, and copulation duration in the hermaphroditic land snail Succinea putris , in which an active individual mates on top of a passive individual. Specifically, we examined (i) whether the entire copulation duration was required to complete reciprocal sperm transfer, (ii) sperm transfer patterns and their relationship with activity role, and (iii) the timing of sperm storage and sperm digestion. We found that reciprocal sperm transfer was completed within the first 5 h of copulation, which is ∼2–3 h before the end of copulation. Sperm transfer was mainly sequential, meaning that one individual donated all his ejaculate before its partner started to reciprocate. The initiation of sperm transfer did not depend on the activity role. The presence of allosperm in the spermatheca before sperm transfer suggests that individuals remate before they are allosperm depleted. No sperm was digested during copulation but sperm digestion took place 0–72 h after copulation. Our results suggest that contact mate guarding is a likely manipulation strategy in S. putris , because partners cannot immediately remate. In addition, staying in copula after sperm transfer is completed seems to prevent the immediate digestion of sperm and therefore may promote sperm displacement and allosperm storage.     

The spermatheca in the land snail, Arianta arbustorum (Pulmonata: Stylommatophora): muscle system and potential role in sexual selection

Abstract. Previous studies have shown variable patterns of paternity after multiple mating, and also variation in sperm storage among individuals of Arianta arbustorum , which suggests that the spermatheca may influence paternity in this promiscuous land snail. To identify possible morphological correlates of sperm manipulation, we investigated arrangement and ultrastructure of the muscles of the spermatheca. The musculature surrounding the 2–9 spermathecal tubules is arranged in a complex three dimensional network. In addition, each tubule has a thin sheath in which longitudinally oriented cells make up the innermost layer. Usually, the smooth muscle cells are enclosed by connective tissue. Only occasionally is direct muscle-muscle contact established through dense plaques. The short thick filaments, their small diameter, the relatively weak development of the tubular system and sarcoplasmic reticulum, and the low density of mitochondria indicate that the muscle cells contract relatively fast but with little strength, that they recover slowly, and have low endurance. A single muscle cell may be innervated by several axons and one axon may contact several muscle cells. Combining evidence of the present paper and a foregoing investigation on the spermathecal epithelium, we suggest that the main function of the spermathecal muscles is to expel sperm stored for fertilization, while the ciliation of the common duct is probably responsible for the distribution of sperm among the tubules.     

Postpartum resumption of ovarian cycling activity in first-calf suckled beef cows exposed to familiar or unfamiliar bulls

The objective of this experiment was to determine if the proportion of first-calf suckled beef cows that resumed ovulatory cycles and the interval to resumption of ovarian cycling activity differ after exposure to either “unfamiliar” bulls or cows on d 35 postpartum, after exposure to either “familiar” bulls or cows for the first 30–32 d after calving. Fifty Angus × Hereford cows were stratified by calving date, calf BW, and calf sex by d 3 postpartum, and assigned to be exposed to familiar epididectomized bulls (BEF; n = 25) or familiar mature ovariectomized (OVX) cows (CEF; n = 25). On d 35 after calving, 12 BEF cows were assigned to be exposed to unfamiliar bulls (BEU); likewise, 12 CEF cows were assigned to be exposed to unfamiliar OVX cows (CEU). Cows were in their treatments for either 95 d (BEF and CEF) or 60 d (BEU and CEU) during the experiment. Blood samples were collected every third d from the beginning to the end of the experiment. A rise in progesterone concentration of >0.5 ng/mL in consecutive samples was used as the criterion for resumption of ovulatory cycles. Exposing cows to bulls on d 5 after calving and then switching a subset of these cows to be exposed to unfamiliar bulls 30–32 d later did not (P > 0.10) alter: (1) the proportion of cows that resumed cycling activity; and (2) postpartum interval to resumption of ovarian cycling activity compared to cows exposed to familiar bulls. However, 32% more (P < 0.05) cows exposed to bulls (BEF and BEU) resumed cycling activity by approximately 14.8 d before cows that were exposed to OVX cows (CEF and CEU). We conclude that the familiarity of first-calf cows to either bulls or ovariectomized cows, did not affect the postpartum occurrence of cycling activity or the interval from calving to resumption of ovulatory cycles. However, bull exposure, whether familiar or unfamiliar, stimulates first-calf cows to resume ovulatory cycles sooner after calving than if they are not exposed to bulls.     

Female Remating Behavior in a Lekking Moth

In polyandrous species, male reproductive success will at least partly be determined by males' success in sperm competition. To understand the potential for post‐mating sexual selection, it is therefore important to assess the extent of female remating. In the lekking moth Achroia grisella, male mating success is strongly determined by female choice based on the attractiveness of male ultrasonic songs. Although observations have indicated that some females will remate, only little is known about the level of sperm competition. In many species, females are more likely to remate if their first mating involved an already mated male than if the first male was virgin. Potentially, this is because mated males are less well able to provide an adequate sperm supply, nutrients, or substances inhibiting female remating. This phenomenon will effectively reduce the strength of pre‐copulatory sexual selection because attractive males with high mating success will be more susceptible to sperm competition. We therefore performed an experiment designed both to provide a more precise estimate of female remating probability and simultaneously to test the hypothesis that female remating is influenced by male mating history. Overall, approximately one of five females remated with a second male. Yet, although females mated to non‐virgin males were somewhat more prone to remate, the effect of male mating history was not significant. The results revealed, however, that heavier females were more likely to remate. Furthermore, we found that females' second copulations were longer, suggesting that, in accordance with theory, males may invest more sperm in situations with an elevated risk of sperm competition.     

Females Remate for Sperm Replenishment in a Seed Bug: Evidence from Offspring Viability

Females of many insects mate multiply but why they do so remains controversial. Here we investigated the effects of multiple matings on female reproductive success of a New Zealand seed bug, Nysius huttoni. We found little evidence for females to gain material (nutritional) benefits through multiple matings because the number of matings did not have significant effect on female fecundity. Females remated to the same males or different males produced similar number of viable offspring, suggesting that females do not obtain genetic benefit from remating in terms of offspring viability. With the increase of the number of matings, however, overall fertility rate significantly increased and daily fertility rate declined significantly slower over time. These results suggest that females remate for the replenishment of sperm. Five matings are sufficient for females to maximize their reproductive success, and additional matings appear to be superfluous. However, the females of this bug mate as many as 68 times if males and females are paired for lifetime. This can be explained by the convenience hypothesis, i.e., females remate superfluously to minimize the costs of harassment by promiscuous males.