Correlated evolution between male ejaculate allocation and female remating behaviour in seed beetles (Bruchidae)

Sperm competition theory suggests that female remating rate determines the selective regime that dictates the evolution of male ejaculate allocation. To test for correlated evolution between female remating behaviour and male ejaculate traits, we subjected detailed experimental data on female and male reproductive traits in seven-seed beetle species to phylogenetic comparative analyses. The evolution of a larger first ejaculate was positively correlated with the evolution of a more rapid decline in ejaculate size over successive matings. Further, as predicted by theory, an increase in female remating rate correlated with the evolution of larger male testes but smaller ejaculates. However, an increase in female remating was associated with the evolution of a less even allocation of ejaculate resources over successive matings, contrary to classic sperm competition theory. We failed to find any evidence for coevolution between the pattern of male ejaculate allocation and variation in female quality and we conclude that some patterns of correlated evolution are congruent with current theory, whereas some are not. We suggest that this may reflect the fact that much sperm competition theory does not fully incorporate other factors that may affect the evolution of male and female traits, such as trade-offs between ejaculate expenditure and other competing demands and the evolution of resource acquisition.     

Males of the seed bug Togo hemipterus (Heteroptera: Lygaeidae) use accessory gland substances to inhibit remating by females

In species in which females mate repeatedly, males can adopt several strategies to reduce the risk of sperm competition with future males. The refractory period of females significantly increased as the mating duration increased in the seed bug Togo hemipterus (Heteroptera: Lygaeidae). To elucidate the mechanisms by which mated females are inhibited from remating, we investigated the effects of male-derived substances on the inhibition of mating receptivity of virgin females by injecting the substances into their abdomens. The length of time from injection to mating in virgin females was significantly longer for females injected with accessory gland B solution than for those injected with seminal vesicle, accessory gland A, or control solutions. This is the first report showing that heteropteran males inhibit female remating by using substances from an accessory gland. We discuss and consider the adoption and evolution of this strategy by T. hemipterus males by focusing on female genitalia structures, oviposition habit, and paternity and comparing these traits with those of other heteropterids.     

Potential for pre-release diet supplements to increase the sexual performance and longevity of male Queensland fruit flies

1 Recent studies have shown that continuous access to a protein source (yeast hydrolysate) can greatly enhance the sexual performance of male Queensland fruit flies ( Bactrocera tryoni ; 'Q-flies'). However, in Sterile Insect Technique programmes used to eradicate or suppress wild populations, mass-reared Q-flies are typically fed only sucrose and water for up to 2 days before release.
2 We investigated whether adding a protein source to the diet of male Q-flies for a 24- or 48-h window after emergence and then removing it is sufficient to enhance mating probability, latency to mate, copula duration, probability of sperm storage, number of sperm stored, female remating tendency and longevity of male Q-flies.
3 Protein-fed males were more likely to mate than males fed only sucrose, especially when young. Protein-fed males also had shorter mating latencies and longer copulations than protein-deprived males.
4 Females mated by protein-fed males were more likely to store sperm, stored more sperm and were less likely to remate than were females mated by protein-deprived males. Females were also less likely to remate if their first mate had been large.
5 Overall, providing male Q-flies access to a protein source for a 24- or 48-h window early on in their adult life was sufficient to greatly enhance all assessed measures of performance. Although 24-h access was sufficient for a notable enhancement, further benefits were evident in males provided 48-h access.
6 The results are discussed in terms of the practical implications for Sterile Insect Technique programs used to eradicate or suppress wild Q-fly populations.     

Male and female effects on sperm precedence in the giant sperm species <Emphasis Type="Italic">Drosophila bifurca</Emphasis>

Sperm competition is expected to be a driving force in sexual selection. In internally fertilized organisms, it occurs when ejaculates from more than one male are present simultaneously within the female’s reproductive tract. It has been suggested that greater sperm size may improve the competitive ability of sperm, but studies provide contradictory results depending on the species. More recently, the role of females in the evolution of sperm morphology has been pointed out. We investigate here the male and female effects that influence sperm precedence in the giant sperm species, Drosophila bifurca Patterson & Wheeler. Females were mated with two successive males, and the paternity outcomes for both males were analyzed after determining sperm transfer and storage. We found very high values of last male sperm precedence, suggesting a strong interaction between rival sperm. However, the data also indicate high frequencies of removal of the sperm of the first male from the female reproductive tract prior to any interaction with the second male. This implies that successful paternity depends mainly on successful sperm storage. Knowing what happens to the sperm within females appears to be a prerequisite for disentangling post-copulatory sexual interactions between males and females.     

Sperm survival in the female reproductive tract in the fly Scathophaga stercoraria (L.)

While sperm competition risk favours males transferring many sperm to secure fertilizations, females of a variety of species actively reduce sperm numbers reaching their reproductive tract, e.g. by extrusion or killing. Potential benefits of spermicide to females include nutritional gains, influence over sperm storage and paternity, and the elimination of sperm bearing somatic mutations that would lower zygote fitness.We investigated changes in sperm viability after in vivo and in vitro exposure to the female tract in the polyandrous fly, Scathophaga stercoraria. Sperm viability was significantly lower in the females' spermathecae immediately after mating than in the experimental males' testes. Males also varied significantly in the proportion of live sperm found in storage in vivo. However, the exact mechanism of sperm degradation remains to be clarified. In vitro exposure to extracts of the female reproductive tract, including female accessory glands, failed to significantly lower sperm viability compared to controls. These results are consistent either with postcopulatory sperm mortality in vivo depending entirely on the male (with individual differences in sperm viability, motility or longevity) or with postcopulatory sperm mortality being subtly affected by female effects which were not detected by the in vitro experimental conditions. Importantly, we found no evidence in support of the hypothesis that female accessory glands contribute to sexual conflict via spermicide. Therefore, female muscular control remains to date the only ascertained mechanism of female influence on sperm storage in this species.     

Bacterially enriched diet improves sexual performance of sterile male Mediterranean fruit flies

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), (medfly) is a polyphagous and cosmopolitan agricultural pest, targeted in many areas for control by the Sterile Insect Technique (SIT). Our objective in the present study was to test the hypothesis that a bacterially enriched diet provided to sterile males will improve their sexual performance in competitive settings that emulate natural conditions. Specifically we determined how feeding on diets enriched with Klebsiella oxytoca affected the ability of sterile males to compete for wild females against wild males, their ability to inhibit female receptivity, and their survival. We found that enriching the sterile male diet with K.oxytoca significantly improved mating competitiveness in the laboratory and in field cages. In addition, bacterially enriched sterile males inhibited female receptivity more efficiently than sugar fed males and survived longer duration of starvation. We conclude that inoculating mass reared sterile flies with bacteria prior to their release is a valid approach to improve the efficacy of SIT.     

Interpopulation variation in female remating is attributable to female and male effects in <Emphasis Type=Italic>Callosobruchus chinensis</Emphasis>

The evolution of female multiple mating is best understood by consideration of male and female reproductive perspectives. Females should usually be selected to remate at their optimum frequencies whereas males should be selected to manipulate female remating to their advantage. Female remating behavior may therefore be changed by variation of male and female traits. In this study, our objective was to separate the effects of female and male strains on female remating for the adzuki bean beetle, Callosobruchus chinensis, for which there is interstrain variation in female remating frequency. We found that interstrain variation in female remating is primarily attributable to female traits, suggesting genetic variation in female receptivity to remating in C. chinensis. Some interstrain variation in female remating propensity was attributable to an interaction between female and male strains, however, with the males of some strains being good at inducing nonreceptivity in females from one high-remating strain whereas others were good at inducing copulation in nonvirgin females from the high-remating strain. There is, therefore, interstrain variation in male ability to deter females from remating and in male ability to mate successfully with nonvirgin females. These results suggest that mating traits have evolved along different trajectories in different strains of C. chinensis.     

The causes of mating abstention in male tsetse flies Glossina morsitans

ABSTRACT. The sexual response of a Glossina morsitans morsitans Westw. male towards a decoy consisting of a dead male dosed with synthetic sex pheromone, was depressed by the presence of the cuticular lipids of the decoy male. However, addition of a C24 paraffin, not normally found on or in the cuticle of male or female G.m.morsitans , to a decoy male from which cuticular lipids were removed had a similar effect. Addition of either the C24 paraffin or an extract of male cuticular lipids to decoy females which possess natural sex pheromone in their cuticles, again depressed the sexual responses of males towards the decoys. The n-hexane solvent used in the application of compounds to decoys, itself depressed responses towards natural pheromone on a dead female at volumes in excess of 100μl, presumably because it mixed the surface waxes which masked the pheromone. Occlusion experiments implied that sex pheromone receptors are present on both the tarsi and tibiae of the male. No evidence was found for the existence of a specific mating deterrent on or in the cuticle of male G.m. morsitans.     

The effect of male mating history on paternal investment, fecundity and female remating in the seed beetle Callosobruchus maculatus

1. In many organisms, males provide nutrients to females via ejaculates that can influence female fecundity, longevity and mating behaviour. The effect of male mating history on male ejaculate size, female fecundity, female longevity and female remating behaviour in the seed beetle Callosobruchus maculatus was determined.
2. The quantity of ejaculate passed to females declined dramatically with successive matings. Despite the decline, a male's ability to fertilize a female fully did not appear to decline substantially until his fourth mating.
3. When females multiply mated with males of a particular mated status, the pattern of egg production was cyclic, with egg production increasing after mating. Females multiply mated to virgins had higher fecundity than females mated to non-virgins, and females mated to twice-mated males had disproportionately increased egg production late in their life.
4. Females that mated to multiple virgins, and consequently laid more eggs, experienced greater mortality than females mated only once or mated to non-virgins, suggesting that egg production is costly, and rather than ameliorating these costs, male ejaculates may increase them by allowing or stimulating females to lay more eggs.
5. Females mating with non-virgin males remated more readily than did females mated to virgins. Females given food supplements were less likely to remate than females that were nutritionally stressed, suggesting that females remate in part to obtain additional nutrients.     

Male and female condition influence mating performance and sexual receptivity in two tropical fruit flies (Diptera: Tephritidae) with contrasting life histories

Recent recognition of widespread polyandry in insects has generated considerable interest in understanding why females mate multiple times and in identifying factors that affect mating rate and inhibit female remating. However, little attention has been paid to understanding the question from both a female and male perspective, particularly with respect to factors that may simultaneously influence female remating rates. Here, we report on a study aimed at ascertaining the possible interactive effects that male and female size and diet, and female access to a host could have on mating latency, probability, and duration and female refractory period using two tropical fruit fly species with contrasting life histories. Of all factors tested, adult diet played the most significant role. Both Anastrepha ludens and Anastrepha obliqua males which had constant access to protein and sucrose mated more often, had shorter copulations and induced longer refractory periods in females than males fed a low quality diet (sucrose offered every third day). Female size and the interaction with male diet determined how quickly female A. ludens mated for the first time. Smaller females mated sooner with low quality fed males than with high quality fed males while there was no difference for large females, suggesting that male choice may be at play if high quality fed males discriminate against smaller females. Copulation duration also depended on both male and female nutritional condition, and the interaction between male diet and female size and diet. Large and high quality fed females had shorter copulations regardless of male condition. Importantly, for A. ludens, female refractory period depended on male size and the nutritional condition of both males and females, which could indicate that for this species, female receptivity does not depend only on the condition of the male ejaculate. For A. obliqua refractory period was associated with the interaction between male size and diet and male diet and host presence. We discuss our results in terms of male ability to inhibit female remating and the relative contribution of female condition to this behavior. We also address the importance of studying effects simultaneously on species with contrasting life histories.     

Theoretical influence of female mating status and remating propensity on male sperm allocation patterns

Theoretical models predict that males should allocate more sperm in matings where the immediate risk of sperm competition is high. It has therefore often been argued that males should invest less sperm in matings with virgin females compared with matings with already mated females. However, with relatively polyandrous females, high sperm competition risk will covary with high sperm competition intensity leading to more unpredictable conditions, as high competition intensity should favour smaller ejaculates. With the use of a genetic algorithm, we found that males should allocate more sperm in matings with virgin females when female mating frequency is relatively high, whereas low remating rates will select for higher effort in matings with nonvirgin females. At higher remating rates, first male sperm precedence favours larger ejaculates in matings with virgin females and second male precedence favours the reverse. These results shed some light on several findings that have been difficult to explain adaptively by the hitherto developed theory on sperm allocation.     

Switchover to the mated state by spermathecal activation in female Anopheles gambiae mosquitoes

Both Aedes aegypti and Anopheles gambiae mosquitoes undergo physiological and behavioral changes after the females mate, but unlike Ae. aegypti, the mated state in An. gambiae is not attained through the action of male accessory gland substances. Experiments in which the spermathecae of mated females were manipulated suggest that a spermatheca filled with sperm is responsible for triggering oviposition behavior. An estimated 48% of the previously mated An. gambiae females mated again when the interval between encounters with males was less than 24h. Unlike male Ae. aegypti that can inseminate up to seven females after their testes are removed, An. gambiae do not store sperm in the vas deferens and without testes would be unable to inseminate any females.     

The ultrastructure of the male accessory reproductive glands of the colorado beetle

Summary The accessory reproductive glands of Leptinotarsa decemlineata males are simple epithelial tubular glands. They are surrounded by a network of slow acting muscle cells. All the gland cells have a very extensive granular endoplasmic reticulum, a prominent Golgi apparatus, but few secretory granules. The secretory product in the lumen, which is a neutral mucopolysaccharide-protein complex, appears as a granular material in which fibrillar structures occur. Neither in the gland cells nor in the lumen were regional differences observed. After allatectomy the gland cells degenerate in a remarkable way. A zone containing microvillous structures appears between the outer and inner cells. While the inner cells degenerate completely, the outer ones do not but become organised for minimal consumption of energy, necessary to survive diapause.Grateful acknowledgment is made for a mandate as Aangesteld Navorser of the National Foundation of Scientific Research in Belgium.We wish to thank Ing. W. Bohijn for operating the EM and Mr. G. Maes for photography.     

Multiple mating reduces male survivorship but not ejaculate size in the polygamous insect Stenomacra marginella (Heteroptera: Largidae)

In polygamous species, successful males should be able to inseminate multiple females, to defeat sperm from previous males, to avoid sperm displacement by other males, and to induce females to use his sperm during fertilization. Since resources are limited, adaptations to perform any of these functions may conflict with each other (and with other life-history traits) and trade-offs are expected to evolve. We studied if males of the polygamous true bug Stenomacra marginella face a trade-off between multiple mating and survivorship, by comparing the survivorship of virgin and multiply mated males. We also looked for physiological costs of ejaculate production by examining ejaculate production in consecutive matings in multiple mated males. Multiply mated males were able to produce ejaculates of similar size in up to six consecutive copulations but they had decreased survivorship in comparison with virgin males. There was no difference in survivorship between males mated three and six consecutive times, suggesting that the negative relation between survivorship and number of copulations is not linear. The decrease in survivorship seems to be a cost of mating and ejaculate production. This cost could favor the evolution of prudence in the allocation of resources to ejaculate production (e.g., cryptic male choice).     

Behavioural switch of female Mediterranean fruit fly, Ceratitis capitata: mating and oviposition activity in outdoor field cages in Hawaii

1 Laboratory-reared normal, and wild female Mediterranean fruit flies, Ceratitis capitata (Wiedemann), were assayed in outdoor field cages to assess the impact of a mating-induced behavioural switch on mating and subsequent oviposition activity. 2 Virgin females preferred interactions with males leading to mating over attraction to, and oviposition in, artificial yellow spheres containing guava odour or green apples hung in a guava tree. Laboratory-reared females previously mated with either laboratory-reared normal males or laboratory-reared irradiated (sterile) males showed little interest in remating with males and instead, were much more likely to be found arrested on artificial and real fruit and ovipositing. Oviposition on artificial fruit was five times greater by females that had mated with either normal or irradiated males than by virgin females. Wild females showed similar qualitative changes in the mating-induced behavioural switch; however, oviposition activity was significantly less than for laboratory-reared females. 3 These results confirm that mating has a profound effect on the behaviour of female Mediterranean fruit flies and that irradiated males are functionally equal with normal males (lab-reared or wild) in their ability to alter female behaviour. These results are discussed in the context of the sterile insect technique for control of Mediterranean fruit flies in the field.     

The developments between gametogenesis and fertilization: ovulation and female sperm storage in Drosophila melanogaster

In animals with internal fertilization, ovulation and female sperm storage are essential steps in reproduction. While these events are often required for successful fertilization, they remain poorly understood at the developmental and molecular levels in many species. Ovulation involves the regulated release of oocytes from the ovary. Female sperm storage consists of the movement of sperm into, maintenance within, and release from specific regions of the female reproductive tract. Both ovulation and sperm storage elicit important changes in gametes: in oocytes, ovulation can trigger changes in the egg envelopes and the resumption of meiosis; for sperm, storage is a step in their transition from being "movers" to "fertilizers." Ovulation and sperm storage both consist of timed and directed cell movements within a morphologically and chemically complex environment (the female reproductive tract), culminating with gamete fusion. We review the processes of ovulation and sperm storage for Drosophila melanogaster, whose requirements for gamete maturation and sperm storage as well as powerful molecular genetics make it an excellent model organism for study of these processes. Within the female D. melanogaster, both processes are triggered by male factors during and after mating, including sperm and seminal fluid proteins. Therefore, an interplay of male and female factors coordinates the gametes for fertilization.     

Spermatophores, female genitalia, and courtship behaviour of two whip spider species, Charinus africanus and Damon tibialis (Chelicerata: Amblypygi)

This paper describes courtship behaviour, spermatophore morphology, and the female genitalia of the African whip spiders Charinus africanus Hansen, 1921 (Charinidae) and Damon tibialis (Simon, 1876) (Phrynichidae). In C. africanus, only the first part of courtship behaviour, up to spermatophore formation, could be observed; though different in detail, it is similar to that of many other species. The small spermatophore of C. africanus contains one large median sperm package. Charinus africanus is one of the Charinus species with thin finger-like gonopods and the first species with such gonopods of which the spermatophore is known. Spermatophores and female genitalia of D. tibialis are similar, though different in details, to those of Trichodamon and Musicodamon. They thus suggest that these two genera are correctly included in the Damoninae.