Morphology of male sex organs and insemination in the grasshopper Taeniopoda eques (Burmeister)

The functional morphology of the male genitalia and the insemination process of Taeniopoda eques were examined using scanning electron microscopy and dissections of mating pairs. Male accessory glands consist of 17 separate tubules belonging to eight categories. Males attach to females via a genital locking mechanism, with special motions of the four aedeagal valves aiding in insertion of the aedeagus. The male passes a series of spermatophores. Each is emptied of its spermatodesm contents, then extracted from the male and female genital tracts through motions of the aedeagal valves, while the pair remain in copulo. This allows the male to keep a strong hold on the female, presumably preventing usurpation by other males, while filling the spermatheca with sperm.     

A Test of Genital Allometry Using Two Damselfly Species does not Produce Hypoallometric Patterns

It is widely admitted that sexual selection is the responsible force behind genital traits. However, the particular mechanisms of genital evolution are still debated. Recently, studies of genital static allometry in insects have been used to elucidate such mechanisms. Insect genital traits are often reported to show negative allometry (i.e., a slope < 1), which has generated a number of ideas on how genital traits are selected. However, many studies that have inferred selection mechanisms have omitted consideration of the function of genital traits, used unreliable indicators of body size, and only rarely included female genitalia in their analysis. We investigated whether negative allometry operates for genitalia in two damselfly species (Protoneura cara and Ischnura denticollis). Damselflies are suitable for genital allometry tests as their genital function and body size indicators (wing length and head width) are relatively well known and established. First, we show that the aedeagus is used to physically remove sperm from both sperm storage organs (bursa and spermatheca) and that wing length and head width correlate positively with other morphological traits for the two study species. Second, we estimated genital allometry by measuring aedeagal length, vaginal length, bursal volume, and spermathecal volume. Our results indicate no consistent allometric pattern. Allometry for aedeagal length and vaginal width was not the same. Thus, there was no support for a negative allometric relationship. We urge researchers investigating allometry to look directly at how genitalia function rather than inferring function from allometric relationships only.     

The morphology of mating plugs and its formation in scorpions: Implications for intersexual participation

Mating plugs have been proposed as a mechanism that has evolved to avoid sperm competition. Their structure and composition vary across taxa and are related to the effectiveness of its function. This effectiveness could be related to different evolutionary interests of the sexes. Urophonius brachycentrus and Urophonius achalensis (Scorpiones, Bothriuridae) are highly suitable species to study mating plugs because both are monandrous species with specific morphological and physiological responses in the female's genitalia. Here, we analyze (a) the morphology and fine structure of the mating plugs of both species, (b) the site of production in males and the formation process of the mating plug, and (c) the changes that it undergoes over time in the female's reproductive tract. In both species, a complex mating plug obliterates the female's genital aperture and fills the genital atrium. We observed considerable interspecific variation in the mating plug morphology. A mating hemi-plug was found surrounding the capsular lobes of the hemispermatophore, which could have a mixed composition (involving portions of the hemispermatophore and glandular products). The glandular portion was transferred in a semi-solid state filling the female's genital atrium and then hardening. Changes that the plug undergoes in the female's genitalia (darkening and increase of the “distal” area of the plug) indicate a participation of the female to the formation of this type of plug. Our study provides new insights into the plugging phenomenon in scorpions, and we discussed the adaptive significance as a post-copulatory mechanism to avoid sperm competition.     

Evolution of a giant intromittent organ in Scydmaeninae (Coleoptera: Staphylinidae): Functional morphology of the male postabdomen in Mastigini

We compared the postabdominal architecture of Mastigini with extremely long (Stenomastigus) or short (Palaeostigus) aedeagus. A novel mode of copulation was discovered: males of Stenomastigus insert a paramere between the female's abdomen and elytra, and the intromission is stabilized by several structures of both sexes. The intrinsic aedeagal mechanism is indicated as responsible for inflating the endophallus, and the long flagellum does not penetrate the ductus spermathecae during copulation. The structure of the flagellum suggests that it is primarily responsible for the sperm transfer. Asymmetrical postabdominal rotators of the aedeagus were only found in Stenomastigus; they presumably facilitate the withdrawal of the genitalia; their origin as bundles separated from larger muscles is postulated. We discuss a scenario in which the evolution of elongated genitalia was facilitated by the lack of structural constraints and existing preadaptations. Benefits of stabilizing the copulation and intromission are indicated as the driving force for the evolution of extremely long aedeagi, while the short aedeagi might have the advantage of freedom of movements facilitating the initiation of copulation by males. Disruptive selection is suggested as a working hypothesis to further investigate mechanisms that have played a role in the evolution of genital structures of Mastigini.     

Isometric patterns for male genital allometry in four damselfly species

Recent studies have found that insect genitalic traits show negative allometry, i.e., are relatively small in relation to body size. One interpretation of this is that males use their genitalia to stimulate females. Thus, given the nature of damselfly copulation in which males physically reach the rival sperm that females have stored from previous matings, male genitalic traits are not expected to show negative allometry. To test this idea, we assessed (a) the rival sperm displacement function by the mating male and (b) allometry of aedeagal length of four damselfly species (Argia anceps, Argia tezpi, Argia extranea, and Enallagma praevarum). Sperm displacement was assessed by inspecting whether the aedeagus reached the rival sperm during copulation in mating pairs for the four species. To have a standard for comparing allometric patterns, allometry of aedeagal was compared to that of two non-genital traits, tibial, and fourth abdominal segment length. In all cases, the aedeagus was found to reach the rival sperm which supports the idea that stimulation is not the mechanism for sperm displacement but physical displacement. Aedeagal length was isometric, and its slope was lower in general compared to that of tibial length and fourth abdominal segment. Given that this isometric pattern is not common for other odonate species, our interpretation of these varying aedeagal scaling patterns in this insect order is that males’ and females’ sexual interests are in conflict (males are evolving an elongated aedeagus to reach rival sperm while females are evolving unreachable sperm storage organs to prevent displacement of stored sperm). This sexual conflict scenario would favor varying scaling patterns for aedeagal length in odonates. A final interpretation is that the risk of interspecific matings in damselflies, may also explain different species-specific, aedeagal allometries.     

Functional association between female sperm storage organs and male sperm removal organs in calopterygid damselflies

Female damselflies in the family Calopterygidae have two sperm storage organs: a spherical bursa copulatrix and a tubular spermatheca. Male flies have a peculiar aedeagus with a recurved head with which to remove bursal sperm, and lateral spiny processes to remove spermathecal sperm. The lateral processes differ among species and populations in terms of their width relative to the spermathecal duct: the narrower processes are physically able to access spermathecal sperm, while the wider ones are not. In the present study, sperm storage patterns and aedeagal structures were compared between two calopterygid species with different spermathecal structures –Calopteryx cornelia and Mnais pruinosa– with respect to not only sperm quantity (number) but also sperm quality (viability), by using a recently developed method based on live/dead dual fluorescence. Calopteryx cornelia is a typical spermathecal sperm remover. In this species, viability was similar between bursal and spermathecal sperm. In contrast, in M. pruinosa, the spermatheca was much smaller than the bursa and often contained no sperm. Even when the spermatheca of this species did contain sperm, a high percentage of it was dead. Although the spermatheca of M. pruinosa has such atrophic tendencies, males have nevertheless developed long and spiny lateral processes similar to those of C. cornelia, suggesting the processes have functions other than spermathecal sperm removal. They possibly function as stoppers or guides for manipulating the aedeagal head to remove the sperm mass from the bursa.     

Determinants of sperm precedence in a noctuid moth Heliothis virescens: a role for male age

1. Females of the noctuid moth Heliothis virescens F. mate more than once. Thus, sperm from two or more males normally compete for fertilisations within the female reproductive tract. The eggs are typically fertilised by sperm from only one male, either the female's last mate or an earlier mate. Twice‐mated females store only one ejaculate's worth of fertilising sperm (eupyrene) but nearly two ejaculates' worth of a nonfertilising sperm morph (apyrene), which is thought to play a role in sperm competition. 2. The mechanism of sperm use in H. virescens was investigated by examining factors that vary with paternity, which was assigned based on allozyme variation. The factors included male and female body masses and ages, male genital characters, the size of the sperm package, and the number of sperm stored by the female. 3. One male typically gained sperm precedence; this was nearly twice as likely to be the second male as it was to be the first. Two factors were found to vary significantly with paternity: female mass and male age. The second male to mate was more likely to gain sperm precedence if the female was larger and if the male was older than the female's first mate. 4. The significance of male age and female mass to several hypothetical models of the mechanism of sperm use is discussed.     

Comparative Analysis of the Male Reproductive System in Bothriuridae Scorpions: Structures Associated with the Paraxial Organs and the Presence of Sperm Packages (Chelicerata, Scorpiones)

The male reproductive system of seven species of the family Bothriuridae are compared. These scorpions are Bothriurus flavidus Kraepelin, B. cordubensis Acosta, B. bonariensis (C. L. Koch), B. chacoensis Maury & Acosta, Brachistosternus ferrugineus (Thorell), Timogenes dorbignyi (Guérin-Méneville), T. elegans (Mello-Leitão) and Urophonius brachycentrus Pocock (Bothriuridae). Additional comparisons are made with the buthid Zabius fuscus (Thorell). Observations on the structures associated with the paraxial organs (testis, seminal vesicle and accessory glands) are given. Sperm obtained from the male reproductive tract and fresh spermatophores as well as from the female's genital atrium and seminal receptacles are examined. Accessory glands occur in six out of eight studied bothriurids and in the buthid Z. fuscus. In most species the distal portion of vas deferens has a developed ampulla. All structures vary in size and shape depending on species. Sperm packages were observed in all bothriurids. In contrast, there is no packaged spermatozoa in Z. fuscus. Each sperm package consists of many spermatozoa surrounded by a common membrane that breaks after the spermatophore capsule is everted into the female genital atrium, releasing the spermatozoa. One hour after insemination, the spermatozoa are found in the atrium and in the seminal receptacles of B. flavidus females, but after 24h spermatozoa are found only in the seminal receptacles. The functional significance of the accessory glands and the presence-absence of sperm packages are discussed.     

Host‐dependent phenotypic plasticity of aedeagus morphology in a pair of cactophilic sibling Drosophila species of the repleta group (Diptera,Drosophilidae)

The rapid evolution of male genital morphology is a characteristic feature of several animal groups. Such rapid divergence makes this trait a useful key for species identification. The aedeagus, the intromittent organ of male genitalia, is considered the main diagnostic trait in the Drosophila repleta group. In this study we analysed phenotypic plasticity and genetic variations associated with aedeagus size and shape in the cactophilic sibling species Drosophila gouveai Tidon‐Sklorz and Sene, 2001 and Drosophila antonietae Tidon‐Sklorz and Sene, 2001. Phenotypic plasticity in aedeagus morphology was evaluated in terms of the response to rearing media prepared with each species’ natural host plant, Pilosocereus machrisii Dawson, 1957 and Cereus hildmannianus Schum, 1890 respectively. Our results show that aedeagal shape differed significantly between species and that both shape and size presented host‐related phenotypic plasticity in both species. Flies reared on P. machrisii had, on average, larger aedeagi than those grown in C. hildmannianus. The general shape of aedeagus also differed significantly between flies that emerged in different host cactus. Patterns of variation in aedeagus morphology are discussed in the light of the current knowledge of evolutionary relationships and host plant use, in the D. buzzatii cluster, an assemblage of species in active cladogenesis.     

Condition dependence of male and female genital structures in the seed beetle Callosobruchus maculatus (Coleoptera: Bruchidae)

Theory predicts that costly secondary sexual traits will evolve heightened condition dependence, and many studies have reported strong condition dependence of signal and weapon traits in a variety of species. However, although genital structures often play key roles in intersexual interactions and appear to be subject to sexual or sexually antagonistic selection, few studies have examined the condition dependence of genital structures, especially in both sexes simultaneously. We investigated the responses of male and female genital structures to manipulation of larval diet quality (new versus once‐used mung beans) in the bruchid seed beetle Callosobruchus maculatus. We quantified effects on mean relative size and static allometry of the male aedeagus, aedeagal spines, flap and paramere and the female reproductive tract and bursal spines. None of the male traits showed a significant effect of diet quality. In females, we found that longer bursal spines (relative to body size) were expressed on low‐quality diet. Although the function of bursal spines is poorly understood, we suggest that greater bursal spine length in low‐condition females may represent a sexually antagonistic adaptation. Overall, we found no evidence that genital traits in C. maculatus are expressed to a greater extent when nutrients are more abundant. This suggests that, even though some genital traits appear to function as secondary sexual traits, genital traits do not exhibit heightened condition dependence in this species. We discuss possible reasons for this finding.     

Possible coevolution of male and female genital form and function in a calopterygid damselfly

In this paper some evolutionary changes of genitalia in the damselfly Calopteryx haemorrhoidalis are investigated by determining their current and past function. Calopteryx haemorrhoidalis males stimulate females by aedeagal frictioning on a set of vaginal sensilla. The aedeagus is considerably variable and positively correlates with volumes of ejected sperm from the spermatheca. Interestingly, females show a significantly reduced sensillum number compared with other family members. Here I explore whether there existed directional selection for aedeagal width at its evolutionary onset; and whether the sensillum reduction evolved to make sperm ejection less effective. Using C. haemorrhoidalis aedeagi in females whose species retained the ancestral conditions (no stimulatory ability and large sensillum numbers), Hetaerina cruentata and C. xanthostoma, my results corroborated these assumptions: variation in aedeagal width inversely correlated with sperm ejection rate while sperm ejection was higher in species with high sensillum numbers. A suggested coevolutionary interpretation of these results in C. haemorrhoidalis is that aedeagal width was favoured which was followed by a sensillum reduction.     

How the length of genital parts affects copulation performance in a carabid beetle: implications for correlated genital evolution between the sexes

To identify factors leading to the correlated evolution of exaggerated male and female genitalia, we studied the effects of the variable dimensions of corresponding functional genital parts (male copulatory piece and female vaginal appendix) on copulatory performance in the polygamous carabid beetle Carabus (Ohomopterus) maiyasanus. We used mating pairs of individuals from two populations to increase the variances in genital dimensions and determined the copulation performance (insemination and spermatophore replacement, and copulation time) in single‐ and double‐mating situations. In single mating, insemination success was not affected by genital dimensions, although the copulation time was significantly shorter when the male aedeagus was longer. In the double‐mating experiment, insemination and replacement of spermatophores by the second male succeeded more frequently when the copulatory piece was shorter and the vaginal appendix was longer, and when the difference between the length of the copulatory piece and the vaginal appendix was smaller. Thus, a matching of the corresponding genital parts between the sexes increases the male's reproductive success in sperm competition, but elongation of the copulatory piece cannot be explained simply by the improvement in male reproductive success. We discuss possible factors for the elongation of genital parts in terms of sexual conflict and reproductive interference through interspecific copulation.     

SPERM COMPETITION AND THE EVOLUTION OF NUPTIAL FEEDING BEHAVIOR IN THE CRICKET,GRYLLODES SUPPLICANS (WALKER)

The pattern of sperm predominance in doubly mated female crickets, Gryllodes supplicans, was investigated using a radiation-sterility technique. Female G. supplicans made significant use of sperm from both males in fertilizing eggs; overall, first males to mate enjoyed a small advantage, fertilizing about 60% of the offspring produced subsequent to the second mating. The combined use of the sperm of both males in fertilizing eggs occurred soon after the second mating; evidently, mixing of ejaculates within a female's spermatheca does occur. Male G. supplicans provide females with a nuptial gift, the spermatophylax, which influences the time at which a female removes the externally attached sperm-ampulla; this in turn determines the quantity of sperm that is transferred. Moreover, the degree of sperm precedence achieved by a male may be positively related to the time at which the female removes his sperm ampulla. Thus males, by feeding females, ensure not only that a sufficient number of sperm are transferred to fertilize all of a female's eggs, but also may increase the certainty of their paternity. In mating systems in which females control sperm transfer and paternity is influenced by numbers of sperm (i.e., numerical sperm competition), an increase in prezygotic investment in females may be an adaptive male response.     

A model to explain spermatophore implantation in cephalopods (Mollusca: Cephalopoda) and a discussion on its evolutionary origins and significance

Male coleoid cephalopods produce spermatophores that can attach autonomously on the female's body during a complex process of evagination called the ‘spermatophoric reaction’, during which the ejaculatory apparatus and spiral filament of the spermatophore are everted and exposed to the external milieu. In some deepwater cephalopods, the reaction leads to the intradermal implantation of the spermatophore, a hitherto enigmatic phenomenon. The present study builds upon several lines of evidence to propose that spermatophore implantation is probably achieved through the combination of (1) an ‘evaginating‐tube’ mechanism performed by the everting ejaculatory apparatus and (2) the anchorage provided by the spiral filament's stellate particles. The proposed theoretical model assumes that, as it is exposed to the external milieu, each whorl of the spiral filament anchors to the surrounding tissue by means of its sharp stellate particles. As the ejaculatory apparatus tip continues evaginating, it grows in diameter and stretches lengthwise, enlarging the diameter of the whorl and propelling it, consequently tearing and pushing the anchored tissue outward and backward, and opening space for the next whorl to attach. After the ejaculatory apparatus has been everted and has perforated tissue, the cement body is extruded, possibly aiding in final attachment, and the sperm mass comes to lie inside the female tissue, encompassed by the everted ejaculatory apparatus tube. It is proposed that this unique, efficient spermatophore attachment mechanism possibly evolved in intimate relationship with the adoption of an active mode of life by coleoids. The possible roles of predation pressure and sperm competition in the evolution of this mechanism are also discussed. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 711–726.     

Anatomy and histology of the male reproductive system of the fire ant,Solenopsis invicta Buren (Hymenoptera : Formicidae)

The male reproductive system of the fire ant, Solenopsis invicta Buren (Hymenoptera : Formicidae), consists of the testes, vasa efferentia, vasa deferentia, seminal vesicles, accessory glands, ejaculatory duct, wedge, aedeagal bladder, and external genitalia. The testes in newly eclosed males appear as 4 large white lobes filled with packets of sperm. Each lobe of the testes contains only one follicle. As the testes degenerate, the maturing sperm migrate through the vasa efferentia and vasa deferentia into the seminal vesicles. The seminal vesicles attach to the accessory glands, which are lined with secretory columnar epithelium. The posterior ends of the accessory glands taper and unite to form the ejaculatory duct. A sclerotized wedge is found at the junction of the accessory glands and the ejaculatory duct. An aedeagal bladder, joining the ejaculatory duct posterior to the wedge, is lined with squamous epithelium enveloped by heavy musculature. The ejaculatory duct continues posteriorly to form a distal aedeagus surrounded by 3 pairs of valves, comprising the external genitalia.     

Fine structure of the male genital system of the predatory mite Rhagidia halophila (Rhagidiidae,Prostigmata, Actinotrichida)

The male genital system of the actinotrichid mite Rhagidia halophila is described and compared with other mites and arachnids. The large testes are composed of germinal and glandular parts and produce numerous small sperm cells. The glandular parts are connected via a testicular bridge. Spermiogenesis occurs in cysts containing spermatids in equal stages of development. Cysts of spermatids are embedded in huge somatic cells. The nuclei of the spermatids loose their envelope. Mature sperm cells are simple exhibiting a ring‐shaped chromatin body and lacking an acrosomal complex. They are most similar to the sperm cells of the related mite Linopodes motatorius. The spermatopositor contains the ejaculatory duct divided into a dorsal channel and a ventral channel that are connected via a narrow passage. At its distal end, the spermatopositor is divided into three eugenital lips. The function of the spermatopositor during deposition of the peculiar thread‐like spermatophores is discussed. Details of the sensilla of the spermatopositor and the progenital lips are reported. The genital papillae located on the inner side of the progenital lips exhibit characteristics of cells performing transport of ions and/or water. The results confirm the overall similarity of actinotrichid genital systems, which is profoundly different from that of anactinotrichid mites. With reference to other Arachnida it is corroborated that testes and sperm structure of Actinotrichida are most similar to that of Solifugae. However, synapomorphies between sperm cells of Rhagidia and Solifugae that could suggest a closer relationship between these two taxa as was suggested in earlier studies were not recognizable. On the contrary, the sperm cells of Rh. halophila being devoid of an acrosomal complex appeared to be more apomorphic than those of many other actinotrichid mites as well as Solifugae. J. Morphol. 276:832–859, 2015. © 2015 Wiley Periodicals, Inc.     

Why Did a Female Penis Evolve in a Small Group of Cave Insects?

The evolution of a female penis is an extremely rare event and is only known to have occurred in a tribe of small cave insects, Sensitibillini (Psocodea: Trogiomorpha: Prionoglarididae). The female penis, which is protrudable and inserted into the male vagina‐like cavity during copulation to receive semen, is thought to have evolved independently twice in this tribe, in the Brazilian Neotrogla and the African Afrotrogla. These findings strongly suggest that there are some factors unique to Sensitibillini that have facilitated female penis evolution. Here, several hypothetical factors are presented that may have enabled the evolution of the female penis in Sensitibillini. The female–female competition for nutritious semen, the oligotrophic environment, and the twin insemination slots with switching valve are considered to be the driving factors for female penis evolution. Additionally, the following factors are considered responsible for relaxing the constraint against female penis evolution: preexistence of the female‐above mating position, the elongated duct connecting the female pre‐penis with the sperm storage organ, and the small male genital cavity accepting the female genital tubercle bearing the opening of this duct. Understanding the factors enabling female penis evolution may also shed light on the evolution of the male penis.     

The male reproductive system and mechanism of sperm transfer in Argulus japonicus (Crustacea: Branchiura)

The histomorphology of the male reproductive system and surface morphology of the “peg-and-socket” in Argulus japonicus are described from serial sagittal and transverse sections and scanning electron micrographs. The prostate complex consists of a glandular part, a reservoir for storing the secretion, and an efferent duct opening into the ejaculatory duct. The openings of both the vas deferens and the prostate duct into the ejaculatory duct are guarded by sphincters. The ejaculatory ducts, which are lined by tall columnar epithelial cells, do not open into the cuticle-lined genital atrium but are blind-ending tubes. This observation and results obtained from observing live specimens, as well as the fact that no spermatophores are formed, suggest that semen could leave the ejaculatory duct only after puncturing of its walls. It is suggested that sperm transfer is accomplished in the following manner: during copulation contraction of the muscular walls of the vas deferens and prostate duct causes semen to be pumped into the ejaculatory duct, which is then closed off by sphincters and a high internal pressure is developed. When a spermathecal spine penetrates the walls of the ejaculatory duct, semen flows from the ejaculatory duct into the spermathecal vesicle due to the higher pressure in the ejaculatory duct. This mechanism is analogous to the sucking up of fluid with a hypodermic syringe. © 1993 Wiley-Liss, Inc.     

Pluviantaenia kassalensis n. g., n. sp. (Davaineidae), a new cestode from the Egyptian plover Pluvianus aegyptius (L.) in the Sudan

Pluviantaenia kassalensis n. g., n. sp. (Davaineidae), from the intestine of Pluvianus aegyptius (Charadriiformes, Glareolidae) in the Kassala area of the Sudan, differs from other davaineid cestodes in having an armed scolex with an inflated pseudorostellum terminating in two fan-shaped lobes, in the absence of suckers, the presence of a very large sucker-like genital atrium provided distally with a powerful sphincter, and the presence of a thin-walled cirrus-sac, smaller than the genital atrium, enclosing a robust, armed cirrus. The genital atrium cirrus-sac complex, but not the female organs, may be duplicated bilaterally and/or additional sets may occur in marginal or submarginal positions.     

Demonstrating sexual selection by cryptic female choice on male genitalia: What is enough?

Rapid divergence in external genital structures occurs in nearly all animal groups that practice internal insemination; explaining this pattern is a major challenge in evolutionary biology. The hypothesis that species‐specific differences in male genitalia evolved under sexual selection as courtship devices to influence cryptic female choice (CFC) has been slow to be accepted. Doubts may stem from its radical departure from previous ideas, observational difficulties because crucial events occur hidden within the female's body, and alternative hypotheses involving biologically important phenomena such as speciation, sperm competition, and male‐female conflicts of interest. We assess the current status of the CFC hypothesis by reviewing data from two groups in which crucial predictions have been especially well‐tested, Glossina tsetse flies and Roeseliana (formerly Metrioptera) roeselii bushcrickets. Eighteen CFC predictions have been confirmed in Glossina and 19 in Roeseliana. We found data justifying rejection of alternative hypotheses, but none that contradicted CFC predictions. The number and extent of tests confirming predictions of the CFC hypothesis in these species is greater than that for other generally accepted hypotheses regarding the functions of nongenital structures. By this criterion, it is reasonable to conclude that some genital structures in both groups likely involved sexual selection by CFC.