Intromittent Organ Research Articles

Mating without intromission in a bat

Copulatory behaviours stand as cornerstones of sexual selection, yet they remain mysterious in many species. Because of their nocturnal and elusive lifestyle, the copulatory behaviours of bats have been mostly overlooked1. Several aspects of bat reproduction differ from other mammals (e.g. prolonged sperm storage2, delayed development3). Here, we show that in serotine bats (Eptesicus serotinus) the penis is used as a 'copulatory arm' rather than an intromittent organ, revealing a novel copulatory behaviour in mammals.

Do male reproductive traits evolve at the intraspecific level in response to the amount of placentotrophy in a genus of viviparous fishes?

Placentotrophy is a particular type of maternal provisioning to developing embryos, in which mothers actively provide nutrients via complex placental structures. Placentotrophy implies less pre-fertilization investment, resulting in a shift from pre- to post-copulatory sexual selection. This change can potentially result in a conflict between females and males. This phenomenon has been demonstrated at the interspecific level in viviparous fishes of the family Poeciliidae, in which males of species that lack placentotrophy have evolved traits related to pre-copulatory sexual selection such as coloration, ornaments, and courtship behavior. Placentotrophic species, on the other hand, have evolved traits associated with post-copulatory sexual selection such as long intromittent organs (gonopodium) and increased sexual coercion behavior. Here we test, for the first time at the intraspecific level, whether there is a similar relationship between a higher degree of female placentotrophy and the evolution of male reproductive traits (larger testes and longer gonopodia) in three species of the genus Poeciliopsis (P. gracilis, P. infans, and P. prolifica). We observed a tendency towards longer gonopodia in males of P. gracilis as well as the largest testes of P. prolifica males in the populations with the highest degrees of placentotrophy. However, the statistical support for these findings was relatively weak. Therefore, we failed to support the hypothesis of a selective effect of female placentotrophy on male gonads and genitalia. We discuss other evolutionary forces that may have driven the observed intraspecific variation in male reproductive traits of Poeciliopsis fishes.

Vaginas

What is the vagina? This seemingly simple question has a rather complex answer, depending on whether we use a functional or a developmental definition. The terminal portion of the female reproductive tract that opens to the environment initially served as a conduit for eggs to be laid, and in species with external fertilization the distal oviduct may be specialized for oviposition but there is no vagina. In animals with internal fertilization, this terminal section of the oviduct interacts with the sperm and the intromittent organ leading to functional specialization of this region that we often call a vagina in insects and some vertebrates. Here we address the evolution, morphology and diverse functions of the vagina and some of the unknown questions that remain to be addressed in the study of this remarkable structure.

Are males just passive? Coupling mechanism of the Brazilian cave insects with inverted genitalia.

Species of the Brazilian cave barklouse genus Neotrogla (Psocodea: "Psocoptera": Trogiomorpha: Prionoglarididae: Sensitibillini) are known to have a "female penis (gynosome)" that functions as an intromittent organ inserted into the membranous pouches in the simple male genital chamber during copulation to receive semen. However, the functions of other male and female genital structures and the copulatory processes of Neotrogla were completely unknown to date. Based on µCT observation of the male and female postabdomen and connected muscles both before and in copula, we clarified the functions of the male and female genital structures. In addition, based on the analyses of the established 3D models, we concluded that precise and rigid contact of multiple genital structures, and step-by-step releases of each holding mechanism achieved by the cooperation of both sexes are involved in the copulatory processes. The coevolution between the male and female genital structures in Neotrogla may provide a new example for the evolution of tolerance traits.

Quantifying Shape Variation in an Antisymmetrical Trait in the Tropical Fish Xenophallus umbratilis

Antisymmetry is a striking, yet puzzling form of biological asymmetry. The livebearing fish Xenophallus umbratilis exhibits antisymmetry in the male intromittent organ and provides a system that is well-suited for studying the nature of variation in antisymmetrical traits. Using geometric morphometrics, we test the hypothesis that because the gonopodium is critical to fitness there will not be significant differences in gonopodium shape between the two gonopodial morphs in this species. Our results are consistent with this prediction, though we found that gonopodium shape differed with gonopodium size.

First evidence of hemiclitores in snakes.

Female genitalia are conspicuously overlooked in comparison to their male counterparts, limiting our understanding of sexual reproduction across vertebrate lineages. This study is the first complete description of the clitoris (hemiclitores) in female snakes. We describe morphological variation in size and shape (n = 9 species, 4 families) that is potentially comparable to the male intromittent organs in squamate reptiles (hemipenes). Dissection, diffusible iodine contrast-enhanced micro-CT and histology revealed that, unlike lizard hemiclitores, the snake hemiclitores are non-eversible structures. The two individual hemiclitores are separated medially by connective tissue, forming a triangular structure that extends posteriorly. Histology of the hemiclitores in Australian death adders (Acanthophis antarcticus) showed erectile tissue and strands/bundles of nerves, but no spines (as is found in male hemipenes). These histological features suggest the snake hemiclitores have functional significance in mating and definitively show that the hemiclitores are not underdeveloped hemipenes or scent glands, which have been erroneously indicated in other studies. Our discovery supports that hemiclitores have been retained across squamates and provides preliminary evidence of differences in this structure among snake species, which can be used to further understand systematics, reproductive evolution and ecology across squamate reptiles.

Loss of Avian Intromittent Organs as a Sperm Competition Strategy: A Race to Be Last.

AbstractThe loss of the intromittent organ (IO) in the majority of birds remains unexplained. Here, I propose that IO loss results from sperm competition in the context of the unique avian sperm storage system. The first stage of fertilization is the movement of sperm through the vagina from the site of ejaculation to the sperm storage tubules (SSTs) at the uterovaginal junction. In a second stage, sperm are released from the SSTs and move through the uterus to the site of fertilization in the infundibulum. Last-male fertilization precedence can occur if sperm that arrive later at the uterovaginal junction occupy uterus-side SSTs, which then have a head start in the race to the infundibulum with each ovulation. Under such "last-in, first-out" conditions, there is strong sperm competition to arrive later at the SSTs. Consequently, the optimal male strategy would be placement of the ejaculate at the cloacal opening to allow any other sperm in the vagina to reach the SSTs first. Cloacal placement is effectively achieved by loss of the IO. The evolution of altricial development in Neoaves, the largest clade that lacks IOs, created conditions that favor IO loss. Specifically, the smaller clutch sizes and hatching asynchrony of altricial birds increase the intensity of sperm competition for fertilization of early eggs in the laying sequence and thus the selective advantage of later arrival at the SSTs. The rarity of IO loss among all animals suggests that the complex mechanism of avian fertilization creates unique conditions for sperm competition.

Genetic assimilation and the evolution of direction of genital asymmetry in anablepid fishes.

Phylogenetic comparative studies suggest that the direction of deviation from bilateral symmetry (sidedness) might evolve through genetic assimilation; however, the changes in sidedness inheritance remain largely unknown. We investigated the evolution of genital asymmetry in fish of the family Anablepidae, in which males' intromittent organ (the gonopodium, a modified anal fin) bends asymmetrically to the left or the right. In most species, males show a 1 : 1 ratio of left-to-right-sided gonopodia. However, we found that in three species left-sided males are significantly more abundant than right-sided ones. We mapped sidedness onto a new molecular phylogeny, finding that this left-sided bias likely evolved independently three times. Our breeding experiment in a species with an excess of left-sided males showed that sires produced more left-sided offspring independently of their own sidedness. We propose that sidedness might be inherited as a threshold trait, with different thresholds across species. This resolves the apparent paradox that, while there is evidence for the evolution of sidedness, commonly there is a lack of support for its heritability and no response to artificial selection. Focusing on the heritability of the left : right ratio of offspring, rather than on individual sidedness, is key for understanding how the direction of asymmetry becomes genetically assimilated.

Environmentally relevant arsenic exposure affects morphological and molecular endpoints associated with reproduction in the Western mosquitofish, Gambusia affinis

Arsenic (As) exposure, even at low environmentally relevant levels, may cause detrimental health outcomes through developmental toxicity and by acting as an endocrine disrupting compound (EDC). Although several studies indicate that wildlife bioaccumulate As, few evaluate the health impact on fish species in their natural environment. In the U.S., As has a drinking water regulatory limit of 10 μg/L. In many parts of Arizona, surface water and groundwater have naturally elevated levels of As from geologic deposits and contamination is exacerbated by anthropogenic activity. In aquatic environments, the Western mosquitofish, Gambusia affinis, is a good bioindicator for EDC exposure because of the distinct androgen-related development of an intromittent organ, the gonapodium, in males. We evaluated morphological and reproductive outcomes in mosquitofish exposed to As. In a laboratory experiment, juvenile male mosquitofish were exposed to sodium arsenite (0 μg/L, 0.75 μg/L, 7.50 μg/L, and 75 μg/L) for 30 days, and in a field study, populations of adult male mosquitofish were collected in Arizona waterways with As levels above and below the World Health Organization's regulatory limit. In both studies, higher As exposure was significantly associated with altered hepatosomatic indices, altered fish morphology, shortened gonopodia, and lower gonopodia-somatic indices. In the field experiment, populations from surface water with higher As concentrations exhibited lower condition factors, lower gonadal-somatic indices, distinct gonopodia shapes, and altered estrogen receptor alpha and vitellogenin gene expression; androgen receptor expression was unchanged. Together, laboratory and field results suggest that As exposure at environmentally-relevant levels affects general growth and reproductive development in mosquitofish. Observed effects may further influence individual health, mobility, or reproductive function, and because G. affinis is a species known to tolerate and adapt to a wide range of environments, it serves as a local bioindicator species as well as a model organism for parallel field and laboratory studies.

Avian reproductive disorders

Reproductive disorders are common in birds and are frequently encountered in avian practice. They often involve emergency presentations and may be life threatening. In female birds, chronic egg laying, egg binding, dystocia, ovarian disease, oviductal disease and egg coelomitis are all examples of common reproductive disorders. In male birds, reproductive disorders include testicular diseases and prolapse of the phallus in those species which possess an intromittent organ. Depending on the disorder, medical or surgical management may be indicated. The use of deslorelin (a gonadotropin-releasing hormone agonist) implants has become popular in avian medicine over the last decade and some studies have explored the efficacy of this treatment in some species, although a lot more research is needed to establish the efficacy in a wider range of species, in both sexes and with repeated use.

Frequent origins of traumatic insemination involve convergent shifts in sperm and genital morphology.

Traumatic insemination is a mating behavior during which the (sperm) donor uses a traumatic intromittent organ to inject an ejaculate through the epidermis of the (sperm) recipient, thereby frequently circumventing the female genitalia. Traumatic insemination occurs widely across animals, but the frequency of its evolution, the intermediate stages via which it originates, and the morphological changes that such shifts involve remain poorly understood. Based on observations in 145 species of the free‐living flatworm genus Macrostomum, we identify at least nine independent evolutionary origins of traumatic insemination from reciprocal copulation, but no clear indication of reversals. These origins involve convergent shifts in multivariate morphospace of male and female reproductive traits, suggesting that traumatic insemination has a canalizing effect on morphology. We also observed sperm in both the sperm receiving organ and within the body tissue of two species. These species had intermediate trait values indicating that traumatic insemination evolves through initial internal wounding during copulation. Finally, signatures of male‐female coevolution of genitalia across the genus indicate that sexual selection and sexual conflict drive the evolution of traumatic insemination, because it allows donors to bypass postcopulatory control mechanisms of recipients.

Preliminary assessment of mating duration and prolonged post-copulatory associations in Arrenurus water mites (Actinotrichida: Parasitengonina: Hydrachnidiae)

Conflicts of interest between the sexes over control of mating can be reflected in various aspects of morphology and behaviour, including structure of genitalia and copulation duration. In Arrenurus water mites (Hydrachnidia: Arrenuridae) there are two main patterns of sperm transfer that differ in degree of potential male control of sperm uptake by females. In some species, males are equipped with an intromittent organ (the ‘petiole’) that is used to forcefully insert sperm into the female reproductive tract. In others, males lack an intromittent organ and females appear to push sperm into their reproductive opening themselves. Theory suggests that the amount of time spent in courtship after sperm transfer should differ between males with and without an intromittent structure. We predicted that male Arrenurus able to push sperm into the female’s reproductive tract (petiolate males) should spend less time courting females after transferring sperm than apetiolate males, which may have to ‘convince’ females to take up their sperm. Here, we examined durations of mating for 11 species of Arrenurus with males that differ in genital morphology: seven species with males equipped with a well-developed petiole (= ‘petiolate’ species) and four species with males that either completely lack a petiole or have a minute peg-like petiole that does not appear to function as an intromittent organ (= ‘apetiolate’ species). We tested whether males of petiolate species spend less time in the stage of courtship that takes place after sperm transfer (= ‘post-transfer courtship’) than apetiolate males. In contrast to our prediction, we found that species with well developed petioles spent significantly more time in post-transfer behaviours than species lacking petioles. The possible function of protracted post-transfer courtship in the genus Arrenurus is discussed.

Micro-Computed Tomography Reveals a Remarkable Twin Intromittent Organ in Spiders – A Novelty for Arachnids With Direct Sperm Transfer

The modification of male pedipalps into secondary sexual intromittent organs is one of the hallmark characteristics of spiders, yet understanding the development and evolution of male genitalia across the order remains a challenging prospect. The embolus – the sclerite bearing the efferent spermatic duct or spermophor, and used to deliver sperm directly to the female genitalia during copulation – has always been considered the single unambiguously homologous palpal sclerite shared by all spider species, fundamental to the bauplan of the order and to the evolution and functional morphology of spider reproductive systems. Indeed, after two centuries of comparative research on spider reproduction, the presence of a single spermophor and embolus on each of a male spider’s two pedipalps remains a central tenet of evolutionary arachnology. Our findings challenge this premise, and reveal a remarkable twin intromittent organ sperm transfer system in a lineage of Australian palpimanoid spiders, characterized by a bifurcate spermophor and the presence of two efferent ducts leading to a pair of embolic sclerites on each pedipalp. This is the first time such a remarkable conformation has been observed in any group of arachnids with direct sperm transfer, complicating our understanding of palpal sclerite homologies, and challenging ideas about the evolution of spider genitalia.

External reproductive structures of male Surfperches (Embiotocidae): Urogenital papilla.

Fishes exhibit a dynamic array of reproductive diversity. Among the various forms of reproduction, internal fertilization has evolved at least 13 times and has been observed in 2-3% of teleost species, half of which are marine. Internal fertilization has led to the evolution of male intromittent organs to aid in sperm transfer during copulation. The intromittent organs of poeciliids have been widely investigated, but little research has focused on the intromittent organs of marine fishes. The surfperches are internally fertilizing, viviparous fishes with a ventrally located papilla. In this article, the external morphology of 19 Embiotocidae species was examined. Additionally, paraffin histology was used to investigate the walleye surfperch (Hyperprosopon argenteum), black perch (Embiotoca jacksoni), and dwarf perch (Micrometrus minimus) papilla to aid in the identification of tissue composition to determine its function. External morphology was documented via macro photography and males were dissected to observe the relationship between the testes, spermatic duct, and papilla. We provide histological evidence that the urogenital papilla is directly connected to the spermatic duct and testis and likely involved in spermatozeugmata transfer. Our research also determined that the papillae were composed of both urinary and reproductive tissues, thus we propose the papilla be referred to as a urogenital papilla. Our study contributes to an overall understanding of intromittent organs of marine fishes, particularly in the surfperches, which has not been previously documented.

A review of sperm storage methods and post-copulatory sexual selection in the Cephalopoda

Abstract In many animal phyla, females have a unique sperm storage organ (SSO). Post-copulatory sexual selection is a powerful driving force of SSO evolution. SSOs are generally considered to have evolved through sexual antagonistic coevolution between male genitalia and the SSO and/or cryptic female choice (CFC). In cephalopods, sperm transfer and fertilization are conducted through complex processes, and sperm storage methods show inter-species variation. In some species, males implant spermatangia superficially under the female skin, and then sperm released from the spermatangia are transferred into a seminal receptacle (SR). Deep-sea cephalopods, which lack a SR, have instead evolved a deep-implanting method by which the spermatangium is embedded deep in the musculature of the mantle wall of the female. In some species, the female stores whole spermatangia within a spermatangium pocket. Because the males of most species do not insert an intromittent organ into the female when transferring sperm, SSO evolution may have been influenced by CFC alone. This review summarizes the sperm storage methods and the mechanisms of post-copulatory sexual selection in cephalopods and it is proposed that these diverse methods evolved as adaptive mechanisms through post-copulatory sexual selection.

Helminth Parasites in the Gastrointestinal Tract of Oreochromis niloticus (Nile Tilapia) in Dadin Kowa Dam, Yamaltu Deba Local Government Area, Gombe State, Nigeria

Aim: The aim of the study is to identify helminth parasites in the gastrointestinal tract of Oreochromis niloticus in Dadin Kowa Dam, Yamaltu Deba Local Government Area, Gombe State.
 Study design: Oreochromis niloticus fish species were randomly obtained from the local fishermen in the landing sites; between 7:00am-10:00am weekly.
 Place and duration of study: The study was carried out at Dadin Kowa Dam in Yamaltu Deba Local Government Area of Gombe State between the months of March to June, 2019.
 Methodology: 60 Oreochromis niloticus (Nile Tilapia) were obtained and examined for gastro intestinal parasites. Sex of the fish was determined by the presence and absent of an intromittent organ. The total length, standard length and body weight was measured using standard method. Dissection was done using standard procedure and the Parasites recovered were identified using the helminth parasites of fish identification guide.
 Results: Out of the 60 fish species examined. (23 female and 37 males), 21 (35.00%) were infected and were observed to habour six (6) endoparasites; viz nematodes, Cestodes and trematodes. The results show that there was a significant difference between infection rate, total length and weight of fish with female species having relatively higher percentage prevalence 11(43.50%) than the males 10 (29.70%). It was observed that the prevalence of parasitic infection increase with length and weight of the fish.
 Conclusion: Presence of these fish helminth parasites may lead to devaluation in protein constant in the body of the fish. The work therefore recommends water quality assessment of Dams as it is one of the factors that bring about the increase in parasitic infection.

Penetration mechanics of elongated female and male genitalia of earwigs

We unveiled the penile penetration mechanics of two earwig species, Echinosoma horridum, whose intromittent organ, termed virga, is extraordinarily long, and E. denticulatum, whose virga is conversely short. We characterised configuration, geometry, material and bending stiffness for both virga and spermatheca. The short virga of E. denticulatum has a material gradient with the stiffer base, whereas the long virga of E. horridum and the spermathecae of both species are homogeneously sclerotised. The long virga of E. horridum has a lower bending stiffness than the spermatheca. The virga of E. denticulatum is overall less flexible than the spermatheca. We compared our results to a previous study on the penetration mechanics of elongated beetle genitalia. Based on the comparison, we hypothesised that the lower stiffness of the male intromittent organ comparing to the corresponding female structure is a universal prerequisite for the penetration mechanics of the elongated intromittent organ in insects.

Does Asymmetrical Gonopodium Morphology Predict Lateralized Behavior in the Fish Xenophallus umbratilis?

Why bilaterally symmetrical organisms express handedness remains an important question in evolutionary biology. In some species, anatomical asymmetries have evolved that accompany behavioral handedness, yet we know remarkably little about causal links between asymmetric morphological traits and behavior. Here, we explore if a dextral or sinistral orientation of the male intromittent organ predicts side preferences in male behaviors. Our study addresses this question in the Costa Rican livebearing fish, Xenophallus umbratilis. This fish has a bilaterally symmetrical body plan, with one exception—the male anal fin (gonopodium), used to inseminate females, terminates with a distinct left- or right-handed corkscrew morphology. We used a detour assay to test males for side biases in approach behavior when exposed to four different stimuli (predator, potential mate, novel object, empty tank control). We found that left morph males preferred using their right eye to view potential mates, predators, and the control, and that right morph males preferred to use their left eye to view potential mates and predators, and their right eye to view the control. Males of both morphs displayed no eye bias when approaching the novel object. Our results suggest that there is a strong link between behavior and gonopodium orientation, with right and left morph males responding with opposite directional behaviors when presented with the same stimuli. This presents the intriguing possibility that mating preferences—in this case constrained by gonopodial morphology—could be driving lateralized decision making in a variety of non-mating behaviors.

Sharply Angled Lateral Curvature Modification in Anal Fin of a Viviparous Fish, Xenotoca variata (Cyprinodontiformes: Goodeidae).

Almost all viviparous species possess male external genitalia; for example, the mammalian penis is an intromittent organ. Some live-bearing bony fish use their anal fins to assist in mating and internal fertilization. We previously reported a male-specific asymmetric curvature at the posterior end of the anal fin in Xenotoca eiseni, a viviparous fish of the family Goodeidae. However, three other goodeid species, Xenotoca melanosoma, Zoogoneticus quitzeoensis, and Chapalichthys pardalis, examined in that study possessed lesser anal fin curvature modifications as compared to those in the anal fin of X. eiseni. Here, we report the second case of acute-angled curvature modification of the male anal fin in the family Goodeidae. We obtained a dead specimen of the goodeid species Xenotoca variata from a city zoo in Japan, and the morphological and histological analyses indicated an acute-angled asymmetric curvature of the posterior end of the anal fin in X. variata, similar to that observed in X. eiseni in the previous study. However, in our previous report, obtuse-angled modification was only observed in one other Xenotoca species, X. melanosoma, and two species belonging to other genera, Z. quitzeoensis and C. pardalis. Therefore, our findings suggest that the acute-angled curvature in the male anal fin has been developed in the genus Xenotoca.

Rapid evolution and plasticity of genitalia.

Genital morphology exhibits tremendous variation and is intimately linked with fitness. Sexual selection, nonmating natural selection and neutral forces have been explored as potential drivers of genital divergence. Though less explored, genitalia may also be plastic in response to the developmental environment. In poeciliid fishes, the length of the male intromittent organ, the gonopodium, may be driven by sexual selection if longer gonopodia attract females or aid in forced copulation attempts or by nonmating natural selection if shorter gonopodia allow predator evasion. The rearing environment may also affect gonopodium development. Using an experimental introduction of Trinidadian guppies into four replicate streams with reduced predation risk, we tested whether this new environment caused the evolution of genitalia. We measured gonopodium length after rearing the source and introduced populations for two generations in the laboratory to remove maternal and other environmental effects. We split full-sibling brothers into different rearing treatments to additionally test for developmental plasticity of gonopodia in response to predator cues and food levels as well as the evolution of plasticity. The introduced populations had shorter gonopodia after accounting for body size, demonstrating rapid genital evolution in 2-3years (8-12 generations). Brothers reared on low food levels had longer gonopodia relative to body size than those on high food, reflecting maintenance of gonopodium length despite a reduction in body size. In contrast, gonopodium length was not significantly different in response to the presence or absence of predator cues. Because the plastic response to low food was maintained between the source and introduced populations, there was no evidence that plasticity evolved. This study demonstrates the importance of both evolution and developmental plasticity in explaining genital variation.