Copulatory behaviour increases sperm viability in female spiders

One remarkable reproductive feature in animals with internal fertilization is a reduction in sperm viability over time in females. Whether this reduction is driven by male-male competition and/or cryptic female choice is unclear. From the perspective of cryptic female choice, we postulated that sperm viability is affected by a particular male copulatory behaviour. In this study, we investigated the following aspects: (1) sperm viability in mated females vs. males; (2) whether sperm viability varies temporally after mating; and (3) whether male copulatory behaviour covaries positively with sperm viability within females. We used the spider Holocnemus pluchei, whose males use several copulatory behaviours to court females. We found that females that stored sperm for 4 or 15 days showed no difference in sperm viability but had lower sperm viability compared with males, and males that performed a longer post-insemination behaviour had higher sperm viability inside the female. It is unclear how sperm viability is reduced and how male post-insemination behaviour affects this. It is possible that extending copulation allows males to induce females to keep sperm alive for longer. This result is predicted by theory whereby males induce females to facilitate sperm to reach and fertilize eggs based on male postcopulatory behaviour.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Costa Schmidt, Luiz Ernesto. Universidade Federal de Pelotas; Brasil. Universidade do Vale do Rio dos Sinos, Rio Grande do Sul; BrasilFil: Córdoba-Aguilar, Alex. Universidad Nacional Autónoma de México. Instituto de Ecología; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Condition-dependent male copulatory courtship and its benefits for females

Postcopulatory sexual selection has shaped the ornaments used during copulatory courtship. However, we know relatively little about whether these courtship ornaments are costly to produce or whether they provide indirect benefits to females. We used the mealworm beetle, Tenebrio molitor, to explore this. We challenged males using an entomopathogenic fungus and compared their courtship (frequency of leg and antennal contacts to the female), copulation duration, number of eggs laid, and hatching rate against control males. Infected males copulated for longer yet they reduced their leg and antennal contacts compared to control males. However, there was no obvious relation between infection, copulation duration, and courtship with egg production and hatching success. In general, our results indicate that the ornaments used during postcopulatory courtship are condition-dependent. Moreover, such condition dependence cannot be linked to male fitness.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Reyes Ramírez, Alicia. Universidad Nacional Autónoma de México; MéxicoFil: Cristancho, Shara. Universidad El Bosque; ColombiaFil: Sandoval García, Iván A.. Universidad Nacional Autónoma de México; MéxicoFil: Rocha Ortega, Maya. Universidad Nacional Autónoma de México; MéxicoFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Palacino, Freddy. Universidad El Bosque; ColombiaFil: Córdoba Aguilar, Alex. Universidad Nacional Autónoma de México; Méxic

Copulatory behaviour and genital mechanics suggest sperm allocation by a non-intromittent sclerite in a pholcid spider

The male genitalia of pholcid spiders, which is one of the most species-rich spider families, are characterized by a procursus, which is a morphologically diverse projection of the copulatory organ. It has been shown that the procursus interacts with the female genitalia during copulation. Here, we investigate the function of the procursus in Gertschiola neuquena, a species belonging to the early branched and understudied subfamily Ninetinae, using behavioural and morphological data. Although many aspects of the copulatory behaviour of G. neuquena follow the general pattern described for the family, males use only one pedipalp during each copulation. Based on our micro-CT analysis of cryofixed mating pairs using virgin females, we can show that the long and filiform procursus is inserted deeply into the unpaired convoluted female spermatheca, and the intromittent sclerite, the embolus, is rather short and stout only reaching the most distal part of the female sperm storage organ. Histological data revealed that sperm are present in the most proximal part of the spermatheca, suggesting that the procursus is used to allocate sperm deeply into the female sperm storage organ. This represents the first case of a replacement of the sperm allocation function of the intromittent sclerite in spiders.Fil: Izquierdo, Matías Andres. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentin

Genital movements are not restricted to spermatozoa transfer in a haplogyne spider

Internally fertilized animals are characterized by the transfer of their spermatozoa during copulation. However, the duration of copulation is highly variable, which suggests that they may serve for other functions apart from spermatozoa transfer. For example, during copulation, males can stimulate the female by using genitalic movements and/or by positioning their spermatozoa adequately within the female genitalia. In the spider Holocnemus pluchei, males perform strong squeezes and torsion movements with their pedipalps (part of their genitalia actively related to copulation) inside the female genitalia and transfer their spermatozoa during Phase I of copulation. However, the exact moment in which such transfer occurs and its relation to the pedipalp movements are still unknown. Herein, we first have identified the precise moment when the spermatozoa transfer occurs during phase I and its relation to genital movements by interrupting mating couples at the beginning, at middle and at end of this phase. Subsequently, the spermatozoa number remaining in male genitalia and stored in female genitalia were counted. We detected no relation between the number of genital movements and the spermatozoa number present in both, female and male genitalia and body size. As stated above, male genital movements serve another role during copulation in addition to spermatozoa transfer in a context of post-copulatory sexual selection; most likely, they stimulate the female in a cryptic female choice framework.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Copulatory behaviour and genital mechanics suggest sperm allocation by a non-intromittent sclerite in a pholcid spider

The male genitalia of pholcid spiders, which is one of the most species-rich spider families, are characterized by a procursus, which is a morphologically diverse projection of thecopulatory organ. It has been shown that the procursus interacts with the female genitalia during copulation. Here, we investigate the function of the procursus in Gertschiola neuquena, a species belonging to the early branched and understudied subfamily Ninetinae, using behavioural andmorphological data. Although many aspects of the copulatory behaviour of G. neuquena follow the general pattern described for the family, males use only one pedipalp during each copulation. Based on our micro-CT analysis of cryofixed mating pairs using virgin females, we can show that the longand filiform procursus is inserted deeply into the unpaired convoluted female spermatheca, and the intromittent sclerite, the embolus, is rather short and stout only reaching the most distal part of the female sperm storage organ. Histological data revealed that sperm are present in the most proximalpart of the spermatheca, suggesting that the procursus is used to allocate sperm deeply into the female sperm storage organ. This represents the first case of a replacement of the spermallocation function of the intromittent sclerite in spiders.Fil: Izquierdo, Matías Andres. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Dederichs, Tim M.. Ernst Moritz Arndt Universität Greifswald. Institut fur Geographie und Geologie; AlemaniaFil: Cargnelutti, Franco Ignacio. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Michalik, Peter. Ernst Moritz Arndt Universität Greifswald. Institut fur Geographie und Geologie; Alemani

Patterns of sperm transfer behavior in a pholcid spider with two distinct copulatory phases

Sexual selection is the responsible force for the evolution and maintenance of genital diversity and function. This is the case for example, of genital movements performed by males during mating and copulation duration. Spiders perform ritualized copulations whereby males carry out different types of movements using their pedipalps with varying duration. The function and duration of these pedipalp movements is unclear. In the pholcid spider, Holocnemus pluchei males that copulate with virgin females perform two copulatory phases: phase I in which the pedipalps move and phase II in which pedipalps remain motionless. Using H. pluchei as a study species, our study aims were: 1) to assess if sperm transfer occurs when pedipalps move or are still and quantify the number of sperm in male bulbs and in the female uterus externus after copulation; and, 2) to determine if amount of sperm transferred to females is associated with duration of each copulatory phase. Two experimental groups (i. e. complete copulation and interrupted copulation) were established in which the amount of sperm remaining in the male bulbs and the amount of sperm stored by females were determined. Our results show that sperm transfer occurs during phase I, that males transfer almost all sperm from their bulbs while the females store only 20% of that male amount. There was no relation between the amount of sperm transferred or stored and the duration of the copulatory phases. These results support the hypothesis that while both phases may serve a copulatory courtship, only phase I (when pedipalps move) serves for sperm transfer.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Córdoba-Aguilar, Alex. Universidad Nacional Autónoma de México; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Successive matings affect copulatory courtship but not sperm transfer in a spider model

Previous studies have reported that males augment their reproductive success by increasing the number of femaleswith which they copulate, and that such copulations are not energetically demanding in terms of trivial spermproduction costs. However, we now know that males do pay reproductive costs. As males mate successively, a reductionin the performance of copulatory behaviours would be expected, as well as in the number of sperm transferred.Here we compared the duration of courtship, mating and post-insemination phase, the number of genital and nongenitalcopulatory courtship occurrences, and the number of sperm transferred in successive matings in Holocnemuspluchei spider males. As matings increased in males, there was no effect on the duration of courtship, mating orpost-insemination phase. Interestingly, genital copulatory courtship varied in successive copulations dependingon male size, but there was no change in the number of sperm transferred. In addition, the occurrence of nongenitalcopulatory courtship decreased along successive copulations. The negative effects of successive matings oncopulatory courtship indicate that these behaviours are costly for males, except for the number of sperm transferred.Our research lays the foundation for future studies on male costs as a function of mating history in spiders.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Córdoba Aguilar, Alex. Universidad Nacional Autónoma de México; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Female secondary sexual traits in spiders: Adaptive interpretations of the sternum projection in the pholcid Holocnemus pluchei

Sexually dimorphic traits are the outcome of a network of selective pressures acting upon each sex. While male secondary sexual traits are frequently restricted to intrasexual competition, female ornaments or weapons are mostly associated to social interactions for the access to limited resources. Here we investigate a sexually dimorphic trait expressed within the female's sternum of the spider species Holocnemus pluchei (Pholcidae), and its adaptive maintenance under the three leading hypotheses: sexual receptivity signaling, mate choice and sexual conflict hypothesis. We provide fine-scaled behavioral descriptions of the copulatory behavior of Holocnemus pluchei, corroborating that mating interactions are mediated by the female's sternum projection. The single moment of contact between the female projection and male body occurs during sperm transfer, always followed by a reduction on the intensity of the male genital movement. Additionally, biometrical properties of the female sternum projection corroborate our functional interpretation for the trait as restricting the intensity of males’ copulatory movements. We claim that female sternum projection is a sexually selected trait mediating sexual conflict, with several adaptive consequences upon the sexes.Fil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Costa Schmidt, Luiz Ernesto. Universidade Do Vale Do Rio Dos Sinos; Brasi

Sex and burrowing behavior and their implications with lytic activity in the sand-dwelling spider Allocosa senex

The immune response can be costly. Studies in several arthropod species have indicated a trade-off between immunity and other life-history traits, including reproduction. In sexually dimorphic species in which females and males largely differ in their life history strategies and related energetic demands, we can expect to find sex differences in immune functions. Sex differences in immunity are well documented in vertebrates; however, we largely lack data from invertebrate systems. Lytic activity, the immune system’s ability to lysate bacteria and viruses, has been widely used as a proxy for the strength of the immune response in several invertebrates. With this in mind, we used the burrowing wolf spider Allocosa senex to test differences in lytic activity between females and males. We also studied whether digging behavior affects the immune responses in this species. While females of A. senex construct simple refuges where they stay during the day, males construct deep burrows, which they donate to females after copulation. In accordance with our hypothesis, females showed higher lytic activity compared with males, and those males who dug showed higher levels of lytic activity than those that did not dig. Furthermore, male body condition and lytic activity did not correlate with burrow length, a trait under female choice in this species. Our results show sexual dimorphism in lytic activity responses, which are likely related to differences in life-history strategies and energetic requirements of each sex in A. senex spiders.Fil: Albín, Andrea. Universidad de la República; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Simó Núñez, Miguel. Universidad de la República; UruguayFil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Aisenberg Olivera, Anita Diana. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin

Sperm viability in spiders: A first approach using Holocnemus pluchei (Scopoli, 1763) (Synspermiata: Pholcidae)

Several studies have shown that sperm viability (SV) is a key trait during sexual competition. However, this has not yet been tested in spiders as no protocol has been developed to quantify SV. Here, we describe a methodology for estimating SV using the pholcid spider Holocnemus pluchei (Scopoli, 1763). In this method, male spermatozoa were released from the sperm ducts of copulatory bulbs in 50 μl saline solution and mixed with a vortexer. Finally, 10 μl of the sperm solution were combined with 1 μl of GelRed stain. We counted a total of 200 cells and calculated the percentage that was not stained. Males showed almost 100% of live sperm inside their genital bulbs. GelRed stain demonstrated a high effectiveness to distinguish between dead and living sperm cells in spiders and it is thus a more reliable option compared to the dyes typically used, such as propidium iodide and sybr14. The high SV in H. pluchei may be explained by the multiple mating nature of females, which suggests a selection for enhanced effectiveness during sperm competition within the female genital tract.Fil: Cargnelutti, Franco Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Uñates, Diego Rafael. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Vrech, David Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Bollatti, Fedra Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Calbacho Rosa, Lucía Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; ArgentinaFil: Córdoba Aguilar, Alex. Universidad Nacional Autónoma de México; MéxicoFil: Peretti, Alfredo Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentin