Precopulatory oral sex contact plays an important role in copulatory success in a cryptic desert beetle

Precopulatory courtship plays an essential role in the insemination process and influences postcopulatory behavior between males and females. Male precopulatory oral stimulation of female genitals is rare for invertebrates. Here, we describe an intriguing oral sexual courtship in a cryptic desert beetle Platyope mongolica Faldermann. The males repeatedly contact the female''s genitals using their mouths to gain consent to mate. Furthermore, the rate at which males contact the female''s genitals relates to the copulation success in a series of observations. However, interference in oral sexual contacts decreased the proportion of successful copulation. Further no‐choice tests found homosexual behavior between males with antenna removed. We report the precopulatory oral sexual behavior and its important role for copulation success in P. mongolica for the first time. These findings highlight the significance of oral sexual courtship in sexual selection.     

Female resistance and male force: context and patterns of copulation in the New Zealand stitchbird Notiomystis cincta

The New Zealand stitchbird or hihi Notiomystis cincta is unique in that it has two distinct mating positions, in addition to the male standing on the female's back, as is seen in all other birds, it also copulates face‐to‐face. In this study, 43 male stitchbirds first attracted a female to their territory and supplemented their within‐pair matings by intruding into other territories and attempting forced copulations – often resulting in high levels of female harassment. I recorded the temporal variation of both attempted and successful copulations relative to the female's fertile period in order to understand the function of copulation variation in this species. Each of 105 observed copulations were classified according to whether they were: (1) within‐pair or extra‐pair, (2) forced or unforced, and (3) face‐to‐face or standing. Two copulation categories ‘within‐pair unforced standing’ (50%) and ‘extra‐pair forced face‐to‐face’ (27%) accounted for the majority of all observed copulations, and this supported the previous categorisation of face‐to‐face copulation being forced by extra‐pair males in this species. However, in 10% of copulations the female conceded to copulate with an extra‐pair male in a standing position while displaying only subtle signs of resistance, thus, female stitchbirds may show convenience polyandry when the costs of resistance are high. The peak in copulation frequency centred on two days prior to the laying of the first egg and occurred within a period of six days before and seven days after the first egg was laid. Unsuccessful extra‐pair forced copulation attempts closely followed the distribution of all copulations. Male age and morphometrics did not predict whether a female would resist or accept extra‐pair copulation attempts, and female resistance did not appear to depend on male quality. Despite the current trend focussing on female fitness benefits associated with EPCs, it appears that male stitchbirds gain EPCs through forced copulation and females gain no apparent benefit.     

Copulatory courtship by bushcricket genital titillators revealed by functional morphology, μCT scanning for 3D reconstruction and female sense structures

Genitalia are rapidly evolving morphological structures most likely under sexual selection. Due to their internal nature they are often hidden inside the body, thus morpho-functional studies of animal genitalia are broadly lacking. Males of some bushcricket taxa bear paired genital appendices called titillators, the exact function of which is unknown since they are obscured inside the female body during pairing. To investigate titillator morphology and possible function during copulation, we studied the bushcricket Metrioptera roeselii (Hagenbach, 1822) using a novel combination of independent, yet complementary, techniques. Copulating pairs were snap-frozen and scanned by X-ray micro-computed tomography (μCT) to visualize the coupling of male and female genitalia in situ. Video recordings of copulating pairs also showed rhythmical insertion of male titillators into the female's genital chamber, where they percuss a softened structure on the female's subgenital plate. Movements did not induce damage to the female's structure, which lacks any sclerotized genital counterparts. Instead, scanning electron microscopy and histological sections show the female subgenital plate to be covered with two different types of sensory receptors at the contact zone between the male's titillator and the female genital chamber. We interpret the non-harmful function of the titillator processes, the lack of a genital counter-structure and the presence of sensory cells on the female's subgenital plate as indicators of a copulatory courtship function of titillators, subject to sexual selection by female choice.     

Impact of cryptic female choice on insemination success: Larger sized and longer copulating male squid ejaculate more,but females influence insemination success by removing spermatangia

In polyandrous mating systems, sperm competition and cryptic female choice (CFC) are well recognized as postcopulatory evolutionary forces. However, it remains challenging to separate CFC from sperm competition and to estimate how much CFC influences insemination success because those processes usually occur inside the female's body. The Japanese pygmy squid, Idiosepius paradoxus, is an ideal species in which to separate CFC from sperm competition because sperm transfer by the male and sperm displacement by the female can be observed directly at an external location on the female's body. Here, we counted the number of spermatangia transferred to, removed from, and remaining on the female body during single copulation episodes. We measured behavioral and morphological characteristics of the male, such as duration of copulation and body size. Although males with larger body size and longer copulation time were capable of transferring larger amounts of sperm, females preferentially eliminated sperm from males with larger body size and shorter copulation time by spermatangia removal; thus, CFC could attenuate sperm precedence by larger males, whereas it reinforces sperm precedence by males with longer copulation time. Genetic paternity analysis revealed that fertilisation success for each male was correlated with remaining sperm volume that is adjusted by females after copulation.     

Behaviours of collared and white-lipped peccaries (Tayassu tajacu andT. pecari) in relation to sexual receptivity of the female

Changes in behaviours of the two peccary species,Tayassu tajacu Linnaeus, 1758 andT. pecari Link, 1795 between non-receptive and receptive periods were followed by presenting females to males daily for 15 minutes. InT. tajacu, the rank order of behaviours, similar in both sexes during the non-receptive period, differs during receptivity. Contact behaviours decrease in males, whereas sexual ones progress. The same tendency appears in females. Inhibited bites replace markings of partner as the most common behaviour in both sexes. InT. pecari, the rank order of behaviours always differs between sexes. When females become receptive, the differences from the non-receptive period are neither numerous nor significant. The most common behaviour of males, previously markings of partner, becomes mounts, whereas in females agonistic behaviours reinforce their dominance. In this species, the only behaviours that increase are those leading directly to copulation or those of an agonistic nature. In both species, females show more agonistic behaviours than males (mainly inhibited bites inT. tajacu, truly aggressive ones inT. pecari). When females are receptive, males ofT. pecari become less active, contrary toT. tajacu where both sexes double their activity. InT. tajacu, most behaviours vary significantly in relation to the progesterone level, contrary to the other species. These pecularities appear correlated to herd composition and organisation.     

Evolutionary trends in Mio-Pliocene Leporinae, based onTrischizolagus (Mammalia, Lagomorpha)

New material ofTrischizolagus dumitrescuae from Moldova and Ukraine is described. The variation of p3 inTrischizolagus shows the gradual shift of morphotype frequencies from the ‘Hypolagus’ pattern in Turolian through the mixture of three patterns (including ‘Nekrolagus’ morphotype) in Early Ruscinian to the dominant ‘Alilepus’ pattern in the Late Ruscinian samples. These transformations took place parallel to that of the North AmericanNekrolagus. Probably North AmericanSylvilagus, Brachylagus, andRomerolagus had an North American origin fromNekrolagus, whereas Eurasiatic and AfricanOryctolagus, Caprolagus, Nesolagus, andPoelagus could have originated in the Old World fromTrischizolagus.     

Matriphagy in the Spider Amaurobius ferox (Araneidae,Amaurobiidae): an Example of Mother-Offspring Interactions

In the spider Amaurobius ferox, the mothers are systematically devoured by their young at a fairly constant interval. Observation of broods maintained under normal conditions showed the cannibalistic processes to be achieved within a few hours, in the course of which mothers and offspring appeared to exchange stimulation. In particular, the mothers exhibited ‘solicitation’ behaviours which appeared to activate and synchronize the young. An experiment was carried out to specify the respective role played by the two parties of the dyad in this case of matriphagy. The female's attitude towards the young (‘solicitation’, tolerance or predatory response) appeared to depend on her reproductive state. The attitude of the young towards the female (cannibalism, attraction, or flight) was shown to depend on their stage of development, but also on the female's attitude. These results support the hypothesis that, in A. ferox, matriphagy is regulated by mother-offspring interactions. Such a mechanism could ensure precise tuning of matriphagy in relation to the requirements of the young, and a certain amount of flexibility in reproduction.     

POLYGYNANDRY IN THE DUSKY PIPEFISH SYNGNATHUS FLORIDAE REVEALED BY MICROSATELLITE DNA MARKERS

In the dusky pipefish Syngnathus floridae, like other species in the family Syngnathidae, ‘pregnant’ males provide all post-zygotic care. Male pregnancy has interesting implications for sexual selection theory and the evolution of mating systems. Here, we employ microsatellite markers to describe the genetic mating system of S. floridae, compare the outcome with a previous report of genetic polyandry for the Gulf pipefish S. scovelli, and consider possible associations between the mating system and degree of sexual dimorphism in these species. Twenty-two pregnant male dusky pipefish from one locale in the northern Gulf of Mexico were analyzed genetically, together with subsamples of 42 embryos from each male's brood pouch. Adult females also were assayed. The genotypes observed in these samples document that cuckoldry by males did not occur; males often receive eggs from multiple females during the course of a pregnancy (six males had one mate each, 13 had two mates, and three had three mates); embryos from different females are segregated spatially within a male's brood pouch; and a female's clutch of eggs often is divided among more than one male. Thus, the genetic mating system of the dusky pipefish is best described as polygynandrous. The genetic results for S. floridae and S. scovelli are consistent with a simple model of sexual selection which predicts that for sex role-reversed organisms, species with greater degrees of sexual dimorphism are more highly polyandrous.     

Temporal patterning of courtship behaviour in some parasitic Hymenoptera,with special reference to Melittobia acasta

In many parasitic hymenoptera copulation is preceded by elaborate courtship displays which include species-specific characteristics. Other features, shared by related species, may be used for defining higher taxa. The male's repertoire consists of movements involving the wings, legs, antennae, and mouthparts. These movements are performed continuously, or intermittently, depending on the species involved. The elements of a repertoire are repeated over and over again until the female indicates her readiness to copulate. Temporal patterning of various displays, and the timing of the female response are described. In Melittobia acasta (Walker) (Eulophidae) the male display is composed not only of repeating elements, but also includes new elements, introduced along the way; the display progresses towards a climactic finale. The timing of the female's copulation signal is accurately predictable. The morphology of Melittobia males is discussed in relation to this behaviour. Courtship of a related species, M.chalybii, is compared to the courtship of M.acasta.     

A comparative approach to the organization of vocal signals in noctural prosimians

Vocalizations and corresponding behavior patterns were recorded inGalago demidovii (Lorisidae), Microcebus murinus, andCheirogaleus medius (Cheirogaleidae). Physical characteristics of the calls were analyzed and calls correlated with their behavioral context. Based on these correlations an «advertisement» call can be identified in all three species. Interspecific comparison suggest that within the Prosimian suborder there are at least two different evolutionary pathways in the organization of this call and of the vocal repertoires in general. The Lorisid speciesG. demidovii conveys acoustic information mainly in pulsed calls, leading to an extreme amplitude-modulation. Individual characteristics are in temporal patterns of calls.C. medius andM. murinus, the Cheirogaleid species, use tonal calls and encode individual characteristics in frequency patterns. Despite the fundamental similarity in the call-structure of the two Cheirogaleid species different adaptations can be found to meet different ecological needs.     

Inside Pandora's box: Development of the lethal myrmecopathogenic fungus Pandora formicae within its ant host

Parasites modify their host's appearance, behaviour and physiology to a certain extent. Many of these modifications are seen as purposefully manipulative, serving the parasite's interest. Endgames are particularly intriguing, where the parasite's development requires the host's death. ‘Summit’ or ‘treetop’ disease is one of such spectacular examples of parasite-induced death: the host dies attached to an elevated part of the vegetation. Many organisms induce summit disease including Ophiocordyceps, Entomophthora and Pandora species. We analyzed the development of Pandora formicae inside its ant host's body after the host's death. Our findings suggest that the fungus consumes the host's muscles and central nervous system quite fast, similarly to the related Entomophthora muscae. Unlike in Ophiocordyceps, no conidial anastomosis tubes were observed. Differences and similarities between the three fungi are discussed with regard to the extent of parasitic manipulation since Pandora's manipulative traits seem less pronounced than in Ophiocordyceps.     

Diversity and composition of arboreal beetle assemblages in tropical pasture afforestations: effects of planting schemes and tree species identity

Timber tree plantations are considered for rehabilitating forest biodiversity in the tropics, but knowledge on determinants of faunal diversity patterns in such human-modified forest landscapes is scarce. We quantified the composition of beetle assemblages on three native timber species (Anacardium excelsum, Cedrela odorata and Tabebuia rosea) planted on former pasture to assess effects of tree species identity, tree species diversity, and insecticide treatment on a speciose group of animals in tropical plantations. The beetle assemblage parameters ‘abundance’, ‘species richness’, ‘Chao1 estimated species richness’ and ‘Shannon diversity’ were significantly reduced by insecticide treatment for each tree species. Shannon diversity increased with stand diversification for T. rosea but not for A. excelsum and C. odorata. Species similarity was highest (lowest species turnover) between beetle assemblages on T. rosea, and it was lowest (highest species turnover) for assemblages on insecticide-treated trees of all timber species. Considering trophic guilds, herbivorous beetles dominated on all tree species and in all planting schemes. Herbivores were significantly more dominant on T. rosea and C. odorata than on A. excelsum, suggesting that tree species identity affects beetle guild structure on plantation trees. Insecticide-treated stands harbored less herbivores than untreated stands, but exhibited a high abundance of predator beetle species. Our study revealed that even young pasture-afforestations can host diverse beetle assemblages and thus contribute to biodiversity conservation in the tropics. The magnitude of this contribution, however, may strongly depend on management measures and on the selected tree species.     

Benefits of size dimorphism and copulatory silk wrapping in the sexually cannibalistic nursery web spider,Pisaurina mira

In sexually cannibalistic animals, male fitness is influenced not only by successful mate acquisition and egg fertilization, but also by avoiding being eaten. In the cannibalistic nursery web spider, Pisaurina mira, the legs of mature males are longer in relation to their body size than those of females, and males use these legs to aid in wrapping a female''s legs with silk prior to and during copulation. We hypothesized that elongated male legs and silk wrapping provide benefits to males, in part through a reduced likelihood of sexual cannibalism. To test this, we paired females of random size with males from one of two treatment groups—those capable of silk wrapping versus those incapable of silk wrapping. We found that males with relatively longer legs and larger body size were more likely to mate and were less likely to be cannibalized prior to copulation. Regardless of relative size, males capable of silk wrapping were less likely to be cannibalized during or following copulation and had more opportunities for sperm transfer (i.e. pedipalpal insertions). Our results suggest that male size and copulatory silk wrapping are sexually selected traits benefiting male reproductive success.     

Mitochondrial and nuclear DNA diversity within six species ofPhytophthora

The extent of intraspecific mitochondrial DNA (mtDNA) diversity was investigated in isolates ofPhytophthora capsici,P. citricola, P. citrophthora, P. megakarya, P. palmivora, andP. parasitica that represented a wide range of host plants and geographic origins. Phenograms were constructed following the analysis of restriction fragment patterns that were generated by several endonucleases. The six species showed different degrees of mtDNA diversity. Restriction fragment patterns inP. palmivora andP. parasitica were very uniform. Distinct subgroups could be distinguished inP. megakarya andP. citrophthora that correlated with the geographic origin or the host plant, respectively. These subgroups did not seem to be closely related to each other. High degrees of diversity were also evident inP. citricola andP. capsici. Although some isolates ofP. capsici had identical mtDNA patterns, no distinct subgroups were found that could be correlated with either a specific host plant or geographic origin. InP. capsici andP. parasitica variation in nuclear DNA was much more pronounced as compared to mtDNA. InP. capsici both types of analysis correlated well. Because of very limited variation of mtDNA inP. parasitica a comparison between the two phenograms was difficult.     

A multilocus perspective on phylogenetic relationships in the Namib darkling beetle genus Onymacris (Tenebrionidae)

Tenebrionid beetles, common constituent faunae of arid ecosystems worldwide, are particularly abundant in Africa’s Namib and Kalahari deserts. Within this region, flightless, diurnal members of the tribe Adesmiini are among the more intensively studied of all desert beetles, especially with regard to ecology. Much of this research centers on Onymacris, a psammophilous genus largely endemic to the Namib. Here we present the first molecular phylogenetic analysis conducted for Onymacris, emphasizing relationships among other adesmiines. Our multilocus phylogeny identifies a strongly supported clade containing Onymacris and two other genera, Eustolopus and Physadesmia—an assemblage recovered in earlier morphological analyses. However, Onymacris is not monophyletic; rather, we demonstrate its paraphyly with respect to the genus Physadesmia, identified as the sister taxon to the white-bodied species of Onymacris. In turn, the Physadesmia-‘white’ Onymacris clade is the sister group to the remaining (black-bodied) Onymacris. Non-monophyly of ‘black’ versus ‘white’ Onymacris is corroborated by distribution patterns and nodal age estimates, which suggest separate origins in different dune systems.     

First record of mating behaviour and induced parturition of the Brazilian endemic Lutz's stingray Hypanus berthalutzae

The recently described Lutz's stingray Hypanus berthalutzae is endemic to the Brazilian Province, including oceanic islands. Although it is expected to have life-history traits similar to the southern stingray H. americanus, little is known about its reproductive biology. Here we present the first observations of courtship behaviour (n = 4), copulation (n = 3) and an induced parturition of H. berthalutzae at the Fernando de Noronha Archipelago, an insular Marine Protected Area from the Equatorial Atlantic Ocean. The mating event records included (1) ‘chasing/close following’, (2) ‘biting/precopulatory biting’, (3) ‘insertion/copulation’ and (4) ‘separation’. These results are especially relevant considering that records of reproductive behaviour in the wild are rare for elasmobranchs in general. Mating events occurred in different months, suggesting that the reproductive cycle of H. berthalutzae in this insular system is asynchronous, as observed for other stingray species in regions with favourable environmental conditions and abundant food throughout the year. The opportunistic documentation of the induced parturition allowed for direct nonlethal observation of the two pups at or near full term in late May. Although preliminary, these observations should be considered in future management plans as they provide relevant data about the life-history traits and mating behaviour of this endemic and threatened species.     

True deception during extra‐pair courtship feeding: cheating whiskered tern Chlidonias hybrida females perform better

Males of many bird species feed their mates during the pre‐incubation period. The food provisioned by males during these courtship feedings (CFs) represents the key source of energy for the female during egg formation. Non‐pair males may trade food for extra pair copulations (EPC) with females during extra pair courtship feeding (EPCF), while females may trade copulations for food with non‐pair males to obtain additional resources. Because EPCs can be costly to the females, they are expected to behave in ways that will deceive non‐pair males to obtain additional resources at no cost to themselves. We investigated EPCFs in whiskered terns Chlidonias hybrida breeding in food‐rich conditions, on carp ponds in southern Poland. Almost all CFs (n = 2751) took place during the female's fertile period and peaked just before clutch initiation. 10% of all CFs were performed by non‐pair males. Females tried to obtain food from the non‐pair male during 39% of EPCFs, by swindling (the female solicits a non‐social male for copulation and tries to swindle food with no cloacal contact) or by snatching (the female tries to take the gift without engaging in copulation). In the remaining 61% of EPCFs, females did not react or chased the visiting male away. The probability of a female's obtaining food during EPCF was much higher (0.69, 95% CI: 0.47–0.85) if she swindled rather than snatched (0.08, 95% CI: 0.02–0.22). Only 0.7% of EPCFs were followed by EPCs. The high availability of food in the study area allows males to perform frequent EPCFs, despite the very low probability of obtaining EPCs. This is the first time that ‘true deception’ during EPCFs has been reported in birds: swindling females obtain food from non‐pair males at no immediate detectable cost to themselves.     

Analysis of Multiple Tsetse Fly Populations in Uganda Reveals Limited Diversity and Species-Specific Gut Microbiota

The invertebrate microbiome contributes to multiple aspects of host physiology, including nutrient supplementation and immune maturation processes. We identified and compared gut microbial abundance and diversity in natural tsetse flies from Uganda using five genetically distinct populations of Glossina fuscipes fuscipes and multiple tsetse species (Glossina morsitans morsitans, G. f. fuscipes, and Glossina pallidipes) that occur in sympatry in one location. We used multiple approaches, including deep sequencing of the V4 hypervariable region of the 16S rRNA gene, 16S rRNA gene clone libraries, and bacterium-specific quantitative PCR (qPCR), to investigate the levels and patterns of gut microbial diversity from a total of 151 individuals. Our results show extremely limited diversity in field flies of different tsetse species. The obligate endosymbiont Wigglesworthia dominated all samples (>99%), but we also observed wide prevalence of low-density Sodalis (tsetse''s commensal endosymbiont) infections (<0.05%). There were also several individuals (22%) with high Sodalis density, which also carried coinfections with Serratia. Albeit in low density, we noted differences in microbiota composition among the genetically distinct G. f. fuscipes flies and between different sympatric species. Interestingly, Wigglesworthia density varied in different species (10⁴ to 10⁶ normalized genomes), with G. f. fuscipes having the highest levels. We describe the factors that may be responsible for the reduced diversity of tsetse''s gut microbiota compared to those of other insects. Additionally, we discuss the implications of Wigglesworthia and Sodalis density variations as they relate to trypanosome transmission dynamics and vector competence variations associated with different tsetse species.     

Environmental Cues,Endocrine Factors,and Reproductive Diapause in Male Insects

Environmental cues, mostly photoperiod and temperature, mediated by effects on the neuroendocrine system, control reproductive diapause in female insects. Arrest of oocyte development characterizes female reproductive diapause, which has two major adaptive functions: It improves chances of survival during unfavorable season(s), and/or it confines oviposition to that period of the year that is optimal for survival of the eggs and progeny. Although reproductive diapause is less well studied in male insects, there may be no sex-dependent differences in regard to the first of these functions. The second one, however, is not valid for the male; instead, selection pressure directs the male's reproductive strategy toward maximum chances of fertilization of the female's eggs with minimum waste of energy. Therefore, in species with female reproductive diapause, the males may or may not exhibit diapause, but if they do, their diapause must be adapted to that existing in conspecific females. Male reproductive diapause is defined as a reversible state of inability of the male to inseminate receptive females. In relation to reproductive diapause, there are several patterns of coadaptations between male reproductive strategy and timing of female receptivity, (a) In some insects, the females are receptive in the early part of their diapause; mating occurs during this period and there is no diapause in the male. The male dies shortly after copulation and the female stores the sperms to fertilize the eggs that develop after termination of the female's diapause, (b) In some species, as in the grasshopper Anacridium aegyptium, females are receptive during diapause; though oocyte development is arrested, copulation occurs and the stored sperms fertilize the eggs when the female's diapause ends. Males were claimed to have no diapause, but recent studies have revealed the presence of a reproductive diapause in a proportion of the males. This and other cases show that female receptivity during reproductive diapause may or may not be accompanied by male reproductive diapause. If there is a reproductive diapause in the male, it is controlled by the same endocrine mechanism, the corpora allata (CA), as in the females, (c) In many species females are refractory during their diapause. In these cases, males exhibit reproductive diapause, which may be light, as in the beetle Oulema melanopus, or well established, as in certain grasshoppers, butterflies, and beetles. In the latter cases, male diapause is controlled by similar environmental cues (photoperiod, temperature) and by the same intrinsic mechanism (neuroendocrine system, especially CA) as female diapause. Nevertheless, male diapause is less intense; the environmental cues leading to its termination are less complex and/or less extreme, so male diapause terminates before that of the females. Presumably, male diapause is under two antagonistic selection pressures: A male should not waste energy by courting dia-pausing refractory females, but he should be ready to copulate as soon as the females become receptive, otherwise he may lose in the competition between males for females. Some further strategies, which do not seem to fit the above patterns, are also outlined.