Sperm transport after insemination in the Alpine newt (Triturus alpestris, Caudata, Salamandridae)

The goal of this study was to test if sperm transport to the spermathecae in the Alpine newt (Triturus alpestris) requires active co-operation of the female. Artificial insemination of anaesthetised female newts was conducted using spermatophores collected from courting males and with sperm duct contents collected from sacrificed males. Sperm was present in the spermathecae of 9 out of 10 females inseminated with the spermatophores but in only 1 out of 8 females inseminated with sperm duct contents. The females of both groups laid some eggs after insemination, and a portion of these eggs in group of females inseminated with spermatophores were fertilized. However, the number of eggs produced by the females was much lower than typical egg-production in newts. The presence of sperm in the spermathecae of females inseminated with spermatophores and lack of sperm in the spermathecae of females inseminated with sperm duct contents suggests that sperm transport is either induced by the substances present in spermatophores and/or that sperm from the sperm duct is not fully mobile in comparison with sperm from the spermatophores.     

Breeding places of Phlebotomus ariasi in the cevennes focus of leishmaniasis in the south of France

In a search for sandfly breeding sites in the Cévennes, France, six larvae of Phlebotomus ariasi were found in rotted manure in a cellar housing goats, and six adult flies of the same species were collected from emergence traps, one in the same cellar and five on a disused refuse dump. No sandflies were found emerging from holes of long-tailed field mice (Apodemus spp.) and no sandfly larvae were extracted from 77 kg (dry weight) and 130 kg (undried weight) of samples of soil or litter from different places. Total catches and the proportions of male flies in CDC miniature light traps were much higher in peridomestic than silvatic stations. About 15% of males caught in 3 peridomestic stations had unrotated genitalia. It is concluded that the larval breeding sites of P. ariasi are probably peridomestic and have a high content of organic matter. Observations on colonized P. ariasi show that, although immature stages of this fly need moisture, their requirements are comparatively low. It is suggested that female P. ariasi choose oviposition sites by sensing the presence of immature stages already in an optimal place.     

The phlebotomine sandflies of Portugal. XIII--Occurrence of Phlebotomus sergenti Parrot, 1917 in the Arrabida leishmaniasis focus

In a survey carried out during the summer in 2002 and 2003, in the canine and vulpine leishmaniasis focus of Arrabida, 665 phlebotomine sandflies were caught. 13.83% were P. ariasi, 58.65% P. pemiciosus, 0.45% P. sergenti and 27.07% S. minuta. Despite the low abundance, the finding of the three adults P. sergenti (two males in Aldeia Grande and one female in Quinta da Ramada) confirm the colonization of the Arrabida leishmaniasis focus by this species, which presence had been previously reported but thought accidental. The abundance of P. ariasi observed at this time is significantly different from that in previous years (Pires, 1984 and Fernandes pers. com., 1994). The occurrence of P. sergenti in this region, in association with the decrease in abundance of P. ariasi, may reflect an increased aridness of this region, a consequence of current climate and environmental changes.     

Asymmetric female genitalia and other remarkable morphology in a new genus of cobweb spiders (Theridiidae, Araneae) from Madagascar

Symmetry is such a conspicuous feature of life that asymmetries draw our immediate attention. While not uncommon in bilateral organisms in general, asymmetry in spiders is rare. Here I report the first case of antisymmetry in external female genitalia in spiders, in the new genus Asygyna (Theridiidae: Araneae) from Madagascar. In the nearly 39 000 species of spiders described to date, the external structure of the female genitalia is symmetric. In entelegyne spiders paired external copulatory openings each lead to an internal copulatory duct, whose roughly symmetrical trajectories terminate in paired receptacles, the spermathecae. In Asygyna , here exemplified by two new species, A. huberi and A. coddingtoni , laterality is evident in the internal and external female genitalia. A single copulatory opening leads (either to the left or right depending on the individual) to a single copulatory duct with a distinctly asymmetric trajectory. The duct splits terminally shortly before entering the two spermathecae. The males are symmetric, but possibly only one palp can be used in copulation with each female. If adaptive, the selective forces behind this asymmetry are perplexing, as male access to females seems reduced. However, if males are plentiful, asymmetry may benefit the female by reducing insertion times and thus shortening copulation time, and by tightening her control over which males sire her offspring. Asygyna has a range of other bizarre sex-related morphologies, including prosomal pits and a well developed stridulatory mechanism in both sexes, a male proboscis, and simplified palps. A phylogenetic analysis, including 63 taxa and 242 morphological characters, places Asygyna in Pholcommatinae, sister to the enigmatic genus Carniella .  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 211–232.     

Genital structures in the entelegyne widow spider Latrodectus revivensis (Arachnida; Araneae; Theridiidae) indicate a low ability for cryptic female choice by sperm manipulation

The female genital structures of the entelegyne spider Latrodectus revivensis are described using semithin sections and scanning electron microscopy. Apart from the tactile hairs overhanging the opening of the atrium, the contact zones of the female epigynum are devoid of any sensilla, indicating that the female does not discriminate in favor or against males due to their genital size or stimulation through copulatory courtship. The dumb-bell shape and the spatial separation of the entrance and the exit of the paired spermathecae suggest that they are functionally of the conduit type. Not described for other entelegyne spiders so far, the small fertilization ducts originating from the spermathecae of each side lead to a common fertilization duct that connects the spermathecae to the uterus externus. During oviposition, it is most likely that spermatozoa are indiscriminately sucked out of the spermathecal lumina by the low pressure produced by the contraction of the muscle extending from the epigynal plate to the common fertilization duct. As no greater amounts of secretion are produced by the female during oviposition, and no activated sperm are present within the female genital tract, the secretion produced by the spermathecal epithelium does not serve in displacement or (selective) activation of spermatozoa. These findings suggest that female L. revivensis are not able to exert cryptic female choice by selectively choosing spermatozoa of certain males.     

Phlebotomine sandflies from Madagascar (Diptera: Psychodidae). V--description of Sergentomyia majungaensis n. sp

Sergentomyia majungaensis, a new species of Phlebotomine sandfly, is described from Madagascar. The female is characterised by an unusual antennal formula (1/IV-V to VII; 2/VI to VIII-XV) and smooth and narrow spermathecae, with a common duct, whose shape is intermediate between Sergentomyia and Parrotomyia subgenera. The male has the same antennal formula than Grassomyia (1/IV-XV) but no hairs on the mesanepistern and a longer third antennal segment.     

Cryptic female choice in the yellow dung fly Scathophaga stercoraria (L.)

Both female choice and male-male competition may take place during reproduction in many species. Female choice tends to be less obvious than male-male competition and consequently has received less attention from researchers. The opportunity for cryptic female choice arises after multiple insemination. Through postcopulatory processes, a female could alter the pattern of paternity among her offspring so that it does not directly reflect the different contributions of sperm made by her mates. To be able to determine if a female alters the relative sperm contributions of her mates, the behaviors and influences of the males must therefore be first taken into account. The interest of each male is to father all the offspring, and the interest of each female is to maximize paternal quality. Female yellow dung flies have complex internal reproductive tracts that may give them considerable control over the fertilization success of stored sperm from different males. In laboratory trials to date, the last male to mate has usually been most successful. In the present study, cryptic choice occurred in Scathophaga stercoraria and the pattern of choice was consistent with previously reported results. The fertilization success of a female's second mate (P2) was substantially larger if a female was kept at constant temperature and if the second male was genetically similar to her at the phosphoglucomutase (Pgm) locus. Females from the field normally have three spermathecae, but some have four. Lines were bred to have either three or four spermathecae. Flies from the different lines were crossed to generate females with similar genetic backgrounds that had either three or four spermathecae. P2 was significantly lower for high-quality females, that is, those that laid larger-than-average-clutches, with four spermathecae than for low-quality females with four spermathecae; female quality had no influence on P2 for females with three spermathecae. The results suggest that only large females may benefit from increased spermathecae number by being able to act against male interests. Females may only have three spermathecae, even though genetic variation for more is present, because selection for more spermathecae is weak.     

Distribution of phlebotomine sandflies, of the subgenus Larroussius, in Morocco

Adult sandflies (Diptera: Psychodidae) of the genus Phlebotomus and of the subgenus Larroussius were collected with sticky traps from thirty-four localities in central and south-western Morocco. Four Larroussius species were present in this area, Phlebotomus perniciosus Newstead, being the most widespread and abundant species particularly in the mountainous area, was sympatric with the closely related species Phlebotomus longicuspis Nitzulescu. Only one male of P. perniciosus displayed the bifid copulatory valves characteristic of the species. All specimens showed copulatory valves curved at their apex. The number of coxite hairs is a taxonomical tool that differentiates this morph of P. perniciosus from P. longicuspis. Phlebotomus ariasi Tonnoir was collected from six mountainous stations. Phlebotomus mariae Rioux et al. was scarce; only five males were collected and were often associated with P. ariasi.     

The association of crustacean cardioactive peptide with the spermatheca of the African migratory locust, Locusta migratoria

Crustacean cardioactive peptide (CCAP)-like immunoreactivity was identified in neurons of the VIIIth abdominal ganglion and in axons in the nerves that project to the spermatheca of 3-4 week old adult female locusts. In addition, lightly stained CCAP-like immunoreactive processes were localized over the spermathecae. The amount of CCAP in the spermathecal tissue was quantified using an enzyme-linked immunosorbent assay (ELISA) performed on extracts of the whole spermatheca, and on its constituent parts, namely the sperm sac, coiled duct and straight duct. The spermatheca contains 920+/-273 fmol (mean+/-SE) of CCAP equivalents, with the majority localized in the coiled duct. There are age-related differences in the amount of CCAP present in the spermathecae with less content in spermathecae from 1 to 5 day old and greater content in spermathecae from 3 to 4 week old adults. There was also no difference in CCAP content of spermathecae in mated and virgin 3 to 4 week old adults. Reversed phase-high performance liquid chromatography (RP-HPLC) followed by ELISA further confirmed the presence of CCAP-like material in extracts of locust spermathecae. Physiological assays demonstrated that CCAP increased the basal tonus and frequency of spontaneous contractions of the spermatheca, with thresholds between 10(-10) and 10(-9)M and maxima at 10(-7)M CCAP. CCAP also increases the amplitude of neurally evoked contractions with a threshold less than 10(-11)M and a maximum of 10(-7)M CCAP. The present study suggests that CCAP acts as a neuromodulator/neurotransmitter at the spermathecal visceral tissue of female Locusta migratoria.     

Remating inhibition in the melon fly,Bactrocera (=Dacus) cucurbitae (Diptera: Tephritidae): Copulation with spermless males inhibits female remating

Eight hour copulation of the melon fly,Bactrocera cucurbitae, which usually mates at dusk and finishes copulation at dawn, inhibited female remating, while 3 h copulation did not. Copulation of females with either normal or virgin sterile males inhibited female remating. Sperm-depleted sterile males inhibited female remating at the same rate as normal males when the copulation duration was 8 h, indicating that existence or amount of sperm in females' spermathecae is not important in remating inhibition. Females of a wild strain remated later than females of a mass-reared strain, irrespective of strains of 1st and 2nd males. This suggests that the females may control their own remating, or that there is a difference between wild and mass-reared strains in their sensitivity to a male substance that inhibits females' receptivity.     

Female-mediated differential sperm storage in a fly with complex spermathecae, Scatophaga stercoraria

Multiple spermathecae potentially allow selective sperm use, provided that sperm from rival males are stored differentially, that is, in different proportions across storage compartments. In the yellow dung fly, Scatophaga stercoraria, females have three spermathecae arranged as a doublet and singlet. To test whether females store the sperm of rival males actively and differentially, we mated fixed male pairs to three females. After copulation, females were (1) dissected immediately before they could start laying a clutch of eggs, (2) left awake for 30 min but prevented from oviposition, or (3) anaesthetized with carbon dioxide for 30 min to interfere with the muscular control presumably required for sperm transport from the site of insemination to the spermathecae. For each female, we estimated the proportion of the second male's sperm stored in her spermathecae (S(2)value), using sperm length as a male marker. After copulation, the S(2)values in the singlet and doublet spermathecae differed significantly, indicating differential sperm storage during copulation. Postcopulatory treatment affected differential sperm storage significantly. Females dissected immediately had lower S(2)values in the doublet than in the singlet spermatheca, while females left awake showed the reverse pattern for the same two males. This reversal did not occur when females were treated with carbon dioxide. The results indicate differential storage of sperm from different males during copulation and that female muscular activity can affect storage and separation of competing ejaculates beyond copulation. Copyright 2000 The Association for the Study of Animal Behaviour.     

Influence of female reproductive anatomy on the outcome of sperm competition in Drosophila melanogaster

Females as well as males can influence the outcome of sperm competition, and may do so through the anatomy of their reproductive tracts. Female Drosophila melanogaster store sperm in two morphologically distinct organs: a single seminal receptacle and, normally, two spermathecae. These organs have different temporal roles in sperm storage. To examine the association between sperm storage organ morphology and sperm competition, we used a mutant type of female with three spermathecae. Although the common measure of sperm competition, P(2), did not differ between females with two and three spermathecae, the pattern of sperm use over time indicated that female morphology did affect male reproductive success. The rate of offspring production by females with three spermathecae rose and fell more rapidly than by females with two spermathecae. If females remate or die before using up second male sperm, then second male reproductive success will be higher when they mate with females with three spermathecae. The results indicate that temporal patterns of sperm use as well as P(2) should be taken into account when measuring the outcome of sperm competition.     

Two oncholaimid species from a South African estuary (Nematoda,Oncholaimidae)

Adoncholaimus papillatus Kreis, 1932 and Viscosia erasmi sp.nov. are described from an intertidal area of the Sundays River Estuary, South Africa. Adoncholaimus papillatus females have an, up to present, unknown organization of the demanian system: the main duct is connected with the two spermathecae by means of a strongly curved ductus uterinus. There is only one osmosium situated at the anterior part of the main duct (anteriad of the vagina). The uvette and the ductus entericus are absent. The main duct extends to the swollen part of the tail where it opens in one or two terminal pores. Viscosia erasmi sp.nov. is mainly characterized by the bifid apical part of the buccal cavity, three distinct teeth with the right ventrosublateral one the largest and by the setiform head sensilla.     

日本北海道虻类雌性外生殖器形态(双翅目：虻科)(英文)

【目的】利用日本北海道虻类评估和验证外生殖器在分类学上的意义。【方法】将虻类成虫标本浸渍在生理盐水中并置于双目显微镜下通过针和镊子在培养皿中进行解剖并绘图,观察第9背板、第10背板、尾叶、第8腹板、受精囊、受精囊管及生殖叉器的形态特征。【结果】在日本北海道共记录了虻科(Tabanidae) 3亚科7属38种。我们观察并描述了3亚科其中的6属24种的雌性外生殖器的主要特征。亚科之间存在明显差异;然而在一般情况下属之间很难建立一种方法来确定共同点;种之间只有在斑虻属Chrysops中有相似之处,其他属中则比较多样化。因此,亚科鉴定根据第9背板、第8腹板及受精囊足以进行区分,属及种鉴定需要结合第9背板、第10背板、尾叶、第8腹板、受精囊、受精囊管及生殖叉器各自的特征组合在一起才能区分开来。我们也制作了虻类外生殖器的检索表。【结论】和许多其他昆虫一样,外生殖器是虻科的重要分类特征,对于促进分类学和系统学的发展具有重要意义。本研究首次对分布在日本北海道的虻科雌性外生殖器进行了系统研究。     

Ultrastructure of the Spermathecae of Parafabricia ventricingulata and Three Species of Oriopsis (Polychaeta: Sabellidae)

Parafabricia ventricingulata females have a pair of spermathecae located in the radiolar crown anterio-dorsal to the buccal opening. The spermathecae have three regions; an entrance, 7 μm across, leading into a ciliated ‘atrium’ that is approximately 50 μm long; a connecting piece, 2–5 μm across and 25 μm long, leading from the ‘atrium’ to the sperm receptacle. The sperm receptacle is heavily pigmented and spherical. The sperm lie in a large mass in the receptacle with no particular orientation. Oriopsis bicoloris females have a pair of unpigmented spermathecae in the collar behind the radiolar crown. Each spermatheca is a simple blind duct 100 μm long, with a lumen 8 μm in diameter. Between 30 and 40 sperm lie in the lumen of each spermatheca. Oriopsis brevicollaris females have a pair of spermathecae located in the radiolar crown above the buccal opening. From the opening, 10 μm across, a blind duct runs for 90 μm. Sperm are stored in the distal region of the duct. Sperm lie along the margins of the duct in close contact with microvilli. Up to 10 sperm were found in each spermatheca. Oriopsis mobilis females have a pair of spermathecae located in the radiolar crown above the buccal opening. The opening, 3 μm across, leads into a blind duct that runs for 30 μm. Sperm are stored in the distal region of the spermathecae where they are embedded in spermathecal cells. Between 10 and 20 sperm were found in each spermatheca. Oriopsis dentata was found not to have spermathecae. The homologies of the spermathecae found within the Sabellinae and Fabriciinae (Sabellidae) and the Spirorbinae (Serpulidae) are discussed, but cannot be resolved on present evidence.     

Female control of sperm ejection and retention in the cornsilk fly Euxesta eluta (Diptera: Ulidiidae)

Promiscuous mating systems provide the opportunity for females to bias fertilization toward particular males. However, distinguishing between male sperm competition and active female sperm choice is difficult for species with internal fertilization. Nevertheless, species that store and use sperm of different males in different storing structures and species where females are able to expel all or part of the ejaculates after copulation may be able to bias fertilization. We report a series of experiments aimed at providing evidence of female sperm choice in Euxesta eluta (Hendel), a species of ulidiid fly that expels and consumes ejaculates after copulation. We found no evidence of greater reproductive success for females mated singly, multiply with the same male, or mated multiply with different males. Female E. eluta possesses two spherical spermathecae and a bursa copulatrix for sperm storage, with a ventral receptacle. There was no significant difference in storing more sperm in spermathecae 24 h after copulation than immediately after copulation. Females mated with protein-fed males had greater reproductive success than similar females mated to protein-deprived males. Protein-fed females prevented to consume the ejaculate, retained more sperm when mated to protein-fed males than when mated to protein-deprived males. Our results suggest that female E. eluta can exert control of sperm retention of higher quality males through ejaculate ejection.     

Honeydew sugars in wild-caught Phlebotomus ariasi detected by high performance liquid chromatography (HPLC) and gas chromatography (GC)

Phlebotomus ariasi Tonnoir sandflies were caught in light traps hung in oak trees and in a house in the Cévennes focus of leishmaniasis in the South of France. The flies were cryopreserved either immediately on removal from the traps, or after starvation for 6-7 days, or after 6-7 days starvation followed by exposure to oak infested with the aphid genera Lachnus or Thelaxes. After transportation to the laboratory, the sandflies were thawed and aqueous extracts of the crushed flies were analysed for their carbohydrate content using high performance liquid chromatography (HPLC) and gas chromatography (GC). Starved female sandflies lacked significant amounts of any saccharides. Four types of sugar, melezitose and its hydrolysis products turanose, glucose and fructose, were observed in flies which had been starved previously and then placed with Lachnus infested oak. The results also indicate the presence of hydrolysis products of melezitose: (a) in flies previously starved and placed with Thelaxes infested oak, (b) in P.ariasi cryopreserved direct from the light traps hung in oak trees infested with Lachnus and Thelaxes, and (c) flies caught in a house. Unidentifiably small quantities of a trisaccharide were also detected in the latter groups of flies. In previous tests, sugars were detected in P.ariasi after their exposure to aphid-infested oak (Quercus ilex L.), but not when P.ariasi females were exposed to washed oak leaves without aphids. The results indicate that P.ariasi feed on melezitose and/or turanose, the main local source of which is aphid honeydew. A better understanding of sugar meal sources of sandflies using HPLC and GC techniques will assist in our understanding of sandfly/Leishmania relationships, parasite transmission and epidemiology.     

Morphogenesis of the Genital Ducts and Spermathecae in Clitellio arenarius, Heterochaeta costata, Tubificoides benedii (Tubificidae) and Stylaria lacustris (Naididae) (Annelida, Oligochaeta)

The development of the male genital ducts in Clitellio arenarius, Tubificoides benedii, Heterochaeta costata (Tubificidae) and Stylaria lacustris (Naididae) is studied with the purpose of investigating the homologies between different parts of the ducts. In both Tubificidae and Naididae, the male duct comprises a funnel, a vas deferens and an atrium. There may also be a prostate gland (diffuse or compact) in association with the duct. The funnel and vas deferens, in all four species, originate from the peritoneal (mesodermal) cells in the posterior septa in the testes segment, whereas the atrium develops from an epidermal (ectodermal) invagination. The prostate glands have different origins in different taxa; in S. lacustris it differentiates from the peritoneal (mesodermal) cells surrounding the atrium, but in H. costata and T. benedii it develops by a multiplication of the atrial (ectodermal) cells; C. arenarius lacks a prostate gland. The development of the spermathecae and the female duct is also studied. The spermatheca is an epidermal invagination. The female duct is of mesodermal origin. However, the position of the duct differs; in S. lacustris the female duct is lateral and the pore is in the ovarian segment, while in the tubificids studied the duct is ventral with the pore in the segment behind the one containing the ovary.     

Record of Phlebotomus (Transphlebotomus) mascittii Grassi, 1908 and Phlebotomus (Larroussius) chadlii Rioux, Juminer & Gibily, 1966 female in Algeria

We report for the first time the presence of Phlebotomus mascittii and the female of Phlebotomus chadlii in Algeria. These two species were collected during an entomological study conducted in endemic visceral leishmaniasis focus from the north part of the country, Kabylia.     

Morphology and function of the reproductive tract of the spider crab Libinia spinosa (Crustacea, Brachyura, Majoidea): pattern of sperm storage

Morphology and function of the male reproductive tract, female spermatheca and patterns of sperm storage were assessed in the crab Libinia spinosa using histological methods. Testes are characterized by the presence of peripheral spermatogonia and different sequences of sperm maturity. Spermatophores begin to be packed in the last portion. The vas deferens consists of three sections: anterior, with undeveloped spermatophores and free sperm; median, with well-developed spermatophores; and posterior with granular secretions. Female spermathecae are of the ventral type, with a velum separating dorsal and ventral chambers. Live individuals were kept in the laboratory and arranged in pairs. An experiment was conducted toward the end of the reproductive season, in which males with the right gonopod excised were placed with receptive females. After mating, females were killed and the spermathecae dissected for histological study and observation of the pattern of sperm storage. Spermatozoa were found forming discrete sperm packages. New ejaculates can fill the entire spermatheca or be restricted to the ventral chamber; sperm are rounded, with a distinguishable acrosomal core. Old ejaculates are restricted to the dorsal chamber and are of irregular shape and larger size; an acrosomal core was not distinguishable. The secretions produced by the glandular epithelium of the dorsal chamber of the spermathecae are likely to have a role in the removal of dead sperm.