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

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

Ultrastructural and functional aspects of the spermatheca in the American harlequin bug, Murgantia histrionica (Hemiptera: Pentatomidae)

The spermatheca of Murgantia histrionica (Hahn) was investigated using fluorescence, scanning and transmission electron microscopy. The aim of the study was to elucidate the structure of this organ, pointing out differences between mated and unmated females. Results have shown an elaborated cuticular structure associated with muscular and glandular tissues. The spermatheca is joined with the common oviduct by the spermathecal duct, forming a thin saccular dilation through two consecutive invaginations. The distal part of the organ is formed by a series of two communicating cuticular chambers. The first cylindrical-shaped chamber, corresponding to the coiled region, is wrapped by longitudinal muscular fibers suspended between two cuticular flanges. The contractions of these fibers compress a deformable zone of the cylinder, pumping the sperm toward the spermathecal duct. Without contractions the cylinder results to be isolated from the proximal part of the spermatheca by means of a valve. The second chamber, corresponding to the spermatheca, is made of two parts: a truncated-conical sub chamber, with a constant cuticular thickness, bearing on itself the distal flange, where muscular fibers are attached. The second part is a bulb-like structure wrapped in a glandular epithelium. The secretory units are composed by two cells: a secretory cell and an associated duct cell. Every evacuating duct shows a little reservoir just after the terminal apparatus, and converge inside the distal bulb after a tortuous path. The functional implications of this structure in the reproductive biology of M. histrionica are discussed.     

THE FERTILIZATION CANAL IN PLODIA INTERPUNCTELLA (HÜBNER) (PYRALIDAE: LEPIDOPTERA)

The spermathecal duct of Plodia interpunctella (Hübner) was studied with light and transmission electron microscopy. The lumen in the duct is enclosed by a thin chitinous wall that has a thicker band that spirals along the length of the duct. The thick spiral band pinches off part of the lumen and creates a smaller canal, which it encloses. Although the two canals are not separated, the duct appears to have a double lumen. The thin wall of the main canal provides a flexibility in which the lumen widens or narrows concomitantly with contractions of the spermatheca and the portion of the duct adjoining the spermatheca. Sperm is transferred from the spermatheca to the vestibulum where the egg is fertilized. The distention of the canal and contractions of the spermatheca thus account for the speed at which eggs are fertilized and deposited.     

Functions of the spermathecal muscle of the boll weevil,Anthonomus grandis

In female boll weevils, Anthonomus grandis, spermathecal filling was not affected by severing the spermathecal muscles. Females whose spermathecal muscles were severed 2 to 4 weeks after mating laid infertile eggs and resumed virginal ovipositional behaviour indicating the importance of this muscle in supplying sperm for egg fertilization. The presence of normal active sperm within the spermatheca in no way influenced ovipositional behaviour. In females whose spermathecal muscles had been severed, 22 per cent sperm displacement occurred after a second mating compared with 66 per cent for normal females. The physical displacement of sperm was thus largely dependent on a functional spermathecal muscle.     

Function of the spermathecal muscle in Chelymorpha alternans Boheman (Coleoptera: Chrysomelidae: Cassidinae)

Abstract. When the spermathecal muscle of a virgin female Chelymorpha alternans Boheman (Coleoptera: Chrysomelidae) was cut, the number of spermatozoa transferred in a single mating to the spermatheca and the spermathecal duct was not affected, but their distribution differed. Cutting the spermathecal muscle also reduced egg fertility. Eggs from females with a cut muscle showed a lower average percentage fertilization. Longer delays in oviposition after removal of the spermathecal muscle were associated with higher proportions of infertile eggs.     

Development of eggs, larvae and juveniles of laboratory-reared blue whiting,Sillago parvisquamis (Percoidei: Sillaginidae)

The embryonic, larval and juvenile development of blue whiting,Sillago parvisquamis Gill, are described from a series of laboratory-reared specimens. Mean egg diameter and mean total length (TL) of newly-hatched larvae were 0.71 mm and 1.58 mm, respectively. The eggs were non-adhesive, buoyant and spherical with an oil globule (mean diameter 0.18 mm). Hatching occurred about 20 hours after fertilization at a temperature of 24.0–25.0°C, newly-hatched larvae having 38–40 myomeres. The yolk and oil globule were completely absorbed 3 days after hatching at 2.8–3.2 (mean 3.0) mm TL. Notochord flexion was completed by 7.2–8.2 (7.7) mm TL, and pectoral and caudal fin rays fully developed by approximately 10 mm and 8.5 mm TL, respectively. Completion of fin development occurred in the following sequence: caudal, pectoral, anal and second dorsal, first dorsal and pelvic, the last-mentioned by approximately 11 mm TL. The larvae ofS. parvisquamis andS. japonica, which closely resemble each other in general morphology and pigmentation, could be distinguished as follows. Newly-hatchedS. parvisquamis larvae had more myomeres thanS. japonica (38–40 vs. 32–34) and more melanophores on the dorsal surface of the body (19–28 vs. about 40).Sillago japonica had a vertical band of melanophores on the caudal peduncle, which was lacking in postflexionS. parvisquamis larvae. In addition, juveniles ofS. parvisquamis (larger than 23 mm TL) had melanophores on the body extending anteriorly to below the lateral line to form a midlateral band, whereas no obvious band occurred on similarly-sizedS. japonica juveniles.     

Reproductive biology and morphology of eggs and larvae of Stiphodon percnopterygionus (Gobiidae: Sicydiinae) collected from Okinawa Island

Reproductive biology and morphology of eggs and early larvae of the sicydiine goby Stiphodon percnopterygionus were investigated on Okinawa Island, southern Japan. Spawning season was estimated as being from May to December. Standard length at maturity was approximately 20 mm in both sexes, and batch fecundity was approximately 1000–10 000 per female. The egg masses, guarded by the male, were laid on the undersurface of stones in freshwater. The pyriform eggs had long- and short-axis diameters of 0.54–0.58 mm and 0.49–0.50 mm, respectively. Newly hatched larvae (1.20–1.32 mm notochord length: NL) were poorly developed, with large yolk sacs and unopened mouths. Three days after hatching (1.87–2.05 mm NL), eyes were fully pigmented and mouths were opened. An erratum to this article is available at .     

The rôle of the spermathecal gland of the boll weevil,Anthonomus grandis

Spermatozoa were relatively inactive and did not enter the spermatheca of female boll weevils, Anthonomus grandis, whose spermathecal glands were removed as teneral adults. However, these females were able to lay fertile eggs for a 2 week period. When the spermathecal gland was removed from older females, spermathecal filling occurred, and although the spermatozoa retained their fertilizing capacity for extended periods, spermathecal emptying did not occur. Spermatozoa gradually lost their motility and fertilizing capacity, indicating that spermathecal secretions are effective in very small amounts. Spermatozoa were not activated by any of the materials contained in the normal male ejaculate. These materials alone did not effect spermathecal filling nor were they capable of maintaining the fertilizing capacity of the spermatozoa for very long. Sperm economy is low with less than 1 fertile egg laid per 100 spermatozoa used.     

Spernathecal morphology,sperm transfer and a novel mechanism of sperm displacement in the rove beetle,Aleochara curtula (Coleoptera,Staphylinidae)

Summary The mechanism by which sperm are transferred from the male's spermatophore to the female's storing cage is described for the rove beetle Aleochara curtula, emphasizing a novel mechanism of sperm displacement by competing males. The cuticular, U-shaped spermatheca is equipped with a valve structure and two sclerotized teeth. The tube of the spermatophore extends into the spermathecal duct through the guidance of the flagellum of the male endophallus. Further elongation of the spermatophore tube, however, occurs only after separation of the pair. A primary tube bursts at its tip after passing through the valve. Within the lumen of the primary tube, a second tube passes through the valve and continues to extend up to the apical bulb of the spermatheca, doubles back on itself and swells to form a balloon filling most of the spermatheca. The balloon of the spermatophore is pierced within the spermatheca by tooth-like structures pressed against the spermatophore through contraction of the spermathecal muscle. The same process of spermatophore growing and swelling is also observed in mated females. Sperm from previous copulations are backflushed through the valve and the spermathecal duct, indicative of last-male sperm predominance.Abbreviations ad adhesive secretion covering the sperm - sac am amorphous secretion of the spermatophore - as ascending portion of the spermatophore - ds descending portion of the spermatophore - end parts of the male endophallus - ext extended tube - f flagellum - gs genital segment - lt large tooth - m muscle of the spermatheca - nsc non sclerotized cuticle - op opening of the spermathecal gland - pt primary tube - sc sclerotized cuticle - sd spermathecal duct - se secretion of the spermathecal gland - sf secretion flowing out of the primary tube - sg spermathecal gland - sm sperm - smt small tooth - sp spermatheca - ss sperm sac - st secondary tube - vm vaginal muscle     

Eggs and larvae of <Emphasis Type="Italic">Awaous melanocephalus</Emphasis> (Teleostei: Gobiidae)

The morphology of eggs and larvae of Awaous melanocephalus is described. The eggs measured 0.33–0.35 mm in long-axis diameter and 0.32–0.34 mm in short-axis diameter. Newly hatched larvae (0.90–0.99 mm in notochord length, NL; 0.93–1.04 mm in total length, TL) were poorly developed, lacking a mouth and having a large yolk sac and unpigmented eyes. The mouth opened and the eyes became fully pigmented 3 days after hatching (1.78–2.00 mm NL, 1.88–2.10 mm TL). The yolk sac was completely absorbed 5 days after hatching at a water temperature of 27°–28°C.     

Some reproductive characteristics of endangered Caspian Lamprey (<Emphasis Type="Italic">Caspiomyzon wagneri</Emphasis> Kessler, 1870) in the Shirud River southern Caspian Sea,Iran

Migration and reproduction of the Caspian Lamprey, Caspiomyzon wagneri, in the Shirud River were investigated during late-March to early-May at water temperatures ranging from 11 to 21.25°C. We examined the effect of water temperature on timing of spawning migrations. There was a significant negative relationship between temperature and intensive migration of Caspian Lamprey (p < 0.05). The most intensive migration of lampreys was at night (21:00–3:00 h) and when the water temperatures averaged 16°C (34.43%). The overall sex ratio (male to female) was 1.07 to 1. The individual absolute fecundity was 31 ‘758–51’ 198 eggs (mean±SD—41,924 ± 5,382). The egg diameter was 0.780–1.151 (0.92 ± 0.081) mm. The individual relative fecundity varies from 80.3 to 148.1 (107.2 ± 15.1) eggs per 1 mm of length and from 260.8 to 677.4 (397.6 ± 93) eggs per 1 g of weight. The gonadosomatic index (GSI) of females was 5.83–31.44 (11.22 ± 4.30).     

A newly discovered sperm transport system in the female of Lygaeidae bugs

In the female reproductive system of the relatively large hemipteran, the western conifer seed bug Leptoglossus occidentalis (Heidemann), a cuticle‐lined tube extends medially along the surface of the vagina from the proximal end of the spermathecal complex anteriorly to the base of the common oviduct. This medial tube houses the proximal end of the spermathecal duct, thereby enabling the transport of material from the spermatheca at the distal end of the spermathecal complex, past the vagina (or bursa copulatrix) and directly to the common oviduct. The proximal portion of the spermathecal complex also contains an insemination duct that is separate from the spermathecal duct. The insemination duct allows the male intromittent organ to extend from the vagina to the spermatheca without navigating through the spermathecal duct. The reproductive systems of two previously studied Hemiptera, the milkweed bug Oncopeltus fasciatus (Dallas) and the box elder bug Leptocoris trivittatus (Say), possess a similar cuticle‐lined medial tube housing the spermathecal duct. This new information provides a clearer understanding of sperm transport in the female reproductive system of Lygaeidae bugs, and helps to clarify the path of the male organ during copulation, as well as the movement of sperm during egg laying.     

Mechanical isolation and ultrastructural characterization of viable egg cells in Plumbago zeylanica

 A protocol for isolating viable eggs in Plumbago zeylanica by mechanical dissection is reported. The optimum solution for isolation was 0.8 M mannitol + 10 mM MOPS + 10 mM CaCl₂, (pH 4.5–5.0) with an osmolality of 860–940 mmol/kg. Eggs retain their viability for at least 24 h. Isolated eggs were true protoplasts without cell walls and could tolerate osmolality of 437 mmol/kg to 965 mmol/kg. Observation of the isolated eggs using transmission electron microscopy indicated that they were well preserved and reflected the ultrastructure of physiologically active cells, displaying features similar to those of in vivo egg cells. Notable differences include the absence of a filiform apparatus and the accumulation of dense particles in the plastids, which was most conspicuous in egg cells that were damaged during isolation. Received: 22 July 1997 / Revision accepted: 5 September 1997     

Surface morphology of eggs of Euproctis chrysorrhoea (Linnaeus, 1758)

Candan, S., Suludere, Z. and Bayrakdar, F. 2007. Surface morphology of eggs of Euproctis chrysorrhoea (Linnaeus, 1758). —Acta Zoologica (Stockholm) 88 : 000–000. Filaments covering the egg batches and chorion structure were studied both by light and scanning electron microscopy in the brown‐tailed moth Euproctis chrysorrhoea (Linnaeus, 1758). Females lay eggs in masses on the underside of apple leaves. The egg batches are covered with brown hairs derived from the bodies of the female. Each female lays about 200–400 eggs. The spherical eggs are about 0.84 mm long and 0.47 mm wide. Newly deposited eggs are golden‐yellow and darken after the onset of embryonic development. The micropylar area appears somewhat depressed and has a circular outline. The region is surrounded by a rosette of 10–12 petal‐shaped primary cells, which are completely surrounded by a series of secondary and tertiary cells. The remainder of the egg is largely smooth, but shows aeropyles. These are located in the corners of ill‐defined polygons.     

Structure and function of the spermathecal complex in the phlebotomine sandflyPhlebotomus papatasi Scopoli (Diptera: Psychodidae): II. Post-copulatory histophysiological changes during the gonotrophic cycle

The spermathecal complex ofPhlebotomus papatasi Scopoli (Diptera: Psychodidae) undergoes histological and physiological changes during its gonotropic cycle. The present histochemical study revealed a mucopolysaccharide secretory mass in the spermathecae of the newly emerged sandfly. Sperm competition occurs when two or more males compete to fertilize an ovum in the female reproductive tract. In this study, spermatophores of two or more competing males were deposited at the base of the spermathecal ducts, which originate from the female bursa copulatrix. This suggests that females play a role in sperm displacement, which is defined as any situation in which the last male to mate with a female fertilizes maximum number her eggs. A blood meal ingested by the female for ovary development and egg laying stimulates the release of sperm from the spermatophore. The spermatozoa then migrate to the lumen of the spermatheca. The ultrastructure of spermatozoa comprises a head with double-layered acrosomal perforatorium, an elongate nucleus, and the axoneme with a 9 + 9 + 0 flagellar pattern. This axomene differs from the aflagellate axoneme of other Psychodinae. Morphological changes, such as the casting off of the acrosomal membrane, and histological changes in the spermatophore are also described. Mating plugs that have been described previously in sandflies appear to be artefacts. Females ofP. papatasi may be inseminated more than once during each gonotrophic cycle, and additional inseminations may be necessary for each cycle. The relationships between the volumes of the sperm and the spermatheca were calculated to determine sperm utilization and fecundity ofP. papatasi. As the females ofP. papatasi mate polyandrously, the anatomical and physiological complexity of the spermathecal complex may be related to post-copulatory sexual selection.     

Spawning and biparental egg-care in a temperate filefish,Paramonacanthus japonicus (Monacanthidae)

Synopsis Reproductive habits of a temperate filefish, Paramonacanthus japonicus, were studied on a rocky reef at Tsuyazaki, Fukuoka, Japan, from 1989 through 1990. Males had territories of 30–70m² and defended them from conspecific males and potential egg predators such as another filefish, Stephanolepis cirrhifer. Egg masses were found on the sandy bottom in male territories. Individual discrimination of males and females occurring in three male territories revealed that males and females stayed in stable pairs during one month of observation in 1989. In these stable pairs, males fed only within their territories, but females occasionally foraged outside. The occurrence of egg masses within male territories and biparental egg care showed that fish were reproducing as monogamous pairs. Contrary to this, males tagged in 1990 changed their territories after the disappearance of females, and males and females mated polygamously. Spawning was observed only four times during the study period, between 1633 and 1754h. Prior to spawning, the female prepared a spawning bed on the sandy bottom. The male nuzzled the female and the pair spawned, touching their gonopores on the spawning bed. Spawning was very quick and took only 1–3 seconds. The adhesive eggs were spherical with a diameter of 0.56 mm. They were mixed with sand particles and formed a doughnut-shaped mass of about 4 cm in diameter. One egg mass contained 3300–3800 embryos of similar developmental stage, which hatched 2–3 days later. P. japonicus appears to be monogamous but may also practice polygamy when pair-bonds are unstable.     

Developmental morphology of the Indian cyprinid fish Barilius canarensis

Embryonic and larval development of an Indian cyprinid fish, Barilius canarensis, is described from laboratory-reared specimens. The eggs, measuring 2.1–2.4 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk without oil globules. Hatching occurred 39–45 h after fertilization at 26.8°–27.4°C. The newly hatched larvae, measuring 4.8–5.1 mm in body length (BL) with 22 + 17 = 39 myomeres, were characterized by melanophores already deposited on the eyes. The eggs of B. canarensis resembled those of the related danionin species Candidia barbatus, Opsariichthys uncirostris uncirostris, Zacco platypus, Z. sieboldii, and Z. temminckii. Although the larvae of B. canarensis were also similar to those of the foregoing species in general morphology, they differed in having a straight notochord tip and pigmentation on the eyes at hatching and the almost entire absence of melanophores on the ventral body surface from the yolk sac to postflexion larval stages. Conversely, melanophores occurred on the anterior abdominal and pericardial cavities from the preflexion to postflexion larval stages.     

Pelagic eggs and larvae of Coelorinchus kishinouyei (Gadiformes: Macrouridae) collected from Suruga Bay, Japan

Pelagic eggs and larvae of the macrourid fish Coelorinchus kishinouyei, collected from Suruga Bay, southern Japan and subsequently identified by 16S rRNA gene nucleotide sequences, are described. The spherical eggs, 1.18–1.31 mm in diameter, contained a single oil globule, 0.28–0.33 mm in diameter, and had hexagonally patterned ornamentation on the chorion, 0.017–0.022 mm in width. Melanophores were present on the embryo, yolk and oil globule after the blastopore had closed. Within 1 day after hatching, the body axis of the yolk-sac larvae was bent slightly at the anterior trunk region. During this stage many melanophores formed on the head, trunk, tail, yolk and oil globule, along with small irregular wrinkles on the dorsal and ventral finfolds. Pelagic eggs (after the caudal end of the embryo had detached from the yolk) and yolk-sac larvae also developed xanthophores on the embryo and yolk, and head, trunk, dorsal and ventral finfolds just before tail tip, and yolk, respectively. The pelagic larvae had a short tail, stalked pectoral-fin base and no elongate first dorsal and pelvic-fin rays. Three clusters of melanophores were present on the tail (anterior two embedded to muscle and one just before tail tip subsequently lost with development) and a cluster around the anus (beyond 3.9 mm head length). Nucleotide sequence analyses of comparative adult specimens appeared to confirm a previous proposal that C. productus is a junior synonym of C. anatirostris.     

Descriptive morphology of the eggs,larvae, and juveniles of two cyprinid fishes belonging to the <Emphasis Type="Italic">Zacco temminckii</Emphasis> species' group

 Embryonic, larval, and juvenile development of two cyprinid species belonging to the Zacco temminckii species' group, Z. temminckii (Temminck and Schlegel) and Zacco sp. (type A), are described and compared with each other from laboratory-reared and wild specimens. The eggs of both species were closely similar except in diameter [1.92–2.20 mm in Z. temminckii vs. 1.60–1.75 mm in Z. sp. (type A)], being demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk, and no oil globule. Hatching occurred 40–53 h after fertilization in Z. temminckii and after 47–60 h in Z. sp. (type A). The newly hatched larvae of both species [4.9–5.3 mm in body length (BL) in Z. temminckii and 3.5–4.8 mm BL in Z. sp. (type A)] also resembled each other, having a large transparent pear-shaped yolk and lacking body pigmentation. Myomere counts of Z. temminckii and Z. sp. (type A) larvae and juveniles were 24–27 + 14–17 = 41–42 and 23–27 + 14–17 = 40–41, respectively. The yolk was completely absorbed at 8.3 mm BL in Z. temminckii and at 6.6 mm BL in Z. sp. (type A). Notochord flexion was initiated and completed at 7.8 mm BL and 8.2 mm BL in Z. temminckii and at 6.3 mm BL and 6.6 mm BL in Z. sp. (type A), respectively. Aggregate numbers of all fin rays were completed at 17 mm BL in Z. temminckii and 13 mm BL in Z. sp. (type A). Although the morphology of larvae and juveniles of both species was very similar, differences in body length of each developmental stage, the duration and process of disappearance of the adipose finfold, the anal fin ray counts, and pigmentation on the lateral body surface were clearly recognized. Received: August 10, 2001 / Revised: March 14, 2002 / Accepted: March 27, 2002     

Kin recognition in immatures of <Emphasis Type="Italic">Heliconius erato phyllis</Emphasis> (Lepidoptera; Nymphalidae)

Females of the neotropical butterfly Heliconius erato phyllis lay eggs individually, which, after hatching into caterpillars, can be cannibalistic towards neighboring eggs. Laboratory experiments were conducted to test for kin recognition between first instar caterpillars and eggs. Two experiments were performed, putting three eggs in the vertices of an equilateral triangle made of green paper with side length of 0.5 cm. In the first experiment, two of the three eggs were full siblings; in the second experiment, one group consisted of three eggs from three independent nonrelated females, and in the other group all three eggs were full siblings. The frequency of cannibalism in experiment 1 (SSN, sibling–sibling–nonsibling) was 66% for nonrelated eggs and 34% for sibling eggs (P = 0.0018). In experiment 2, in group NNN (three nonsibling eggs) it was 83%, and for group SSS (three sibling eggs), the frequency of cannibalism was 53% (P < 0.0001). These results indicate kin recognition (caterpillar–egg). This recognition may have been important in the evolution of some of the butterfly’s adaptations, and possibly as an opportunity for kin selection.