Formation and structure of the fertilization envelope in Xenopus laevis

This paper reports the morphological events that occur when the vitelline envelope (VE) of an unfertilized egg of Xenopus laevis is transformed into the fertilization envelope (FE) surrounding the zygote. The VE is about 1 μm thick and is composed of an interlacing network of small filaments. The FE is constructed from the VE plus an electron-dense layer (fertilization layer), about 2–6 μm thick, on the outer surface of the VE, i.e., at the interface between the VE and the innermost jelly-coat layer. The fertilization layer is a stable component of the FE and is not removed by mercaptan solutions used to dejelly eggs. The events of FE formation were observed in the light and electron microscopes after dejellied eggs were activated by pricking. The FE is established when material from the cortical granules is extruded into the perivitelline space. The cortical granule material passes through the VE as the envelope lifts away from the egg surface. Some cortical granule material deposits in the interstices of the VE, but most of it forms the fertilization layer on the outer surface of the envelope. The cortical reaction is completed about 8–9 min after addition of sperm when eggs are fertilized in vitro.     

THE EXTRACELLULAR RELEASE OF ECHINOCHROME

A study was made of the diffusion of the red pigment echinochrome from the eggs of the sea urchin, Arbacia punctulata, into sea water. Unfertilized eggs retained their pigment, over periods of hours. Outward diffusion of pigment from unfertilized eggs normally is entirely negligible, or does not occur at all. Enchancing the calcium or potassium content of the artificial sea water (while retaining isosmotic conditions) did not induce pigment release. Under anaerobic conditions, unfertilized eggs release pigment in small quantities. Fertilization alone brings about echinochrome release. Fertilized eggs invariably released pigment, whether in normal sea water, or sea water with increased calcium or potassium. This diffusion of the pigment began during the first cleavage, possibly soon after fertilization. The pigment release is not a consequence solely of the cell''s permeability to echinochrome (or chromoprotein, or other pigment combination) but is preceded by events leading to a release of echinochrome from the granules in which it is concentrated within the cell. These events may be initiated by activation or by anaerobiosis. The phenomenon was not due to cytolysis.     

Microtubules in the cortical region of the egg of Lymnaea during cortical segregation

The cortical region of the 2-cell stage egg of the gastropod Lymnaea palustris was studied by light and electron microscopy. This region includes (1) a vitelline membrane and perivitelline space which contain membrane-limited dense bodies derived from the cell surface, (2) oolemma with surface coat material and microvilli, and (3) a peripheral zone of cytoplasm (0.5-5.0 μm wide) composed of irregular vesicles, electron dense granules, and cytoplasmic microtubules. Microtubules are most abundant in the equatorial region of the egg, where they form arrays that are parallel and oblique to the egg's surface. Microtubular profiles also occur in the cortical region at the animal and vegetal poles of the egg and in the endoplasm. They may play a role in cortical segregation.     

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM BELIZE: P. NORRISIANUM SP. NOV., P. TROPICALIS SP. NOV., and P. RETICULATUM SP. NOV.1

Three new benthic, photosynthetic dinoflagellate species, Prorocentrum norrisianum, Prorocentrum tropicalis, and Prorocentrum reticulatum, from floating detritus and coral rubble of Central America are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, thecal plate ornamentation, and architecture of the periflagellar area and intercalary band. Cells of P. norrisianum are ovate with a cell size of 20–25 μm long and 13–16 μm wide. The theca is delicate, its surface smooth, pores species specific with 95 to 105 pores per valve. Pores are round with a diameter of about 0.1 μm. The periflagellar area is V-shaped, located on the right valve in a shallow depression. It has no ornamentation. The flagellar and auxiliary pores are unequal in size. The intercalary band is smooth. Prorocentrum tropicalis cells are ovoid, 50–55 μm long and 40–45 μm wide in valve view with maximum width behind the middle region, narrow at the anterior end. The periflagellar area, situated in the right valve, is a V-shaped wide triangle with a deeply indented depression; the left valve exhibits a flat ridge. The periflagellar area is unornamented, and the flagellar and auxiliary pores are unequal in size. The valve surface is rugose with evenly distributed valve poroids. Each poroid appears to have a small dome in the center. The intercalary band is rimlike around the cell margin, granulated, and horizontally striated. Prorocentrum reticulatum cells are oblong in valve view; cells are 55–60 μm long and 40–45 μm wide. Thecal surface is reticulated; it is composed of a labyrinth of ridges with alternating depressions that vary in size and shape. Each depression has a narrow, oblong-kidney-shaped opening about 0.6 μm long. The periflagellar area is a deep, V-shaped triangle. The right valve of P. reticulatum is excavated, and contains a large flagellar pore and a smaller auxiliary pore surrounded by a narrow apical collar. The left valve margin exhibits a curved flat ridge. The intercalary band is smooth.     

The effects of extenders and sperm‐egg ratios on fertilizing ability of cryopreserved testicular sperm of northern pike (Esox lucius L.)

The effects of four different extenders and two sperm‐to‐egg ratios on fertilizing ability of cryopreserved testicular sperm of northern pike (Esox lucius L.) were tested in the present study. Testicular sperm was diluted with each of the four extenders (0.6 m sucrose + 15% DMSO, 0.3 m glucose + 15% DMSO, 0.6 m sucrose + 10% methanol and 0.3 m glucose + 10% methanol, all supplemented with 10% egg yolk and 1.7 g KCl L^?1) and frozen in 0.5‐ml straws at 2 cm above the surface of liquid nitrogen and thawed at 25°C for 30 s. Then 125 μl or 50 μl of frozen‐thawed testicular sperm was poured onto about 1250 eggs for fertilization. The results showed that both sperm‐to‐egg ratio and diluent had no significant influence on cryopreservation efficiency of testicular sperm, whereas cryoprotectant had a significant influence. The highest fertilization rate (92.2%) was obtained from testicular sperm cryopreserved in glucose‐based extender containing 10% methanol at a sperm‐to‐egg ratio of 1 × 10⁶ spermatozoa per egg. The results indicated that glucose‐based extender containing 10% methanol, 10% egg yolk and 1.7 g KCl L^?1 was a useful combination.     

Microfilaments during sea urchin fertilization: Fluorescence detection with rhodaminyl phalloidin

Rhodaminyl-labeled phalloidin is used to demonstrate the distribution of microfilaments during fertilization and early development in eggs of the sea urchins Arbacia punctulata and Lytechinus variegatus. The surface of unfertilized eggs have numerous punctate fluorescence sites at which rhodaminyl phalloidin binds, indicating the presence of actin oligomers or polymers. During fertilization this punctate pattern of fluorescence begins to change. Within thirty seconds of insemination, the fertilization cone is first detectable with this technique as an erect structure on the surface of the egg. The fertilization cone grows to a maximum size by 8–9 minutes, and is resorbed by 16 minutes after insemination. The surface of the fertilized egg displays numerous fluorescent fibers by 10 minutes after insemination rather than the punctate fluorescence observed in unfertilized eggs, indicative of the burst of microfilament assembly resulting in microvillar elongation. The elongated microfilaments persist through cytokinesis. Staining is also detected throughout the cortices of unfertilized, fertilized, and cleaving eggs. Cytochalasin E (10 μM, 30 min) prevents microfilament elongation and cytokinesis and reduces the cortical staining intensity after fertilization. At cleavage, contractile rings, appearing as narrow equatorial bundles of fibers, have been detected in Lytechinus variegatus as transient structures.     

Prefeeding larval development time is not correlated with egg size in regular echinoids (Strongylocentrotus species)

Summary

We compared prefeeding development times, from fertilized egg to prism larva, for Strongylocentrotus embryos from four clutches of eggs (each from a different species) differing in size. Development times did not vary consistently with egg diameter, and trends among eggs of different sizes varied with stage of development. In some cases, development times for eggs of intermediate diameter (S. franciscanus) were longer than those for larger or smaller eggs. Although mean egg diameters in clutches ranged from 84 μm (S. purpuratus) to 162 μm (S. pallidus), differences in development time to the last embryonic stage (prism) were very small. We conclude that the inverse relationship between parental investment in offspring and premetamorphic development time in echinoids depends only on the functional consequences of reduced size of feeding larval stages: effects of egg size on prefeeding development time are not evident.     

ANIMAL/VEGETAL DIFFERENCES IN CORTICAL GRANULE EXOCYTOSIS DURING ACTIVATION OF THE FROG EGG*

One of the more striking morphological events during egg activation is exocytosis of the cortical granules. In the frog egg, the wave of cortical granule exocytosis takes about 100 sec to traverse the animal half, and travels slower in the vegetal half. We examined cortical granule exoctyosis during activation with respect to this animal/vegetal difference. In eggs which were acquiring the ability to be activated (recovering from CO₂-intoxication or undergoing meiotic maturation), animal half cortical granules became capable of responding to activating stimuli prior to vegetal half ones. Since Ca²⁺ is involved in exocytosis, we examined the effect of Ca²⁺ on cortical granule breakdown in vitro. There was no difference in sensitivity to Ca²⁺ of cortical granules from immature vs. mature eggs, but animal half cortical granules were more sensistive to Ca²⁺ than vegetal half ones. Finally, we found that prick-activation of eggs at the vegetal pole was frequently unsuccessful but would occur when external Ca²⁺ was raised. These experiments show that there are regional differences in the frog egg with respect to cortical granule responsiveness, and they suggest that the differences are due to Ca²⁺ sensitivity.     

Fertilization and aqueous development of the puerto rican terrestrial-breeding frog,Eleutherodactylus coqui

The Puerto Rican tree frog, Eleutherodactylus coqui, has internal fertilization and direct development on land. In light of these reproductive adaptations, the events of fertilization and early development were studied. Cytological examination of just-fertilized eggs showed that sperm entry is restricted to about 10% of the surface of these large, yolky eggs, and all nuclear events of the first cell cycle occur near the animal pole. Although the oocytes have cortical granules, a number of polyspermic fertilizations were found. One clutch consisted of eggs with a high frequency of polyspermy and of normal development. This raises the possibility that normal development can occur despite multiple sperm entry, a situation not found in other anuran amphibians. With respect to saline requirements, the sperm and the embryo are similar to those in amphibians with external fertilization and aqueous development. Sperm motility was high in low-tonicity conditions, and the normally terrestrial embryo could develop completely from a fertilized egg to a froglet in a low-tonicity aqueous solution.     

Viviparous development in Botrylloides (Compound Ascidians)

The mode of sexual reproduction and embryogenesis was compared in 3 species of Botrylloides: B. simodensis, B. lenis, and B. violaceus. In all species, a testis and an egg (occasionally 2 eggs), the former being anterior to the latter, mature in the mantle on either side of a zooid. The egg is surrounded by 2 follicular layers and is attached by a vesicular follicle stalk (oviduct) to the atrial brood pouch. The egg is ovulated into the brood pouch, where it is fertilized and undergoes embryogenesis. The egg of B. simodensis is heavily yolked and measures about 180 μm in diameter. The course of embryogenesis in this species is that typical of ascidians. A mature tadpole larva is produced and shed in about 5 days; then, the mother zooid degenerates. The larva is smallest of the three species and has 8 ampullae. The metamorphosed oozooid bears a single bud on the right side only. Extraembryonic nutrition seems to be very limited. Both Botrylloides lenis and B. violaceus are species which display extreme examples of viviparity. Their eggs are devoid of yolk granules, measuring about 90 μm in diameter in the former species and 60 μm in the latter. The course of embryogenesis is similar in these 2 species. The neurula stage is characterized by a spherical vesicular shape owing to precocious differentiation of the embryonic pharynx, whose ectoderm becomes vacuolated. At the posterior end of the neurula, the mesodermal cells are located in a mass, from which the tail is extended later. In B. lenis, embryogenesis takes about 20 days. At the neurula stage of the embryo, the mother zooid becomes a mantle sac as a result of visceral disintegration. During further embryogenesis, the growth of buds of successive generations in the colony is characteristically arrested. A swimming larva of this species is somewhat larger than that of B. simodensis. It has 14–24 ampullae, and the oozooid carries a single bud on its right side. In B. violaceus, the gestation period lasts for more than a month. At the early gastrula stage of the embryo, the body of the mother zooid fully disintegrates. Only the brood pouch bearing the embryo survives and remains connected with the colonial vascular system. In this species, sexual reproduction does not affect the growth of buds in the colony. The swimming larva is gigantic, being furnished with 24–34 ampullae, and the oozooid always bears 3 buds, 2 on the right side and one on the left side.     

Comparative morphology of the egg of the castniid palm borer,Paysandisia archon (Burmeister, 1880) (Lepidoptera: Castniidae)

Paysandisia archon (Burmeister, 1880) is an attractive neotropical castniid moth whose presence in Europe was recently reported. Its larvae are endophagous, feeding inside the trunks and branches of several species of palm trees (Arecaceae). The present paper deals with the morphology and biometry of the egg of this moth, comparing them with those of other castniid species.

The egg is a typical castniid egg, fusiform, upright sensu Döring, light cream or creamy pink when freshly laid, 4.69 ± 0.37 mm long and 1.56 ± 0.11 mm wide. Larvae emerge by gently splitting the chorion along one of the longitudinal ridges, on the half closer to the micropyle. SEM, TEM and LSCM photographs showing ultrastructural details of the egg are shown for the first time. The micropylar rosette (c. 54 μm in diameter) has generally 14–17 cells; in its centre lies the micropylar pit (c. 6 μm in diameter) which bears 12–16 micropylar canal openings (= micropyles) around its periphery. The pathways followed by those canals through the chorion have been figured. Eggs sampled in the wild (so laid by several females) were found to have a slightly variable number of ridges: most bore seven ridges (68.87%), although a significant portion (30.46%) bore eight and 1 egg (0.67%) bore only six; this against the currently accepted rule of five‐ridged eggs for Castniini (i.e. Neotropical castniids) to which Paysandisia archon belongs. It has also been found that the same female specimen has the capability of producing six‐, seven‐ or eight‐ridged eggs. Five types of egg irregularities affecting the longitudinal ridges are also figured and described. Transverse striae on the egg of P. archon are about 122. Aeropyles (c. 4 μm in diameter) occur on the ridges, at the intersections between the latter and two contiguous (left and right of the ridge) transverse striae, amounting to c. 854 on a seven‐ridged egg and to c. 976 on an eight‐ridged egg. Occasionally minute aeropyles ('microaeropyles') (c. 1.96–3.13 μm in diameter) also occur on transverse striae located close to both egg poles.

The chorion of P. archon shows the typical ditrysian fine structure with very thin basal layer (C‐1), 0.3–0.2 μm thick, gas‐filled trabecular layer (C‐2), c. 0.9 μm thick, and lamellar layer (C‐3), its thickness varying between 18.5 and 13 μm due to the bumpy external surface of the chorion. Aeropylar canals, that penetrate layer C‐3, connect the air‐containing inner chorionic meshwork (the trabecular layer C‐2) with the surrounding air; their outer part forms a big bulbous cavity (which opens to the outside through the small opening seen in external SEM images) and, underneath, a narrow canal follows, leading into the trabecular layer (C‐2).     

Fine structural investigation of the insemination response in Urechis caupo.

Electron microscopy of Urechis eggs revealed no changes in the egg cortex or investing layers until 4 min after insemination at 172C. From 4 min to about 30 min after insemination the surface coat gradually elevates, widening the perivitelline space. During this period, microvilli separate from the tightly woven layer of the surface coat, fibrogranular aggregates resembling surface coat material appear in the perivitelline space, and some cortical granules are extruded from the egg cortex into cytoplasmic processes. There is no statistically significant decrease in the number of cortical granules remaining in the egg surface during the first 95 min after insemination; many cortical granules persist in postgastrulae. Most of the cortical granules remain in fertilized eggs after removal of the surface coat with glucose-EGTA. We found no morphological correlates of the polyspermy block which is established within 1 min of insemination (Paul, 1975).     

Studies on the yolk granules of the silkworm,Bombyx mori L.: The morphology of diapause and non-diapause eggs during early developmental stages

Summary The morphological features during development of diapause and non-diapause eggs of the silkworm,Bombyx mori, were investigated by means of light and electron microscopy, with special reference to eggs up to 24 h after oviposition.The blastoderm and yolk cells began to be formed about 6 and 24 h after oviposition, respectively, in both the diapause and non-diapause eggs, indicating that the diapause and non-diapause eggs develop at similar rates at least until 24 h after oviposition.Specific changes in the distribution of yolk granules were observed during early development of the diapause egg. Its yolk granules gradually aggregated into clusters from the periphery toward the inside of the egg during the period of blastoderm formation. Aggregation of yolk granules was most noticeable about 12 h after oviposition and then they dispersed again before yolk cell formation. On the other hand, yolk granules of the non-diapause eggs remained dispersed during development.     

Generational shape shifting: changes in egg shape and size between sexual and asexual generations of a cyclically parthenogenic gall former

Successive generations of multivoltine species experience selection specific to the spatiotemporal environments encountered that may lead to adaptive divergence in reproductive traits among generations. To compare reproductive effort within and between generations, appropriate volumetric models, selected on the basis of the analysis of egg shape, are required to estimate the sizes (volumes) of individual eggs. We assessed the shape and estimated the volume of individual eggs produced by the temporally and spatially segregated sexual and asexual generations of the gall former, Belonocnema treatae Mayr (Hymenoptera: Cynipini: Cynipidae). Egg shape, indexed as the difference between the polar and equatorial axes of the ellipsoidal eggs, was independent of egg size, but differed between generations. The relationship of egg shape and female body size within and between generations confirmed that egg shape is an intrinsic property of each generation. Generational differences in egg shape then informed the selection of volumetric models to estimate egg size. We modeled asexual generation eggs as both spheres and prolate spheroids, and sexual generation eggs as both cylinders and prolate spheroids. Choice of volumetric model changed estimates of egg size within the asexual generation by 23% and within the sexual generation by 50%. Comparisons between generations based on the above models produced estimated differences in egg volume that ranged from 16 to 114%. In both generations, a prolate spheroid was the most parsimonious model of egg volume. Based on this model, sexual generation eggs averaged 43% larger than asexual generation eggs. The increased size of sexual eggs was achieved via conservation of the egg’s equatorial axis and elongation of the polar axis. The shift in egg shape between sexual and asexual B. treatae is the first documented dimorphism in an egg characteristic expressed between generations of a cyclically parthenogenic organism.     

Structure, development and induction of a new diapause stage in rotifers

1. This study shows that females of Synchaeta pectinata Ehrenberg can produce two morphologically and physiologically distinct kinds of amictic eggs that develop parthenogenetically into females. One kind, until now the only one known in monogonont rotifers, is a thin-shelled (about 1.4 μm) subitaneous egg that develops without arrest. The other kind is a thick-shelled (about 9 μm) egg that enters obligatory diapause after 1–3 cleavage divisions before resuming development. The thicker shell of the diapausing egg is due to an expansion of the outer sublayer of the single-layered shell. 2. The mean duration of the diapause at 19 °C (L : D 16 : 8) is about 14 days. This diapause is not broken by low temperature (5 °C), and it is not greatly extended by storage at low temperature (5 °C). 3. Diapausing eggs are induced immediately after a brief starvation period, and seem to be produced at no additional energetic cost. However, a population producing a high proportion of diapausing eggs has a much reduced reproductive potential. 4. Production of diapausing amictic eggs may be a strategy to increase the ability of clones to survive food limitation. 5. Diapausing amictic eggs differ markedly from fertilized resting eggs produced following bisexual reproduction during favourable conditions.     

THREE NEW BENTHIC SPECIES OF PROROCENTRUM (DINOPHYCEAE) FROM CARRIE BOW CAY,BELIZE: P. SABULOSUM SP. NOV., P. SCULPTILE SP. NOV., AND P. ARENARIUM SP. NOV.1

Three new benthic, sand-dwelling dinqflagellate species, Prorocentrum sabulosum, Prorocentrum scuptile, and Prorocentrum arenarium, from coral rubble are described from scanning electron micrographs. Species were identified based on shape, size, surface micromorphology, ornamentation of thecal plates, and architecture of the periflagellar area and intercalary band. Cells of P. sabulosum are oval with a cell size of 48–50 μm long and 41–48 μm wide. The areolae are round to oval and numerous (332–450 per valve) and range from 1 to 1.6 μm in size. The periflagellar area of P. sabulosum bears a wide V-shaped depression with a flat ridge and lacks ornamentation; it accommodates six pores: one large flagellar pore, an adjacent smaller auxiliary pore, and four pores of unknown function. The flagellar and auxiliary pores are surrounded by a narrow apical collar. The intercalary band of P. sabulosum is smooth. Prorocentrum sculptile cells are broadly oval, 32–37 nm long, and 30–32 μm wide in valve view with a deep-sculptured apical area. The valves are smooth and are marked with shallow depressions (856–975 per valve). Some of these depressions have a small round opening (0.13 μm in diameter). The periflagellar area is V-shaped with a deeply indented depression; it accommodates the two flagella and a thin angled apical plate. The intercalary band is smooth. Prorocentrum arenarium cells are nearly round in valve view 30–32 μm in diameter. Thecal surface is smooth with scattered kidney-shaped valve poroids (65–73 per valve) and marginal poroids (50–57 per valve). Length and width of poroids are 0.62 μm and 0.36 μm, respectively. The periflagellar area is an unornamented, broad triangle into which a large flagellar pore and a smaller auxiliary pore are fitted. Both flagella, longitudinal and transverse, protrude from the flagellar pore. The intercalary band is smooth. The presence of a peduncle-like structure (2–3 μm long) in P. arenarium was observed situated in the flagellar pore.     

Induction of cross-fertilization between sea urchin eggs and starfish sperm by polyethylene glycol treatment

Cross-fertilization between sea urchin eggs (Strongylocentrotus nudus) and starfish sperm (Asterina pectinifera) was induced by treatment with polyethylene glycol (PEG). Without treatment with PEG, the denuded egg surface (jelly coat- and vitelline coat-free) engulfed the head of acrosome-reacted sperm; however, sperm penetration did not occur [Kyozuka and Osanai, 1988]. When these eggs were exposed briefly to PEG (molecular weight 3,000) in seawater, the sperm entered the egg by membrane fusion. Cortical granules were discharged, and embryogenesis began following sperm penetration. PEG did not induce parthenogenesis in Strongylocentrotus eggs. Egg activation is thus closely linked with gamete membrane fusion.     

Isolation of protein from Urechis sperm acrosomal granules that binds sperm to eggs and initiates development

Protein was isolated from the ring-shaped acrosomal granules of Urechis sperm which bound tightly to Urechis egg surface coats and also initiated development. The acrosomal protein preparation migrated as a single major band in acetic acid-urea PAGE, had a lysine + arginine content of ∼50%, and lacked carbohydrate. The molecular mass of the acrosomal protein was 25,000–30,000 Da in cetyltrimethylammonium bromide PAGE. Acrosomal rings of acrosomereacted sperm were selectively labeled with fluorescamine and the fluorescence persisted throughout the isolation procedure and was observed in the major band on gels. Although the acrosomal protein and Urechis sperm protamine had similar amino acid contents and migrated similarly in acetic acid-urea gels, acrosomal protein differed from protamine by its relative insolubility in hot 5% trichloroacetic acid and cold 0.25 N H₂SO₄, by its migration in cetyl- and tetratrimethylammonium bromide PAGE and in a major spot on its peptide map following thermolysin digestion. Agglutination of eggs by Urechis acrosomal protein was not species-specific and included various echinoderm eggs and algal zygotes as well as Urechis eggs. Both the acrosomal protein preparation and protein extracted from the major band of gels initiated development of Urechis eggs, causing rounding out, elevation of the surface coat, germinal vesicle breakdown, polar body formation, and establishment of a polyspermy block. Polylysine and calf thymus histones were equally effective in activating Urechis eggs, but Urechis sperm protamine was less effective, and salmon sperm protamine, although highly basic, activated only a small percentage of eggs. Urechis acrosomal protein also induced partial elevation of sea urchin egg fertilization envelopes, similar to the response of sea urchin eggs to Urechis sperm. Sea urchin eggs were not activated by polylysine.     

Life history of a dipteran predator (Scathophagidae: Acanthocnema) of insect egg masses in a northern California stream

1. Little is known about the predators of insect eggs in fresh waters. This study describes aspects of the life history of a scathophagid fly (Acanthocnema sp.), whose larvae are predators of aquatic insect eggs. 2. Because the Acanthocnema predator oviposits its eggs on the surface of aquatic insect egg masses, all insect egg masses were collected regularly within a 200‐m reach of Redwood Creek (California, U.S.A.) between September 2003 and June 2007. Acanthocnema predators were found predominantly within egg masses of the caddifly Neophylax rickeri (Trichoptera: Uenoidae). 3. There was a mean of 0.25 Acanthocnema individuals per N. rickeri egg mass (n = 2367 egg masses). In general, N. rickeri egg masses were more commonly found clustered in aggregations (93.7%) than singly (6.3%), and Acanthonema were found more often within the aggregations of N. rickeri (98.7%) compared to singly laid egg masses (1.3%). 4. The duration of the Acanthocnema predator life stages was: egg 2.9 ± 0.8 (mean ± SD) days, larva 15.6 ± 10.2 days, pupa 80.3 ± 24.9 days and adult 7.2 ± 4.8 days. The short duration of the Acanthocnema egg stage (1–7 days) compared to that of its prey N. rickeri (2–4 weeks) raises the probability that the undeveloped eggs of N. rickeri would be available to the young predators upon hatching. Egg consumption of N. rickeri eggs by Acanthocnema averaged 262.6 eggs per larval period. 5. Acanthocnema had a bivoltine life cycle in which the first generation fed exclusively on N. rickeri egg masses in the winter and the second generation fed on the egg masses of several species, including other Trichoptera (Brachycentridae) and Diptera (Ceratopogonidae, Chironomidae) in the spring. These findings suggest differing feeding strategies by the first and second generations of Acanthocnema in response to the seasonal availability of prey species. This type of autecological information is important for understanding mechanisms of community interactions.     

Spawn and development of the olivid gastropod Olivancillaria carcellesi from north Patagonia,Argentina

Olivancillaria carcellesi occurs in shallow sandy shores from north Patagonia, in intertidal and subtidal sandy bottoms. Females of O. carcellesi exhibited a remarkable specificity for spawning on the shells of living males and females, indiscriminately, of the buccinanopsid Buccinastrum deforme, measuring 26.9 ± 4.7 mm in shell length. The egg capsule was semispherical and attached to B. deforme shells by a small elliptical and wide base. The capsule was translucid when spawned, with a thick and semirigid wall and a hatching aperture of 1.8 ± 0.1 mm (n = 111) in diameter. Each egg capsule contained a single egg that measured 1367 ± 34 μm (n = 5) in diameter before cleavage. The embryo developed a small bilobed velum and an operculum, which were both lost before hatching as a crawling juvenile of 1762 ± 47 μm (n = 28) in shell length. As in other species in the genus, the eggs of O. carcellesi are among the largest in the caenogastropods with direct development. The time from oviposition to hatching is estimated to be approximately 6 months.