Toxicity and palatability of some British ascidians

Bioassays were conducted to test crude extracts of nine temperate-water ascidians for bioactivity against a range of marine organisms. The extracts generally produced moderate to weak effects, with the exception of those from the ascidian Clavelina lepadiformis, which consistently produced high mortalities against invertebrates. Some of these were then tested in simple palatability assays for feeding-deterrent effects. In general, no strong effects were detected for assays with shore crabs (Carcinus maenas) and fishes (the goby Pomatoschistus minutus and the grey mullet Chelon labrosus). Feeding-deterrent effects were observed with Carcinus maenas for the extracts of Aplidium proliferum, Botryllus schlosseri and Dendrodoa grossularia. It is postulated that these ascidians are unpalatable, but no toxic effects are evident.     

Spermatogenesis inPyrosoma atlanticum (Tunicata: Thaliacea: Pyrosomatida): Implications for tunicate phylogeny

Colonies ofPyrosoma atlanticum were collected by submersible in October 1988 in the Caribbean Sea, and testes were studied by electron microscopy. Spermatogonia, spermatocytes and early spermatids have two centrioles. The proximal centriole subsequently disappears, its remains apparently persisting in the spermatozoon as dense material adjacent to the distal centriole, which gives rise to the axoneme. At the tip of early spermatids are several 50 nm proacrosomal vesicles, which disappear leaving no trace in early elongating spermatids. The spermatozoon lacks an acrosome and has a head 35µm long. The head is differentiated into a bulbous posterior portion 5µm long × 1µm wide, a thinner anterior portion 25µm long tapering from a width of 0.7µm to a width of 0.4µm, and a very thin anterior extension 5µm long × 0.5µm wide. At the start of elongation, the anterior extension begins to form just lateral to the proacrosomal vesicles as a spiral projection comprising part of the nucleus, covered by a thin sheath of cytoplasm. This sheath of cytoplasm undergoes a complex differentiation. Ultimately, the nucleus in the anterior extension is overlain by two membrane-bound sheaths of cytoplasm connected by a spiral flange of cytoplasm. Between these two sheaths is a spiral space, open to the exterior through a subterminal pore near the sperm tip. In early spermatids the mitochondria fuse into a single mitochondrion, which remains lateral to the nucleus. The cristae become modified late in spermatogenesis. Throughout elongation of the spermatid there are patches of dense material between the nucleus and mitochondrion. A manchette of microtubules transiently encircles the thin anterior portion of the nucleus during the last phase of elongation. A manchette is not present during most of elongation. In the spermatozoon the mitochondrion, which has reticulate cristae, spirals a few times about the nucleus and extends from the junction between the bulbous portion and the thinner anterior portion of the nucleus to the junction between the thinner anterior portion and the nuclear extension. Spermatogenesis inP. atlanticum, compared to that in other tunicates, most closely resembles that in colonial ascidians, and supports the majority view that pyrosomes arose from aplousobranch ascidians that lost their attachment to the substratum. Pyrosome sperm are more highly derived than doliolid sperm, which have an acrosome that is probably capable of exocytosis. When salp and pyrosome sperm are compared, both are highly derived, but neither shares any apomorphies with the other that it does not share with at least one other tunicate order. Thus, sperm morphology does not support the majority view that pyrosomes gave rise to doliolids and neither confirms nor denies the idea that pyrosomes are intermediate between aplousobranch ascidians and salps. Therefore, it is likely that the class Thaliacea is polyphyletic, with doliolids arising very early from the ascidian lineage and with salps and pyrosomes arising somewhat later.     

Acrosome differentiation and the acrosome reaction in ascidian spermatozoa:<Emphasis Type="Italic"> Ascidiella aspersa</Emphasis> and<Emphasis Type="Italic"> Ascidia mentula</Emphasis> with some implications for tunicate phylogeny

The spermatozoa of both Ascidiella aspersa and Ascidia mentula have architectural features characteristic of ascidian spermatozoa that have previously been described. They have an elongated head (7 µm long for A. aspersa and 4 µm long for A. mentula), a single mitochondrion that is applied laterally to the nucleus and lacks a midpiece. The acrosome of A. aspersa spermatozoa is a flattened vesicle, about 200 nm×100 nm×40 nm (length, width and height). The acrosome of A. mentula spermatozoa consists of multiple vesicles; they are about 50 nm×50 nm×40 nm (length, width and height). During spermiogenesis in both species, several proacrosomal vesicles (50–70 nm in diameter) appear in a blister at the future apex of the spermatid. In A. aspersa, these vesicles fuse with each other to form a single acrosomal vesicle, while in A. mentula these vesicles do not fuse with each other, and form multiple acrosomal vesicles. In A. aspersa spermatozoa, calcium ionophore A23187 induces the acrosome reaction in which membrane fusion between the acrosomal apical membrane and the overlying sperm plasma membrane occurs along the peripheral margin of the acrosome, resulting in the release of a hybrid, membrane-bound, small vesicle. In A. mentula, multiple acrosomal vesicles disappear by releasing small vesicles after treatment with the calcium ionophore A23187; this also appears to be an acrosome reaction. This paper discusses the way in which acrosome structure and function may have changed during the evolution of the Tunicata.Communicated by T. Ikeda, Hakodate     

Ethologie alimentaire d'invertébrés benthiques filtreurs (ascidies). II. Variations des taux de filtration et de digestion en fonction de l'espèce

Rates of filtration and digestion of 4 species of ascidians (Clavelina lepadiformis), Müller, Ciona intestinalis (Linné, 1767), Halocynthia papillosa (Linné, 1767) and Microcosmus sabatieri (Roule, 1885) from a rocky shore at banyuls-sur-Mer, France have been studied using the unicellular algae Monochrysis lutheri as food and the methods previously applied to Phallusia mammillata (Fiala-Médioni, 1973). Eleven experiments of 24 h each were performed under constant conditions of temperature, pH, salinity, oxygen and food concentration. No recognizable feeding rhythm emerged; filtration is irregular, varying around a mean value. Observed filtration rates averaged 2489 ml/h/g organ dry weight in Clavelina lepadiformis, 3515 in Ciona intestinalis, 6349 in Halocynthia papillosa and 6909 in Microcosmus sabatieri. The higher rates are related to larger size and higher complexity of the gills. No pseudo-faeces are formed; a very small part of the faecal material is discharged within 24 h. The mean rates of digestion, in mg albumin equivalent/24 h/g organ dry weight, are: Clavelina lepadiformis, 3.05 mg; Ciona intestinalis, 4.74 mg; Halocynthia papillosa, 9.25 mg; Microcosmus sabatieri, 10.41 mg. The amount of digested algae corresponds to 83–92% (mean=85%) of algae filtered. This high percentage indicates good assimilation of Monochrysis lutheri by ascidians.

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Ethologie alimentaire d'invertébrés benthiques filtreurs (ascidies). II. Variations des taux de filtration et de digestion en fonction de l'espèce

     

Filtration rate in seven Scandinavian ascidians: implications of the morphology of the gill sac

The ascidian branchial basket is functionally responsible for pumping water through the animal. Within Ascidiacea the branchial basket and the stigmata are morphologically variable and these features are important taxonomical characters. However, the functional significance of this morphological variation has not been studied in great detail. Here, we have measured filtration rates in seven ascidians: Clavelina lepadiformis (Order Aplousobranchiata); Ciona intestinalis, Corella parallelogramma, Ascidia virginea (Order Phlebobranchiata); Boltenia echinata, Molgula manhattensis, Pyura tessellata (Order Stolidobranchiata) and compared these rates with the morphological characteristics of the branchial basket and the stigmata. Filtration rates were measured in the laboratory and determined from an exponential reduction in algal cell (Rhodomonas sp.) concentration as a function of time. The branchial baskets of the experimental ascidians were dissected free and photographs were taken of the entire basket and of the stigmata. It was found that filtration rates in the investigated ascidians are determined primarily by the area of the branchial basket and the length of the ciliary band lining the stigmatal openings, and that the form of the stigmatal opening has no impact on filtration rates.     

Ultrastructural investigations on sperm penetration and gamete fusion in the ascidians Boltenia villosa and Phallusia mammillata

Sperm-egg interactions during fertilization were studied by scanning and transmission electron microscopy in the two ascidian species, Boltenia villosa (collected in 1990 near Friday Harbor, Washington, USA) and Phallusia mammillata (collected in 1990 near Brest, France). Emphasis was put on sperm-egg vitelline coat (VC) and sperm-egg plasma membrane interaction. In both species the sperm activation as revealed by translocation of the mitochondrion was exhibited by sperm binding firmly to or penetrating the VC. Vesicles are present in the sperm apex of both species. Their persistence in late stages of VC penetration suggests for them a function rather in Sperm-egg fusion than in VC penetration. In P. mammillata sperm which are about to make contact and to start fusion with the egg plasma membrane, two types of vesicles, one located at the tip and the other slightly behind the apical cap, were detected. Function and fate of these prominent vesicles remain to be determined. Membrane fusion starts between the sperm membrane of the cap-like structure and short microvilli of the egg surface and then proceeds simultaneously at several sites of the apical region. In B. villosa conspicuous microvilli which are formed at the contact site and interdigitate with the corkscrew-like sperm represent fusion sites on the egg. In both ascidian species sperm are incorporated into the egg at an acute angle. Once fully embodied the sperm lie close and almost parallel to the egg membrane. Our findings are compared with observations in Ciona intestinalis, the only other ascidian species in which Sperm-egg interactions have been thoroughly investigated so far. We suggest that observed differences in the fusion processes are based on morphological differences in sperm structure which reflect phylogenetic modifications.     

Spermiogenesis and spermatozoa in the relict bivalve genus Neotrigonia: relevance to trigonioid relationships,particularly Unionoidea

Spermiogenesis and spermatozoa in the relict trigonioid genus Neotrigonia are examined ultrastructurally. Mature Neotrigonia spp. spermatozoa exhibit the following features: (1) a blunt-conical nucleus; (2) an acrosomal complex composed of discoidal vesicles (9 to 15) arranged as a thin layer over the nuclear apex; (3) five (rarely four) spherical mitochondria positioned in depressions at the base of the nucleus; (4) proximal and distal centrioles (surrounded by the mitochondria); (5) a satellite complex anchoring the distal centriole and flagellum to the plasma membrane; and (6) a single flagellum. Spermatozoa of unionoids examined to date (Unionidae and Hyriidae only) exhibit similar features, including a nucleus with a blunt apex capped by a thin acrosomal complex (showing up to three vesicles). Among the Bivalvia, only Neotrigonia spp. and unionoid sperm possess an acrosomal complex composed of multiple vesicles. These data suggest that the Trigonioidea and Unionoidea are phylogenetically linked, but further work particularly on primitive unionoids is required to determine if this relationship is one of common ancestry or whether the Unionoidea have in fact been derived from the Trigonioidea.     

Fine structure of spermatids and sperm of Dolioletta gegenbauri and Doliolum nationalis (Tunicata: Thaliacea): implications for tunicate phylogeny

In Dolioletta gegenbauri and Doliolum nationalis, collected in 1987 in the bay of Villefranche-sur-mer (French Mediterranean Sea), spermiogenesis is essentially the same. Early spermatids have a round head, a flagellum arising from a single centriole with short microtubules at 45° to its base, several mitochondria, and an acrosome 50 nm thick and 250 nm long with its long axis parallel to the plasma membrane. The acrosomal contents are dense, with a central denser plate. The nuclear envelope next to the acrosome is thickened and concave. In elongating nuclei, strands of chromatin become oriented parallel to the length of the nucleus and then twist helically. Although the mitochondria surround the nucleus, they remain relatively short and do not fuse into a single mitochondrion as in sperm of other tunicates. In very late doliolid spermatids, the acrosome undergoes exocytosis, and exposes fibrous material that stays associated with the tip of the sperm; no acrosomal tubule forms. Exocytosis at this stage may be triggered by fixation. If so, exocytosis probably occurs naturally at some time before fusion of sperm and egg. Sperm have elongate heads (1 m×10 m), with the anterior two-thirds of the nucleus surrounded by mitochondria. Spermiogenesis in doliolids, compared to that in other tunicates, is most like that in solitary members of the class Ascidiacea, except that in the latter the sperm mitochondria fuse and the acrosome appears incapable of exocytosis. In contrast, previous work has shown that salps (class Thaliacea) and colonial didemnid ascidians have an acrosomeless sperm with a spiral mitochondrion, while the class Appendicularia has a sperm with a midpiece, a compact head and an acrosome capable of exocytosis and acrosomal tubule formation. By outgroup comparison with echinoderms and acraniates, appendicularian sperm are plesiomorphic within the Tunicata. Thus, gamete morphology indicates that (1) solitary ascidians and doliolids had a common ancestor, (2) the popular idea that doliolids gave rise to appendicularians is incorrect, and (3) the Thaliacea are polyphyletic, doliolids having arisen very early from the ascidian lineage and salps having arisen later.     

Storage of exogenous sperm by the compound ascidianDiplosoma listerianum

Experimental evidence is presented for the storage of exogenous sperm by zooids of the simultaneously hermaphroditic didemnid ascidianDiplosoma listerianum. Cross-fertilized zygotes can be produced up to 1 mo after a ramet is placed in reproductive isolation. Previously published observations, considered in the light of the present data, suggest that the lumen of the ovary is the site of sperm storage. This is believed to be the first report of post-transmission storage of sperm by an ascidian. Existing records of storage of exogenous sperm in other taxa rarely involve aquatic animals in which spermatozoa are transmitted between mates singly through the surrounding water, as inD. listerianum. The potential significance of sperm storage in the mating systems of sessile aquatic invertebrates is discussed.     

Cryptic species of Clavelina (Ascidiacea) in two different habitats: harbours and rocky littoral zones in the northwestern Mediterranean

Marinas and harbours provide ideal sites for the study of population genetics of marine invertebrates with restricted dispersal capabilities. They combine a confinement effect, particular ecological conditions (pollution, turbidity), and the possibility of high gene flow through ship-borne propagules, which greatly increases the natural dispersal capability of sexual and asexual propagules in many species with short-lived larvae. We studied the genetic structure of populations of the ascidian Clavelina lepadiformis living inside and outside harbours in the north-western Mediterranean. A 500-bp segment of the cytochrome c oxidase subunit I (COI) mitochondrial gene was sequenced in three populations from inside harbours (interior form) and in three populations from the rocky littoral (exterior form). Two congeneric Mediterranean species, Clavelina sp. and C. dellavallei, were used for comparison. We found that the interior and exterior forms of C. lepadiformis belong to two distinct clades, with a genetic divergence of 5%. Gene-flow values among these forms were insignificant. The lack of gene flow and the genetic divergence suggest that the interior and exterior forms of C. lepadiformis are in fact cryptic species rather than differentiated populations of the same species. Levels of gene flow were higher among interior habitats than among exterior habitats, a pattern likely maintained by genetic exchange through ships. We discuss the possible origins of the present-day distribution of these cryptic species. We contend that the study of species living both inside and outside these particular habitats will reveal more instances of genetic discontinuities allowing local adaptations.     

Morphology of the reproductive system and spermatozoa of Mesodasys adenotubulatus (Gastrotricha: Macrodasyida)

Mesodasys adenotubulatus Hummon, Todaro and Tongiorgi 1993 is a simultaneous hermaphrodite. Two club-shaped testes extend along the sides of the anterior intestine and show spermatogenetic stages maturing in a caudo-cephalic and centripetal direction. Sperm ducts join together dorsally to the intestine before opening into a pear-like caudal organ. An unpaired, dorsal ovary lies anteriorly to the confluence of sperm ducts. Oocytes mature in a caudo-cephalic direction; no oviduct nor female pore is present. The spermatozoa are fusiform, with a very short tail. The anterior end of the sperm is a small, conic hood which corresponds to the acrosome. The following, spindle-shaped corkscrew region contains a nuclear helix surrounding a central axis made of a single, giant mitochondrion. The 9+2 axoneme is enclosed by a bilayered, obliquely striated cylinder only in the proximal tract. The morphology of the reproductive system and spermatozoa of Mesodasys species is discussed, and the hypothesis of hypodermic impregnation in this genus is supported. Received: 20 November 1998 / Accepted: 6 July 1999     

Spermatogenesis,sperm storage and comparative sperm morphology in nine species of Capitella,Capitomastus and Capitellides (Polychaeta: Capitellidae)

Light microscopy and transmission electron microscopy (TEM) were used to describe spermatogenesis and the morphology of mature sperm and sperm storage organs in five sibling species of Capitella, three species in the related genus Capitomastus, and one species in the genus Capitellides. These capitellids lack a well-developed testis, but young males have a few specialized regions of the peritoneum in the eighth setiger, where germ cells proliferate and spermatogonia are released into the coelom, and spermiogenesis is completed. Mature sperm are stored in the central regions of paired genital ducts (coelomoducts), which lie between the seventh and eighth setigers. The cells forming the walls of the coelomostome and central region of the duct are ciliated and have large glycogen deposits. The lumenal borders have extensive microvilli and there is evidence that they secrete glycogen-containing materials into the duct. All species have modified primitive sperm with a conical acrosome, elongated nucleus, and long middle piece extending along the proximal portion of the flagellum. A single ring-shaped mitochondrion encircles the centriolar region of the middle piece and the cytoplasm is filled with glycogen. The sperm of all nine species differ significantly in the lengths of their middle pieces, acrosomes and especially in their nuclear lengths. The nuclear lengths have a twofold range among the sibling species of Capitella and Capitomastus. Subtle differences in the shape and volume of the acrosomal vesicle and acrosomal space characteristic of the Capitella sibling species seem to correlate with a basic division of these species into those with diploid chromosome numbers of 20 or 26. Spermiogenesis, the number of sperm produced, and the method of sperm storage are appropriate for efficient sperm utilization in fertilization. No evidence indicates that spermatophores are formed and transferred between individuals and the method of sperm transfer is not understood. The differences in the dimensions and acrosome morphology of mature sperm, and the previously demonstrated specializations in the egg envelopes in the Capitella sibling species, are characteristic features of the reproductive isolation that exists among these capitellid species.     

Sexual reproduction in the compound ascidian Diplosoma listerianum (Tunicata). I. Metamorphosis,storage and phagocytosis of sperm in female duct

In the colonial ascidian Diplosoma listerianum (collected in the Lagoon of Venice in 1986 and 1991), ovulating eggs detach themselves from the ovary wall and segregate in the tunic without exposure to seawater. With the aim of finding evidence of the pathway followed by spermatozoa to approach the oocyte, histological and ultrastructural observations were made. The results showed that the hollow ovary elongates in a fertilization canal which flanks the sperm duct and opens externally, next to the anus. Intercellular extended tight junctions isolate the lumen of this canal from the blood. Sperm were found in the fertilization canal, both free and in intracellular vacuoles of wandering cells (phagocytes). These sperm showed differences in comparison to those from the sperm duct, which are possible signs of a sperm reaction. The main modifications were shown by the dense groove, a narrow invagination of the plasmalemma bound to the nuclear envelope by dense material. In non-modified sperm, the groove runs spirally all along the head, while in those in the fertilization canal, it coils in the anterior half of the head, pressing back the long mitochondrion and endoplasmic tubules. These modifications were interpreted to be result of the release of the dense groove, considered to be a sort of stretched, contractile spring. The presence of sperm in the fertilization canal and particularly in phagocytes is discussed in relation to the capacity for exogenous sperm storage by zooids and to the necessity of waste clearance in order for successive waves of fresh sperm to be able to move towards fully grown oocytes. The origin of the female fertilization canal, the role of its tight junctions, and the complex sperm morphology are discussed as adaptations to internal fertilization in D. listerianum.     

Chemical defense and aposematic coloration in larvae of the ascidian Ecteinascidia turbinata

In the coastal waters of Florida (USA) tadpole larvae of the colonial ascidian Ecteinascidia turbinata contain chemicals which make them unpalatable to planktivorous juvenile pinfish Lagodon rhomboides. Experiments demonstrate that the bright organe color of E. turbinata tadpoles is aposematic. Fish that have recently tasted larvae of E. turbinata will not attack the palatable tadpoles of Clavelina oblonga when the latter are dyed organe to resemble larvae of E. turbinata. Tadpoles of E. turbinata that have been mouthed and rejected by fish generally survive to complete a normal metamorphosis. Individual selection explains the evolution of aposematic coloration in E. turbinata better than kin selection. The identity of the defensive chemical is unknown. The unpalatable substance in larvae of E. turbinata is removed by dialysis, indicating that it has a molecular weight less than 14000 d. Larvae are not acidic, nor is the active substance denatured by doiling.     

Light-enhanced growth of the ascidian Didemnum molle/Prochloron sp. symbiosis

To obtain a direct measurement of the importance of Prochloron sp. to the ascidian Didemnum molle, ascidian colonies from Lizard Island Lagoon, Great Barrier Reef, were grown for 9 d at 0, 10, 40 and 100% sunlight in situ using unidirectional flow chambers. Growth (wet weight) was enhanced up to 40% of full sunlight, at which point growth appears to have been light-saturated. Colonies in 10 and 40% sunlight responded by (1) climbing up the sides of the growth chambers, and (2) flattening out to a more encrusting morphology; also (3) the chlorophyll content of three colonies in zero sunlight decreased by >80%, yet the ascidians remained healthy and did not lose weight. These data show that although the symbiosis may not be obligatory for D. molle, the ascidian's growth is enhanced by Prochloron sp., and the morphology of the ascidian colony is affected by its photobiology.Contribution No. 344 of the Australian Institute of Marine ScienceCommunicated by G. F. Humphrey, Sydney     

Three new species of free-living marine nematodes from sublittoral sediments in Liverpool Bay

Three new species of free-living marine nematodes are described from sublittoral sediments in Liverpool Bay, UK. Pselionema longiseta n.sp. (Ceramonematoidea) is closest to P. longissimum Gerlach, 1952, from which it is distinguished by the longer cephalic setae and the larger amphids; Laimella filicaudata n.sp. (Chromadoroidea) is closest to L. longicauda Cobb, 1920, from which it is mainly distinguished by the much greater curvature of the spicules and the longer caudal apophyses of the gubernaculum; Pseudonchus decempapillatus n.sp. (Choanolaimoidea) is closest to P. deconincki Warwick, 1969 and is distinguished by the presence of 10, rather than 13, pre-anal supplements in the male, by the longer cephalic setae, and by the presence of numerous caudal setae.     

Beat frequency of cilia in the branchial basket of the ascidian Ciona intestinalis in relation to temperature and algal cell concentration

To elucidate the effects of temperature and algal cell concentration on pumping of water in the ascidian Ciona intestinalis a number of different experiments were performed. Beat frequency of the lateral cilia in the openings of the branchial sac was measured in intact specimens using a microprojection objective and a monochrome CCD video camera. At constant low algal cell concentration, beat frequencies increased linearly with temperature from 4.0 Hz (±0.5) at 7.4 °C to 13.6 Hz (±1.6) at 20.1 °C. At a constant temperature of 15 °C, beat frequency decreased with increasing algal cell concentration from approximately 3000 to >10 000 Rhodomonas sp. cells ml⁻¹. The decrease was observed both in experiments where the ascidians had been acclimated to a fixed algal cell concentration and in experiments with changing concentrations. Effect of algal cell concentration on squirting/siphon closure and flow velocity in the exhalent siphon was measured using a thermistor. At low algal cell concentrations, flow velocity in the exhalent siphon was stable, apart from a few short squirts. At very high algal cell concentrations, the flow velocity was reduced and much less stable, with prolonged squirting. The effect of gut content on filtration was studied in experiments with specimens acclimated to high algal cell concentrations. Results showed a close relation between gut clearance and filtration rate. From the experimental results and a qualitative analysis of the Ciona-pump it was concluded that the ciliary beat frequency is proportional to the water flow through the sea squirt and that changes in pumping caused by temperature or algal cell concentration are under nervous control or governed by enzyme kinetics, rather than being a result of physico-mechanical properties, i.e. pump efficiency versus flow resistance, of the ascidian pump. Received: 6 October 1997 / Accepted: 8 October 1998     

Quantitative differences in mitochondrial ultrastructure of a thiobiotic and an oxybiotic turbellarian

Specimens of the thiobiotic acoel Solenofilomorpha funilis Crezée, 1975 and the oxybiotic acoel Otocelis sp. were collected from Mount Desert Island, Maine, USA in spring and summer of 1982, and the ultrastructure and distribution of their mitochondria were compared. The volume density and the surface density of the mitochondria are significantly higher in the epidermis and parenchyma of S. funilis than in the corresponding tissues of Otocelis sp. Surface densities of mitochondrial cristae are higher in all tissues studied (epidermis, muscle, and parenchyma) of S. funilis than in Otocelis sp. The differences in mitochondrial ultrastructure of these two species suggest that mitochondria in anaerobic tissues differ in function from their aerobic counterparts.     

Comparative fertilization biology of gymnolaemate bryozoans

Locations and oogenic stages of eggs at the time of sperm-egg fusion were determined for nine gymnolaemate species exhibiting different reproductive strategies with respect to site of embryonic development (e.g., water column, introvert, embryo sac, or ovicell) larval type, and zooid sexuality. Ovarian, coelomic, and recently spawned oocytes obtained from freshly collected colonies between 1988 and 1995 were stained with aceto-orcein and examined for the presence of a sperm nucleus. All nine species are shown to have a similar fertilization biology in which sperm fuse with eggs in or near ovaries and egg activation is delayed until eggs are spawned. In species such as the etenostomeAlcyonidium sp. and the cheilostomeElectra pilosa (Linnaeus, 1767), which produce marierons oocytes that are spawned into the water column and have cyphonautes larvae, sperm fuse with late growth stage ovarian oocytes after germinal vesicle breakdown (GVBD) at or near ovulation. In species that produce few oocytes, brood embryos, and release coronate larvae, sperm fuse with late growth stage ovarian primary oocytes before GVBD, as in the ctenostomeBowerbankia gracilis Leidy, 1855, or with very early growth stage ovarian primary oocytes, as in the cheilostomesCribrilina corbicula (O'Donoghue, 1923),Dendrobeania lichenoides (Robertson, 1900),Hippodiplosia insculpta (Hincks, 1882),Schizoporella serialis (Heller, 1867),T ricellaria gracilis (Smitt, 1867), andWatersipora arcuata Banta, 1969. Fertilization success was high in all nine species, with 83 to 100% of oocytes at an appropriate oogenic stage containing a sperm nucleus or early male pronucleus. Gymnolaemate bryozoans may maintain high levels of fertilization success by: (1) concentrating spawned sperm from the water column using colony feeding currents; (2) localizing sperm-egg encounters in or near the ovary, (3) using sperm when available by permitting sperm to fuse with early growth stage oocytes; and (4) selfing.     

In vitro effects of lead on fatty acid composition,oxidative stress biomarkers and quality of ram spermatozoa

The sperm quality and several parameters associated with oxidative stress were evaluated in ram (Ovis aries) spermatozoa suspensions incubated with 0, 50 or 500 ng/mL Pb during 0, 1, 3 or 6 h. The presence of Pb during incubation, reduced the integrity of the acrosome, % sperm motility and integrity, and the functionality of membrane. On the contrary, % polyunsaturated fatty acids and the activities of glutathione peroxidase and superoxide dismutase in the spermatozoa suspensions were not affected by Pb. Moreover, glutathione peroxidase activity was decreased and the fatty acid composition changed due to the relative increase in % stearic acid during the incubation time independently on Pb presence. Data showed that several effects of Pb on sperm quality usually observed in vivo also occurred in vitro, but without any relationship with oxidative stress biomarkers.