Occurrence and coexistence in Nereis diversicolor O.F. Müller (Annelida Polychaeta) of substances immunologically related to vertebrate neuropeptides

Summary Numerous immunochemical and immunohistochemical studies have shown a wide distribution of several families of neuropeptides in invertebrates as well as vertebrates. There are relatively few data available for Annelida: Polychaeta. Therefore, we undertook an immunohistochemical investigation in the marine worm Nereis.Among the vertebrate type antibodies tested, those against met-enkephalin, LH-RH, vasopressin, oxytocin and ACTH had negative or only very slight effects. Slight to moderate reactions were obtained for VIP, SRIF, CRF, GRF, and leu-enkephalin. Moderate to very strong responses were found with anti-CCK/gastrin, -substance P, and --MSH sera. Immunopositive reactions were usually observed in the entire CNS (except, until now, in neurosecretory cells, type II, in nuclei 20, and in nerve fibres located in the infracerebral neurohemal area). The immunoreactivity was, however, more or less abundant according to different CNS regions. For example, it appeared that the immunostaining for CRF is more important in the VNC while the leu-enkephalin family is more abundant in the brain (particularly in fuchsinophilic neurosecretory cells, type I, in nuclei 20). Moreover, several vertebrate type peptides (such as CRF/GRF and CCK/gastrin) may coexist in a single neurone. Several antisera may elicit a positive reaction in some specific area (for example, substance P in the nuchal organ; SRIF in oocytes; CCK/gastrin in the gastrointestinal tract).Nothing is known about the role of the different substances immunologically detected in Nereis. It is suggested that CCK/gastrin-, -MSHand substance P-like materials transmit external stimuli to neurosecretory centres located in the caudal part of the brain.Principal Abbreviations used ACTH adrenocorticotropic hormone - CCK cholecystokinin - CRF corticotropin-releasing factor - GRF growth hormone-releasing factor - LH-RH luteinizing hormone-releasing factor - MSH melanocyte-stimulating hormone - sP substance P - SRIF somatotropin release-inhibitory factor - VIP vasoactive intestinal peptide - AE anterior eye - AG anterior group - CA corpora allata - CC corpora cardiaca - CNS central nervous system - CP corpora pedunculata - EP epidermis - ICR infracerebral region - MG medial group - n nerve - NO nuchal organ - NP neuropile - OC optic commissure - Oo oocyte - PE posterior eye - PF paraldehyde fuchsin - PF⁺ fuchsinophilic - PF^- not fuchsinophilic - Pn palpal nerve - S septum - VNC ventral nerve cord The roman and arabic numerals respectively refer to different nerves and nuclei     

Observations on the clypeasteroid Echinocyamus pusillus (O. F. Müller)

The tiny echinoid Echinocyamus pusillus (O. F. Müller) is equipped with specialized external structures that suit it for a wide variety of environments. Special features include the ability to burrow in sediments of fine sand to shell gravel and to climb vertically.Specimens dredged off the west coast of Scotland were observed in aquaria and with the SEM. E. pusillus is characterized by three kinds of spines, and by two of pedicellariae. In contrast to sand dollars, the spines play a passive rôle in the feeding and burrowing operation, probably retaining a defensive nature as in the regular urchins. It is the podia that are chiefly involved in climbing, burrowing, righting, and probably feeding. Surface ciliary currents transport particles, but not to the mouth; they may have a respiratory or cleansing function. Experimental animals did not burrow in either very fine or very coarse sand, probably because a certain relationship exists between particle weight and podia size.E. pusillus shares behavioural and structural characteristics with regular and irregular urchins. It is not a true sand dollar, but may illustrate an evolutionary stage towards such a form.     

Immunohistochemical demonstration of a CCK-like peptide in the nervous system of a marine annelid worm,Nereis diversicolor O.F. Müller

Summary Perikarya and nerve fibers containing a substance immunologically related to CCK-8 were detected in the nervous system of Nereis, a marine annelid worm. The most noteworthy immunostaining was seen in cell bodies, localized at the periphery of the brain, within nuclei 5, 6, 7, 9, 10, 13, 14, 15, 17, 20, 23–24. Immunoreactive fibers were also found in the neuropile without any particular grouping. Numerous other positive perikarya occur in the medio-ventral portion of the ventral nerve cord, and in the ventral and dorsal parts of the suboesophageal ganglion. In addition to the cell bodies in the cerebral external layer, immunoreactive axons were abundantly observed in the connectives between the ganglia. Moreover, our results demonstrate CCK-like staining in neurons showing variations in size and shape, and in affinity for paraldehyde fuchsin. The present results support the hypothesis that this peptide may exert a role as neurotransmitter or neuromodulator in annelids.     

Effects of tissue extract of adults on metamorphosis in Ascidia mentula O.F. Müller and Ascidiella scabra (O.F. Müller)

Unhatched, but fully developed larvae of two solitary ascidians Ascidia mentula O.F. Müller and Ascidiella scabra (O.F. Müller) were incubated in a variety of adult tissue extracts, which an earlier study had shown to accelerate metamorphosis. Our study confirmed this observation and showed, furthermore, that tissue extracts induce tail resorption in unhatched larvae of Ascidiella. For both species, the most active induction of metamorphosis was obtained with tunic tissue extracts. The results indicate that chemical stimulation without the presence of tactile cues may initiate metamorphosis. We suggest that the ability of tunic extracts to induce metamorphosis may explain juvenile establishment on adult ascidians and their aggregated distribution found in nature.     

Effect of water temperature on the excystment of the dinoflagellate Ceratium hirundinella (O. F. Müller) Bergh

The effect of water temperature on the excystment of the dinoflagellate Ceratium hirundinella (O. F. Müller) Bergh was studied in the laboratory. Excystment was observed between 15–30 °C and was 2% at an optimum water temperature of 20 °C–25°C. Little excystment occurred between 5 and 10 °C. The results at low temperatures are not in accordance with those obtained in an English lake. This disagreement suggests an adaptability of excystment to the temperature regime of the lake.     

Influence of the age of algae fed to rotifers (Brachionus plicatilis O.F. Müller) on the expression of mixis in their progenies

Summary The sequence of the appearance of mixis in the rotifer Brachionus plicatilis was followed among the descendents of amictic rotifers transferred from a high salinity media (40 S) to a low one (9 S). All the neonates that hatched from the amictic eggs, after being transferred to a low salinity, were amictic. Each one of these neonates was cultured individually and its offspring removed periodically every 8–10 h. It was observed that throughout their reproductive phase, these parental females retained their potential to produce either mictic or amictic offspring. All the first produced neonates developed into amictic females, but among those produced later, three patterns were prevalent. The prevalent pattern (type A) was one in which the probability of a neonate being mictic increased towards the middle of the parents' reproductive phase and was followed by a slow decline. In the second pattern (type B), the probability of a daughter being mictic was constant throughout the parents' reproductive phase. It is suspected that the quality of food supplied to the rotifers determines the appearance of patterns, A, B or C. It is postulated that the innate capacity of rotifers to undergo mixis is genetically controlled, while its expression is modulated by environmental conditions.     

A review of the biology, ecology and potential use of the common ragworm Hediste diversicolor (O.F. Müller) (Annelida: Polychaeta)

The common ragworm Hediste diversicolor inhabits the shallow marine and brackish waters in the North Temperate Zone of the Atlantic. This infaunal species builds U or Y-shaped burrows in the soft sediments. H. diversicolor is gonochoristic and remains atokous throughout its life. Due to the facility of harvesting of individuals by hand and their maintenance, this species represents a good biological material for experimental studies. In consequence, the common ragworm served as a model for endocrine control of reproduction in Polychaeta and played a fundamental role in the study of immune defence in Polychaeta.Life history characteristics (longevity, spawning season, feeding tactics and growth) and population dynamics (sex ratio, density and biomass) vary greatly according to geographical location of the populations. In addition, inter-population morphological, biochemical and physiological differences have been noticed in individuals from different areas and different environmental conditions in the same areas and could be related to the limited dispersal capacity of the species. H. diversicolor interacts with its environment. The construction of burrows by individuals increases the sediment–water interface. When they ventilate their burrows, individuals verticalize oxic zones into the sediment and promote microbial and meiofaunal growth alongside their burrows. Moreover, the common ragworm is a species of commercial interest because of its use as bait in recreational fishing and as food in aquaculture. In order to alleviate environmental pressure caused by excessive demand over optimal sustainable yield of bait fisheries, intensive aquaculture has been proposed. The common ragworm could be also used as biomonitor of pollution in estuarine ecosystems.     

The reproductive morphology of Dumontia incrassata (O. F. Müller) Lamouroux

The reproductive morphology of Dumontia incrassata (O. F. Müller) Lamouroux is described from New Hampshire, U.S.A. populations. Mature carpogonial and auxiliary branches are usually 5 and 6 celled, respectively, but both may range from 4 to 9 cells. Auxiliary branches are more numerous than carpogonial branches; however, the ratio of carpogonial to auxiliary branches increases towards the apex of the frond. A linear relationship exists between the position of the generative auxiliary cell and the number of cells in the auxiliary branches. In most cases the third cell from the top of the auxiliary branch becomes the generative auxiliary cell. The probabilities for different positions of the generative auxiliary cells are summarized. During spermatangial development, four mother cells are initially produced per cortical cell initial; subsequently, successive spermatangia are cut off obliquely and on opposite sides of each mother cell. Monoecious asexual plants of D. incrassata are described, as well as an irregular tetraspore development.Scientific Contribution Number from the New Hampshire Agriculture Experiment Station; also issued as contribution No. 81 of the Jackson Estuarine LaboratoryScientific Contribution Number from the New Hampshire Agriculture Experiment Station; also issued as contribution No. 81 of the Jackson Estuarine Laboratory     

Versuche über die Phototaxis von Nereïs diversicolor O. F. Müller

Ohne Zusammenfassung     

Distribution pattern of the dinoflagellateCeratium hirundinella (O. F. Müller) Bergh in a reservoir

Horizontal distribution of the dinoflagellateCeratium hirundinella (O. F. Müller) Bergh in the Ishitegawa Reservoir, Ehime Prefecture, Japan, was investigated. Water quality was also surveyed. It was observed that the population ofC. hirundinella exponentially decreased in number from the head of the reservoir to the dam site. Further investigation proved thatC. hirundinella initiated growth at the head of the reservoir, and later gradually expanded downstream. It was found during the period of increase in water temperature that the cell density ofC. hirundinella at the uppermost station exponentially increased.     

Analysis of the emergence of the Scarce Chaser Libellula fulva O.F. Müller, 1764 (Odonata: Libellulidae)

The Scarce Chaser (Libellula fulva) is widespread in Europe and in many regions is relatively common. It is also abundant in the lake districts of western and northern Poland. Although the species frequently occurs in large numbers its biology is poorly known. Emergence of L. fulva was investigated in this study as a contribution to the life history of the species. Studies were carried out in the vicinity of Chojno (52°41′ N, 16°12′ E) on the edge of the Notecka Forest in 2003 and 2004. The stem habitat of the population is a stream with the adjacent swamps. Libellula fulva belongs to the spring species sensu Corbet (1999). The emergence of the studied population lasted 19 days (between 18 May and 5 June) and 31 days (between 13 May and 12 June) in 2003 and 2004, respectively. It was characterised by synchronous and, especially in the first phase, mass emergence. The high synchronisation is reflected in the fact that 50% of the population had emerged by the seventh (in 2003) and the fifth (in 2004) days of the process. This is vital to the imagines, which mature and return to the water body as soon as possible to reproduce. Timing of the end of emergence depends on atmospheric conditions during its duration and the conditions preceding the start of emergence on a given day. In the daily course there is a tendency for emergence to finish as soon as possible. Extension of the emergence is synonymous with increased mortality in the dragonfly population caused by predators.     

Smelling Danger – Alarm Cue Responses in the Polychaete Nereis (Hediste) diversicolor (Müller, 1776) to Potential Fish Predation

The harbour ragworm, Nereis (Hediste) diversicolor is a common intertidal marine polychaete that lives in burrows from which it has to partially emerge in order to forage. In doing so, it is exposed to a variety of predators. One way in which predation risk can be minimised is through chemical detection from within the relative safety of the burrows. Using CCTV and motion capture software, we show that H. diversicolor is able to detect chemical cues associated with the presence of juvenile flounder (Platichthys flesus). Number of emergences, emergence duration and distance from burrow entrance are all significantly reduced during exposure to flounder conditioned seawater and flounder mucous spiked seawater above a threshold with no evidence of behavioural habituation. Mucous from bottom-dwelling juvenile plaice (Pleuronectes platessa) and pelagic adult herring (Clupea harengus) elicit similar responses, suggesting that the behavioural reactions are species independent. The data implies that H. diversicolor must have well developed chemosensory mechanisms for predator detection and is consequently able to effectively minimize risk.     

Studies on the ovotestis of the slug Agriolimax reticulatus (Müller)

Summary The follicle cells, nurse cells and germinal epithelia, which are closely associated with the oocyte of Agriolimax reticulatus (Müller) during its development in the ovotestis, have been studied using light and electron microscopy. The various secretory, digestive and phagocytic activities of these cells have also been investigated using electron cytochemical tests for oxidisable polysaccharide, acid phosphatase and electron-opaque tracer molecules. The oocyte lies initially between the germinal epithelia and a layer of nurse cells but, as oocyte vitellogenesis proceeds, it becomes encapsulated by a layer of follicle cells. Both the follicle and the nurse cells are active in secretion and digestion and contain Golgi apparatus, granular endoplasmic reticulum and acid phosphatase-rich digestive vacuoles. The significance of these activities is discussed in relation to oocyte vitellogenesis, secondary envelope formation and the digestion and recycling of cellular material.     

General morphology and ultrastructure of the radula of Testudinalia testudinalis (O. F. Müller, 1776) (Patellogastropoda,Gastropoda)

The radular morphology of the patellid species Testudinalia testudinalis (O. F. Müller, 1776) from the White Sea was studied using light, electron, and confocal microscopy. The radula is of the docoglossan type with four teeth per row and consisting of six zones. We characterize teeth formation in T. testidinalis as follows: one tooth is formed by numerous and extremely narrow odontoblasts through apocrine secretion; this initially formed tooth consists of numerous vesicles; the synthetic apparatus of the odontoblasts is localized in the apical and central parts of the cells throughout the cytoplasm and is penetrated by microtubules which are involved in the transport of the synthesized products to the apical part of the odontoblast; the newly formed teeth consist of unpolymerized chitin. Mitotic activity is located in the lateral parts of the formation zone. The first four rows contain an irregular arrangement of teeth, but the radular teeth are regularly arranged after the fifth row. The irregularly arranged teeth early on could be a consequence of the asynchronous formation of teeth and the distance between the odontoblasts and the membranoblasts. The morphological data obtained significantly expands our knowledge of the morphological diversity of the radula formation in Gastropoda.     

The morphology and fine structure of the colourless colonial flagellate Anthophysa vegetans (O. F. Müller) Stein

A strain of Anthophysa vegetans (O. F. Müller) Stein (Chrysophyceae or Chrysomonadida) was grown in culture and examined in detail by light and electron microscopy. It was shown to be a typical, though apochlorotic, member of its class; features of interest include a well-developed leucoplast and a starch-like reserve product. Aspects of the largely inorganic stalk are also discussed.     

Physiological adaptations in the survival of rotifers (Brachionus plicatilis,O. F. Müller) at low temperatures

The biology of the rotifer Brachionus plicatilis has been studied extensively in recent years, due to its importance in aquaculture. Today, the culture of several marine fish species relies completely on the daily production of live rotifers.In the present paper we explore the conditions that facilitate maintaining live rotifers for extensive periods at low temperatures. In addition to its possible contribution in providing reserve stocks for commercial application, these studies may be of ecological importance. They could explain some of the physiological adaptive mechanisms that are involved in the survival of rotifers under adverse environmental conditions.Experimental results showed relatively high survival rates (82–85%) in rotifers that were cultured at 25 °C and exposed later to -1 °C for 12–14 days. During this period, rotifers were kept without food and their media were not changed. The survival was found to depend on the rotifer culture conditions, prior to exposure to -1 °C. These included the type of food fed to rotifers (yeast or algae), the salinity in which they were cultured, and an essential acclimation period of 2–6 day at 4 °C or 10 °C. The acclimation period was associated with the synthesis of at least one specific protein and accumulation of lipids. Profiles of protein synthesis in rotifers incubated at 10 °C revealed a 94 kD protein, which did not appear in rotifers cultured at 25 or 37 °C. Immunoisolation, using a polyclonal antibody that was prepared against HSP60, revealed that this protein was synthesized in rotifers kept at 10, 25 or 37 °C. However, this antibody did not react with the 94 kD peptide.In addition, rotifers kept at 10 °C accumulated substantial amounts of lipids, including eicosapentaenoic acid (EPA), which is found in the algae fed to them. These results support the hypothesis of specific adaptations to survival at low temperatures during an acclimation period.     

Studies on the ovotestis of the slug Agriolimax reticulatus (Müller)

Summary The ovarian oocytes of Agriolimax reticulatus (Müller) have been studied by light and electron microscopy and electron cytochemistry. The development of the oocyte in the ovotestis may be divided into three stages.During Stage I the oocyte cytoplasm contains mainly ribosomes and also strands of endoplasmic reticulum, scattered mitochondria and Golgi systems. The nucleus contains both a paranucleolus and an eunucleolus. By Stage II the oocyte has enlarged, especially in a plane parallel to the basement membrane. In addition to the above mentioned organelles, the cytoplasm contains lipid, glycogen and early yolk platelets. During Stage III, the oocyte continues to enlarge, but mainly in a plane perpendicular to the basement membrane. A considerable degree of cytoplasmic differentiation has also taken place. The plasma membrane of the oocyte has become specialized with the appearance of a polysaccharide-rich glycocalyx, microvilli and pinocytotic tubules. Elsewhere, much of the background cytoplasm, containing Golgi-derived, polysaccharide and acid phosphatase-rich multivesiculate bodies, lipid and glycogen, is sequestered by smooth membranes and ultimately fuses with the growing yolk platelets. The nucleus contains an amphinucleolus, characteristic of many gastropods.The findings of this study are discussed in relation to results from other studies on oogenesis.