Zschokkella mugilis N. Sp. (Myxosporea: Bivalvulida) from Mullets (Teleostei: Mugilidae) of Mediterranean Waters: Light and Electron Microscopic Description

A new Myxosporea, Zschokkella mugilis n. sp., was found in the gall bladder of several mugilids. It is distinguished from all previously reported Zschokkella from mugilids by the absence of valve ornamentation and from other species by the size of the spores. Prevalence of infection was 62.5% for Mugil capito, 70% for M. cephalus and 64.3% for Liza saliens. Plasmodia with rhizoids attached to epithelial cells appeared to compromise the host as evidenced in light and transmission electron micrographs presented herein. General ultrastructure and disporous sporogenesis of Z. mugilis resembled other Myxosporeans, but sporogonic cells did not seem to be formed in pansporoblasts.     

Homologies between hemerythrins of sipunculids and cadmium-binding metalloprotein (MP II) from a polychaete annelid, Nereis diversicolor

The determination of the first 33 amino acids of the Cd-binding-protein (MP II) of Nereis diversicolor (Annelida, Polychaeta) shows a homology of 79 and 61% with 2 respiratory proteins of sipunculids, respectively the myohemerythrin and the hemerythrin. The positive reaction obtained by immunocytochemistry over the hemerythrocytes of Sipunculus nudus using antibodies raised against MP II and the presence of iron on the MP II reinforce this similarity.     

Expression and evolution studies of ets genes in a primitive coelomate,the polychaete annelid,Hediste (Nereis) diversicolor

The Ets family includes numerous proteins with a highly conserved DNA-binding domain of 85 amino acids named the ETS domain. Phylogenetic analyses from ETS domains revealed that this family could be divided into 13 groups, among them are ETS and ERG. The ets genes are present in the Metazoan kingdom and we have previously characterized the Nd ets and Nd erg genes in the polychaete annelid Hediste diversicolor. Here, we isolated a fragment encoding the ETS domain from Nd Ets, by genomic library screening. By Northern blot analysis, we showed that this gene was transcribed as one major mRNA of 2.6 kb and one minor mRNA of 3.2 kb. By in situ hybridization, we observed that Nd ets was expressed in the intestine and oocytes and that Nd erg was expressed in cellular clumps present in the coelomic cavity, in an area of proliferating cells situated between the last metamere and the pygidium. Finally, we showed that Nd erg shared the expression pattern of Nd ets in oocytes. Molecular modeling studies have revealed that the spatial structure of ETS domain of Nd Ets and Nd Erg was conserved, in comparison to the murine Ets-1 and human Fli-1 proteins, respectively.     

Ultrastructural aspects of Ellipsomyxa mugilis (Myxozoa: Ceratomyxidae) spores and developmental stages in Nereis diversicolor (Polychaeta: Nereidae)

The ultrastructure of the spores and developmental stages of Ellipsomyxa mugilis in Nereis diversicolor were studied by transmission electron microscopy. The ultrastructure features and the developmental stages show many similarities with the general pattern described for other actinospores. However, several new features are definitely worth noting. For example, tetranucleated cells precede the formation of the initial pansporocyst, which preserves the 2 original enveloping cells until the end of sporogony. In the initial stages of sporogony, the future sporoplasm cell acquires the first secondary cell by an engulfment process. In the final stage of sporogony, spores are formed by a sporoplasm with 2 secondary cells and 1 somatic nucleus, and the polar capsule has a polar filament with a helicoidal arrangement possessing 7-8 coils.     

Development of Myxobolus macrocapsularis (Myxosporea: Myxobolidae) in an oligochaete alternate host,Tubifex tubifex

The development of Myxobolus macrocapsularis Reuss, 1906, a myxosporean parasite of the gills of common bream Abramis brama L., was studied in experimentally infected oligochaetes. In 3 experiments uninfected Tubifex tubifex Muller and Limnodrilus hoffmeisteri (Claparéde) were exposed to mature myxospores of M. macrocapsularis. In all experiments, typical triactinospores developed in T. tubifex specimens but no infection was found in L. hoffmeisteri. Triactinospores were released from oligochaetes 66 to 99 d after initial exposure. At that time pansporocysts containing 8 triactinospores were located in the gut epithelium of experimental oligochaetes, but free actinosporean stages were also found in the gut lumen of the oligochaetes. Each triactinospore had 3 pyriform polar capsules and a barrel-shaped sporoplasm with 32 secondary cells. The spore body joined the 3 caudal projections with a stout style.     

Assessment of the influence of confounding factors (weight,salinity) on the response of biomarkers in the estuarine polychaete Nereis diversicolor

The influences of salinity and body size on biochemical (activities of glutathione-S-transferase, lactate dehydrogenase (LDH), acetylcholinesterase and digestive enzymes amylase and CMCase), physiological (feeding and egestion rates, energy reserves) and behavioural (burrowing speed) biomarkers were examined in the infaunal polychaete Nereis diversicolor. Only a few biomarkers were affected, including increased egestion rate and activities of CMCase and LDH at higher salinity, and higher egestion rate in larger worms. These findings reinforce the status of N. diversicolor as a robust sentinel species for estuaries which are environments that are particularly productive but also particularly at risk.     

Synopsis of the species of Thelohanellus Kudo, 1933 (Myxozoa: Myxosporea: Bivalvulida)

A synopsis of the species of Thelohanellus Kudo, 1933 (Myxozoa: Myxosporea: Myxobolidae) is presented. It includes a total of 108 nominal species. For each species, the most relevant morphological and morphometric characteristics are provided, together with data on the type-host and locality, the site of infection within the host and the original references.     

Synopsis of the species of the genus Zschokkella Auerbach, 1910 (Myxozoa: Bivalvulida: Myxidiidae)

Systematic Parasitology - A synopsis of the species of Zschokkella Auerbach, 1910 (Myxozoa: Bivalvulida: Myxidiidae) is presented, including 94 nominal species from piscine hosts and 3 additional...     

Transport of carbon dioxide and ammonium in bioturbated (Nereis diversicolor) coastal, marine sediments

Two identical experiments with sieved and homogenized sandy and muddy sediment were conducted to determine transport enhancement of porewater solutes (TCO₂ and NH₄ ⁺) by the presence of the polychaeteNereis diversicolor (1000–1500 m^–2). Flux measurements showed thatN. diversicolor enhanced the release of CO₂ and NH₄ ⁺ 1.5–5 times. Accordingly, porewater concentrations of these compounds were reduced considerably in the bioturbated zone of both types of sediments. Two different diagenetic models, effective (eddy) diffusion and nonlocal exchange, were used to describe solute profiles in the bioturbated sediments. In permeable sandy sediments advective porewater movements may occur more readily than in more cohesive muddy sediments. The effective diffusion model (with D_e=1.6–2.0 cm² d^–1) provided an excellent fit to the measured concentrations of both solutes below the bioturbated zone in permeable sandy sediment, whereas this model overestimated the concentration in the bioturbated zone. However, in the less permeable muddy sediment the effective diffusion model overestimated the NH₄ ⁺ profile considerably at all depths. The nonlocal exchange model (with=0.17–0.29 d^–1), on the other hand, provided an excellent fit in the less permeable muddy sediment, suggesting that solute profiles here were controlled by molecular diffusion, even in the presence of burrow irrigation. For the permeable sediment, the nonlocal exchange model (with=0.14 d^–1) underestimated the measured NH₄ ⁺ profile. Accordingly, linear slopes from plots of porewater TCO₂ as a function of porewater NH₄ ⁺ revealed that eddy diffusion (or advective porewater movements) was important in the bioturbated zone of this sediment type. However, combined with the generally more realistic shape of profiles derived by the nonlocal exchange, these evidences suggest that both eddy and molecular diffusion must operate in the bioturbated zone of permeable sediments.     

楔形两极虫(黏体门,双壳目)的形态学重描述及其近缘种的分子系统学研究

两极虫属Myxidium和楚克拉虫属Zschokklella在形态上非常相似,形态鉴定界限模糊。为进一步厘清两者的分类学关系,本研究对采自涪江重庆市潼南县江段的楔形两极虫M.cuneiforme Fujita,1924进行了形态学重描述,并对其分子系统学进行了研究。楔形两极虫孢子壳面观呈长条形,中部稍凹陷或孢子一边突出,壳瓣上有6~8条与缝嵴平行的条纹;缝嵴直,缝面观呈梭形。孢子长12.5μm±0.3μm,宽5.4μm±0.3μm(n=20)。极囊2个,呈梨形,分布于孢子两极端;极囊长4.3μm±0.2μm,宽3.1μm±0.2μm(n=20),极丝细长且明显,盘曲5~6圈。以18S rDNA为分子标记,对楔形两极虫及其近缘种进行了保守区变异、遗传距离、序列相似度和分子系统发育分析,结果表明:两极虫属和楚克拉虫属均非单系发生,两属间物种相互交叉聚支;两极虫属与楚克拉虫属物种间具有很近的亲缘关系。本文基于形态和分子数据的研究结果支持将两极虫属和楚克拉虫属合并为一个属的观点。     

Transmission of Thelohanellus hovorkai Achmerov, 1960 (Myxosporea: Myxozoa) to common carp Cyprinus carpio through the alternate oligochaete host

Thelohanellus hovorkai (Myxosporea: Myxozoa) was transmitted to common carp Cyprinus carpio by exposing fish to Aurantiactinomyxon spores collected from the oligochaete Branchiura sowerbyi. The morphological characteristics of the actinosporean stage are described in detail. B. sowerbyi were exposed to T. hovorkai spores isolated from the experimentally infected carp, and after 3 and 4 months the worms exhibited prevalences of the actinosporean stage at 19.47% (7/36) and 14.6% (6/41), respectively. Control, unexposed worms were negative for the actinosporean infection. This is the first report of an Aurantiactinomyxon transforming into a myxosporean belonging to the suborder Platysporina.     

Cellular immunity in an annelid (Nereis diversicolor,Polychaeta): production of melanin by a subpopulation of granulocytes

Summary We attempted to identify the nature and origin of the pigment produced by the marine worm Nereis diversicolor in order to isolate, in inert brown capsules, foreign objects introduced into its body cavity. This brown pigment, characterized by cytochemical techniques, could be a melanin. The activity of the enzyme phenoloxidase responsible for melanin biosynthesis was detected by enzyme cytochemistry techniques in vacuoles and the Golgi apparatus of coelomocytes activated by the presence of foreign bodies. Morphological techniques combined with a monoclonal immunological probe enabled us to establish that the G₂ granulocytes contain both the precursor of the pigment in dense bodies and the capacity for phenoloxidase synthesis when activated to encapsulate foreign bodies. The G₂ granulocyte may therefore be compared to a melanocyte in which melanin is not stored as in mammals, but immediately extruded following synthesis in the form of a thick fluid.Abbreviations ACTH adrenocorticotropic hormone - Dopa L-3,4 dihydroxyphenylalanine - FITC fluorescein isothiocyanate - G ₁, G ₂, G ₃ granulocyte of types 1, 2, 3 - MSH melanocyte-stimulating hormone - proPo prophenoloxidase     

Syntheses of nucleic acids during spermatogenesis in Nereis diversicolor (annelida polychaeta): a quantitative autoradiographic study

The development of DNA and RNA synthesis in the germ cell population was studied after a 3H-thymidine or 3H-uridine pulse at each stage of spermatogenesis. The autoradiographic results show that the first sign (after 3 days in vitro) of cellular changes is an increase in RNA synthesis which reaches a maximum at day 5. DNA replication (premeiotic S phase) occurred at day 7, then cells entered meiotic prophase (day 9). Meiotic divisions and spermiogenesis occurred after 11 days. Silver grain counts permit the conclusion that RNA synthesis is clearly higher during premeiotic interphase (days 3-7) than during spermatogonial proliferation (day 0). It appears therefore that male meiotic differentiation in Nereidae is accompanied by increased RNA synthesis.     

Two species of myxozoan parasites (Myxosporea: Bivalvulida), including a new genus,fromPatagonotothen sima (Richardson, 1845) (Pisces: Teleostei) in the southwest Atlantic

A new genus and species of myxozoan parasite,Renispora simae, is described from the gall-bladder of the nototheniid fishPatagonotothen sima (Richardson). Four of 12 fish caught in Stanley harbour in the Falkland Islands in May, 1993 were infected. The new genus is placed in the family Alatosporidae and differs from other genera in this family by the shape of the spore body and the shape and site of adhesion of the membranous extensions to the spore valves. To accommodate the new genus, we propose an amendment to the definition of the family Alatosporidae to include forms with reniform spores.Myxidium baueri Kovaleva & Gaevskaya, 1982 is reported fromP. sima for the first time. It was found in the gall-bladder of one of the same 12 fish caught in Stanley harbour.     

Exchanges of sodium and chloride at low salinities by Nereis diversicolor (Annelida, Polychaeta).

1. Experiments to compare the exchange (total influx) of sodium and chloride in the polychaete Nereis diversicolor in steady-state adaptation to very low salinities are reported. 2. The Na-uptake mechanism shows a high affinity for sodium, reaching half the maximal uptake rate at an external Na-concentration of 8-10 mM/liter (ca. 2% SW), and becomes "saturated" or reaches a plateau of uptake at concentrations of 40-50 mM/liter (ca. 10% SW) up to ca. 350 mM/liter (75% SW), above which Na-exchange is proportional to the external concentration. 3. The Cl-uptake curve differs from the Na-uptake curve in showing a relative depression at very low salinities before reaching "saturation" at Cl-concentrations of 50-60 mM/liter (ca. 10% SW). Cl-uptake becomes proportional to external concentration in salinities of 50% SW or greater, suggestive of passive diffusion in the ionic and osmotic conforming range. 4. It is shown that the permeability of the body wall, both to Na and to Cl, is reduced at very low salinities, thus destroying one of the assumptions upon which a previously-presented balance-sheet for chloride exchanges in N. diversicolor was based (Smith, 1970a). 5. Attempts to demonstrate an activation of the Na-uptake mechanism at very low salinities were inconclusive; reduction of body-wall permeability to sodium masks any possible activation. 6. It is suggested that the inside-negative body-wall potential is related to the depression of the Cl-uptake curve in salinities below 10% SW.     

Research on polychaete annelid osmoregulatory peptide(s) by immunocytochemical and physiological approaches. Computer reconstruction of the brain and evidence for a role of angiotensin-like molecules in Nereis (Hediste) diversicolor OF Müller

Summary— Immunohistochemical and physiological studies were carried out on Nereis (Hediste) diversicolor OF Müller in order to obtain evidence concerning the neuroendocrine control of polychaete osmoregulation. The occurrence in this animal of peptides immunologically related to mammalian angiotensin II and I (AII and AI) and oxytocin (OT) was demonstrated in the brain and the ventral nerve cord (VNC) perikarya and nerve fibres as well as in a few peripheral structures (peripheral nerves, epithelial cells, nuchal organ, intestine and nephridia). The exact localization of immunoreactive cells was achieved by serial sections of brain and ventral nerve cord followed by a three-dimensional reconstruction of brain ganglionic nuclei using the CATIA (‘Conception Assistée Tridimensionnelle Inter Active’) Dassault system program. Injections of polyclonal antisera against AII or OT provoked a partial inhibition of the increase in body weight in Nereis exposed to hypo-osmotic medium. The effect of a-AII seemed more pronounced than that of a-OT. In a subsequent test, injections of synthetic AII and AII-amide (peptide recently isolated from an achaete (Salzet et al (1995) J Biol Chem 270, 1575–1582) enhanced the increase in body weight and, therefore, strenghthened the hypothesis of the neuroendocrine control of Nereis osmoregulation. The antidiuretic effect of both synthetic peptides in this study was indicative of the exact role of Nereis endogenous melecule(s). AII was less potent than its amidated form. If AI-like can easily be struck off the list of putative endogenous osmoregulaory factors, the role of OT-like substance in Nereis osmoregulation, which is partially demonstrated in this study, needs to be clarified by further physiological experiments using injection of synthetic peptide(s) or endogenous substance(s). All these results are discussed and compared to those recently obtained in an achaete annelid (Salzet et al (1993) Brain Res 631, 247–255; Salzet et al (1993) Brain Res 601, 173–184; Salzet et al (1995) J Biol Chem 270, 1575–1582.     

Cytochemical localization of acid phosphatase in light- and dark-adapted eyes of a polychaete worm,Nereis limnicola

Summary The amount and distribution of the lysosomal enzyme acid phosphatase in light- and dark-adapted eyes of the brackish-water annelid Nereis limnicola were studied by standard cytochemical techniques. Precipitate from the acid phosphatase reaction was observed in Golgi-endoplasmic reticulum-lysosomal complexes, primary lysosomes, and secondary lysosomes, formed by fusion of primary lysosomes with phagocytic and pinocytic vesicles containing products of presumed rhabdomeric degradation. The acid phosphatase reaction occurred in these organelles in both sensory and supportive cells of both light- and darkadapted ocelli. Secondary lysosomes were more abundant in sensory cells of illuminated ocelli than in those maintained in the dark. Sparse reaction product was found in Golgi cisternae, none in rough endoplasmic reticulum. We suggest that the increase of lysosomal activity in light-adapted eyes is correlated with the breakdown of photosensory microvilli upon exposure to light. A diagram of our interpretation of recycling of photoreceptoral membrane in N. limnicola is presented.