Genotoxic damage in polychaetes: A study of species and cell-type sensitivities

The marine environment is becoming increasingly contaminated by environmental pollutants with the potential to damage DNA, with marine sediments acting as a sink for many of these contaminants. Understanding genotoxic responses in sediment-dwelling marine organisms, such as polychaetes, is therefore of increasing importance. This study is an exploration of species-specific and cell-specific differences in cell sensitivities to DNA-damaging agents in polychaete worms, aimed at increasing fundamental knowledge of their responses to genotoxic damage. The sensitivities of coelomocytes from three polychaetes species of high ecological relevance, i.e. the lugworm Arenicola marina, the harbour ragworm Nereis diversicolor and the king ragworm Nereis virens to genotoxic damage are compared, and differences in sensitivities of their different coelomic cell types determined by use of the comet assay. A. marina was found to be the most sensitive to genotoxic damage induced by the direct-acting mutagen methyl methanesulfonate (MMS), and showed dose-dependent responses to MMS and the polycyclic aromatic hydrocarbon benzo(a)pyrene. Significant differences in sensitivity were also measured for the different types of coelomocyte. Eleocytes were more sensitive to induction of DNA damage than amoebocytes in both N. virens and N. diversicolor. Spermatozoa from A. marina showed significant DNA damage following in vitro exposure to MMS, but were less sensitive to DNA damage than coelomocytes. This investigation has clearly demonstrated that different cell types within the same species and different species within the polychaetes show significantly different responses to genotoxic insult. These findings are discussed in terms of the relationship between cell function and sensitivity and their implications for the use of polychaetes in environmental genotoxicity studies.     

Effect of crowding on nauplii production during mating time inTisbe clodiensis andT. holothuriae (Copepoda,Harpacticoida)

Tisbe clodiensis andT. holothuriae females and males were maintained at different densities, from sexual maturity to the extrusion of the first egg sac, in order to study the effect of crowding on the number of nauplii produced by the first egg sac. They were tested in pure cultures at densities of 1, 2, 4, 6 and 8 couples per 20 ml of sea water, and in mixed cultures (half of the individuals belonging to each of the two species) with 2, 4, 6 and 8 couples per 20 ml, without food limitation. At the highest crowding level, and forT. holothuriae only, the effect of daily renewal of the culture water was examined. The mean number of nauplii per female decreases with increased density. A significant linear regression of number of nauplii on number of couples was found. The regression slopes appear to be the same for both species, regardless of whether they are cultured singly or together. The daily renewal of water diminishes the effect of crowding. The possibility that a complex chemical compound, produced by the animals, which enables the latter to perceive and to react to crowding, is discussed.     

Taxonomic status of three nominal species of Indo-Pacific symphurine tonguefishes (Symphurus: Cynoglossidae: Pleuronectiformes)

The taxonomic status of three rarely captured, nominal species of Indo-Pacific symphurine tonguefishes,Symphurus trifasciatus (Alcock, 1894),S. microrhynchus (Weber, 1913), andS. holothuriae Chabanaud, 1948, is resolved.Symphurus microrhynchus, previously known only from three specimens, is redescribed based on 12 specimens.Symphurus holothuriae, previously reported only from the holotype, is a junior subjective synonym ofS. microrhynchus. Symphurus trifasciatus is redescribed based on six specimens, including the lectotype designated in this study.Symphurus microrhynchus is distinguished from congeners by a combination of meristic features, head and opercle shape, dorsalfin position, and features of its pigmentation. The species occurs in relatively shallow tropical waters from Zanzibar through French Polynesia (the only reported occurrence of a shallow-waterSymphurus from a locality on the Pacific Plate), including Indonesia, Papua New Guinea, Vietnam, and northwestern Australia.Symphurus trifasciatus, known from deep waters along both coasts of the Indian subcontinent and adjacent waters, has an overall similarity in body shape and meristic features reminiscent of those inS. microrhynchus, but differs notably in having a longer and wider head, a distinctive opercle shape, number of scale rows on the head, and pigmentation pattern.     

Experimental study of 110mAg transfer from sediment to biota in a simplified freshwater ecosystem

Radioecological field studies show that most of the ^110m in contaminated freshwater ecosystems occurs in the sediment, which therefore represents a potential source of radioactive pollution. Laboratory experiments were carried out to quantify ^110mAg uptake and retention by three species of bottom-dwelling organisms, a limicolous midge larva (Chironomus luridus), a benthic crustacea (Gammarus pulex) and an omnivorous fish (Cyprinus carpio), and to assess the potential incorporation of this radionuclide in freshwater food webs. The rate of contamination of each organism was described using a two-compartment model. Concentration factors (^110mAg in organism: ^110mAg in water) reached a maximum (1100 ww) in 30 and 60 d respectively for gammarid and midge larva. The concentration factor for Cyprinus carpio increased as a function of time to reach a maximum value of 106 after 180 d. ^110mAg uptake from the sediment suggests a real possibility of direct transfer from this compartment to the three organisms. The maximum value of the transfer factor (^110mAg in organism: ¹¹⁰Ag in sediment) was about 1.9, 0.17 and 0.022, for gammarid, midge larva and carp, respectively. The ingestion of contaminated gammarids and midge larvae by carp results in a ^110mAg accumulation corresponding to a trophic transfer factor (^110mAg in predator: ^110mAg in prey) of 0.023 and 0.135, respectively. Comparison of the different factors that influence the ^110mAg transfer suggests that water is the most important contamination vector. However, taking into account their ^110mAg concentration, prey and sediment can lead to a greater level of contamination in consumer organisms than uptake from water.     

Preparation and testing of a polyvalent conjugate for direct fluorescent-antibody detection ofLegionella pneumophila

A polyvalent conjugate forLegionella pneumophila, the Legionnaires’ disease bacterium, was prepared by combining monospecific antibodies for the four recognized serogroups ofL. pneumophila. Pure cultures ofL. pneumophila and other bacteria representing 18 genera and 50 species of heterologous organisms were used in evaluating the reagent. A total of 358 specimens from patients suspected of having Legionnaires’ disease also were tested. The results show the practicality and advantages of using a polyvalentL. pneumophila conjugate for screening clinical specimens.     

Determination and localization of oil components in living benthic organisms by fluorescence microscopy

Microspectrofluorometric measurements are made to determine uptake and distribution of oil in marine organisms after exposure to crude oil. Equipment combining fluorescence microscopy with spectral analysis of the fluorescence emission is described. After contamination with oil, the intestine content ofLumbricillus lineatus, Nereis diversicolor andAnaitides mucosa shows a fluorescence emission at long wavelengths with a maximum at about 550 nm; this is in contrast to the fluorescence emission of these organisms without oil contamination. There is evidence that aromatic hydrocarbons are metabolized in the intestine of the worms studied.     

Antimicrobial susceptibility of anaerobic bacteria isolated from clinical specimens by using an agar dilution procedure

An agar dilution procedure was used to evaluate the antimicrobial susceptibility of 2,126 anaerobic bacteria isolated from a broad spectrum of properly obtained clinical specimens. Significant numbers ofBacteroides species,Fusobacterium species, anaerobic Gram-positive cocci, and anaerobic nonsporeforming Gram-positive bacilli would not grow on Wilkins-Chalgren agar used in the procedure. The failure to obtain growth from theBacteroides species was of special concern since these organisms generally show a higher degree of resistance to antibiotics than the other anaerobic bacteria, which failed to grow or grew poorly on that medium. When compared to other investigations, the data indicated an emerging resistnace to clindamy a forB. thetaiotaomicron andB. distasonis. IndividualClostridium species showed a wide divergence in susceptibility.     

Guanine plus cytosine content of theMycobacterium avium complex and other mycobacteria by high-performance liquid chromatography analysis of deoxyribonucleotides

DNA was extracted from 12 clinical isolates of theMycobacterium avium-M. intracellulare-M. scrofulaceum serocomplex (including serovars 1, 2, 3, 4, 8, 42A, and 43), as well as from nine reference strains of other mycobacterial species. Rapid nitrogen decompression was used to weaken the cells for subsequent DNA extraction by a modified Marmur procedure. The DNA was hydrolyzed with phosphodiesterase, and the concentration of released deoxyribonucleotides was determined by reversed-phase high-performance liquid chromatography. The data were used to calculate the mole percentage guanine plus cytosine. Values ranged from 58.6 forM. leprae to 71.5 forM. avium serovar 3. The lowest value for theM. avium-M. intracellulare-M. scrofulaceum serocomplex was 66.3 forM. scrofulaceum serovar 42A. The data indicate that there is greater genetic variability for this group of organisms than is suggested by diagnostic clinical tests and seroagglutination reactions.     

Effect of culture media onLactobacillus hydrophobicity and electrophoretic mobility

The hydrophobicity of six strains representing three species ofLactobacillus was measured using dextran-polyethylene glycol contact angle measurements. These ranged from 123.6° forLactobacillus casei douche to 26.2° forL. casei RC-17 under identical growth conditions. The results indicated that the nutritional environment affected bacterial hydrophobicity. Electrophoretic mobilities of the lactobacilli were also determined and found to be negative for all specimens, and to vary with growth media, especially when sugars were added to urine. The electrophoretic mobility histograms showed one main peak for all strains, exceptLactobacillus acidophilus T-13 which had two peaks, suggesting two morphological sizes or types within its population. In addition, strain T-13 was more positively charged than the other five strains after growth in agar, urine, and supplemented urine. The use of contact angle and electrophoretic mobility techniques allows examination of cell surface properties of lactobacilli that may have importance in the colonization of mucosal epithelia.     

Feeding habits of young predatory fishes in marsh creeks situated along the salinity gradient of the Schelde estuary, Belgium and The Netherlands

Fish and macrobenthos were sampled in four different marshes along the salinity gradient of the Schelde estuary, Belgium/Netherlands, to investigate the importance of marsh creeks as foraging grounds for the dominant, larger fish species. The total density and biomass of all the main macrobenthic taxa (Corophium volutator, Nereis diversicolor, Oligochaeta, Macoma baltica and Heteromastus fliliformis) were measured. The feeding habits of the larger predatory fishes (Platichthys flesus, Dicentrarchus labrax) were investigated. Qualitative and quantitative stomach analyses included the calculation of different indices, showing the niche breadth (as diet diversity) and the niche overlap (as similarity between the predators diet) for this habitat. These analyses showed that the two most important benthic prey species for P. flesus were C. volutator and N. diversicolor. D. labrax preyed upon a wider range of species, including C. volutator, N. diversicolor, Crangon crangon, Carcinus maenas and Orchestia spp. The stomach diversity of D. labrax and P. flesus showed differences between the marshes although there was no consistent pattern in diet composition, reflecting the opportunistic nature of feeding by these large predators. The fullness indices of both flounder and sea bass did not differ significantly along the salinity gradient and the estimated minimum consumption by these predators did not indicate a top-down control of the macrobenthic community. The salt marsh creeks seem to provide excess food for the visiting fish species. The benthic prey was present in very high abundances, which may suggest that the typical nursery species such as C. crangon and C. maenas, and early juveniles of P. flesus, D. labrax and Pomatoschistus microps were not preyed upon significantly. This supports the hypothesis that salt marsh creeks provide good refuge areas for nursery species against predation by larger fish.     

Competition betweenCulex quinquefasciatus Say andAnopheles arabiensis patton in the Khartoum area,Sudan

The range of environmental temperatures that allow of survival would appear to be greater forAnopheles arabiensis than forCulex quinquefasciatus. Nevertheless the latter is the more common species in the Khartoum (Sudan) area.This relative abundance is suggested to be attributed largely to the success ofCulex quinquefasciatus in exploiting a wide range of breeding sites, even those which are obviously contaminated in various ways.     

Nitrogen acquisition from inorganic and organic sources by boreal forest plants in the field

A wide range of recent studies have indicated that organic nitrogen may be of great importance to plant nitrogen (N) nutrition. Most of these studies have, however, been conducted in laboratory settings, excluding important factors for actual plant uptake, such as competition, mycorrhizal associations and soil interactions. In order to accurately evaluate the importance of different N compounds to plant N nutrition, field studies are crucial. In this study, we investigated short- as well as long-term plant nitrogen uptake by Deschampsia flexuosa, Picea abies and Vaccinium myrtillus from ¹⁵NO₃^–, ¹⁵NH₄⁺ and (U-¹³C, ¹⁵N) arginine, glycine or peptides. Root N uptake was analysed after 6 h and 64 days following injections. Our results show that all three species, irrespective of their type of associated mycorrhiza (arbuscular, ecto- or ericoid, respectively) rapidly acquired similar amounts of N from the entire range of added N sources. After 64 days, P. abies and V. myrtillus had acquired similar amounts of N from all N sources, while for D. flexuosa, the uptake from all N sources except ammonium was significantly lower than that from nitrate. Furthermore, soil analyses indicate that glycine was rapidly decarboxylated after injections, while other organic compounds exhibited slower turnover. In all, these results suggest that a wide range of N compounds may be of importance for the N nutrition of these boreal forest plants, and that the type of mycorrhiza may be of great importance for N scavenging, but less important to the N uptake capacity of plants.     

Diversity and indicator species as measures of water pollution in a subarctic lake

Benthic invertebrates were collected from a subarctic lake during 1976 to assess the effectiveness of diversity indices and indicator species as measures of heavy metal pollution. Collections were made near an operating metal mine, where sediments were contaminated with high levels of arsenic (up to 2,500 mg/kg dry weight), mercury (500 µg/kg), lead (850 mg/kg), copper (750 mg/kg) and zinc (950 mg/kg). A total of 25 species and a diversity index of 2.4—2.9 were recorded in this heavily impacted area. Chironomids (Procladius denticulatus, Heterotrissocladius changi, Chironomus decorus) were most common in the sediments, followed in importance by molluscs (Pisidium casertanum) and oligochaetes (Lumbriculus variegatus). There were 23 and 25 species in the areas of moderate and negligible contamination, respectively. The diversity indices ranged from 2.4–2.6 and 2.4–2.8 and the main species were generally similar to those found in the heavily impacted area. While diversity indices and indicator species were therefore ineffective in monitoring metal contamination, the strong negative correlation between the concentration of metals and the abundance of benthic organisms provided a much more realistic assessment of the level of contamination.     

Effects of Copper on the Burrowing Behavior of Estuarine and Coastal Invertebrates,the Polychaete Nereis diversicolor and the Bivalve Scrobicularia plana

Alterations of the burrowing behavior of two benthic invertebrates living in intertidal mudflats, the polychaete Nereis diversicolor and the bivalve Scrobicularia plana, were studied in individuals exposed to soluble copper. The design of the contamination procedure took into account the results assessing the influence of some natural factors potentially able to influence burrowing (artificial vs. natural sediment, lighting, size of the shell for bivalves). Animals were exposed for 4 d to concentrations ranging from 25 to 150 μ g Cu l^{? 1}. At the end of exposure, the burrowing kinetics in clean sediment were determined after 1 and 2 d, then the animals were frozen until acetylcholinesterase (AChE) activity determination. Even at the lowest tested concentrations, copper caused hypoactivity in organisms belonging to both species studied. Behavioral impairments were not related to AChE activity inhibition. Metabolical or physiological disturbances could be the cause of these impairments. Concentrations affecting burrowing behavior were below those responsible for lethality in these species.     

Sensitivity of thermophilic methanogenic bacteria to heavy metals

The effects of Cd, Cu, and Ni on pure cultures of thermophilic methanogenic bacteria were studied. The bacteria used wereMethanobacterium thermoautotrophicum and TAM, a thermophilic, acetate-decarboxylating, methanogenic bacterium. Much lower concentrations of heavy metals were needed to cause initial inhibition of TAM (1 mg/liter Cu and Cd; 5 mg/liter Ni) compared withM. thermoautotrophicum (10 mg/liter Cu and Cd; and 100 mg/liter Ni). No growth of TAM occurred at 5 mg/liter Cu and 25 mg/liter Ni, while the corresponding values forM. thermoautotrophicum were 50 mg/liter Cu and 200 mg/liter Ni. Cd (50 mg/liter) was totally inhibitory toM. thermoautotrophicum but allowed minimal growth of TAM. Ni stimulated both organisms at an optimal concentration of 5 mg/liter forM. thermoautotrophicum and 1 mg/liter for TAM. The toxicity of Cd and Cu was found to depend upon the presence of Ni in the medium.     

Remedial habitat creation: does Nereis diversicolor play a confounding role in the colonisation and establishment of the pioneering saltmarsh plant, Spartina anglica?

Increasing concerns over global warming and expected sea level rises have led to the adoption of new coastal management strategies around the south-east coast of England. This paper explores the role played by the estuarine invertebrate Nereis diversicolor in limiting the colonisation and establishment of the invasive pioneering salt marsh plant, Spartina anglica. The biology of N. diversicolor is briefly reviewed and data from field experiments are presented demonstrating significant negative effects of worm abundance on transplanted S. anglica biomass. Laboratory-based experiments demonstrated significant negative effects of N. diversicolor abundance on the survival of S. anglica seeds transplanted to sediment cores. The importance of estuarine invertebrates in engineering the mudflat habitat may confound the foreseen ecosystem services and function provided by saltmarsh management schemes. Received: 15 February 1999 / Received in revised form: 4 June 1999 / Accepted: 11 June 1999     

Alien fungal species distribution: the study case of Favolaschia calocera

Invasive organisms (especially plants and animal species) are considered to be one of the main causes of global biodiversity loss. Up to now, few papers have dealt with the spreading of fungi. The establishment of the geographic origin of alien organisms could be useful to assess their impact on the environment. Favolaschia calocera is a basidiomycete species which was first described from Madagascar, and successively observed in New Zealand in 1969, where it has currently been recorded in more than 200 stands. It has recently also been reported in Australia, Thailand, China, Kenya, and Reunion Island. F. calocera was found in Genoa, Italy, in 1999: this recording represented the first in Europe. Till now, Favolaschia specimens have been collected in six areas around Genoa. F. calocera was observed growing on debris of various vascular plant species (Pteridophytes, Conifers, Mono- and Dicotyledons), thus showing to be a polyphagous species. Because it is spreading, it needs to be monitored. The main goal of our research is to investigate, through molecular phylogeographic analysis, the origin of the Italian strains. The sequencing of the ribosomal DNA ITS region of the Italian specimens followed by Neighbour-joining analysis showed that they cluster with the specimens from New Zealand, Kenya, Norfolk Island and Réunion Island. Hypotheses on the origin and introduction way as well as on its mechanisms of spreading are provided.     

Arnoglossus macrolophus Alcock (Pleuronectiformes: Bothidae); a valid species distinct fromA. tapeinosomus (Bleeker)

The holotype of a bothid flounder,Arnoglossus tapeinosomus (Bleeker, 1866), was re-examined and found to bear none of the diagnostic characters ascribed by many authors to the species. In addition, the shape of the prevomer was clearly different between the holotype and 18 specimens supposedly“A. tapeinosomus.” A. macrolophus Alcock, 1889, which was synonymized underA. tapeinosomus by Weber and de Beaufort (1929), is considered as a valid replacement name for“A. tapeinosomus,” because of the elongated anterior rays in the dorsal fin and a large, dark spot on the posterior dorsal and anal fin bases.A. tapeinosomus is redescribed from the holotype.     

Biosorption of americium using biomasses of various Rhizopus species

Biomasses from eight different Rhizopus species were tested for the sorption of americium from nitric acid medium. Rhizopus arrhizus NCIM 997 showed maximum sorption at pH 2. Laboratory scale experiments were carried out using this biomass in packed columns for the sorption of -activity from an americium spiked low level waste stream of PUREX process. The biomass was found to be an excellent sorbent for remediation of low level waste streams on once through basis.     

Iron uptake by the ichthyotoxic Chattonella marina (Raphidophyceae): impact of superoxide generation1

Significant production of superoxide, a known reductant of both inorganic and organically complexed iron(III), occurs in natural systems by both biotic and abiotic pathways. We have investigated the generation of superoxide by Chattonella marina (Subrahman.) Y. Hara et Chihara, a phytoplankton taxon known to produce high levels of this reactive oxygen species, and examined the role of superoxide in the acquisition of iron by this organism. Additionally, a generalized model for iron acquisition by C. marina has been developed, which includes three pathways of iron acquisition from organically complexed iron(III): nondissociative reductive uptake, dissociative reductive uptake, and nonreductive dissociative uptake. The model is shown to be particularly useful in ascertaining the relative importance of these various iron‐uptake pathways as a function of solution parameters including concentration and iron‐binding strength of the organic ligand and superoxide concentration. Our results suggest that superoxide can participate in the C. marina iron‐uptake process when iron is complexed to weak ligands, such as citrate, but plays only a minor role when iron is bound to a strong ligand. It thus appears that facilitation of iron acquisition is not the sole purpose of superoxide production by these organisms.