Flow cytometric assessment of the effects of chlorine,chloramine, and UV on bacteria by using nucleic acid stains and 5-cyano-2,3-ditolyltetrazolium chloride

Flow cytometry (FCM) has been widely used in multi-parametric assessment of cells in various research fields, especially in environmental sciences. This study detected the metabolic activity of Escherichia coli and Staphylococcus aureus by using an FCM method based on 5-cyano-2,3-ditolyltetrazolium chloride (CTC); the accuracy of this method was enhanced by adding SYTO 9 and 10%R2A broth. The disinfection effects of chlorine, chloramine, and UV were subsequently evaluated by FCM methods. Chlorine demonstrated stronger and faster destructive effects on cytomembrane than chloramine, and nucleic acids decomposed afterwards. The metabolic activity of the bacteria persisted after the cytomembranewas damaged as detected using CTC. Low-pressure (LP) UV or medium-pressure (MP) UV treatments exerted no significant effects on membrane permeability. The metabolic activity of the bacteria decreased with increasing UV dosage, and MP-UV was a stronger inhibitor of metabolic activity than LP-UV. Furthermore, the membrane of Gram-positive S. aureus was more resistant to chlorine/chloramine than that of Gram-negative E. coli. In addition, S. aureus showed higher resistance to UV irradiation than E. coli.

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Environmental modeling of uranium interstitial compositions of non-stoichiometric oxides: experimental and theoretical analysis

Study of uranium interstitial compositions of non-stoichiometric oxides UO_2+x (x ∈ 0.1–0.02) in gas and condense phases has been presented, using various soft-ionization mass spectrometric methods such as ESI-, APCI-, and MALDI-MS at a wide dynamic temperature gradient (∈ 25–300 °C). Linearly polarized vibrational spectroscopy has been utilized in order to assign unambiguously, the vibrational frequencies of uranium non-stoichiometric oxides. Experimental design has involved xUO_2.66·yUO_2.33, xUO_2.66·yUO_2.33/SiO₂, xUO_2.66·yUO_2.33/SiO₂ (NaOH) and SiO₂/x′NaOH·y′UO₂(NO₃)₂·6H₂O, multicomponent systems (x = 1, y ∈ 0.1–1.0 and x′ = 1, y′ ∈ 0.1–0.6) as well as phase transitions UO₂(NO₃)₂·6H₂O → {U₄O₉(UO_2.25)} → U₃O₇(UO_2.33) → U₃O₈(UO_2.66) → {UO₃}, thus ensuring a maximal representativeness to real environmental conditions, where diverse chemical, geochemical and biochemical reactions, including complexation and sorption onto minerals have occurred. Experimental factors such as UV-irradiation, pH, temperature, concentration levels, solvent types and ion strength have been taken into consideration, too. As far as uranium speciation represents a challenging analytical task in terms of chemical identification diverse coordination species, mechanistic aspects relating incorporation of oxygen into UO _2+x form the shown full methods validation significantly impacts the field of environmental radioanalytical chemistry. UO₂ is the most commonly used fuel in nuclear reactors around the globe; however, a large non-stoichiometric range ∈ UO_1.65–UO_2.25 has occurred due to radiolysis of water on UO₂ surface yielding to H₂O₂, OH^·, and more. Each of those compositions has different oxygen diffusion. And in this respect enormous effort has been concentrated to study the potential impact of hazardous radionuclide on the environment, encompassing from the reprocessing to the disposal stages of the fuel waste, including the waste itself, the processes in the waste containers, the clay around the containers, and geological processes. In a broader sense, thereby, this study contributes to field of environmental analysis highlighting the great ability of various soft-ionization MS methods, particularly, MALDI-MS one, for direct assay of complex multicomponent heterogeneous mixtures at fmol–attomol concentration ranges, along with it the great instrumental features allowing, not only meaningful quantitative, but also structural information of the analytes, thus making the method indispensable for environmental speciation of radionuclides, generally.     

Accelerated degradation of orange G over a wide pH range in the presence of FeVO<Subscript>4</Subscript>

In this study, FeVO₄ was prepared and used as Fenton-like catalyst to degrade orange G (OG) dye. The removal of OG in an aqueous solution containing 0.5 g·L^–1 FeVO₄ and 15 mmol·L^–1 hydrogen peroxide at pH 7.0 reached 93.2%. Similar rates were achieved at pH 5.7 (k = 0.0471 min^–1), pH 7.0 (k = 0.0438 min^–1), and pH 7.7 (k = 0.0434 min^–1). The FeVO₄ catalyst successfully overcomes the problem faced in the heterogeneous Fenton process, i.e., the narrow working pH range. The data for the removal of OG in FeVO₄ systems containing H₂O₂ conform to the Langmuir–Hinshelwood model (R² = 0.9988), indicating that adsorption and surface reaction are the two basic mechanisms for OG removal in the FeVO₄–H₂O₂ system. Furthermore, the irradiation of FeVO₄ by visible light significantly increases the degradation rate of OG, which is attributed to the enhanced rates of the iron cycles and vanadium cycles.

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Zur Beständigkeit von Aluminiumchloridlösungen mit tridekameren Kationen (Al<Subscript><Emphasis Type="Bold">13</Emphasis></Subscript>)

Background, aim, and scopeThe focus of popular interest on the subject of acid rain has been on the effects, which it has on aquatic ecosystems. Aluminium toxicity and the bioavailability depend on the species of aluminium present. The cationic aluminium species are important for the phytotoxocity and for the precipitation of aluminium containing precipitate.Materials and methodsSpeciation diagrams of aqueous aluminium chloride solutions with tridecameric cations (Al₁₃) at room temperature as function of concentration (1·10^–1–1·10^–5 mol·l^–1 Al) and aging time (9 month) have been obtained from a kinetic method; these Ferron method offers a simple and inexpensive alternative for identification and quantification of aluminium cations.ResultsThe Al₁₃-cation stability decrease with decreasing concentration and increasing aging time.DiscussionPolynuclear aluminium species can be formed in nature by the dissolution of Al-containing minerals, as a consequence of surface water acidification, followed by neutralization process. The question of the occurrence of the tridecameric polycation (Al₁₃) in natural soil solutions and stream waters is of considerable interest.ConclusionsThe results are important on the occurrence of the Al₁₃ in natural waters, for the precipitation of aluminium species by acidification and for the aluminium phytotoxicity.Recommendations and perspectivesThis research suggests strongly that Al₁₃ should be negligible in natural soil and surface waters. The toxicity of Al₁₃ to plants and aquatic organism in natural conditions may be considered to be very low.     

Diurnal changes in photosynthesis of Antarctic fast ice algal communities determined by pulse amplitude modulation fluorometry

The aim of this project was to determine both the diurnal changes in photosynthetic activity of Antarctic sea ice algae and also the protective mechanisms they use to mitigate the effects of in situ UV radiation. Changes in the diurnal photosynthetic parameters of fast ice algal communities at McMurdo Sound were measured in situ, using a custom designed monitoring pulse amplitude modulation fluorometer. The sea ice microalgae were able to adapt rapidly to either increasing or decreasing ambient irradiances. ΔF/F_m' values were between 0.2 and 0.51, while E_k varied between 2.1 and 18 μmol photons m^-2 s^-1. ΔF/F_m', E_k, and relative electron transfer rate (rETR) all varied sequentially over the course of a day. rETR and E_k were highest at midday at the highest irradiances, when there was apparent midday down regulation of photosynthesis, while ΔF/F_m' was highest at midnight. The effects of natural UV radiation on sea ice were examined, but it was not possible to detect the effect of either UVB or UVA and UVB on photosynthesis. This was considered to be largely because of the large spatial and temporal heterogeneity of the under ice community, changing irradiances throughout the day and the relatively small change caused by UV.     

Selection and characterization of eight freshwater green algae strains for synchronous water purification and lipid production

The objective of this study is to select and characterize the candidate for synchronous water purification and lipid production from eight freshwater microalgae strains (Chlorella sp. HQ, C. emersonii, C. pyrenoidosa, C. vulgaris, Scenedesmus dimorphus, S. quadricauda, S. obiquus, Scenedesmus sp. LX1). The strains Chlorella sp. HQ, C. pyrenoidesa, and S. obliquus showed superiority in biomass accumulation, while the top biomass producers did not correspond to the top lipid producers. S. quadricauda achieved higher lipid content (66.1%), and Chlorella sp. HQ and S. dimorphus ranked down in sequence, with lipid content above 30%. Considering nutrient removal ability (total nitrogen (TN): 52.97%; total phosphorus (TP): 84.81%), the newly isolated microalga Chlorella sp. HQ was the possible candidate for water purification coupled with lipid production. To further investigate the lipid producing and nutrient removal mechanism of candidate microalga, the ultra structural changes especially the lipid droplets under different water qualities (different TN and TP concentrations) were characterized. The results elucidate the nutrient-deficiency (TN: 3.0 mg·L^–1; TP: 0.3 mg·L^–1) condition was in favor of forming lipid bodies in Chlorella sp. HQ at the subcellular level, while the biomass production was inhibited due to the decrease in chloroplast number which could further suppress the nutrient removal effect. Finally, a twophase cultivation process (a nutrient replete phase to produce biomass followed by a nutrient deplete phase to enhance lipid content) was conducted in a photo-bioreactor for Chlorella sp. HQ to serve for algae-based synchronous biodiesel production and wastewater purification.     

Patterns of gamete incompatibility between the blue mussels<Emphasis Type="Italic"> Mytilus edulis</Emphasis> and<Emphasis Type="Italic"> M</Emphasis>.<Emphasis Type="Italic"> trossulus</Emphasis>

Previous research on gametic incompatibility in marine invertebrates suggests that for highly dispersive marine invertebrate species, barriers to fertilization among closely related taxa are often incomplete and sometimes asymmetric. The nature of these barriers can dramatically affect the patterns of gene flow and genetic differentiation between species, and thus speciation. Blue mussels, in the genus Mytilus, are genetically distinct in allopatry yet hybrids are present wherever any two species within the group co-occur. The present study sampled M. edulis (L.) and M. trossulus (Gould) in May and June 2001 from the East Bay section of Cobscook Bay, Maine, USA (latitude 44°56′30″N; longitude 67°07′50″W), where the two species are sympatric. Gamete incompatibility was investigated in a series of laboratory fertilizations carried out in July 2001. The proportion of fertilized eggs typically exceeded 80% at sperm concentrations of 10³–10⁴ ml^?1 among intraspecific matings (n=18), but was <30% even at sperm concentrations in excess of 10⁵–10⁶ ml^?1 for interspecific matings (n=13). Further analysis indicated that approximately 100- to 700-fold higher sperm concentrations were required to achieve 20% fertilization in interspecific matings relative to intraspecific matings, indicating strong barriers to interspecific fertilization. The proportion of fertilized eggs did not follow this general pattern in all matings, however. The eggs from two (out of five) M. edulis females were almost as receptive to M. trossulus sperm as they were to M. edulis sperm. In contrast, the eggs from all M. trossulus females (n=3) were unreceptive to M. edulis sperm, suggesting that fertilization barriers between these species may be asymmetric. Given the experimental design employed in this study, the results are also consistent with a strong maternal or egg effect on the level of interspecific gamete compatibility in M. edulis.     

Physiological adaptations of the invasive cordgrass<Emphasis Type="Italic"> Spartina anglica</Emphasis> to reducing sediments: rhizome metabolic gas fluxes and enhanced O<Subscript>2</Subscript> and H<Subscript>2</Subscript>S transport

Cordgrasses of the genus Spartina form dense monospecific stands worldwide, profoundly influencing the ecology of estuaries. One species, Spartina anglica, originated by allopolyploidy in the 1800s and has been particularly prolific as an invasive species worldwide. S. anglica tolerates low-lying estuarine mudflats that its progenitor species and other coastal halophytes cannot. However, very little is known of the physiology of S. anglica. In the present study, an automated flow-through respirometry system was used to quantify metabolic gas fluxes (O₂, H₂S, CO₂, and NH₃) of S. anglica rhizomes. Enhanced physiological mechanisms to transport O₂ and H₂S in both directions between the rhizosphere and the atmosphere were exhibited by S. anglica, but not by the native North American species S. alterniflora. These results suggest that tolerance of anoxia and H₂S may assist S. anglica in colonizing extensively flooded environments. Enhanced sediment oxygenation by S. anglica may be potentially useful for phytoremediation of contaminated sediments, since microbial degradation of organic pollutants is often limited by O₂ availability.     

Microalgae cultivation and culture medium recycling by a two-stage cultivation system

Nutrients and water play an important role in microalgae cultivation. Using wastewater as a culture medium is a promising alternative to recycle nutrients and water, and for further developing microalgae-based products. In the present study, two species of microalgae, Chlorella sp. (high ammonia nitrogen tolerance) and Spirulina platensis (S. platensis, high growth rate), were cultured by using poultry wastewater through a two-stage cultivation system for algal biomass production. Ultrafiltration (UF) or centrifuge was used to harvest Chlorella sp. from the first cultivation stage and to recycle culture medium for S. platensis growth in the second cultivation stage. Results showed the two-stage cultivation system produced high microalgae biomass including 0.39 g·L^–1Chlorella sp. and 3.45 g·L^–1S. platensis in the first-stage and second-stage, respectively. In addition, the removal efficiencies of NH₄⁺ reached 19% and almost 100% in the first and the second stage, respectively. Total phosphorus (TP) removal reached 17% and 83%, and total organic carbon (TOC) removal reached 55% and 72% in the first and the second stage, respectively. UF and centrifuge can recycle 96.8% and 100% water, respectively. This study provides a new method for the combined of pure microalgae cultivation and wastewater treatment with culture medium recycling.

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Efficient photoelectrochemical oxidation of rhodamine B on metal electrodes without photocatalyst or supporting electrolyte

We designed photoelectrochemical cells to achieve efficient oxidation of rhodamine B (RhB) without the need for photocatalyst or supporting electrolyte. RhB, the metal anode/cathode, and O₂ formed an energy-relay structure, enabling the efficient formation of O ₂ ^– species under ultraviolet illumination. In a single-compartment cell (S cell) containing a titanium (Ti) anode, Ti cathode, and 10 mg·mL^–1 RhB in water, the zero-order rate constant of the photoelectrochemical oxidation (k_PEC) of RhB was 0.049 mg·L^–1·min^–1, while those of the photochemical and electrochemical oxidations of RhB were nearly zero. k_PEC remained almost the same when 0.5 mol·L^–1 Na₂SO₄ was included in the reactive solution, regardless of the increase in the photocurrent of the S cell. The k_PEC of the illuminated anode compartment in the two-compartment cell, including a Ti anode, Ti cathode, and 10 mg·mL^–1 RhB in water, was higher than that of the S cell. These results support a simple, eco-friendly, and energysaving method to realize the efficient degradation of RhB.

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Genetic structure of<Emphasis Type="Italic"> Linckia laevigata</Emphasis> and<Emphasis Type="Italic"> Tridacna crocea</Emphasis> populations in the Palawan shelf and shoal reefs

Allozyme variation of 10 populations of Linckia laevigata at 8 polymorphic loci and 13 populations of Tridacna crocea at 6 polymorphic loci were analyzed to compare genetic variability and genetic affinities among reefs in Palawan, Philippines. Two to five populations were sampled from each of four regions: the shelf reefs in (1) northern Palawan and (2) southern Palawan and the offshore reefs in (3) the Kalayaan island group (KIG) in the South China Sea and (4) the Tubbataha shoals in the Sulu Sea. Heterozygosity was highest in populations of L. laevigata from the south shelf of Palawan and populations of T. crocea from the Tubbataha shoals of the Sulu Sea. The lowest heterozygosity estimates were from the reefs of the KIG in the South China Sea, for both species. Overall F _ST values for both species were significant, with an estimated average number of effective migrants per generation (N _EM) of 4.85 (~5 individuals) for L. laevigata and 3.54 (~4 individuals) for T. crocea. Within-region comparisons showed N_EM ranging from 6.29 to 92.34 for L. laevigata and from 3.40 to 6.30 for T. crocea. The higher gene flow among L. laevigata populations relative to T. crocea is consistent with the greater dispersal potential of the former species. Finer scale genetic structuring was evident in T. crocea populations. For both species, the Tubbataha reefs in the Sulu Sea have higher genetic affinity with the populations from the southern shelf of Palawan, while the reefs in the KIG had higher affinity with the northern Palawan shelf reefs. The north and south shelf populations have the least genetic affinity. Genetic patchiness among reefs within regions suggests the importance of small-scale physical factors that affect recruitment success in structuring populations in small island and shoal reef systems in Palawan.     

Growth and production of<Emphasis Type="Italic"> Ophiocten gracilis</Emphasis> (Ophiuroidea: Echinodermata) on the Scottish continental slope

Growth and production of the bathyal ophiurid brittle star Ophiocten gracilis was studied from skeletal growth bands and disc size frequencies of specimens collected in sled and trawl samples taken on the continental slope off Scotland. Growth bands showed up in SEM examination as ring-like zones in surface relief and texture of the stereom microstructure of the intervertebral muscle insertions on the arm ossicles. Seasonal variability in somatic growth, presumed to underlie this growth pattern, may reflect reproduction and/or a possible non-feeding period during gonad maturation. Disc size-at-age was back-calculated from size-at-age interpreted from growth-band series on the vertebral ossicles from arms of O. gracilis. Pooled growth-band frequency data and normal-distribution mixtures based on size-at-age data were used to test for overgrowth of early growth bands on the ossicles from larger individuals. Von Bertalanffy and Gompertz growth models were fitted to the finalised back-calculated disc size-at-age data. These were used along with the modal structure of the observed disc size frequencies to develop a demographic model based on normal-distribution mixtures constrained by the growth model. These and other defining parameters were fitted by non-linear regression to size structure observed in a sample of the breeding population from 997 m depth on the Hebrides Terrace. Recruitment was estimated according to available data from sediment-trap time series. A ratio of somatic production/biomass, P_S/B, in the range of 0.43–0.54 was estimated using a fitted size/mass relationship and the increment summation method (ISM) applied to the fitted growth models. A narrower, but otherwise similar, range in estimated P_S/B ratios (0.48–0.49) was obtained in a parallel approach using the mass-specific growth rate method (MSGRM), whereby the same size/mass relationship was applied to the observed frequencies and growth parameters fitted to growth banding. Using previously obtained data on population density, a standing crop of 4.8 g wet weight (~0.58 mg AFDW) m^?2 would provide annual wet weight production in the range of 1.9–2.4 g (~0.23–0.29 mg AFDW) m^?2 in the population between ca. 700–1000 m depth. Somewhat greater production estimates (P_S/B=0.73–0.98) were obtained from MSGRM by pooling the sample with size frequencies from other large samples in which postlarval sizes were more numerous, but larger sizes less numerous. Similarly high production was estimated by MSGRM from a box corer sample from the Wyville-Thomson Ridge. Explanations for variability in size structure are discussed, but even the lower estimates are comparable to boreal shallow-water brittle stars. The high rate of growth and production by accepted deep-sea standards may be related to a capability for interface feeding.     

New beach in a shallow estuarine lagoon: a model-based <Emphasis Type="Italic">E. coli</Emphasis> pollution risk assessment

A 3D hydrodynamic model has been applied to the Curonian Lagoon to study the pollution impact of E. coli on a new beach that might be opened in the lagoon. Through a field survey the E. coli inputs were measured and then used in the numerical model, and through laboratory experiments the decay rate of E. coli was established. The model has been calibrated and validated for the year 2015, and several scenarios have been studied, such as sewage system breakdown, severe weather conditions or high river loads. The model has then been run for a period of 12 years to obtain a robust statistics for the pollution on the planned beach. Results show that the decay rate of E. coli is between 0.55 days and 2.3 days and the modeled decay times are compatible with these numbers. The only scenario that would create a risk for the bathing waters of the beach is a breakdown of the sewage system on the Curonian Spit. In this case the hours (and days) over legally allowable bathing threshold were computed in order to estimate the number of days the beach could be closed. These results have been confirmed by the 12 year simulations. With an influence map analysis the two most critical sewage systems could be identified.     

Enhanced disinfection of <Emphasis Type="Italic">Escherichia coli</Emphasis> and bacteriophage MS2 in water using a copper and silver loaded titanium dioxide nanowire membrane

Titanium dioxide (TiO₂) is a widely used photocatalyst that has been demonstrated for microorganism disinfection in drinking water. In this study, a new material with a novel structure, silver and copper loaded TiO₂ nanowire membrane (Cu-Ag-TiO₂) was prepared and evaluated for its efficiency to inactivate E. coli and bacteriophage MS2. Enhanced photo-activated bactericidal and virucidal activities were obtained by the Cu-Ag-TiO₂ membrane than by the TiO₂, Ag-TiO₂ and Cu-TiO₂ membranes under both dark and UV light illumination. The better performance was attributed to the synergies of enhanced membrane photoactivity by loading silver and copper on the membrane and the synergistic effect between the free silver and copper ions in water. At the end of a 30 min test of deadend filtration under 254 nm UV irradiation, the Cu-Ag-TiO₂ membrane was able to obtain an E. coli removal of 7.68 log and bacteriophage MS2 removal of 4.02 log, which have met the US EPA standard. The free metal ions coming off the membrane have concentrations of less than 10 ppb in the water effluent, far below the US EPA maximum contaminant level for silver and copper ions in drinking water. Therefore, the photo-activated disinfection by the Cu-Ag-TiO₂ membrane is a viable technique for meeting drinking water treatment standards of microbiological water purifiers.

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Titanium dioxide nanoparticles induce bacterial membrane rupture by reactive oxygen species generation

Nano-titania is widely used in the food industry due to its efficient antimicrobial activity. However, the mechanism of microbial toxicity of nano-titania is poorly known. Here, nano-TiO₂ has been fabricated by microwave-irradiation chemistry, a new method, and then tested for antimicrobial activity. Mutagenicity of nano-TiO₂ was evaluated using Salmonella typhimurium histidine-auxotrophic strains. The reactive oxygen generation test was performed using 2,7-dichlorofluorescein diacetate dye. To test membrane permeabilization, E. coli cultures were grown in nutrient broth at an optical density of 0.3–0.5 at 610 nm, harvested by centrifugation at 11,000g for 10 min, washed and resuspended in 0.5 % NaCl solution. We also analyzed superoxide formation and membrane integrity, and we used scanning electron microscopy. Results show that nano-TiO₂ has a minimum inhibitory concentration of 15 µg/mL, and a minimum bactericidal concentration of 20 µg/mL for E. coli. The bacterial inner wall was ruptured, and cytoplasmic content was released after 5 min of treatment in a dose-dependent manner. Notably, superoxide formation was not observed, which establishes the fact that reactive oxygen generation and alteration of membrane integrity, as well as permeability, is the major mechanism of antimicrobial activity of nano-TiO₂.     

Ecology of a corallivorous gastropod,<Emphasis Type="Italic"> Coralliophila abbreviata</Emphasis>, on two scleractinian hosts. I: Population structure of snails and corals

Despite their potential importance in structuring reef communities, invertebrate corallivores and their population structures are poorly understood. We found distinct differences in the population structures (length-frequency distribution and sex ratio) of the corallivorous gastropod Coralliophila abbreviata residing on two coral-host taxa, Montastraea spp. and Acropora palmata, in the Florida Keys. In each of two survey years, around 50% of the Montastraea spp. colonies were infested, with a mean snail density of eight snails per infested colony (range 1–45), while around 20% of A. palmata colonies harbored three snails per infested colony (range 1–23). Variation in patterns of snail occurrence was also observed within a host taxon. A. palmata occurred in low- and high-density stands (0.4 and 1 colony m^–2, respectively, at the initial survey) at different sites. Hurricane Georges struck the area in September 1998. When resurveyed in 1999, density of colonies in low-density stands had decreased by 75% to 0.1 colonies m^–2. This decrease was accompanied by a doubling in the proportion of colonies infested with snails (from 19% to 46%) and an increase in snail density per infested colony (from 3.7±3.3 SD to 5.4±4.6 SD) as snails apparently concentrated on surviving A. palmata. In contrast, sites with high density A. palmata stands (thickets) retained colony densities of about ~1 colony m^–2 among years, while snail infestation increased only from 9% to 14% of colonies surveyed and snail density essentially remained unchanged (from 2.7±1.8 to 2.9±1.9 snails per infested colony). Snails collected from Montastraea spp. were shorter than those from A. palmata in low-density stands and were longest on A. palmata in thickets. On both host taxa, female snails were longer than males. The sex ratio of snails on Montastraea spp. hosts was even (1:1), while that of snails on A. palmata was skewed (70% males). Factors that could explain observed differences in size structure and sex ratio between Coralliophila populations on the two coral host taxa include: differential susceptibility to predators, influence of host tissue nutritional quality and/or secondary metabolite content, and genetic differences (cryptic species). The host-specific characteristics of C. abbreviata populations imply that the impact of gastropods on reef communities will vary with the coral species composition.     

Buffer sensitivity of photosynthetic carbon utilisation in eight tropical seagrasses

Some of the mechanisms involved in inorganic carbon (Ci) acquisition by tropical seagrasses from the western Indian Ocean were described by Björk et al. (Mar Biol 129:363–366, 1997). However, since then, it has been found that an additional, buffer-sensitive, system of Ci utilisation may operate in some temperate seagrasses (Hellblom et al. in Aquat Bot 69:55–62, 2001, Hellblom and Axelsson in Photos Res 77:173–191, 2003); this buffer sensitivity indicates a mechanism in which electrogenic H⁺ extrusion may form acidic diffusion boundary layers, in which either HCO ₃ ^? –H⁺ is co-transported into the cells, or where HCO ₃ ^? is converted to CO₂ (as catalysed by carbonic anhydrase) prior to uptake of the latter Ci form. Because a buffer was used in the 1997 study, we found it important to reinvestigate those same eight species, taking into account the direct effect of buffers on this potential mode of Ci acquisition in these plants. In doing so, it was found that all seagrass species investigated except Cymodocea serrulata were sensitive to 50 mM TRIS buffer of the same pH as the natural seawater in which they grew (pH 8.0). Especially sensitive were Halophila ovalis, Halodule wrightii and Cymodocea rotundata, which grow high up in the intertidal zone (only ca. 50–65% of the net photosynthetic activity remained after the buffer additions), followed by the submerged Enhalus acoroides and Syringodium isoetifolium (ca. 75% activity remaining), while Thalassia hemprichii and Thalassodendron ciliatum, which grow in-between the two zones, were less sensitive to buffer additions (ca. 80–85% activity remaining). In addition to buffer sensitivity, all species were also sensitive to acetazolamide (AZ, an inhibitor of extracellular carbonic anhydrase activity) such that ca. 45–80% (but 90% for H. ovalis) of the net photosynthetic activity remained after adding this inhibitor. Raising the pH to 8.8 (in the presence of AZ) drastically reduced net photosynthetic rates (0–14% remaining in all species); it is assumed that this reduction in rates was due to the decreased CO₂ concentration at the higher pH. These results indicate that part of the 1997 results for the same species were due to a buffer effect on net photosynthesis. Based on the present results, it is concluded that (1) photosynthetic Ci acquisition in six of the eight investigated species is based on carbonic anhydrase catalysed HCO ₃ ^? to CO₂ conversions within an acidified diffusion boundary layer, (2) C. serrulata appears to support its photosynthesis by extracellular carbonic anhydrase catalysed CO₂ formation from HCO ₃ ^? without the need for acidic zones, (3) H. ovalis features a system in which H⁺ extrusion may be followed by HCO ₃ ^? –H⁺ co-transport into the cells, and (4) direct, non-H⁺-mediated, uptake of HCO ₃ ^? is improbable for any of the species.     

Phytotoxic effects of phenolic acids from <Emphasis Type="Italic">Merostachys riedeliana,</Emphasis> a native and overabundant Brazilian bamboo

Merostachys riedeliana Rupr. is a native and overabundant bamboo species in the Brazilian Atlantic Forest. Moderate to strong allelopathic activity may be one mechanism that explains this super-dominance and the changes in structure and composition of forest areas occupied by bamboo. This study evaluated the phytotoxic effect of M. riedeliana extracts and fractions and identified their putative allelochemicals. We investigated the presence of allelochemicals in soil collected from stands occupied by M. riedeliana. Furthermore, we evaluated the putative effect of tree allelochemicals, individually and combined, on germination and growth. The aqueous extract of leaves and its ethyl acetate fraction presented the highest inhibitory effects on seed germination and seedling growth. The effect of the extracts and fractions on the target species was species-specific. Neither the individual nor the combined phenolic acids significantly inhibited seed germination; however, a pronounced growth inhibition was observed in M. bimucronata seedlings treated with vanillic acid and in E. verna and M. bimucronata seedlings treated with combined phenolic acids. Isovitexin, vitexin, isoorientin, orientin, and their O-glycoside derivatives, the lactonic dimer of the p-hydroxybenzoic acid and 3,4-methylenedioxymandelic acid were identified in the aqueous extracts and ethyl acetate fraction by Liquid Chromatography-Diode Array Dectector/Electrospray Ionization/Mass Spectrometry (LC-DAD/ESI–MS/MS). The Gas Chromatography-Mass Spectrometer (GC–MS) profile of the same extract and fraction showed the presence of benzoic, benzeneacetic, salicylic, p-hydroxybenzoic, p-hydroxyphenylacetic, vanillic, p-coumaric, protocatechuic, syringic, gallic, m-coumaric vanillylmandelic, 4-methylmandelic, 3,4-methylenedioxymandelic and trans-ferulic acids. The p-benzoic acid and the apigenin 6-C-glucoside (isovitexin) were identified in the soil extract collected from under bamboo-growing areas. Even though laboratory bioassays are not completely predictive of the allelopathic effects that occur in nature, the results of this study provide preliminary evidence of allelopathy as a possible species-specific inhibition mechanism of native species that explain the impoverishment of floristic richness and the functional groups in areas where M. riedeliana is overabundant.     

Phytotoxicity in seven higher plant species exposed to di-<Emphasis Type="Italic">n</Emphasis>-butyl phthalate or bis (2-ethylhexyl) phthalate

We investigated phytotoxicity in seven plant species exposed to a range of concentrations (0–500 mg·kg^?1 soil) of di-n-butyl phthalate (DnBP) or bis (2-ethylhexyl) phthalate (DEHP), two representative phthalate esters (PAEs) nominated by USEPA as priority pollutants and known environmental estrogens. We studied seed germination, root elongation, seedling growth, biomass (fresh weight, FW) and malondialdehyde (MDA) content of shoots and roots of wheat (Triticum aestivum L.), alfalfa (Medicago sativa L.), perennial ryegrass (Lolium perenne), radish (Raphanus sativus L.), cucumber (Cucumis sativus L.), oat (Avena sativa) and onion (Allium cepa L.), together with monitoring of plant pigment content (chlorophyll a, b and carotinoids) in alfalfa, radish and onion shoots. Root elongation, seedling growth and biomass of the test species were generally inhibited by DnBP but not by DEHP, indicating a lower level of phytotoxicity of DEHP than of DnBP. MDA contents of four species were promoted by PAE exposure, but not in alfalfa, ryegrass or onion shoots, indicating lower sensitivity of these three species to PAE pollutants. Plant pigment contents were clearly affected under the stress of both pollutants, implying the potential damage to the photosynthetic system of test plants, mainly by decreasing the content of chlorophyll a and b. Results of DnBP and DEHP phytotoxicity to the primary growth of test plants has provided information for the assessment of their environmental risk in the soil and also forms a basis for the further analysis of their toxic effects over the whole growth period of different plant species.     

Differences in the species- and size-composition of fish assemblages in three distinct seagrass habitats with differing plant and meadow structure

Fish faunas were sampled seasonally using a large and a small beam trawl in three seagrass habitats comprising predominantly Amphibolis griffithii or Posidonia sinuosa or Posidonia coriacea, which differ in seagrass and meadow structure. Amphibolis griffithii and P. sinuosa both produce a relatively dense leaf canopy, but the former exhibits a distinct architecture, with the leaf canopy overlying relatively open spaces surrounding woody stems, compared to the uniformly dense blade-like leaves of P. sinuosa which emerge directly from the sediment. In comparison, P. coriacea provides a landscape of patchy seagrass amongst areas of bare sand. Since the latter seagrass habitat contains large areas of sand, fish were also sampled in adjacent unvegetated areas. The number of species and density of fish were greater (P<0.05) in P. sinuosa than in either A. griffithii or P. coriacea. The mean number of species caught using the large trawl ranged from 16 to 24 in the first of these habitats compared to 14–21 and 9–15 in the last two habitats, respectively, and the mean densities ranged between 78 and 291 fish 1000 m^?2 in P. sinuosa compared to 31–59 fish 1000 m^?2 in A. griffithii and 31–59 fish 1000 m^?2 in P. coriacea. The biomass of fish was greater (P<0.05) in both P. sinuosa and A. griffithii than in P. coriacea (4.2–5.3 kg and 3.3–6.2 kg versus 0.7–1.9 kg 1000 m^?2, respectively). Furthermore, the size-structure of fish differed among these habitats, where the median weight of fish was 72.1 g in A. griffithii, compared to 7.5 g and 19.8 g in P. sinuosa and P. coriacea, respectively. Ordination and ANOSIM demonstrated that the species-composition differed markedly among the three seagrass habitats (P<0.001), suggesting that fish species display a distinct preference for particular seagrasses characterised by different architecture. Differences in species-composition among the seagrass habitats partly reflected the size-composition of fish in each habitat. The open space below the canopy of A. griffthii is likely to allow larger fish to occupy this habitat, whereas only small fish would be able to penetrate the dense foliage of P. sinuosa. Differences in species- and size-composition of fish among these habitats may be the result of settlement-sized larvae discriminating between particular seagrass and meadow structures, or fish being subject to different levels of predation and/or accessibility to food or space. The species-composition in P. coriacea was highly dispersed and did not differ from that of unvegetated areas. While several species were associated with both P. coriacea and bare-sand habitats, some species did display a high affinity with the seagrass P. coriacea. This may reflect an association with the sparse and narrower leaves of this seagrass or with the patchy occurrence of the seagrass Heterozostera tasmanica, which commonly occurs as an understorey in this habitat.