Control of microbial growth on eelgrass (Zostera marina L.: Spermatophyta) by leaf-derived metabolites

The role of minor components of the leaves ofZostera marina L. in altering the activity of micro-organisms directly and indirectly by affecting amphipod grazers was investigated in laboratory experiments. Water-soluble extracts of eelgrass leaves inhibited the growth of many marine bacteria and a micro-alga (Platymonas sp.) at concentrations equivalent to as little as 10 g dry wt. of leaf.ml^–1. The potency of leaf extracts was higher in young, actively metabolizing tissue than in older leaves, and was higher in leaves collected in summer during rapid growth than in winter. Water-soluble inhibitors may explain the low biomass of epiphytes on actively growing leaves. Phenolic acids, previously identified in eelgrass leaves, may account for part of the antimicrobial activity. Three free phenolic acids inhibited the test micro-organisms at concentrations as low as 2 M; caffeic acid was more potent than either protocatechuic or gentisic acid. Extracts of young leaves also inhibited grazing by amphipods on dead leaves. The time required for leaching of soluble inhibitors may account for the delay observed between the loss of leaves from the plants and the onset of grazing. Thus, interactions among the biotic components of the detritus ecosystem may be significantly modified by minor components of the macrophyte tissue.     

Chemotaxis of rhizoplane bacteria to amino acids comprising eelgrass (Zostera marina L.) root exudate

Scanning electron microscopy showed a large bacterial population on Zostera marina L. roots. The five dominant motile rhizoplane bacteria isolated were chemotactically attracted to Z. marina root exudate. Dansylation and thin-layer chromatography analysis of root exudate identified the presence of serine, threonine, glycine, alanine, and valine. When gentamicin sulfate was added during exudate collection, serine, and threonine were not detected. Exudate amino acids were shown to chemotactically attract motile rhizoplane isolates.     

Using light-permeable grating to mitigate impacts of residential floats on eelgrass Zostera marina L. in Puget Sound, Washington

This study evaluated whether light-permeable deck grating could mitigate impacts of residential mooring floats constructed over eelgrass (Zostera marina L.) in Puget Sound, Washington. Eelgrass shoot densities in undisturbed control areas and underneath and adjacent to 11 residential floats (16–50% of each float was grated) were monitored prior to float installation and annually for 3 years following installation. Using linear regression analysis, a decline in eelgrass shoot densities relative to controls was detected underneath three floats (eelgrass was eliminated under only one float) and adjacent to two floats. When control data were used to represent 100% grated, there was a weak relationship between eelgrass bed quality and percent of the deck grated (r = 0.46, p = 0.032), but no relationship when the range of grating was 16–50% (p = 0.90). The percent of a float deck grated did not contribute significantly to a multiple regression model examining change in eelgrass density that included five other dependent variables associated with the design of the floats. We conclude that either there was no effect of grating up to 50% of a float deck or we could not detect an effect. We hypothesize that the large number of site and landscape scale variables associated with a float influenced the effect (and our ability to detect it) of any one variable (such as grating). Consequently, we recommend that managers manipulate as many attributes of a float as possible (including grating) in order to reduce risks to eelgrass.     

Growth of Hydrilla verticillata (L.f.) Royle under controlled conditions

Aquatic macrophytes are important resources for the maintenance of trophic chains and in biogeochemical processes, but they can also be deleterious for several uses if present in excess. Hydrilla verticillata was found in the Paraná River (Brazil) after 2005, which requires monitoring owing to the invasive potential of this species. In this study, we measured the growth of H. verticillata under controlled conditions and compared the growth dynamics for the two development strategies (branch and tuber). We show that this species has great potential to develop in tropical (Brazilian) aquatic ecosystems. The parameters from the modelling of the growth kinetics indicated a doubling time of 19.8 days for H. verticillata growing from stems; however, the growth from tubers were much faster, with doubling times ranging from 2.5 to 11 days. The delay for the tubers to sprout caused a decrease in the number of branches of H. verticillata stems. From the growth parameters obtained from the experiments under controlled conditions, we concluded that the high temperature and light availability in most South American reservoirs (including the Porto Primavera Reservoir where it was first recorded) are suitable for H. verticillata to compete and probably displace other native aquatic macrophytes, such as Egeria najas, Egeria densa, and Cerathophyllum demersum. This is a matter of concern because these and other submersed species are commonly found in several natural and man-made South-American aquatic ecosystems, where they are key for biodiversity maintenance.     

Growth and biomass turnover ofHydrocharis dubia L. cultured under different nutrient conditions

Growth of a floating-leaved plant,Hydrocharis dubia L., was examined under varying nutrient conditions between 0.3 and 30 mgN l⁻¹ total inorganic nitrogen.H. dubia plants cultured under the most nutrient-rich condition showed the highest maximum ramet density (736 m⁻²), the highest maximum biomass (80.4 g dry weight m⁻²), and the highest total net production (185 g dry weight m⁻² in 82 days). Plants under nutrient-poor conditions had a relatively large proportion of root biomass and a small proportion of leaves with a long life span. Compared with other floating-leaved and terrestrial plants, the maximum biomass ofH. dubia was relatively small. This, and the rapid biomass turnover, was related to the short life span of leaves (13.2–18.7 days) and large biomass distribution to leaves.     

Oviposition in Culicoides impunctatus under laboratory conditions

We present a laboratory-based examination of oviposition preference in the Scottish biting midge, Culicoides impunctatus (Diptera: Ceratopogonidae). A variety of oviposition substrates were screened in no-choice and choice bioassays for efficacy in eliciting an egg-laying response. Both upper-layer photosynthetic Sphagnum spp. moss and Juncus articulatus infusions were identified as producing increased oviposition under no-choice conditions. During choice trials against a control of damp cottonwool, upper-layer Sphagnum spp. moss produced a significantly greater egg-laying response. These conclusions are interpreted in terms of possible cues involved in oviposition site selection and assessed for future use in colonisation of this troublesome species.     

Carbon and nitrogen metabolism in the seagrass, Zostera marina L.: Environmental control of enzymes involved in carbon allocation and nitrogen assimilation

This study experimentally examined influences of environmental variables on the activities of key enzymes involved in carbon and nitrogen metabolism of the submersed marine angiosperm, Zostera marina L. Nitrate reductase activity in leaf tissue was correlated with both water-column nitrate concentrations and leaf sucrose levels. Under elevated nitrate, shoot nitrate reductase activity increased in both light and dark periods if carbohydrate reserves were available. When water-column nitrate was low, glutamine synthetase activity in leaf tissue increased with environmental ammonium. In contrast, glutamine synthetase activity in belowground tissues was statistically related to both nitrate and temperature. At the optimal growth temperature for this species (ca. 25 °C), increased water-column nitrate promoted an increase in glutamine synthetase activity of belowground tissues. As temperatures diverged from the optimum, this nitrate effect on glutamine synthetase was no longer evident. Activities of both sucrose synthase and sucrose-P synthase were directly correlated with temperature. Sucrose-P synthase activity also was correlated with salinity, and sucrose synthase activity was statistically related to tissue ammonium. Overall, the enzymatic responses that were observed indicate a tight coupling between carbon and nitrogen metabolism that is strongly influenced by prevailing environmental conditions, especially temperature, salinity, and environmental nutrient levels.     

Copulation behaviour of Lygocoris pabulinus under laboratory conditions

As a prerequisite to the elucidation of the sex pheromone of the green capsid bug Lygocoris pabulinus (L.), we studied the bug's reproductive development and behaviour. When kept under long-day conditions (L18:D6) at 25 °C, both males and females start to copulate 4 days after the final moult. Second matings occur in some females, even on consecutive days. The premating period is 10 min on average and copulation lasts 1 to 2 min. At long range, when males are attracted to traps baited with live females, we did not observe a specific calling position of the females. At short range, a characteristic courtship behaviour of the females was not observed either. Males showed a characteristic vibration of the abdomen, which was repeated several times before copulation. This behaviour can be used as a bioassay to test potential sex pheromone compounds.     

Specific growth rate as a determinant of the carbon isotope composition of the temperate seagrass Zostera marina

The seasonal variability of specific growth rate and the carbon stable isotope ratio (δ¹³C) of leaf blades (δ¹³C_leaf) of a temperate seagrass, Zostera marina (within 10 days old) were measured simultaneously, together with the δ¹³C of dissolved inorganic carbon (δ¹³C_DIC) at three sites in the semi-closed Akkeshi estuary system, northeastern Japan, in June, September, and November 2004. The δ¹³C_leaf ranged from −16.2 to −6.3‰ and decreased from summer to winter. The simultaneous measurement of the δ¹³C_leaf, growth rate, and morphological parameters (mean leaf length and width, mean number of leaves per shoot, and sheath length) of the seagrass and δ¹³C_DIC in the surrounding water allowed us to compare directly the δ¹³C_leaf and specific growth rate of seagrass. The difference in the δ¹³C of seagrass leaves relative to the source DIC (Δδ¹³C_{leaf − DIC}) was the least negative (−11 to −7‰) in June at all three sites and became more negative (−17 to −8‰) as the specific growth rate decreased. This positive correlation between Δδ¹³C_{leaf − DIC} and specific growth rate can be used to diagnose the growth of seagrasses. Δδ¹³C_{leaf − DIC} changed by −1.7 ± 0.2‰ when the leaf specific growth rate decreased by 1% d⁻¹.     

Susceptibility of Cydia pomonella to Bacillus thuringiensis under laboratory and field conditions

The delta-endotoxin of 12 strains in 10 subspecies of Bacillus thuringiensis Berliner was highly active against Cydia pomonella (L.) when assayed under laboratory conditions on artificial diet. These results could not be confirmed in the field.The disappointing results obtained under field conditions are due to the behaviour of the target insect. C. pomonella larvae do not ingest food during penetration of the fruit. The larva bites pieces of the epidermis and deposits them without ingestion on top of the entry hole.

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Zusammenfassung Das delta-Endotoxin von B.t. war in Laborversuch auf Kunstmedium gegenüber den Larven des Apfelwicklers, C. pomonella, sehr aktiv. Die hohe Aktivität konnte aber unter Feldbedingungen nicht mehr bestätigt werden.Es wurde nachgewiesen, dass die unbefriedigenden Resultate von B. thuringiensis unter Feldbedingungen auf das Verhalten der Junglarven zurückzuführen sind: Die Larven nehmen während dem Eindringen in den Apfel keine Nahrung auf, sondern deponieren die herausgebissenen Epidermistücke über der Einbohrstelle.

     

Anopheles culicifacies: The occurrence of multiple insemination under laboratory conditions

Multiple insemination was detected under laboratory conditions in 38 of 399 females of Anopheles culicifacies Giles using a sex-linked, recessive eye-color marker system. Multiple mating occurred less frequently at low pair density and was not detected in a large cage. Remating after a 24-hr delay was detected in 5 of 131 families. The high incidence of multiple mating was attributed to interrupted copulations resulting in the transference of inadequate quantities of male accessory gland substance and/or by remating before insemination barriers were established after the initial insemination.

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Anopheles culicifacies giles: La presence d'inseminations multiples dans les conditions de laboratoire

Résumé Les inséminations multiples ont été mises en évidence dans les conditions de laboratoire chez 38 sur 399 femelles d'Anopheles culicifacies en utilisant un marqueur récessif colorant l'il, lié au sexe. Les copulations répétées sont moins fréquentes aux faibles densités et n'ont pas été observées dans de grandes cages. La recopulation après un délai de 24 h a été observée dans 5 familles sur 131. La haute fréquence de copulations multiples est envisagée comme conséquence de copulations interrompues, empéchant le transfert de quantités adéquates de sécrétions des glandes accessoires mâles et/ou par la recopulation avant que les barrières de non réceptivité aient été établies par l'insémination initiale.

     

Partitioning of food resources among Sillago japonica, Ditremma temmincki, Tridentiger trigonocephalus, Hippocampus japonicus and Petroscirtes breviceps in an eelgrass, Zostera marina, bed

We carried out dietary analysis on five numerically abundant fishes, Sillago japonica, Ditremma temmincki, Tridentiger trigonocephalus, Hippocampus japonicus and Petroscirtes breviceps in an eelgrass bed in Kwangyang Bay, Korea. Comparisons between species demonstrated that the dietary composition of each fish species was significantly different from that of every other species. Although gammarid amphipods and caprellid amphipods were consumed by all species, their individual contributions to each species' diet varied. Furthermore, polychaetes contributed to the diets of S. japonica and T. trigonocephalusand crab larvae were consumed byD. temmincki. Algae and eelgrass were not consumed by four fish species and made only a minimal contribution to the diet of P. breviceps. The diet of each fish species except H. japonicus underwent size-related changes; smaller fishes consumed gammarid amphipods, mysids and copepods, while larger fishes ate polychaetes, gastropods, isopods and other fishes. Differences in the prey organisms consumed of each individual species could be often related to differences in mouth length and width. S. japonica, D. temmincki, T. trigonocephalus, and P. breviceps underwent also a significant diel changes that could be related to differences in foraging behavior and/or prey availability. Thus, use of vision to detect prey would account for the greater daytime consumption of copepods by S. japonica and of crab larvae by D. temmincki, whereas the nocturnal emergence of gammarid amphipods, polychaetes and isopods from the substrate explained their greater consumption by S. japonica, D. temmincki, T. trigonocephalusand P. breviceps at night. Dietary breadth was greater for species with larger mouth dimensions.     

Growth and biomass allocation of sweet flag (Acorus calamus L.) under different nutrient conditions

Hydrobiologia - The antropogenically-induced process of eutrophication is accompanied by changes in the structure and function of the whole wetland ecosystem. Since the shift in a species ability...     

Biology of Dermacentor silvarum (Acari: Ixodidae) under laboratory conditions

The minimum life cycle of Dermacentor silvarum Olenev had a mean duration of 87.5 days (range 74–102 days) under laboratory conditions [(27±1 °C), 70% RH, 6 L: 18 D]. The mean time in (days) for the different stages of its cycle was as follows: incubation period of eggs was 15.3 days; prefeeding, feeding and premoulting periods of larvae and nymphs averaged 5.5, 4.0 and 7.3 days, and 5.2, 5.0 and 14.6 days, respectively; prefeeding, feeding, preoviposition and oviposition periods of female adults lasted for 7.8, 4.5, 4.3 and 14.0 days, respectively. There existed a highly significant correlation between engorged body weight of females and egg masses laid (r = 0.9877, p<0.001). the= reproductive= efficiency= index= (rei)= and= reproductive= fitness= index= (rfi)= in= females= were= 11.09= and= 9.58,= respectively.= no= relationship= between= nymphal= engorged= body= weight= and= resultant= sexes= was= observed.= delayed= feeding= and= non-oviposition= (in= june= and= july)= existed= in= females,= and= low= temperature= (−10 °c)= treatment= for= 45 days= could= terminate= oviposition= diapause.= however,= the= egg= masses= laid= by= post-diapause= females= were= significantly= smaller= than= those= laid= by= females= engorged= in= march,= april= and=>     

Regional variation in eelgrass (Zostera marina) morphology, production and stable sulfur isotopic composition along the Baltic Sea and Skagerrak coasts

Morphology, total sulfur content and stable sulfur isotopic composition of Zostera marina were examined in the Baltic Sea–Skagerrak transition zone through surveys. The seagrass meadows were denser and less productive at the low salinities in the Baltic Sea (salinity 6–7 psu), and total sulfur accumulations in plants were lower and δ³⁴S values were higher compared to the west coast of Sweden (salinity 21–29 psu). The δ³⁴S values of the three plant compartments (leaves, rhizomes, roots) indicated lower sulfide invasion at low salinities, which was mainly due to environmental conditions (e.g. low epiphytic biomass, low sediment organic matter and low sulfate concentration) and plant characteristics (productivity, shoot morphology). Between 13% and 63% of the sulfur in the plants was derived from sediment sulfides with highest percentages in the roots (27–63%) and lower in rhizomes (13–50%) and leaves (14–51%). The high sulfide invasion on the west coast of Sweden was coincident with high sediment organic matter, probably increasing sulfide pressure on the plants, and high epiphytic biomass, probably constraining the oxygen dynamics in the plants and enhancing sulfide invasion. Regional and spatial variability in the δ³⁴S were extensive, emphasizing the need for detailed analysis of local sources when applying stable sulfur isotopes in food web analyses. The observed invasion of sulfides suggests sulfide as a contributing factor to reported declines of Z. marina in the Skagerrak region.     

Removal of Prymnesium parvum (Haptophyceae) cells under different nutrient conditions by clay

In this study, we have shown that the ichthyotoxic Prymnesium parvum (haptophyte) can be successfully removed by spraying the surface with a phosphatic clay/polyaluminium chloride (PAC) mix (final concentration, 4 g l⁻¹ and 5 ppm, respectively). The alga was grown in non-axenic batch cultures with nitrogen deficient, phosphorus deficient and nutrient sufficient media. Sub-samples of the nutrient sufficient culture were diluted to obtain cell abundances equal to those in the nutrient deficient cultures. Clay/PAC removed up to 100% of the cells in the low cell nutrient sufficient treatment after 72 h, but removal in the other treatments was lower (up to 84%). The nitrogen deficient cells were found to be the most toxic, measured as haemolytic activity of the cells (HE₅₀), just prior to the start of the experiment. However, the toxicity of the cells in all treatments was found to fluctuate during the incubation time with a general increase in toxicity towards the end, suggesting that the cells became stressed during sedimentation and/or when trapped in the clay. But the amount of released toxins was always below the detection limit of the haemolytic assay, and the abundance of free-living bacteria, derived from flow cytometer counts, increased throughout the experiment. This suggests that released toxins were either trapped by the clay particles or effectively degraded by the bacterial community. The study shows that the clay method can be efficient in mitigation of P. parvum blooms, but further studies have to be conducted where optimum clay concentrations are determined to ensure that the efficiency is high against nutrient deficient cells and at high cell abundances, i.e. conditions found during blooms in eutrophic coastal waters, and to determine the fate of the cells and their toxins during and after sedimentation.     

Growth of phototrophic biofilms from limestone monuments under laboratory conditions

In the current study, five phototrophic biofilms from different Southern Europe limestone monuments were characterised by molecular techniques and cultivated under laboratory conditions. Phototrophic biofilms were collected from Orologio Tower in Martano (Italy), Santa Clara-a-Velha Monastery and Ajuda National Palace, both in Portugal, and Seville and Granada Cathedrals from Spain. The biofilms were grown under laboratory conditions and periodically sampled in order to monitor their evolution over a three-month period. Prokaryotic communities from natural samples and cultivated biofilms were monitored using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments in conjunction with clone sequencing and phylogenetic analysis. DNA-based molecular analysis of 16S rRNA gene fragments from the natural green biofilms revealed complex and different communities composition with respect to phototrophic microorganisms. The biofilms from Orologio Tower (Martano, Italy) and Santa Clara-a-Velha Monastery (Coimbra, Portugal) were dominated by the microalga Chlorella. The cyanobacterium Chroococcidiopsis was the dominating genus from Ajuda National Palace biofilm (Lisbon, Portugal). The biofilms from Seville and Granada Cathedrals (Spain) were both dominated by the cyanobacterium Pleurocapsa. The DGGE analysis of the cultivated biofilms showed that the communities developed differently in terms of species establishment and community composition during the three-month incubation period. The biofilm culture from Coimbra (Portugal) showed a remarkable stability of the microbial components of the natural community in laboratory conditions. With this work, a multiple-species community assemblage was obtained for further stone colonisation experiments.     

Reproductive phenology of the Mediterranean seagrasses Posidonia oceanica (L.) Delile, Cymodocea nodosa (Ucria) Aschers., and Zostera noltii Hornem.

Investigations on the sexual reproduction of Posidonia oceanica (L.) Delile, Cymodocea nodosa (Ucria) Aschers. and Zostera noltii Hornem. were conducted in situ from 1979 to 1988 in beds around the island of Ischia. Seed germination and phenological features of seedlings of P. oceanica and C. nodosa were observed in situ and in the laboratory. The reproductive cycles showed, for each species, different timing of start and duration of flowering, fruiting and germination. Posidonia oceanica flowers always appeared in September in shallow stands and in November in stands deeper than 15 m. This time delay seems to be related to the different maximum summer temperatures found at those depths. This species did not show dormancy. Cymodocea nodosa flowering was always recorded after the minimum winter temperature (April–May); germination of the seeds followed after 8–10 months of dormancy. Zostera noltii flowers were always recorded in July, with fruiting in August. The life cycles of these species seem to be related to the annual temperature range rather than to other environmental parameters. The different reproductive strategies adopted by each species are discussed.     

Growth responses of the mixotrophic dinoflagellates, Cryptoperidiniopsis sp. and Pfiesteria piscicida, to light under prey-saturated conditions

Recent research emphasis on the ecology of Pfiesteria spp. (Dinophyceae) has led to recognition of several morphologically similar heterotrophic dinoflagellates that often co-occur with Pfiesteria spp. in estuaries along the United States Atlantic coast. These include cryptoperidiniopsoid dinoflagellates, which resemble Pfiesteria spp. in having complex life cycles that include zoospores capable of kleptoplastidy. To examine and compare the role of kleptoplastidy in Cryptoperidiniopsis sp. and Pfiesteria piscicida, we tested the effects of irradiance on growth under prey-saturated (Storeatula major, Cryptophyceae) conditions. Growth of Cryptoperidiniopsis was strongly influenced by light intensity while no major effects were observed in P. piscicida. In Cryptoperidiniopsis, highest cell numbers and specific growth rates, but lowest specific cryptophyte consumption rates, were found at the highest light intensity tested (100 μmol photons m⁻² s⁻¹). A growth model was developed and used to estimate that the average half-life of chloroplasts ingested by Cryptoperidiniopsis decreased 3.4-fold from 12.6 h at high light to 3.7 h in the dark. These results show that light strongly enhances specific growth rate and growth efficiency of Cryptoperidiniopsis feeding on cryptophytes, and suggest that retained kleptochloroplasts may play a quantitatively significant role in carbon and energy metabolism of this organism. Differences in the effects of light between Cryptoperidiniopsis and P. piscicida may reflect different nutritional strategies, and allow these closely related dinoflagellates to occupy different niches and co-exist.