Chara hispida beds as a sink of nitrogen: Evidence from growth,nitrogen uptake and decomposition

Chara hispida forms dense beds (0.78–0.95 kg DW m⁻²) in Colgada Lake. The ability of Chara meadows to act as a nitrogen source or sink was evaluated by the following methods: (1) investigating Chara growth, (2) nitrogen incorporation and decomposition laboratory experiments and (3) relating experimental results to field conditions. Sediment oospores were germinated in large aquaria and observed growth rates were 0.001 m day⁻¹ (shoot length) and 0.0002 g day⁻¹ (dry weight). Nitrogen uptake rates were determined by addition of K¹⁵NO₃ during two different periods of Chara growth and the rates were 1.21 and 3.86 μM g DW⁻¹ h⁻¹ when charophytes were 166 days old (not sexually mature) and 323 days old (sexually mature), respectively. After the uptake experiments, the same charophytes were allowed to decompose within two types of litter bags (3 mm-pore litter bags and entire, non-porous plastic litter bags). Decomposition rates of Ch. hispida were 0.016 and 0.009 day⁻¹ in perforated and non-perforated bags, respectively, and fit a negative exponential model. The nitrogen release rate, calculated as the disappearance of N content from Chara tissues, was 0.012 day⁻¹ and there were no statistically significant differences between the values from the two different bag types. The dissolved organic nitrogen concentrations in aquarium and non-perforated litter bags waters increased linearly with time due to the leaching of soluble compounds from Chara. The rate of N loss from Chara tissues, total nitrogen and dissolved organic nitrogen release rates and the decrease in initial dry weight rate were all lower than the daily rate of Chara N uptake. By extrapolating laboratory data to field situations, we determined that approximately 38% of the N taken up by charophytes in Colgada Lake during the growth period is retained. Given the high charophyte biomass in the lake, its ability to incorporate nitrogen, its low decomposition rate and its ability to over-winter, we conclude that Chara beds could be acting as nitrogen sinks in this ecosystem.     

Seasonal variations in photosynthetic irradiance response curves of macrophytes from a Mediterranean coastal lagoon

The main photosynthesis and respiration parameters (dark respiration rate, light saturated production rate, saturation irradiance, photosynthetic efficiency) were measured on a total of 23 macrophytes of the Thau lagoon (2 Phanerogams, 5 Chlorophyceae, 10 Rhodophyceae and 6 Phaeophyceae). Those measurements were performed in vitro under controlled conditions, close to the natural ones, and at several seasons. Concomitantly, measurements of pigment concentrations, carbon, phosphorous and nitrogen contents in tissues were performed. Seasonal intra-specific variability of photosynthetic parameters was found very high, enlightening an important acclimatation capacity. The highest photosynthetic capacities were found for Chlorophyceae (e.g. Monostroma obscurum thalli at 17 °C, 982 μmol O₂ g⁻¹ dw h⁻¹ and 9.1 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹, respectively for light saturated net production rate and photosynthetic efficiency) and Phanerogams (e.g. Nanozostera noltii leaves at 25 °C, 583 μmol O₂ g⁻¹ dw h⁻¹ and 2.6 μmol O₂ g⁻¹ dw h⁻¹/μmol photons m⁻² s⁻¹ respectively for light saturated net production rate and photosynthetic efficiency). As expected, species with a high surface/volume ratio were found to be more productive than coarsely branched thalli and thick blades shaped species. Contrary to R_d (ranging 6.7–794 μmol O₂ g⁻¹ dw h⁻¹, respectively for Rytiphlaea tinctoria at 7 °C and for Dasya sessilis at 25 °C) for which a positive relationship with water temperature was found whatever the species studied, the evolution of P/I curves with temperature exhibited different responses amongst the species. The results allowed to show summer nitrogen limitation for some species (Gracilaria bursa-pastoris and Ulva spp.) and to propose temperature preferences based on the photosynthetic parameters for some others (N. noltii, Zostera marina, Chaetomorpha linum).     

Characteristics of simultaneous organic and nutrient removal in a pilot-scale vertical submerged membrane bioreactor (VSMBR) treating municipal wastewater at various temperatures

A pilot-scale vertical submerged membrane bioreactor (VSMBR) with anoxic and oxic zones in one reactor was operated in an attempt to reduce the problems concerning effective removal of organic matter and nutrients from municipal wastewater. Source water with total chemical oxygen demand (TCOD)/total nitrogen (TN) ratio of 5.5 was treated at various temperatures (13–25 °C) over an interval of about 1 year. As a result, total suspended solid (TSS) and TCOD were removed by 100% and higher than 98%, respectively. Moreover, the average removal efficiencies of TN and total phosphorus (TP) were found to be 74% and 78% at 8 h-hydraulic retention time (HRT) and 60-days sludge retention time (SRT). Under these conditions, the specific removal rates (SRR) of TN and TP were found to be 0.093 kg N m⁻³ day⁻¹ and 0.008 kg P m⁻³ day⁻¹, and the daily production of excess sludge (DPES), 0.058 kg TSS day⁻¹.     

Evaluation of the effects of light intensity on growth of the benthic dinoflagellate Ostreopsis sp. 1 using a newly developed photoirradiation-culture system and a novel regression analytical method

Benthic dinoflagellates of the genus Ostreopsis are found all over the world in temperate, subtropical, and tropical coastal regions. Our recent studies revealed that a putative “cryptic” species of Ostreopsis ovata is present widely along Japanese coasts. This organism, Ostreopsis sp. 1, possesses palytoxin analogs and thus its toxic blooms may be responsible for potential toxification of marine organisms. To evaluate the bloom dynamics of Ostreopsis sp. 1, the present study examined the growth responses of Ostreopsis sp. 1 strain s0716 to various light intensities (photon flux densities: μmol photons m⁻² s⁻¹) using a newly devised photoirradiation-culture system. This novel system has white light-emitting diodes (LEDs) capable of more closely simulating the wavelength spectrum of light entering the oceanic water column than do fluorescent tubes and halogen lamps. In this system, the light intensity of the white LEDs was reduced through two polarizing filters by varying the rotation angles of the filters. Thereby, the new system was capable of culturing microalgae under well-controlled light intensity conditions. Ostreopsis sp. 1 grew proportionally when light intensity was increased from 49.5 to 199 μmol photons m⁻² s⁻¹, but its growth appeared to be inhibited slightly at ≥263 μmol photons m⁻² s⁻¹. The relationship between observed growth rates and light intensity was calculated at R > 0.99 (P < 0.01) using a regression analysis with a modified equation of the photosynthesis-light intensity (P-L) model. The equation determined the critical light intensities for growth of Ostreopsis sp. 1 and the organism's growth potential as follows: (1) the threshold light intensity for growth: 29.8 μmol photons m⁻² s⁻¹; (2) the optimum light intensity (L_m) giving the maximum growth rate (μ_max = 0.659 divisions day⁻¹): 196 μmol photons m⁻² s⁻¹; (3) the optimum light intensity range (L_opt) giving ≥95% μ_max: 130–330 μmol photons m⁻² s⁻¹; (4) the semi-optimum range (L_sopt) giving ≥80% μ_max: 90 to over 460 μmol photons m⁻² s⁻¹. The L_sopt represents 4.5–23% ambient light intensity present in surface waters off of a temperate region of the Japanese coast, Tosa Bay; putatively, this semi-optimum range of light intensity appears at depth of 12.9–27.8 m. Considering these issues, our data indicate that Ostreopsis sp. 1 in coastal environments may form blooms at ca. ∼28 m depth in regions along Japanese coasts.     

Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria

The relationship between light intensity, nitrogen availability and pigmentation was investigated in mixotrophic and heterotrophic cultures of the unicellular red alga Galdieria sulphuraria 074G, a potential host for production of the blue pigment, phycocyanin (PC). During the exponential growth phase of batch cultures, G. sulphuraria 074G contained 2–4 mg phycocyanin per g dry weight. In carbon-limited and nitrogen-sufficient batch cultures grown in darkness, this value increased to 8–12 mg g⁻¹ dry weight during the stationary phase, whereas the phycocyanin content in nitrogen-deficient cells decreased to values below 1 mg g⁻¹ dry weight during stationary phase. Light intensities between 0 and 100 μmol photons m⁻² s⁻¹ had no influence on phycocyanin accumulation in mixotrophic cultures grown on glucose or fructose, while light stimulated phycocyanin synthesis in cultures grown on glycerol, in which the phycocyanin content in stationary phase was increased from 10 mg g⁻¹ dry weight in darkness to 20 mg g⁻¹ dry weight at a light intensity of 80 μmol photons m⁻² s⁻¹. At higher light intensities, less phycocyanin accumulated than at lower intensities, irrespective of the carbon substrate used. In carbon-limited continuous flow cultures grown on glucose or glycerol at a dilution rate of 0.63 day⁻¹, corresponding to 50% of the maximum specific growth rate, the highest steady-state phycocyanin content of 15–28 mg g⁻¹ dry weight was found at 65 μmol photons m⁻² s⁻¹. In contrast to the apparent glucose repression of light-induced PC synthesis observed in batch cultures, no glucose repression of the light stimulation was observed in continuous flow cultures because the glucose concentration in the culture supernatant always remained at limiting levels. Despite the fact that G. sulphuraria 074G contains less phycocyanin than some other microalgae and cyanobacteria, the ability of G. sulphuraria 074G to grow and synthesize phycocyanin in heterotrophic or mixotrophic cultures makes it an interesting alternative to the cyanobacterium, Spirulina platensis presently used for synthesis of phycocyanin.     

Effects of intraspecific competition on growth and photosynthesis of Atriplex prostrata

We investigated the effect of intraspecific competition on growth parameters and photosynthesis of the salt marsh species Atriplex prostrata Boucher in order to distinguish the effects of density-dependent growth inhibition from salt stress. High plant density caused a reduction of 30% in height, 82% in stem dry mass, 80% in leaf dry mass, and 95% in root dry mass. High density also induced a pronounced 72% reduction in leaf area, 29% decrease in length of mature internodes and 50% decline in net photosynthetic rate. The alteration of net photosynthesis paralleled growth inhibition, decreasing from 7.6 ± 0.9 μmol CO₂ m⁻² s⁻¹ at low density to 3.5 ± 0.4 μmol CO₂ m⁻² s⁻¹ at high density, indicating growth inhibition caused by intraspecific competition is mainly due to a decline in net photosynthesis rate. Plants grown at high density also exhibited a reduction in stomatal conductance from 0.7 ± 0.1 mol H₂O m⁻² s⁻¹ at low density to 0.3 ± 0.1 mol H₂O m⁻² s⁻¹ at high density and a reduction in transpiration rate from 6.0 ± 0.3 mmol H₂O m⁻² s⁻¹ at low density to 4.3 ± 0.3 mmol H₂O m⁻² s⁻¹ at high density. Biomass production was inhibited by an increase in plant density, which reduced the rate of photosynthesis, stomatal conductance and leaf area of plants.     

Leaf production and shoot dynamics of Thalassia testudinum by a direct census method

The annual leaf growth and shoot dynamics of Thalassia testudinum were examined in a meadow located near Havana City, Cuba, using direct censuses between January 1995 and January 1996. The net rate of shoot population change, specific shoot recruitment and mortality rates were calculated as the difference between the densities of shoots (tagged or untagged) in consecutive sampling events. The leaf biomass, the daily production, the turnover rate and the rate of leaf biomass loss were also estimated. The estimated mean dry leaf biomass (124.9 ± 9.5 g m⁻²), daily dry leaf production (3.3 ± 0.2 g m⁻² day⁻¹) and turnover rate (2.7 ± 0.1% day⁻¹) were comparable to values previously reported for this species in Cuba and elsewhere. The production of leaves and shoots were higher in spring, declined towards mid summer, and showed the minimum values in January. Shoot recruitment prevailed over shoot mortality from January to March and from July to August, whereas most of the annual shoot mortality occurred between May and July and between August and October. The meadow examined was in close demographic balance along the study period. The results demonstrate that direct census provides reliable estimates of rapid shoot dynamics in T. testudinum.     

Growth and loss of distal tissue in blades of Lessonia nigrescens and Lessonia trabeculata (Laminariales)

Meristematic growth and loss of distal tissue from blades of two ecologically important species in the south-east Pacific, Lessonia nigrescens and Lessonia trabeculata, was evaluated during 1 year. Comparative growth was determined by a hole-punch method, loss of distal tissue from the blades was determined by subtracting final blade length (with loss) from expected blade lengths (without loss); growth and tissue loss were transformed to fresh biomass units for calculation of inter-algae differences. The results showed that blade elongation rate increased at the beginning of spring, and declined towards the end of summer, with mean values between 0.40 and 0.08 cm day⁻¹ for L. nigrescens, and 0.65–0.17 cm day⁻¹ for L. trabeculata. Loss of distal tissue varied seasonally when examined as length units for both species; with mean values between 0.24 and 0.10 cm day⁻¹ for L. nigrescens, and 0.51–0.25 cm day⁻¹ for L. trabeculata. Variations in fresh biomass units were only observed in Lessonia trabeculata, increasing in spring, with mean values to 0.13 g (fresh weight) day⁻¹. Annual growth and loss of distal tissue were higher in L. trabeculata (0.41 and 0.39 cm day⁻¹, respectively) than in L nigrescens (0.19 and 0.15 cm day⁻¹). When growth and tissue loss were considered as fresh biomass, monthly gains significantly outweighed loss of distal tissue in both species, but parallel results based on length data followed a different trend. L. trabeculata released about 50% of its growth biomass as particulate organic matter, while the comparative value for L. nigrescens was about 20%.     

The effects of NH4+ and NO3− on growth,resource allocation and nitrogen uptake kinetics of Phragmites australis and Glyceria maxima

The effects of NH₄⁺ or NO₃⁻ on growth, resource allocation and nitrogen (N) uptake kinetics of two common helophytes Phragmites australis (Cav.) Trin. ex Steudel and Glyceria maxima (Hartm.) Holmb. were studied in semi steady-state hydroponic cultures. At a steady-state nitrogen availability of 34 μM the growth rate of Phragmites was not affected by the N form (mean RGR = 35.4 mg g⁻¹ d⁻¹), whereas the growth rate of Glyceria was 16% higher in NH₄⁺-N cultures than in NO₃⁻-N cultures (mean = 66.7 and 57.4 mg g⁻¹ d⁻¹ of NH₄⁺ and NO₃⁻ treated plants, respectively). Phragmites and Glyceria had higher S/R ratio in NH₄⁺ cultures than in NO₃⁻ cultures, 123.5 and 129.7%, respectively.Species differed in the nitrogen utilisation. In Glyceria, the relative tissue N content was higher than in Phragmites and was increased in NH₄⁺ treated plants by 16%. The tissue NH₄⁺ concentration (mean = 1.6 μmol g fresh wt⁻¹) was not affected by N treatment, whereas NO₃⁻ contents were higher in NO₃⁻ (mean = 1.5 μmol g fresh wt⁻¹) than in NH₄⁺ (mean = 0.4 μmol g fresh wt⁻¹) treated plants. In Phragmites, NH₄⁺ (mean = 1.6 μmol g fresh wt⁻¹) and NO₃⁻ (mean = 0.2 μmol g fresh wt⁻¹) contents were not affected by the N regime. Species did not differ in NH₄⁺ (mean = 56.5 μmol g⁻¹ root dry wt h⁻¹) and NO₃⁻ (mean = 34.5 μmol g⁻¹ root dry wt h⁻¹) maximum uptake rates (V_max), and V_max for NH₄⁺ uptake was not affected by N treatment. The uptake rate of NO₃⁻ was low in NH₄⁺ treated plants, and an induction phase for NO₃⁻ was observed in NH₄⁺ treated Phragmites but not in Glyceria. Phragmites had low K_m (mean = 4.5 μM) and high affinity (10.3 l g⁻¹ root dry wt h⁻¹) for both ions compared to Glyceria (K_m = 6.3 μM, affinity = 8.0 l g⁻¹ root dry wt h⁻¹). The results showed different plasticity of Phragmites and Glyceria toward N source. The positive response to NH₄⁺-N source may participates in the observed success of Glyceria at NH₄⁺ rich sites, although other factors have to be considered. Higher plasticity of Phragmites toward low nutrient availability may favour this species at oligotrophic sites.     

Escape protein supplementation of growing steers grazing stargrass

A grazing trial utilizing 35 individually supplemented growing steers (211±42 kg initial body weight (BW)) was conducted to study the effect of supplemental escape protein on the performance of steers grazing on stargrass (Cynodon plectostachyus) during the dry season. N in supplements was 100%, 50%, or 0% natural protein (bloodmeal, coconut meal, and soybean meal), and 0%, 50% or 100% urea. All steers received 2 kg of supplement dry matter (DM) (2.2% N) daily during the 90 days of the experiment. Steers fed the urea supplement had the lowest ADG (0.97 kg day⁻¹). There was a linear (P<0.05) response in ADG to the natural protein level (50 and 100%) in supplements containing bloodmeal (1.11 and 1.21 kg day⁻¹) and coconut meal (1.05 and 1.21 kg day⁻¹), but no response was observed with soybean meal (1.01 and 1.0 kg day⁻¹). Forage intake was not affected by supplementation. As a result of the growth response observed for supplements containing bloodmeal and coconut meal above the urea-based and soybean meal supplements, it was concluded that growing ruminants grazing stargrass in the dry season were deficient in escape protein. ©1997 Elsevier Science B.V.     

β-carotene production from soap stock by loofa-immobilized Rhodotorula rubra in an airlift photobioreactor

In this study, the soap stock as a sole carbon source was used for growing a carotenoid producing yeast (Rhodotorula rubra). The application of soap stock resulted in increase of carotenoids yield up to 5.36 folds when compared with the grown cultures on glucose. On the best Monod equation fitted on the specific growth rate (μ) data, the maximum specific growth rate (μ_m) and half-saturation concentration (K_S) were respectively determined at 0.064 h⁻¹ and 3.26 g L⁻¹ for total fatty acids presented in soap stock. Further tests on the carotenogenesis process were carried out in a cell-immobilized airlift photobioreactor where the natural loofa sponge was used for immobilization of the cells. The performance of the bioreactor was statistically studied by the response surface methodology (RSM) where aeration rate of 0.11 vvm and light irradiation intensity of 2517 Lx provided an optimum condition for producing β-carotene with a specific production rate of 22.65 mg g_cell⁻¹ day⁻¹.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.     

Increasing nitrogen depositions can reduce lichen viability and limit winter food for an endangered Chinese monkey

Increasing economic growth and industrial development in China is starting to impact even remote areas such as the Shennongjia nature reserve, where nitrogen pollution is becoming a major environmental threat. The epiphytic lichen flora is particularly rich in this area and is one of the components of this habitat most sensitive to nitrogen pollution. Since lichens represent an important food resource for the endangered monkey species Rhinopithecus roxellana, a reduction in lichen availability would have harmful consequences for the conservation of its habitat in the Shennongjia Mountains. To investigate the effects of increased nitrogen availability on the local lichen communities, so far scarcely considered, we conducted a one-year field experiment measuring growth, survival, and phosphomonoesterase activity of the widespread species Usnea luridorufa in response to nitrogen (up to 50 kg N ha⁻¹ year⁻¹ deposition) and phosphorus supply. Growth and survival of thalli and propagules of U. luridorufa decreased when treated with N deposition >12.05 kg N ha⁻¹ year⁻¹ and >2.14 kg N ha⁻¹ year⁻¹, respectively. The important role of phosphorus availability in relation to nitrogen supply was demonstrated by the increase in phosphomonoesterase activity with increasing nitrogen availability until a nitrogen toxicity threshold was reached. However, the high concentration of phosphorus in rainwater showed that phosphorus is not a limiting nutrient in the area.The results make a contribution to the knowledge of the negative effects of increased N deposition in the Shennongjia forest ecosystem.     

Two ways to achieve an anammox influent from real reject water treatment at lab-scale: Partial SBR nitrification and SHARON process

A comparative study to produce the correct influent for Anammox process from anaerobic sludge reject water (700–800 mg NH₄⁺-N L⁻¹) was considered here. The influent for the Anammox process must be composed of NH₄⁺-N and NO₂⁻-N in a ratio 1:1 and therefore only a partial nitrification of ammonium to nitrite is required. The modifications of parameters (temperature, ammonium concentration, pH and solid retention time) allows to achieve this partial nitrification with a final effluent only composed by NH₄⁺-N and NO₂⁻-N at the right stoichiometric ratio. The equal ratio of HCO₃⁻/NH₄⁺ in reject water results in a natural pH decrease when approximately 50% of NH₄⁺ is oxidised. A Sequencing batch reactor (SBR) and a chemostat type of reactor (single-reactor high activity ammonia removal over nitrite (SHARON) process) were studied to obtain the required Anammox influent. At steady state conditions, both systems had a specific conversion rate around 40 mg NH₄⁺-N g⁻¹ volatile suspended solids (VSS) h⁻¹, but in terms of absolute nitrogen removal the SBR conversion was 1.1 kg N day⁻¹ m⁻³, whereas in the SHARON chemostat was 0.35 kg N day⁻¹ m⁻³ due to the different hydraulic retention time (HRT) used. Both systems are compared from operational (including starvation experiments) and kinetic point of view and their advantages/disadvantages are discussed.     

Germination fluctuation of toxic Alexandrium fundyense and A. pacificum cysts and the relationship with bloom occurrences in Kesennuma Bay,Japan

While cyst germination may be an important factor for the initiation of harmful/toxic blooms, assessments of the fluctuation in phytoplankton cyst germination, from bottom sediments to water columns, are rare in situ due to lack of technology that can detect germinated cells in natural bottom sediments. This study introduces a simple mesocosm method, modeled after previous in situ methods, to measure the germination of plankton resting stage cells. Using this method, seasonal changes in germination fluxes of toxic dinoflagellates resting cysts, specifically Alexandrium fundyense (A. tamarense species complex Group I) and A. pacificum (A. tamarense species complex Group IV), were investigated at a fixed station in Kesennuma Bay, northeast Japan, from April 2014 to April 2015. This investigation was conducted in addition to the typical samplings of seawater and bottom sediments to detect the dinoflagellates vegetative cells and resting cysts. Bloom occurrences of A. fundyense were observed June 2014 and February 2015 with maximum cell densities reaching 3.6 × 10⁶ cells m⁻² and 1.4 × 10⁷ cells m⁻², respectively. The maximum germination fluxes of A. fundyense cysts occurred in April 2014 and December 2014 and were 9.3 × 10³ cells m⁻² day⁻¹ and 1.4 × 10⁴ cells m⁻² day⁻¹, respectively. For A. pacificum, the highest cell density was 7.3 × 10⁷ cells m⁻² during the month of August, and the maximum germination fluxes occurred in July and August, reaching 5.8 × 10² cells m⁻² day⁻¹. Thus, this study revealed the seasonal dynamics of A. fundyense and A. pacificum cyst germination and their bloom occurrences in the water column. Blooms occurred one to two months after peak germination, which strongly suggests that both the formation of the initial population by cyst germination and its continuous growth in the water column most likely contributed to toxic bloom occurrences of A. fundyense and A. pacificum in the bay.     

Seasonal changes of the introduced Caulerpa racemosa var. cylindracea (Caulerpales,Chlorophyta) at the northwest limit of its Mediterranean range

A study of the meadows of the invasive Caulerpa racemosa var. cylindracea (Sonder) Verlaque, Huisman et Boudouresque was carried out over one year at Marseilles (Provence, France) where the alga is thriving, probably since 1994, in the cold waters of the north western Mediterranean Sea. At an early phase of colonisation, the C. racemosa meadow is characterized by a patchy distribution pattern. Several years are necessary to obtain a dense and continuous meadow. In one area colonized for more than 4 years, C. racemosa has developed a continuous meadow with wide seasonal variations. Maximum development was reached in autumn (biomass: 82 ± 3 g DW m⁻²; length of stolons: 1162 ± 86 m m⁻²; number of apices: 8360 ± 405 m⁻²; number of erect axes: 20955 ± 1499 m⁻²) and the minimum from winter to early spring (respectively, 0.3 ± 0.1 g DW m⁻²; 3 ± 1 m m⁻²; 220 ± 55 apices m⁻²; 35 ± 15 erect axes m⁻²). Seasonal variations in the growth rate were highly significant. The season of high growth lasted from June to October. The apical growth rate of a stolon reached a maximum of 7.5 ± 0.3 mm day⁻¹ in early October, then began to decrease significantly from the end of October to December, before becoming nearly nil from January to early May. Annual net production rate expressed in terms of stolon length and biomass was estimated as 5801 m m⁻² a⁻¹ and 612 g DW m⁻² a⁻¹, respectively. During the growth period, the turnover rate of the C. racemosa stolons was estimated at from 25 to 46 days. The growth rate was closely correlated to the seawater temperature (R² = 0.83), whereas no significant correlation was found between growth and irradiance. During the growth period, a decrease in temperature rapidly affects the growth rate, which soon recovers its earlier level when the temperature rises again. In winter, the growth rate decreased rapidly with the seasonal drop in the seawater temperature. Grazing by fish (Sarpa salpa and Boops boops) can also affect the growth rate from September to December by consumption of the erect axes and stolon apices, enhancing the ramification of stolons. Seasonal changes at Marseilles are much sharper than those reported for warmer Mediterranean localities (French Riviera, Italy, Croatia): in winter and early spring C. racemosa meadows decreased and locally disappeared, leaving a barren substrate. C. racemosa survives the lower winter seawater temperatures of the north-western Mediterranean Sea probably in the form of zygotes and/or small fragments (rhizoids, stolons, propagules).     

Diurnal exchanges of CO2 and CH4 across the water–atmosphere interface in a water chestnut meadow (Trapa natans L.)

CH₄ and CO₂ fluxes across the water–atmosphere interface were measured over a 24 h day–night cycle in a shallow oxbow lake colonized by the water chestnut (Trapa natans L.) (Lanca di Po, Northern Italy). Only exchanges mediated by macrophytes were measured, whilst gas ebullition was not considered in this study. Measurements were performed from 29 to 30 July 2005 with short incubations, when T. natans stands covered the whole basin surface with a mean dry biomass of 504 ± 91 g m⁻². Overall, the oxbow lake resulted net heterotrophic with plant and microbial respiration largely exceeding carbon fixation by photosynthesis. The water chestnut stand was a net sink of CO₂ during the day-light period (−60.5 ± 8.5 mmol m⁻² d⁻¹) but it was a net source at night (207.6 ± 6.1 mmol m⁻² d⁻¹), when the greatest CO₂ efflux rate was measured across the water surface (28.2 ± 2.4 mmol m⁻² h⁻¹). The highest CH₄ effluxes (6.6 ± 1.8 mmol m⁻² h⁻¹) were determined in the T. natans stand during day-time, whilst CH₄ emissions across the plant-free water surface were greatest at night (6.8 ± 2.1 mmol m⁻² h⁻¹). Therefore, we assumed that the water chestnut enhanced methane delivery to the atmosphere. On a daily basis, the oxbow lake was a net source to the atmosphere of both CO₂ (147.1 ± 10.8 mmol m⁻² d⁻¹) and CH₄ (116.3 ± 8.0 mmol m⁻² d⁻¹).     

Microbial protein synthesis in growing-finishing bulls estimated from the urinary excretion of purine derivatives

In the frame of a feeding experiment with three periods, balance trials were carried out with 18 double-muscled Belgian White–blue bulls, allocated to one of four feeding regimes. The ration consisted of maize silage and concentrate in the ratio of 35:65 on DM basis. Six concentrates were formulated to supply three levels of protein and energy. The three periods corresponded to distinct live weight intervals of 360–460, 460–570 and 570–680 kg. In the first feeding regime, low protein and intermediate energy level were given during the whole trial; in the second regime, protein level decreased at a constant intermediate energy level; in the third regime, the energy level increased at a constant high protein level; in the fourth regime, protein level decreased simultaneously with an increase in the energy level. Total daily intake varied from 6.2 to 9.7 kg for dry matter (DM), from 65 to 106 MJ for metabolisable energy (ME) and from 719 to 1326 g for crude protein (CP) (50 animal observations). At the end of each period, the excretion of the purine derivatives (PD), allantoin and uric acid, was measured after total urine collection during 4 days to estimate microbial nitrogen supply to the duodenum (MN_PD). The effect of the intake of DM, organic matter (OM), digestible OM (DOM), digestible carbohydrates (DCHO), total digestible nutrients (TDN), rumen fermentable OM (FOM), metabolisable energy (ME), fermentable ME (FME), CP, digestible CP (DCP) and rumen degradable protein (RDP) on PD and MN_PD was examined. Further, the relationship of MN_PD to MN, calculated according to different systems, was examined. The amount (mean±SD) of allantoin excreted in urine was 147±23 mmol day⁻¹ and of uric acid 11±3 mmol day⁻¹. The MN_PD amounted to 97±21 g day⁻¹, varying from 57 to 154 g day⁻¹, and significantly increased with all measures of nutrient intake. The correlation coefficients, ranging from 0.45 for DCP to 0.57 for DOM and FME, were, however, not significantly different. MN_PD showed a larger variation and was on average lower than the potential MN values calculated from the intake of FOM (108±13 g day⁻¹), DCHO (141±16 g day⁻¹) and FME (109±13 g day⁻¹), similar to that calculated from the intake of RDP (99±14 g day⁻¹) and higher than the MN value calculated from the intake of TDN (84±11 g day⁻¹). The correlations of MN_PD to the calculated MN values ranged from 0.47 for MN_FOM to 0.59 for MN_FME, but were not significantly different.     

Partitioning of latent heat flux at a northern peatland

The partitioning of latent heat flux (Q_E) to vascular plant and moss surface components was assessed for a Sphagnum-dominated bog with a hummock–hollow surface having a sparse canopy of low shrubs. Results from porometry and eddy covariance measurements of Q_E showed evaporation from the moss surface ranged from greater than 50% of total Q_E early in the growing season to less than 20% after a dry period toward the end of the growing season. Both soil moisture and vapour pressure deficit (D_a) affected this partitioning with drier moss and peat, lower water table, and smaller D_a all reducing moss Q_E. Daily maximum moss Q_E ranged from greater than 200 W m⁻² early in the growing season to less than 100 W m⁻² during a dry period. In contrast, vascular contribution to total Q_E increased over the season from a daily maximum of about 150 W m⁻² to 250 W m⁻² due to increase in leaf area by leaf replacement and emergence and to drying of the moss surface. Porometry results showed average daily maximum conductance from bog shrubs was near 8 mm s⁻¹. These conductance values were smaller than those reported for vascular plants from more nutrient-rich wetlands. The effect of increases in D_a on vascular Q_E were moderated by decreases in stomatal conductance. At constant available energy, vascular leaf conductance was reduced by as much as 2 mm s⁻¹ and moss surface conductance was enhanced by up to 3 mm s⁻¹ by large D_a. Considering vascular and non-vascular water transport characteristics and frequency of water table position and given the observed variations of Q_E partitioning with water table location and moss and peat water content, it is suggested that modelling efforts focus on how dry hummocks and wet hollows each contribute to Q_E, especially as related to D_a and soil moisture dynamics.     

Evaluation of zinc tolerance and accumulation potential of the coastal shrub Limoniastrum monopetalum (L.) Boiss.

The coastal shrub Limoniastrum monopetalum is capable of growth in soil containing extremely high concentrations of heavy metals. A greenhouse experiment was conducted in order to investigate the effects of a range of Zn concentrations (0–130 mmol l⁻¹) on growth and photosynthetic performance, by measuring relative growth rate, total leaf area, plant height, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations. We also determined the total zinc, nitrogen, phosphorus, sulphur, calcium, magnesium, sodium, potassium, iron and copper concentrations in the plant tissues. The study species demonstrated hypertolerance to Zn stress, since survival was recorded with leaf concentrations of up to 1700 mg Zn kg⁻¹ dry mass when treated with 130 mmol Zn l⁻¹. L. monopetalum exhibited little overall effects on photosynthetic function at Zn levels of up to 90 mmol l⁻¹. At greater external Zn concentration, plant growth was negatively affected, due in all probability to the recorded decline in net photosynthetic rate, which may be linked to the adverse effect of the metal on photosynthetic electron transport. Growth parameters were virtually unaffected by leaf tissue concentrations as high as 1400 mg Zn kg⁻¹ dry mass thus indicating that this species could play an important role in the phytoremediation of Zn-polluted areas.