Survival of juvenile Ecklonia radiata sporophytes after canopy loss

The understorey beneath a canopy of the kelp Ecklonia radiata often contains juvenile sporophytes of the same species. When canopy disturbance occurs, these juvenile sporophytes are exposed to new environmental conditions. If these juvenile sporophytes survive these new conditions, then they become a ready source of kelps to rapidly form a new canopy. This study investigated the potential of pre-existing juvenile sporophytes of E. radiata to survive post-disturbance conditions and contribute to the rapid formation of a new canopy. The potential of canopy recovery by recruitment of kelp from zoospores was also investigated. These processes were studied at different times during the summer and on reefs ranging in topographic complexity from simple, flat reefs to highly complex, rugose reefs. By tagging juvenile sporophytes after the adult kelp canopy was removed and monitoring them through time, it was demonstrated that most juveniles (> 50%) survived the change in conditions after canopy loss, with some juveniles going on to become members of a new canopy. Approximately 6-47% of tagged sporophytes died within 3-4 days after canopy removal possibly due to excessive photoinhibition and photostress as demonstrated by changes in photosynthetic performance (decreased alpha values) of juveniles. The potential contribution of juvenile sporophytes to the rapid formation of a new canopy appears to be dependent on the timing of canopy removal with late summer-autumn canopy loss favouring faster recovery. Topographically complex reefs had less short-term (7 days) survival of juvenile sporophytes than topographically simple reefs; however this difference was not carried through to the long-term (6 months) abundance of adult kelp in experimental clearings, which was greater on topographically complex reefs. Clearly, juvenile sporophytes in arrested development under existing canopies of the small kelp E. radiata are important for the rapid recovery of the kelp canopy once adults are lost through physical disturbance.     

Effect of pre-incubation irradiance on survival of cryopreserved gametophytes of Undaria pinnatifida (Phaeophyta) and morphology of sporophytes formed from the gametophytes

Gametophyte strains originating from indigenous sporophytes of Undaria pinnatifida (Harvey) Suringar in Iwate Prefecture, Northeast Japan, were maintained for 9–10 months at 45 μmol photons m⁻² s⁻¹. Before cryopreservation in liquid nitrogen for more than 12 h (1–14 days) using a two-step cooling method with a mixture of cryoprotectants (10% l-proline and 10% glycerol), these were pre-incubated for 2, 4 and 8 months at 15 μmol photons m⁻² s⁻¹. After 1 week of thawing, no surviving gametophytes were detected in the strains without pre-incubation, but both the female and male gametophytes, pre-incubated for more than 4 months, showed high survival rates (43–60% for females and 64–100% for males). This revealed the induction of freezing tolerance by incubation at low irradiance. Thereafter, sporophytes derived from cryopreserved gametophytes and subcultured gametophytes, stored under pre-incubation conditions, were formed from the strain, and a morphological comparison was conducted with 10 characters (stipe length, stipe wet weight, blade length, blade wet weight, blade width, incision depth, blade thickness, sporophyll length, sporophyll wet weight, and sporophyll width). The morphology of the sporophytes formed from the cryopreserved gametophytes corresponded well with that of the subcultured gametophytes from the same strain. The results suggest that the cryopreservation method is applicable for preserving culture stocks of U. pinnatifida to be used in mariculture.     

Elk Kelp, Pelagophycus porra, distribution limited due to susceptibility of microscopic stages to high light

Elk Kelp, Pelagophycus porra (Leman) Setchell, is commonly observed in deep (20-50 m) water along the outer edge of Giant Kelp, Macrocystis pyrifera (C. Agardh), forests in southern California, USA and northern Baja California, Mexico, but rarely occurs in shallower water or within the adjacent M. pyrifera beds. To investigate the factors that limit P. porra from establishing populations within these shallower habitats, juvenile P. porra sporophytes were transplanted from a deep (20 m) water P. porra zone to shallower (15 and 8 m) sites within and inside an adjacent M. pyrifera beds. Transplanted P. porra exhibited no differences in survival or growth among depths, although reproductive maturity was observed only at the two shallower sites. When P. porra propagules were experimentally introduced to areas under dense M. pyrifera canopies and areas cleared of the M. pyrifera canopies within the shallower M. pyrifera bed, P. porra still failed to recruit even though this procedure resulted in a four-fold increase in recruitment in the natural P. porra zone. Laboratory-based culturing of P. porra microscopic stages revealed that they grew and survived better under low light conditions characteristic of the natural P. porra zone (2-4 μmol photons m^− 2 s^− 1) than under higher light conditions characteristic of the M. pyrifera beds(18-20 μmol photons m^− 2 s^− 1). Pulse-amplitude modulated (PAM) fluorometry indicated that while naturally-occurring P. porra's adult sporophytes were able to photoacclimate to increasing irradiance as they grew from the benthos towards the surface, P. porra's microscopic stages were unable to photoacclimate to increased irradiances, and subsequently exhibited 100% mortality under these higher light conditions. Altogether, our study suggests that vulnerability of P. porra microscopic stages to higher irradiances appears to be the primary factor inhibiting P. porra from establishing populations in shallower water and stresses the importance of a multiple life-history approach when investigating species distributions.     

Primary production,respiration and calcification of the temperate free-living coralline alga Lithothamnion corallioides

Calcification and primary production responses to irradiance in the temperate coralline alga Lithothamnion corallioides were measured in summer 2004 and winter 2005 in the Bay of Brest. Coralline algae were incubated in dark and clear bottles exposed to different irradiances. Net primary production reached 1.5 μmol C g⁻¹ dry wt h⁻¹ in August and was twice as high as in January–February. Dark respiration showed significant seasonal variations, being three-fold higher in summer. Maximum calcification varied from 0.6 μmol g⁻¹ dry wt h⁻¹ in summer 2004 to 0.4 μmol g⁻¹ dry wt h⁻¹ in winter 2005. According to P–E curves and the daily course of irradiance, estimated daily net production and calcification reached 131 μg C g⁻¹ dry wt and 970 μg CaCO₃ g⁻¹ dry wt in summer 2004, and 36 μg C g⁻¹ dry wt and 336 μg CaCO₃ g⁻¹ dry wt in winter 2005. The net primary production of natural L. corallioides populations in shallow waters was estimated at 10–600 g C m⁻² y⁻¹, depending on depth and algal biomass. The mean annual calcification of L. corallioides populations varied from 300 to 3000 g CaCO₃ m⁻². These results are similar to those reported for tropical coralline algae in terms of carbon and carbonate productivity. Therefore, L. corallioides can be considered as a key element of carbon and carbonate cycles in the shallow coastal waters where they live.     

Biomass and primary production of a 8–11 m depth meadow versus <3 m depth meadows of the seagrass Cymodocea nodosa (Ucria) Ascherson

Current knowledge about the abundance, growth, and primary production of the seagrass Cymodocea nodosa (Ucria) Ascherson is biased towards shallow (depth <3 m) meadows although this species also forms extensive meadows at larger depths along the coastlines. The biomass and primary production of a C. nodosa meadow located at a depth of 8–11 m was estimated at the time of maximum annual vegetative development (summer) using reconstruction techniques, and compared with those available from shallow meadows of this species. A depth-referenced data base of values at the time of maximum annual development was compiled to that end. The vegetative development of C. nodosa at 8–11 m depth was not different from that achieved by shallow (depth <3 m) meadows of this species. Only shoot density, which decreased from 1637 to 605 shoots m⁻², and the annual rate of elongation of the horizontal rhizome, which increased from 23 to 71 cm apex⁻¹ year⁻¹, were different as depth increased from <3 to 8–11 m. Depth was a poor predictor of the vegetative development and primary production of C. nodosa. The biomass of rhizomes and roots decreased with depth (g DW m⁻² = 480 (±53, S.E.) − 32 (±15, S.E.) depth (in m); R² = 0.12, F = 4.65, d.f. = 35, P = 0.0381) which made total biomass of the meadow to show a trend of decrease with depth but the variance of biomass data explained by depth was low. The annual rate of elongation of the horizontal rhizome showed a significant positive relationship with depth (cm apex⁻¹ year⁻¹ = 18 (±5.1, S.E.) + 5.0 (±1.33, S.E.) depth (in m); R² = 0.50, F = 14.07, d.f. = 14, P = 0.0021). As shoot size and growth did not change significantly with depth, the reduction of shoot density should drive any changes of biomass and productivity of C. nodosa as depth increases. The processes by which this reduction of C. nodosa abundance with depth occur remain to be elucidated.     

Understanding the potential effects of water regime and salinity on recruitment of Melaleuca ericifolia Sm.

Although many emergent wetland plants may readily tolerate rapid changes in flooding and drying under freshwater conditions, their tolerance to dynamic water regimes may be compromised by salinity. Melaleuca-dominated woodlands occur naturally in Australia, south-east Asia and New Caledonia. Coastal wetlands dominated by Swamp paperbark (Melaleuca ericifolia) (Myrtaceae), native to south-east Australia, are commonly degraded as a consequence of altered water regime and salinity. This study simulates the release of M. ericifolia seeds from the aerial canopy under a range of water regime and salinity scenarios to determine conditions limiting sexual recruitment. Plant growth and survival were examined following seed release under two static water regimes (moist and flooded sediment) and two dynamic water regimes (simulated drawdown—“flooded-moist” and simulated re-flooding—“moist-flooded”). All water regimes, excluding the continuously flooded regime, were examined at three salinities: 0.1 dS m⁻¹ (fresh), 8 dS m⁻¹ and 16 dS m⁻¹, over a 50-day period commencing 44 days after the seeds were sown. The flooded treatment was examined at 0.1 dS m⁻¹ only, to confirm that flooding prohibits establishment of M. ericifolia. Seed and seedlings were positively buoyant and establishment was limited to moist soil. Flotation of seedlings in the flooded-moist treatment, however, did not inhibit subsequent establishment upon moist soil, even at the highest salinity of 16 dS m⁻¹. Growth, but not survival, was reduced by salinities of 8 dS m⁻¹ and 16 dS m⁻¹ in the moist treatment. Flotation of seedlings in saline water in the flooded-moist treatment did not reduce growth or survival compared with fresh water. Survival of seedlings in the moist-flooded treatment was lower in the freshwater and 16 dS m⁻¹ treatment compared with the moist treatment, but not at 8 dS m⁻¹. These findings suggest that water regime influences establishment of young M. ericifolia plants more strongly than does salinity, at least up to ∼1/3 seawater and in the short term (<2 months). Seedlings are likely to establish during a drawdown where the soil is exposed at salinities of ≤16 dS m⁻¹. In contrast, premature re-flooding of seedlings, even with fresh water, will compromise survival.     

Community metabolism and buffering capacity of nitrogen in a ruppia cirrhosa meadow

Fluxes of oxygen, inorganic nitrogen (DIN) and denitrification (isotope pairing) were measured from January 1997 to February 1998 via intact cores incubation in a shallow brackish area within the eutrophic Valli di Comacchio (northern Adriatic coast, Italy). Rates were measured in the light and in the dark in sediments colonized by the rooted macrophyte Ruppia cirrhosa and in adjacent sediments with benthic microalgae. Ruppia biomass (25-414 g DW m^− 2) exhibited a seasonal evolution whilst that of microphytobenthos (12-66 mg chl a m^− 2) was more erratic. Net (NP) and gross (GP) primary productivity was 1.15 and 6.89 mol C m^− 2y^− 1 for bare and 25.4 and 51.7 mol C m^− 2y^− 1 for Ruppia vegetated sediments. Nitrogen pools in Ruppia standing stock varied from 43.6 to 631.4 (annual average 201.2) mmol N m^− 2; the macrophyte N content was correlated with DIN concentration in the water column. Estimated N pool in microphytobenthos was one order of magnitude lower (from 2.4 to 14.5 mmol N m^− 2, annual average 7.2). Theoretical DIN assimilation calculated from NP was 127.8 and 1112.6 mmol N m^− 2y^− 1 whilst that calculated from GP was 765 and 2282 mmol N m^− 2y^− 1 for microphytobenthos and Ruppia respectively. Measured annual fluxes of DIN were 974.6 and − 577 mmol N m^− 2y^− 1 in bare and Ruppia vegetated sediments meaning that the two sites were a source and sink for DIN and that from 25 to 50% of Ruppia annual DIN requirements came from the water column. During the period of this study total denitrification was lower in the macrophyte colonized (92.3 mmol N m^− 2y^− 1) compared to bare sediments (163.3 mmol N m^− 2y^− 1) as a probable consequence of higher competition between denitrifiers and phanerogams. At both sites the ratio between denitrification of water column nitrate (D_W) and denitrification coupled to nitrification (D_N) was >1.6 due to little oxygen penetration in reducing sediments (< 1.2 mm) and scarce nitrification activity. D_W (0-35 µmol N m^− 2h^− 1) was significantly correlated with water column NO₃⁻  (2-16 µM). Theoretical DIN assimilation to denitrification ratio varied from 12.0 to 24.8 for Ruppia vegetated and from 0.8 to 4.7 for unvegetated sediments.At Valle Smarlacca, Ruppia may influence nitrogen cycling by incorporating large DIN pools in biomass which is scattered in surrounding areas and fuels intense bacterial activity. With increasing anthropogenic nutrient input and insignificant organic matter export in the open sea the already severe eutrophic conditions are enhanced and may accelerate the decline of the macrophyte meadow.     

The effects of algae species and densities on the population growth of the marine rotifer, Colurella dicentra

Colurella dicentra clones isolated from bay water in the Mississippi Gulf Coast were cultured with artificial seawater. Experiments were conducted to determine the effects of six algae species (Nannochloropsis oculata, Tetraselmis chuii, Chaetoceros gracilis, Rhodomonas salina, Isochrysis galbana, and Prorocentrum micans), six C. gracilis densities, and six N. oculata densities (25,000, 50,000, 100,000, 250,000, 500,000, and 1,000,000 cells ml^− 1) on C. dicentra population growth. Algae type influenced rotifer production (p < 0.0001). C. gracilis treatment (9120 ± 3351SD) produced the highest number of rotifers followed by N. oculata (5760 ±2232SD). P. micans had the lowest number of rotifers, although not significantly different from numbers in T. chuii, R. salina, and I. galbana treatments (p > 0.05).The population growth rate (r) varied with algae species treatment. The highest values were recorded for C. gracilis treatment (0.22 to 0.26 d^− 1), followed by N. oculata (0.21 to 0.24 d^− 1), and the lowest for P. micans (− 0.19 to 0.14 d^− 1). C. gracilis and N. oculata densities had significant effects (p < 0.0001) on C. dicentra population growth. The highest rotifer production was recorded at a C. gracilis density of 100,000 cells ml^− 1, followed by 250,000 cells ml^− 1 and 50,000 cells ml^− 1. Algae densities of 500,000 cells ml^− 1 and above produced the lowest rotifer numbers. Population growth rate (r) varied with C. gracilis densities. The highest values were observed for C. gracilis concentrations of 100,000 cells ml^− 1 (0.17 to 0.19 d^− 1), and the lowest for concentrations of 500,000 cells ml^− 1 and above (− 0.19 to 0.09 d^− 1). The 100,000 cells ml^− 1N. oculata density gave the highest rotifer production followed by 50,000, 250,000, 25,000, and 500,000 cells ml^− 1. Algae densities of 1,000,000 cells ml^− 1 produced the lowest rotifer numbers. Population growth rate (r) varied with N. oculata densities, with the highest values obtained for algae densities of 100,000 cells ml^− 1 (0.35 to 0.40 d^− 1), and the lowest for concentrations of 1,000,000 cells ml^− 1 (0.05 to 0.012 d^− 1). This is the first report of C. dicentra in Mississippi Coastal waters, and perhaps the smallest marine rotifer species (93 by 49 μm) ever cultured successfully.     

Shoot age as a confounding factor on detecting the effect of human-induced disturbance on Posidonia oceanica growth performance

The response of orthotropic rhizome elongation and primary production of Posidonia oceanica to anthropogenic perturbations and potential confounding effects of shoot age were assessed using a Linear Multilevel Model (LMM). This model examined the confounding effect of age by comparing the estimates of impact and variance components obtained by excluding and including Age as an explanatory variable. Age had a negative effect on rhizome elongation and primary production with an annual decrease of 0.6 mm y^− 1 and 7 mg dw y^− 1 respectively. According to the LMM when age effect was omitted, the differences between disturbed and control locations in rhizome elongation and primary production were 2.62 mm y^− 1 and 0.044 g dw y^− 1 respectively. These effects were statistically not significant. On the contrary, when age effect was included in the statistical model, impacts became evident for both variables, with significant differences between disturbed and control locations of 5.85 mm y^− 1 and 0.081 g dw y^− 1 for rhizome elongation and primary production, respectively. Thus, particular attention should be paid to the potential confounding effect of shoots age in analyses of impacts on growth performance of P. oceanica.     

The effects of bee (Apis mellifera) venom phospholipase A2 on Trypanosoma brucei brucei and enterobacteria

The potential role of phospholipases in trypanosomiasis was investigated using bee venom phospholipase A2 (bvPLA2) as a model. The effects of bvPLA2 on the survival of Trypanosoma brucei brucei, 2 h and 12 h cultures of Enterobacter cloacae, Escherichia coli, Citrobacter freundii were studied. About 1 mg ml⁻¹ bvPLA2 was trypanocidal after 30 min. Some growth occurred at lower concentrations up to 2 h after treatment but viability decreased up to 8 h. Even very low concentrations of bvPLA2 (10⁻¹² mg ml⁻¹) had some trypanocidal activity. Bee venom PLA2 was bactericidal to 2 h bacterial cultures but bacteriostatic to 12 h ones. Minimum bactericidal concentrations were 10⁻⁵-10⁻⁶ mg ml⁻¹. The results showed that bvPLA2 had significant trypanocidal and antibacterial effects on Gram-negative bacteria. The relationship to events occurring during infection is discussed. Phospholipases may play a role in increased endotoxin levels in trypanosomiasis.     

Inter-population variability of leaf morpho-anatomical and terpenoid patterns of Pistacia atlantica Desf. ssp. atlantica growing along an aridity gradient in Algeria

Three Algerian populations of female Pistacia atlantica shrubs were investigated in order to check whether their terpenoid contents and morpho-anatomical parameters may characterize the infraspecific variability. The populations were sampled along a gradient of increasing aridity from the Atlas mountains into the northwestern Central Sahara.As evidenced by Scanning Electron Microscopy, tufted hairs could be found only on seedling leaves from the low aridity site as a population-specific trait preserved also in culture. Under common garden cultivation seedlings of the high aridity site showed a three times higher density of glandular trichomes compared to the low aridity site. Increased aridity resulted also in reduction of leaf sizes while their thickness increased. Palisade parenchyma thickness also increases with aridity, being the best variable that discriminates the three populations of P. atlantica.Analysis of terpenoids from the leaves carried out by GC-MS reveals the presence of 65 compounds. The major compounds identified were spathulenol (23 μg g⁻¹ dw), α-pinene (10 μg g⁻¹ dw), verbenone (7 μg g⁻¹ dw) and β-pinene (6 μg g⁻¹ dw) in leaves from the low aridity site; spathulenol (73 μg g⁻¹ dw), α-pinene (25 μg g⁻¹ dw), β-pinene (18 μg g⁻¹ dw) and γ-amorphene (16 μg g⁻¹ dw) in those from medium aridity and spathulenol (114 μg g⁻¹ dw), α-pinene (49 μg g⁻¹ dw), germacrene D (29 μg g⁻¹ dw) and camphene (23 μg g⁻¹ dw) in leaves from the high aridity site. Terpene concentrations increased with the degree of aridity: the highest mean concentration of monoterpenes (136 μg g⁻¹ dw), sesquiterpenes (290 μg g⁻¹ dw) and total terpenes (427 μg g⁻¹ dw) were observed in the highest arid site and are, respectively, 3-, 5- and 4-fold higher compared to the lower arid site. Spathulenol and α-pinene can be taken as chemical markers of aridity. Drought discriminating compounds in low, but detectable concentrations are δ-cadinene and β-copaene. The functional roles of the terpenoids found in P. atlantica leaves and principles of their biosynthesis are discussed with emphasis on the mechanisms of plant resistance to drought conditions.     

Continuous culture of the microalgae Schizochytrium limacinum on biodiesel-derived crude glycerol for producing docosahexaenoic acid

Crude glycerol is a major byproduct of the biodiesel industry; previous research has proved the feasibility of producing docosahexaenoic acid (DHA, 22:6 n − 3) through fermentation of the algae Schizochytrium limacinum on crude glycerol. The objective of this work is to investigate the cell growth kinetics, substrate utilization efficiency, and DHA production of the algae through a continuous culture. Steady-state biomass yield, biomass productivity, growth yield on glycerol, specific glycerol consumption rate, and fatty acid composition were investigated within the range of dilution rate (D) from 0.2 to 0.6 day⁻¹, and the range of feed crude glycerol concentration (S₀) from 15 to 120 g/L. The maximum specific growth rate was determined as 0.692 day⁻¹. The cells had a true growth yield of 0.283 g/g but with a relatively high maintenance coefficient (0.2216 day⁻¹). The highest biomass productivity of 3.88 g/L-day was obtained at D = 0.3 day⁻¹ and S₀ = 60 g/L, while the highest DHA productivity (0.52 g/L-day) was obtained at D = 0.3 day⁻¹ and S₀ = 90 g/L due to the higher DHA content at S₀ = 90 g/L. The biomass and DHA productivity of the continuous culture was comparable to those of batch culture, while lower than the fed-batch culture, mainly because of the lower DHA content obtained by the continuous culture. Overall, the results show that continuous culture is a powerful tool to investigate the cell growth kinetics and physiological behaviors of the algae growing on biodiesel-derived crude glycerol.     

In silico optimization and low structured kinetic model of poly[(R)-3-hydroxybutyrate] synthesis by Cupriavidus necator DSM 545 by fed-batch cultivation on glycerol

Glycerol was utilized by Cupriavidus necator DSM 545 for production of poly-3-hydroxybutyrate (PHB) in fed-batch fermentation. Maximal specific growth rates (0.12 and 0.3 h⁻¹) and maximal specific non-growth PHB production rate (0.16 g g⁻¹ h⁻¹) were determined from two experiments (inocula from exponential and stationary phase). Saturation constants for nitrogen (0.107 and 0.016 g L⁻¹), glycerol (0.05 g L⁻¹), non-growth related PHB synthesis (0.011 g L⁻¹) and nitrogen/PHB related inhibition constant (0.405 g L⁻¹), were estimated. Five relations for specific growth rate were tested using mathematical models. In silico performed optimization procedures (varied glycerol/nitrogen ratio and feeding) has resulted in a PHB content of 70.9%, shorter cultivation time (23 h) and better PHB yield (0.347 g g⁻¹). Initial concentration of biomass 16.8 g L⁻¹ and glycerol concentration in broth between 3 and 5 g L⁻¹ were decisive factors for increasing of productivity.     

Alkyl-methylimidazolium ionic liquids affect the growth and fermentative metabolism of Clostridium sp

In this study, the effect of ionic liquids, 1-ethyl-3-methylimidazolium acetate [EMIM][Ac], 1-ethyl-3-methylimidazolium diethylphosphate [EMIM][DEP], and 1-methyl-3-methylimidazolium dimethylphosphate [MMIM][DMP] on the growth and glucose fermentation of Clostridium sp. was investigated. Among the three ionic liquids tested, [MMIM][DMP] was found to be least toxic. Growth of Clostridium sp. was not inhibited up to 2.5, 4 and 4 g L⁻¹ of [EMIM][Ac], [EMIM][DEP] and [MMIM][DMP], respectively. [EMIM][Ac] at <2.5 g L⁻¹, showed hormetic effect and stimulated the growth and fermentation by modulating medium pH. Total organic acid production increased in the presence of 2.5 and 2 g L⁻¹ of [EMIM][Ac] and [MMIM][DMP]. Ionic liquids had no significant influence on alcohol production at <2.5 g L⁻¹. Total gas production was affected by ILs at ?2.5 g L⁻¹ and varied with type of methylimidazolium IL. Overall, the results show that the growth and fermentative metabolism of Clostridium sp. is not impacted by ILs at concentrations below 2.5 g L⁻¹.     

Vertical tubular photobioreactor for semicontinuous culture of Cyanobium sp

We evaluated the kinetic culture characteristics of the microalgae Cyanobium sp. grown in vertical tubular photobioreactor in semicontinuous mode. Cultivation was carried out in vertical tubular photobioreactor for 2 L, in 57 d, at 30 °C, 3200 Lux, and 12 h light/dark photoperiod. The maximum specific growth rate was found as 0.127 d⁻¹, when the culture had blend concentration of 1.0 g L⁻¹, renewal rate of 50%, and sodium bicarbonate concentration of 1.0 g L⁻¹. The maximum values of productivity (0.071 g L⁻¹ d⁻¹) and number of cycles (10) were observed in blend concentration of 1.0 g L⁻¹, renewal rate of 30%, and bicarbonate concentration of 1.0 g L⁻¹. The results showed the potential of semicontinuous cultivation of Cyanobium sp. in closed tubular bioreactor, combining factors such as blend concentration, renewal rate, and sodium bicarbonate concentration.     

Influence of ammonium sulphate feeding time on fed-batch Arthrospira (Spirulina) platensis cultivation and biomass composition with and without pH control

Previous work demonstrated that a mixture of NH₄Cl and KNO₃ as nitrogen source was beneficial to fed-batch Arthrospira (Spirulina) platensis cultivation, in terms of either lower costs or higher cell concentration. On the basis of those results, this study focused on the use of a cheaper nitrogen source mixture, namely (NH₄)₂SO₄ plus NaNO₃, varying the ammonium feeding time (T = 7-15 days), either controlling the pH by CO₂ addition or not. A. platensis was cultivated in mini-tanks at 30 °C, 156 μmol photons m⁻² s⁻¹, and starting cell concentration of 400 mg L⁻¹, on a modified Schlösser medium. T = 13 days under pH control were selected as optimum conditions, ensuring the best results in terms of biomass production (maximum cell concentration of 2911 mg L⁻¹, cell productivity of 179 mg L⁻¹ d⁻¹ and specific growth rate of 0.77 d⁻¹) and satisfactory protein and lipid contents (around 30% each).     

Effects of Bacillus thuringiensis toxin Cry1Ac and Beauveria bassiana on Asiatic corn borer (Lepidoptera: Crambidae)

In this study, interactions between Cry1Ac, a toxic crystal protein produced by Bacillus thuringiensis (Berliner), and Beauveria bassiana on the mortality and survival of Ostrinia furnacalis was evaluated in the laboratory. The results showed that Cry1Ac is toxic to O. furnacalis. Not only were larval growth and development delayed, but pupation, pupal weight and adult emergency also decreased when larvae were fed on artificial diet containing purified Cry1Ac toxin. When third instars O. furnacalis were exposed to combination of B. bassiana (1.8 × 10⁵, 1.8 × 10⁶ or 1.8 × 10⁷ conidia ml⁻¹) and Cry1Ac, (0.2 or 0.8 μg g⁻¹), the effect on mortality was additive, however, the combinations of sublethal concentrations showed antagonism between Cry1Ac (3.2 or 13 μg g⁻¹) and B. bassiana (1.8 × 10⁵ or 1.8 × 10⁶ conidia ml⁻¹). When neonates were reared on sublethal concentrations of Cry1AC until the third instar, and survivors exposed B. bassiana conidial suspension, such treatments showed additive effect on mortality of O. furnacalis except for the combination of Cry1Ac (0.2 μg g⁻¹) and B. bassiana (1.8 × 10⁶ conidia ml⁻¹) that showed antagonism.     

Nitrogen and phosphorus effect on Typha domingensis Presl. rhizome growth in a matrix of Schoenoplectus americanus (Pers.) Volkart ex Schinz and Keller

In an outdoor mesocosm experiment of 80 weeks, the effect of nitrogen and phosphorus addition was tested on growth of Typha domingensis Presl. rhizomes in a matrix of Schoenoplectus americanus (Pers.) Volkart ex Schinz and Keller, under loading rates of 0.23 gm⁻² d⁻¹ of nitrogen, 0.17 gm⁻² d⁻¹ of P, both nutrient together and control conditions, to assess the potential expansion of T. domingensis in response to nutrient inputs.     

Long-term effects on the nitrogen budget of a short-rotation grey alder (Alnus incana (L.) Moench) forest on abandoned agricultural land

Short-rotation energy forestry is one of the potential ways for management of abandoned agricultural areas. It helps sequestrate carbon and mitigate human-induced climate changes. Owing to symbiotic dinitrogen (N₂) fixation by actinomycetes and the soil fertilizing capacity and fast biomass growth of grey alders, the latter can be suitable species for short-rotation forestry. In our study of a young grey alder stand (Alnus incana (L.) Moench) on abandoned arable land in Estonia we tested the following hypotheses: (1) afforestation of abandoned agricultural land by grey alder significantly affects the soil nitrogen (N) status already during the first rotation period; (2) input of symbiotic fixation covers an essential part of the plant annual N demand of the stand; (3) despite a considerable N input into the ecosystem of a young alder stand, there will occur no significant environmental hazards (N leaching or N₂O emissions). The first two hypotheses can be accepted: there was a significant increase in N and C content in the topsoil (from 0.11 to 0.14%, and from 1.4 to 1.7%, respectively), and N fixation (151.5 kg N ha⁻¹ yr⁻¹) covered about 74% of the annual N demand of the stand. The third hypothesis met support as well: N₂O emissions (0.5 kg N ha⁻¹ yr⁻¹) were low, while most of the annual gaseous N losses were in the form of N₂ (73.8 kg N ha⁻¹ yr⁻¹). Annual average NO₃-N leaching was 15 kg N ha⁻¹ yr⁻¹ but the N that leached from topsoil accumulated in deeper soil layers. The soil acidifying effect of alders was clearly evident; during the 14-year period soil acidity increased 1.3 units in the upper 0-10 cm topsoil layer.     

Sorgoleone,a sorghum root exudate: Algicidal activity and acute toxicity to the ricefish Oryzias latipes

The discovery of natural and natural-based compounds has resulted in its application as an alternative to synthetic algicides to control harmful algae in aquatic systems. Of the many natural-product-based algicides, sorgoleone, a natural plant product from Sorghum bicolor root exudates has been investigated for its controlling effect on different algal species and its acute fish toxicity. Growth of the blue green algal species Microcystis aeruginosa Kützing was completely inhibited by the crude methanol extract of sorghum root at 20 μg mL⁻¹. The most noticeable inhibition was observed in the bioassay of n-hexane soluble extract, where 98% growth inhibition occurred in M. aeruginosa at the concentration of 1.25 μg mL⁻¹. Sorgoleone very effectively controlled blue green algae inhibiting 97% of M. aeruginosa at 0.5 μg mL⁻¹ and 99% of Anabaena affinis Lemmermann at 4 μg mL⁻¹. In contrast, inhibition of the green algae species Chlorella vulgaris Beijerinck and Scenedensmus spp. at 16 μg mL⁻¹ sorgoleone was 87 and 68%, respectively. There were no mortalities or adverse effects observed in any of the fish exposed to water control, solvent control, and a nominal concentration of 1 μg mL⁻¹ during the test period. The no observed effect concentration (NOEC) value was 1.5 μg mL⁻¹ for the tested fish (O. latipes). Sorgoleone can be considered as an effective and an ecologically and environmentally sustainable approach to controlling harmful algae.