Molybdate reduction by Pseudomonas sp. strain DRY2

Aims: To isolate and characterize a potent molybdenum‐reducing bacterium. Methods and Results: A minimal salt medium supplemented with 10 mmol l^?1 molybdate, glucose (1·0%, w/v) as a carbon source and ammonium sulfate (0·3%, w/v) as a nitrogen source was used in the screening process. A molybdenum‐reducing bacterium was isolated and tentatively identified as Pseudomonas sp. strain DRY2 based on carbon utilization profiles using Biolog GN plates and partial 16S rDNA molecular phylogeny. Strain DRY2 produced 2·4, 3·2 and 6·2 times more molybdenum blue compared to Serratia marcescens strain DRY6, Enterobacter cloacae strain 48 and Eschericia coli K12, respectively. Molybdate reduction was optimum at 5 mmol l^?1 phosphate. The optimum molybdate concentration that supported molybdate reduction at 5 mmol l^?1 phosphate was between 15 and 25 mmol l^?1. Molybdate reduction was optimum at 40°C and at pH 6·0. Phosphate concentrations higher than 5 mmol l^?1 strongly inhibited molybdate reduction. Inhibitors of electron transport system such as antimycin A, rotenone, sodium azide and cyanide did not inhibit the molybdenum‐reducing enzyme activity. Chromium, copper, mercury and lead inhibited the molybdenum‐reducing activity. Conclusions: A novel molybdenum‐reducing bacterium with high molybdenum reduction capacity has been isolated. Significance and Impact of the Study: Molybdenum is an emerging global pollutant that is very toxic to ruminants. The characteristics of this bacterium suggest that it would be useful in the bioremediation of molybdenum pollutant.     

Blood lactate loads of redthroat emperor Lethrinus miniatus associated with angling stress and exhaustive exercise

Baseline, post‐angling and maximum attainable blood lactate concentrations were measured for the fishery species redthroat emperor Lethrinus miniatus to gain insight into the condition of fish released following c. 30 s angling and <45 s air exposure. Mean ± s.d . baseline blood lactate was 1·5 ± 0·6 mmol l^?1, which increased and plateaued around 6 mmol l^?1 at 15–30 min post‐angling. These values were significantly lower than those obtained from fish maximally exhausted with a prolonged chase and air exposure protocol following capture (10·9 ± 1·8 mmol l^?1), suggesting that L. miniatus is not maximally exhausted during standard angling practices.     

Comparison of supplements* to enhance recovery of heat‐injured Salmonella from egg albumen

Aims: The purpose of this study was to determine the proficiency of supplements to enhance the recovery of Salmonella from heat‐treated liquid egg albumen on solid agar media. Methods and Results: Salmonella‐inoculated albumen, heated at 53·3°C for 4 min, was plated on 39 combinations of solid media with or without the addition of 12 supplements. Greater numbers of Salmonella (P < 0·05) recovered with the addition of 1·0 g l^?1 ferrous sulfate (FeSO₄) than with any other supplements, except for 0·5 or 1·0 g l^?1 3′3′‐thiodipropionic acid (TDP), which recovered equivalent populations. Addition of 1·0 g l^?1 sodium pyruvate or 6·0 g l^?1 yeast extract plus 1·0 g l^?1 sodium pyruvate supported greater resuscitation than unsupplemented tryptic soy agar (TSA) or supplementing with 0·01 or 0·1 g l^?1 N‐propyl gallate, 10 g l^?1 activated charcoal, 0·1 g l^?1 KMnO₄ or 50 mg l^?1 ethoxyquin. The remaining supplements supported recovery of equivalent numbers of Salmonella, which were fewer cells than recovered with 1·0 g l^?1 FeSO₄, yet greater populations than recovered with 50 mg l^?1 ethoxyquin. Conclusion: Supplementation of plating media with FeSO₄, TDP or sodium pyruvate enhanced recovery of sublethally injured Salmonella from albumen. Significance and Impact of the Study: Pasteurizing albumen impedes recovery of pathogens. These results suggest that the addition of supplements to plating media may assist resuscitation and colony development of heat‐injured salmonellae.     

Arthrospira platensis biomass with high protein content cultivated in continuous process using urea as nitrogen source

Aims: Arthrospira platensis has been studied for single‐cell protein production because of its biomass composition and its ability of growing in alternative media. This work evaluated the effects of different dilution rates (D) and urea concentrations (N₀) on A. platensis continuous culture, in terms of growth, kinetic parameters, biomass composition and nitrogen removal. Methods and results: Arthrospira platensis was continuously cultivated in a glass‐made vertical column photobioreactor agitated with Rushton turbines. There were used different dilution rates (0·04–0·44 day^?1) and urea concentrations (0·5 and 5 mmol l^?1). With N₀ = 5 mmol l^?1, the maximum steady‐state biomass concentration was1415 mg l^?1, achieved with D = 0·04 day^?1, but the highest protein content (71·9%) was obtained by applying D = 0·12 day^?1, attaining a protein productivity of 106·41 mg l^?1 day^?1. Nitrogen removal reached 99% on steady‐state conditions. Conclusions: The best results were achieved by applying N₀ = 5 mmol l^?1; however, urea led to inhibitory conditions at D ≥ 0·16 day^?1, inducing the system wash‐out. The agitation afforded satisfactory mixture and did not harm the trichomes structure. Significance and Impact of the Study: These results can enhance the basis for the continuous removal of nitrogenous wastewater pollutants using cyanobacteria, with an easily assembled photobioreactor.     

Vanadate enhances insulin-receptor binding in gestational diabetic human placenta

Although vanadium is found abundantly in animal and plant kingdoms its biological effects are not clear. Vanadate compounds have been shown to normalize blood glucose levels in streptozotocin treated rats, enhance glucose oxidation and improve the sensitivity to insulin by enhanced receptor binding in rat adipocytes. The aim of the present study was to investigate the effect of vanadate, at high (0–8 mmol l^?1) and low (0–1·0 mmol l^?1) physiological concentrations, on [¹²⁵I]-insulin binding in the placenta of three groups of pateints, namely from normal (N) controls, gestational diabetics (GDM) and women with risk factors in their medical history for developing diabetes mellitus (RF). Vanadate at low concentrations (0·2–0·6 mmol l^?1) enhanced the maximal binding 2-fold in GDM placenta but only increased (up to 1·2-fold) the binding slightly at high cncentrations (5 mmol l^?1). However with placenta from normal or women at risk, vanadate increased the [¹²⁵I]-insulin binding up to 1·2-fold both at low and high concentrations. Thus it appears that vanadate augements insulin binding in the placenta from women with gestational diabetes mellitus.     

Study of laccase production by Pleurotus ostreatus in a 5 l bioreactor and application of the enzyme to determine the antioxidant concentration of human plasma

Aims: To achieve high laccase production from Pleurotus ostreatus in a bench top bioreactor and to utilize the enzyme for determination of the total antioxidant concentration (TAC) of human plasma. Methods and Results: Laccase production by P. ostreatus studied in a benchtop bioreactor was as high as, 874·0 U ml^?1 in presence of copper sulfate. The enzyme was used to replace metmyoglobin and hydrogen peroxide for the estimation of TAC in human plasma. The trolox equivalent antioxidant concentrations determined by the laccase‐based method and metmyoglobin method ranged from 1·63 ± 0·011 to 1·80 ± 0·006 mmol l^?1 and from 1·41 ± 0·004 to 1·51 ± 0·008 mmol l^?1 plasma, respectively. Conclusions: Pleurotus ostreatus produced high amount of extracellular laccase in a benchtop bioreactor. The enzyme can be used to assay TAC of blood plasma without the interference encountered with the hydrogen peroxide and metmyoglobin mediated assay method. Significance and Impact of the Study: Laccase production by P. ostreatus obtained in this study was the highest among all reported laccase producing white‐rot fungi. Moreover, an accurate laccase‐based assay method was developed for detection of TAC in human plasma.     

Effect of the cathode potential and sulfate ions on nitrate reduction in a microbial electrochemical denitrification system

Recently, bioelectrochemical systems have been demonstrated as advantageous for denitrification. Here, we investigated the nitrate reduction rate and bacterial community on cathodes at different cathode potentials [?300, ?500, ?700, and ?900 mV vs. standard hydrogen electrode (SHE)] in a two-chamber microbial electrochemical denitrification system and effects of sulfate, a common nitrate co-contaminant, on denitrification efficiency. The results indicated that the highest nitrate reduction rates (3.5 mg L^?1 days^?1) were obtained at a cathode potential of ?700 mV, regardless of sulfate presence, while a lower rate was observed at a more negative cathode potential (?900 mV). Notably, although sulfate ions generally inhibited nitrate reduction, this effect was absent at a cathode potential of ?700 mV. Polymerase chain reaction–denaturing gradient gel electrophoresis revealed that bacterial communities on the graphite-felt cathode were significantly affected by the cathode potential change and sulfate presence. Shinella-like and Alicycliphilus-like bacterial species were exclusively observed on cathodes in reactors without sulfate. Ochrobactrum-like and Sinorhizobium-like bacterial species, which persisted at different cathode potentials irrespective of sulfate presence, were shown to contribute to bioelectrochemical denitrification. This study suggested that a cathode potential of around ?700 mV versus SHE would ensure optimal nitrate reduction rate and counteract inhibitory effects of sulfate. Additionally, sulfate presence considerably affects denitrification efficiency and microbial community of microbial electrochemical denitrification systems.     

Microbial sulfate reduction at low (8 °C) temperature using waste sludge as a carbon and seed source

Biological treatment of sulfate and metal-containing wastewater (such as acid mine drainage) is a viable option due to lower cost and better sludge quality compared to conventional chemical treatment. Although several substrates can be used as carbon source, a low-cost substrate is required for large scale applications. This study was conducted to investigate the suitability of waste sludge as a carbon and seed source for sulfate reduction at 8 °C in batch bioassays. Around 7 mmol of sulfate was reduced when the waste sludge mixture (WS) (6700 mg SS l^?1) from primary and secondary settling tank was supplemented as a carbon and seed source. However, only 1.6 mmol of sulfate was reduced with anaerobic digester effluent (ADS) (5300 mg SS l^?1). The produced H₂S from 1 g VSS l^?1 WS and ADS oxidation can theoretically precipitate around 90 and 35 mg Fe²⁺, respectively. Both WS and ADS oxidized ethanol to acetate at similar rates. It appears that WS is a good candidate for carbon and start-up seed source of sulfate reduction at 8 °C, whereas sulfidogenic acetate oxidation was the limiting step. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed that both sludge sources contain Desulfomicrobium apsheronum strain.     

Isolation and characterization of butachlor‐catabolizing bacterial strain Stenotrophomonas acidaminiphila JS‐1 from soil and assessment of its biodegradation potential

Aims: Isolation, characterization and assessment of butachlor‐degrading potential of bacterial strain JS‐1 in soil. Methods and Results: Butachlor‐degrading bacteria were isolated using enrichment culture technique. The morphological, biochemical and genetic characteristics based on 16S rDNA sequence homology and phylogenetic analysis confirmed the isolate as Stenotrophomonas acidaminiphila strain JS‐1. The strain JS‐1 exhibited substantial growth in M9 mineral salt medium supplemented with 3·2 mmol l^?1 butachlor, as a sole source of carbon and energy. The HPLC analysis revealed almost complete disappearance of butachlor within 20 days in soil at a rate constant of 0·17 day^?1 and half‐life (t_½) of 4·0 days, following the first‐order rate kinetics. The strain JS‐1 in stationary phase of culture also produced 21·0 μg ml^?1 of growth hormone indole acetic acid (IAA) in the presence of 500 μg ml^?1 of tryptophan. The IAA production was stimulated at lower concentrations of butachlor, whereas higher concentrations above 0·8 mmol l^?1 were found inhibitory. Conclusions: The isolate JS‐1 characterized as Stenotrophomonas acidaminiphila was capable of utilizing butachlor as sole source of carbon and energy. Besides being an efficient butachlor degrader, it substantially produces IAA. Significance and Impact of the Study: The bacterial strain JS‐1 has a potential for butachlor remediation with a distinctive auxiliary attribute of plant growth stimulation.     

Role of casein on induction and enhancement of production of a bacterial milk clotting protease from an indigenously isolated Bacillus subtilis

Aims: To isolate and enhance the yield of a bacterial milk clotting protease (MCP) through process optimization and scale up. Materials and Results: Bacillus subtilis was isolated as MCP producer with good milk clotting activity (MCA) per proteolytic activity (PA) index. The enzyme production was inducible with casein and enhanced with fructose and ammonium nitrate resulting in 571·43 U ml^?1 of enzyme. Conclusions: Medium containing 4% fructose, 0·75% casein, 0·3% NH₄NO₃ and 10 mmol l^–1 CaCl₂, pH 6·0, inoculated with 4% (v/v) inoculum, incubated at 37°C, 200 rev min^?1 for 72 h gave maximum production. A 6·67‐fold increase in MCP yield with very high MCA per PA index was observed after final optimization indicating similarity to rennets. Significance and Impact of the Study: Mostly fungal MCPs have been reported. The MCA and MCA per PA index of this bacterium is comparable to that of many fungal reports and better than quite a few bacterial MCPs. Thus, this enzyme by B. subtilis has good probability of successful use in cheese production.     

Modelling the inhibitory effect of copper sulfate on the growth of Penicillium expansum and Botrytis cinerea

Aims: This study aimed to investigate the effect of copper sulfate (from 0 to 8 mmol kg^?1) on radial growth rate and lag time of two moulds responsible for vine grapes spoilage: Penicillium expansum strain 25·03 and Botrytis cinerea, strains BC1 and BC2. Methods and results: A new model was developed to describe tailing and shoulders in the inhibition curves. Because of tailing, the minimum inhibitory concentration (MIC), was not defined as the concentration at which no growth was observed, but as the concentration at which the lag time was infinite. The concentrations at which μ = μ_opt/2, (Cu₅₀), were in the range of 2·2–2·6 mmol kg^?1. Radial growth rate of P. expansum and the reciprocal of the lag time were linearly correlated (r = 0·84). In contrast, in the range 0–4 mmol kg^?1, an inhibition of growth of B. cinerea was observed whereas germination remained unaffected (i.e. the lag time was constant). In the range 4–8 mmol kg^?1, the radial growth rate of B. cinerea was almost constant (c. 1 mm day^?1), but germination was inhibited (i.e. the lag time was increased). Conclusions: The MIC values were 4·7 mmol kg^?1 for P. expansum, 8·2 and 7·3 mmol kg^?1 for B. cinerea strain BC1 and BC2, respectively, demonstrating that some isolates of these moulds are resistant to copper. Significance and Impact of the Study: Copper concentrations at 4 mmol kg^?1 would be sufficient to control the development of these isolates, but the toxicity of copper should be extended to other isolates and evaluated in vineyards.     

Production of propionic acid by microbiological way. Part 1. Influence of the initial sugars and product concentrations

The best yields and productivities of 0.38 g · g^?1 and 0.35 g · l^?1 h^?1, respectively, for the propionic acid production in a batchsystem using sugar-cane final molasses as carbon source were obtained when an initial TRS concentration of 50 g · l^?1 was used. It was obvious that this process is severely inhibited by the acids produced and the most drastic effect (μ = 0) was at a TVA concentration near to 250 mmol · l^?1, independently of the initial TRS concentration employed. A generalizated equation of noncompetitive inhibition was adjusted: and kinetic inhibition constants for each initial TRS concentration studied were estimated.     

Isolation and characterization of actinomycetes from healthy goat faeces

Aims: To isolate and characterize actinomycetes with probiotic activities from healthy goat faeces. Methods and Results: Faecal actinomycetes were isolated by dilution methods and identified by 16S rRNA gene sequence analysis. The hydrolytic enzyme activities were analysed by clear zone formation. The antimicrobial activities and resistance to heavy metals were tested by growth inhibition methods. The isolates belong to a small group of actinobacterial genera, including Streptomyces, Nocardiopsis and Oerskovia. The Oerskovia was the most widely distributed genus among the cultures. The proportion of streptomycete‐like strains producing amylase or protease is significantly higher than those of other actinomycetes (P < 0·05). Compared with streptomycete‐like strains, a higher proportion of (α‐ or β‐) galactase‐producing other actinomycetes was found in goat faeces. More than 50% of streptomycete‐like strains showed activities against test fungi. Streptomycetes could tolerate 0·25 mmol l^?1 Cr₂O₇^2?, 2 mmol l^?1 Ni²⁺; however, other actinomycetes are liable to 40 mmol l^?1 Fe³⁺ and 0·25 mmol l^?1 Cr₂O₇^2? and resistant to 5 mmol l^?1 Ni²⁺ and 2 mmol l^?1 Cu²⁺. Conclusions: The different physiological characteristics of the actinomycetes suggested that the cooperation in the actinomycetes might be involved in their association with goat. Significance and Impact of the Study: Probiotic mixtures based on faecal actinomycetes showed potentials in animal production.     

Inhibitory effects of silver ions on Legionella pneumophila grown on agar, intracellular in Acanthamoeba castellanii and in artificial biofilms

Aims: We undertook a series of experiments to investigate the susceptibility of Legionella pneumophila grown under extracellular and intracellular conditions and other water‐related bacteria to silver ions. Methods and Results: In this study, the antimicrobial effect of silver ions to intra‐ and extra‐cellular grown Legionella bacteria was investigated. The minimal inhibitory concentration (MIC) after 24 h exposure, leading to a 5 log reduction, was c. 64 μg l^?1 AgNO₃ for extracellular grown Legionella and other tested Gram‐positive and Gram‐negative bacteria. In contrast, the MIC for intracellularly grown Legionella was up to 4096 μg l^?1 AgNO₃ after 24 h. Furthermore, the heterotrophic bacteria grown within a biofilm model were killed at a concentration of 4–16 μg l^?1 AgNO₃. In contrast, biofilm‐associated Legionella were less sensitive (MIC 128–512 μg l^?1 AgNO₃). Conclusion: Intracellularly and biofilm‐grown legionellae are less sensitive against silver compared with agar‐grown bacteria. Significance and Impact of the Study: The reduced sensitivity of Legionella grown in amoebae might explain why the effect of silver decontamination requires an extended exposure in field trials.     

Agrobacterium-mediated transformation of Eruca sativa

An Agrobacterium tumefaciens—mediated transformation system was developed for Eruca sativa (eruca). Hypocotyl explants were co-cultivated with bacterial cells carrying a plasmid harboring a uidA:nptII fusion gene along a phosphinothricin acetyl transferase (PAT) gene cassette, for a period of 2 days. These were grown on a high cytokinin/auxin medium containing 5.0 mg l^?1 6-benzyladenine (BA), 1.0 mg l^?1 indole-3-acetic acid (IAA), and 0.1 mg l^?1 α-naphthaleneacetic acid (NAA). Explants were then transferred to a lower cytokinin/auxin medium containing 2.0 mg l^?1 BA and 0.1 mg l^?1 NAA along with 5.0 mg l^?1 silver nitrate and 300 mg l^?1 Timentin®. Upon transfer to a selection medium containing either 20 mg l^?1 kanamycin or 2 mg l^?1 L-phosphinothricin (L-ppt), shoot regenerants were observed. Expression of the transgenes in putative transformants was confirmed using a histochemical GUS assay. Presence of the PAT transgene in GUS-positive T0 plants was confirmed by Southern blot analysis. Moreover, spot tests of T1 seedlings were conducted using the L-ppt herbicide. A transformation frequency of 1.1% was obtained with more than 60% of transgenic lines containing single copies of the transgenes.     

In situ ammonia production as a sanitation agent during anaerobic digestion at mesophilic temperature

Aim: To measure the sanitizing effect of mesophilic (37°C) anaerobic digestion in high ammonia concentrations produced in situ. Methods and Results: Indicator organisms and salmonella were transferred to small‐scale anaerobic batch cultures and D‐values were calculated. Batch cultures were started with material from two biogas processes operating at high (46 mmol l^?1) and low (1·6 mmol l^?1) ammonia concentration. D‐values were shortened from c. 3 days to <1 day for the bacteria. MS2 had the same D‐value (1·3 days) independent of ammonia concentration whereas ΦX174 and 28B were faster inactivated in the control (1·1 and 7·9 days) than in the high ammonia (8·9 and 39 days) batch cultures. Conclusion: Running biogas processes at high levels of ammonia shortens the time to meet EU regulation concerning reduction of salmonella and enterococci (5 log). Unless a minimum retention time of 2 days, post‐treatment digestion is needed to achieve sufficient sanitation in continuous biogas processes. Significance and Impact of the Study: Running mesophilic biogas processes at high ammonia level produces residue with a high fertilizer value. With some stipulations concerning management parameters, such processes provide a method of bacterial sanitation without preceding pasteurization of the incoming organic waste.     

Tolerance and Immobilization of Cobalt by Some Bacteria from Ferromanganese Crusts of the Afanasiy Nikitin Seamounts

Bacteria isolated from cobalt–enriched ferromanganese crusts on the Afanasiy Nikitin Seamounts in the Equatorial Indian Ocean were examined for their ability to tolerate, and immobilize cobalt in unamended seawater and seawater amended with 0.01% glucose. Retrievable bacterial counts in the form of CFU (colony forming units) on media supplemented with 1 mmol Co l^?1 (58 mg Co l^?1) and 1 mmol Mn l^?1 (54 mg Mn l^?1) were in the range of 1.71 × 10⁴ to 1.05 × 10⁵ gm^?1 (wet wt) of crust, respectively. Most of the isolates (14/24) were pigmented and showed taxonomic affinities to Flavobacterium sp. Two representative isolates were tested for their tolerance of cobalt. We observed that in amended medium, the isolates tolerated up to 1 mmol Co l^?1, whereas in unamended medium they tolerated upto 10 mmol Co l^?1. Microscopic observations of cultures incubated with 10 mmol Co l^?1 showed the occurrence of an extracellular slime layer, which may be responsible for immobilizing the cobalt from the liquid phase. In the unamended medium, the tolerance and stimulation in total cell counts was similar to that in amended medium or sometimes greater. Total cell counts peaked at 100 μmol Co l^?1 for incubations in unamended medium (1.1–2.5 × 10¹¹ cells l^?1) and at 0.1–1 μmol Co l^?1 for incubations in amended medium (1.5–2.6 × 10¹¹ cells l^?1). Counts of formazan-stained respiring cells of both the isolates in the unamended medium reached up to a maximum of 2.9–7.8 × 10¹⁰ l^?1 after incubation for 10 days at 23(±1)°. In the amended medium cell counts of respiring cells attained a maximum in the range of 4.6–15.8 × 10¹⁰ l^?1 at 100 μmol Co l^?1. The Co immobilization rate was on average 82 (± 87.9, n = 24) μmol of Co d^?1. Since the isolates were naturally occurring bacteria from crusts, they could be more environmentally acceptable and safe if used for metal recovery and bio-leaching.     

Selective production of acetone during continuous synthesis gas fermentation by engineered biocatalyst Clostridium sp. MAceT113

Aims: To engineer acetogen biocatalyst capable of fermenting synthesis gas blend to acetone as the only liquid carbonaceous product. Methods and Results: The metabolic engineering comprised inactivation of phosphotransacetylase via integration of a cassette comprising synthetic genes erm(B), thiolase and HMG‐CoA synthase. Acetaldehyde dehydrogenase was inactivated via integration of a cassette consisting of synthetic genes cat, HMG‐CoA lyase and acetoacetate decarboxylase. The engineered biocatalyst Clostridum sp. MAceT113 lost production of 253 mmol l^?1 ethanol and 296 mmol l^?1 acetate and started producing 1·8 mol l^?1 acetone in single‐stage continuous syngas fermentation. Conclusions: The acetone concentration in culture broth is economical for bulk manufacture because it is about twenty times of that achieved with known acetone–butanol–ethanol fermentation of sugars. Significance and Impact of the Study: The process shows the opportunity to produce acetone from synthesis gas at concentrations comparable with production of acetone from products of petroleum cracking. This is the first report on elimination of acetate and acetaldehyde production and directing carbon flux from Acetyl‐CoA to acetone via a non‐naturally occurring in acetogen acetone biosynthesis pathway identified in eukaryotic organisms.     

Nitrate and nitrite in interstitial waters of eelgrass beds in relation to the rhizosphere

The distribution of nitrate and nitrite in the interstitial water of the sediment of eelgrass (Zostera marina) bed of Izembek Lagoon, Alaska, were investigated. Their concentrations were relatively high (0 to 9.8 μg-at.N·1^?1, average 4.8 for nitrate; 0 to 4.0 μ-at.N·1^?1, average 1.9 for nitrite) although the sediments were anoxic and contained hydrogen sulphide. The rates of bacterial denitrification measured by ¹⁵N tracer technique ranged from 0.49×10^?10 to 1.2 × 10^?9 g-atN·g^?1·h^?1. When a steady state is maintained, the loss of nitrate and nitrite must be balanced by their production by bacterial nitrification. Experimentally determined rate of nitrification in the sediment was of the same order. A model experiment demonstrated that oxygen is transported from leaves to rhizomes and roots of eelgrass and released into the sediment. The oxygen is used for nitrification in the rhizosphere in anoxic sediments.     

Inorganic carbon uptake by phytoplankton in Vineyard Sound,Massachusetts. II. Comparative primary productivity and nutritional status of winter and summer assemblages

Using time-course, natural-light incubations, we assessed the rate of carbon uptake at a range of light intensities, the effect of supplemental additions of nitrogen (as NH₄⁺ or urea) on light and dark carbon uptake, and the rates of uptake of NH₄⁺ and urea by phytoplankton from Vineyard Sound, Massachusetts from February through August 1982. During the winter, photoinhibition was severe, becoming manifested shortly after the start of an incubation, whereas during the summer, there was little to no evidence of photoinhibition during the first several hours after the start of an incubation. At light levels which were neither photoinhibiting nor light limiting, rates of carbon uptake normalized per liter were high and approximately equal during winter and summer (22–23 μg C·l^?1 · h^?1), and low during spring (<10 μgC·l^?1· h^?1). In contrast, on a chlorophyll a basis, rates of carbon fixation were as high during spring (15–20μg C·μg Chl a^?1·h^?1), when concentrations of chlorophyll a were at the yearly minimum (<0.5 μg · l^?1) as during the summer, when chlorophyll a concentrations were substantially higher (0.8–1.3 μg · l^?1). Highest rates of NH₄⁺ and urea uptake were observed during summer, and at no time of the year was there evidence for severe nitrogen deficiency, although moderate nitrogen nutritional stress was apparent during the summer months.