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Long-term exposure to primary air pollutants, such as sulphur dioxide (SO2) and nitrogen oxides (NOx), alters the structure and functions of forest ecosystems. Many biochemical and biogeochemical processes discriminate against the heavier isotopes in a mixture; thus, the values of δ13C and δ15N (i.e. the ratio of stable isotopes 13C to 12C and that of 15 N to 14 N, respectively) may give insights into changes in ecosystem processes and identify the immediate drivers of these changes. We studied sources of variation in the δ13C and δ15N values in the foliage of eight boreal forest C3 plants at 10 sites located at the distance of 1–40 km from the Monchegorsk nickel-copper smelter in Russia. From 1939‒2019, this smelter emitted over 14,000,000 metric tons (t) of SO2, 250,000 t of metals, primarily nickel and copper, and 140,000 t of NOx. The δ13C value in evergreen plants and the δ15N value in all plants increased near the smelter independently of the plant mycorrhizal type. We attribute the pollution-related increase in the foliar δ13C values of evergreen species mainly to direct effects of SO2 on stomatal conductance, in combination with pollution-related water stress, which jointly override the potential opposite effect of increasing ambient CO2 concentration on δ13C values. Stomatal uptake of NOx and root uptake of 15N-enriched organic N compounds and NH4+ may explain the increased foliar δ15N values and elevated foliar N concentrations, especially in the evergreen trees (Pinus sylvestris), close to Monchegorsk, where the soil inorganic N supply is reduced due to the impact of long-term SO2 and heavy metal emissions on plant biomass. We conclude that, despite the uncertainties in interpreting δ13C and δ15N responses to pollution, the Monchegorsk smelter has imposed and still imposes a great impact on C and N cycling in the surrounding N-limited subarctic forest ecosystems.

peer-reviewed Opportunities for exploiting the inherent value of protein-rich meat processing co-products, in the context of increased global demand for protein and for sustainable processing systems, are discussed. While direct consumption maybe the most profitable route for some, this approach is influenced greatly by local and cultural traditions. A more profitable and sustainable approach may be found in recognizing this readily available and under-utilised resource can provide high value components, such as proteins, with targeted high value functionality of relevance to a variety of sectors. Applications in food & beverages, petfood biomedical and nutrition arenas are discussed. Utilization of the raw material in its entirety is a necessary underlying principle in this approach to help maintain minimum waste generation. Understanding consumer attitudes to these products, in particular when used in food or beverage systems, is critical in optimizing commercialization strategies. This work forms part of the ReValueProtein Research Project (Grant Award No. 11/F/043) which is supported by the Irish Department of Agriculture, Food and the Marine (DAFM) and the Food Institutional Research Measure (FIRM) both funded by the Irish Government under the National Development Plan 2007–2013. Department of Agriculture, Food and the Marine

Osmoregulation is by no means an energetically cheap process, and its costs have been extensively quantified in terms of respiration and aerobic metabolism. Common products of mitochondrial activity are reactive oxygen and nitrogen species, which may cause oxidative stress by degrading key cell components, while playing essential roles in cell homeostasis. Given the delicate equilibrium between pro- and antioxidants in fueling acclimation responses, the need for a thorough understanding of the relationship between salinity-induced oxidative stress and osmoregulation arises as an important issue, especially in the context of global changes and anthropogenic impacts on coastal habitats. This is especially urgent for intertidal/estuarine organisms, which may be subject to drastic salinity and habitat changes, leading to redox imbalance. How do osmoregulation strategies determine energy expenditure, and how do these processes affect organisms in terms of oxidative stress? What mechanisms are used to cope with salinity-induced oxidative stress? This Commentary aims to highlight the main gaps in our knowledge, covering all levels of organization. From an energy-redox perspective, we discuss the link between environmental salinity changes and physiological responses at different levels of biological organization. Future studies should seek to provide a detailed understanding of the relationship between osmoregulatory strategies and redox metabolism, thereby informing conservation physiologists and allowing them to tackle the new challenges imposed by global climate change.

The formation of acrylamide during the frying of potatoes is nowadays one of the major concerns of the potato-processing agriculture industry. Much research has been carried out to identify the acrylamide precursors in raw potatoes in order to minimize its formation during frying. Raw potatoes that give rise to an excess of acrylamide can currently not be detected in a fast, sensitive and non-destructive way. Therefore, we investigate the use of spatially-resolved spectroscopy to optically identify raw potatoes with high acrylamide precursors concentrations, on basis of their internal scattering properties. To obtain potatoes that induce high acrylamide concentrations, we stored the potatoes in a fridge at 4 °C. Measurements of the potatoes after different storage times (11, 15, 21 and 28 weeks), corresponding with different acrylamide concentrations in the French fries, show an evolution of the scatter pattern. Furthermore, when comparing the scatter properties at different wavelengths for potatoes giving rise to low (600 ppb) acrylamide concentrations during frying, an optimized classification of the potato batches was obtained at 1444 nm. We can conclude that the internal scatter properties of peeled, raw potatoes can be used to monitor the acrylamide precursors, enabling a non-destructive exclusion of the potatoes that are not suited for French fries production.

International audience; To sustain eel aquaculture, development of reproduction in captivity is vital. The aim of this review is to assess our current knowledge on the nutrition of broodstock eels in order to improve the quality of broodstock under farming conditions, drawing information from wild adult eels and other marine pelagic spawners. Freshwater eels spawn marine pelagic eggs with an oil droplet (type II), and with a large perivitelline space. Compared with other marine fish eggs, eel eggs are at the extreme end of the spectrum in terms of egg composition, even within this type II group. Eel eggs contain a large amount of total lipids, and a shortage of neutral lipids has been implied a cause for reduced survival of larvae. Eel eggs have higher ARA but lower EPA and DHA levels than in other fish. Too high levels of ARA negatively affected reproduction in the Japanese eel, although high levels of 18:2n-6 in the eggs of farmed eels were not detrimental. The total free amino acid amount and profile of eel eggs appears much different from other marine pelagic spawners. Nutritional intervention to influence egg composition seems feasible, but responsiveness of farmed eels to induced maturation might also require environmental manipulation. The challenge remains to succeed in raising European eel broodstock with formulated feeds and to enable the procurement of viable eggs and larvae, once adequate protocols for induced maturation have been developed

The quality of commercial cocoa beans, the principal raw material for chocolate production, relies on the combination of factors that include the type of planting material, the agricultural practices, and the post-harvest processing. Among these, the fermentation of the cocoa beans is still the most relevant since it is the process whereby the precursors of the cocoa flavor arise. The formation of these precursors depends on the activity of different microbial groups on the beans pulp. A comparison of fermentations in different countries showed that a well-defined microbial succession does not always take place and that the role of Bacillus spp. in this process remains unclear. Considering the overriding importance of the fermentation to achieve high quality commercial cocoa beans, we discuss the need of addressing the impact of the farming system, the ripeness state of the pods, and the role of microbial interactions on the fermentation in future research. In addition, the problem of high acidification cocoa beans, aspects dealing with the volatile fraction of the flavor, and the cocoa butter properties, all were identified as critical aspects that need further investigation. The standardization of the microbiological methods and the application of metagenomic approaches would magnify the knowledge in this domain.

This report produced by an independent Expert Group is the second commissioned by the ILSI Europe Task Force on Process-related Compounds. The first report, which reviewed human exposure and internal dose assessments of acrylamide in foods, stressed the need for comparison of added risks or reduced benefits ν. the benefit of acrylamide reduction. This second Expert Group was established to examine and report on the considerations for risk-benefit analysis of the mitigation measures on acyrylamide in commodities which contribute significantly to acrylamide exposure, and their impact on safety, quality, nutritional and organoleptic parameters; to position the relevance of the suspected side effects and to quantify their potential impact. The Expert Group was asked to focus on the impact of mitigation on exposure scenarios and the risk to human health using multidimensional risk assessment approaches. The report has four main objectives: 1. To summarise the impact of pre-harvest, post-harvest and processing conditions on acrylamide formation in potatoes, cereals and coffee. 2. To evaluate the impact of pre-harvest, post-harvest and processing conditions on the formation of acrylamide in these commodities. 3. To consider the nutritional value and beneficial health impact of consuming these commodities. 4. To calculate the impact of mitigation using probabilistic risk-benefit modelling to demonstrate the principle of this approach. Animal studies have shown that acrylamide is a genotoxic carcinogen. Under the premise that for genotoxic carcinogens no threshold levels of effect exists, exposure is to be minimized as far as possible. Minimization strategies therefore have been and consistently are developed, although direct evidence of carcinogenic effects in humans as a result of acrylamide intake is not available. The margin of exposure (MOE), representing the ratio between a defined point on the relevant dose-response curve, the bench mark dose lower limit (BMDL, 300 μg/kg body weight (bw) per day) and average (1 μg/kg bw per day) or high (4 μg/kg bw per day) consumer exposure has been estimated at 300 and 75, respectively. According to the European Food Safety Authority (EFSA) and the Joint FAO/WHO Expert Committee on Food Additives (JECFA) a MOE of ≥ 10 000 would indicate low concern, underlining that the relatively low MOE for acrylamide calls for rapid and effective mitigation measures in order to lower consumer exposure. Acrylamide production in foods is a consequence of the Maillard reaction between asparagine and reducing sugars. Therefore, factors affecting the concentration of these precursors in food, together with processing conditions during food preparation and storage will affect the final concentration of acrylamide in food. The commodities giving the highest exposure to acrylamide in the western diet are potatoes, cereals and coffee. The first two of these are staple foods providing for the majority of carbohydrate and some protein in the diet. Coffee is not an essential food, but has a high level of intake as a beverage, chosen not only for its flavour and hedonic value but also as a provider of caffeine. Acrylamide formation in foodstuffs depends on the concentrations and availability of the precursor molecules asparagine and reducing sugar(s).

3-N-oxalyl-L-2,3-diaminopropanoic acid (beta-ODAP) is a neuroexcitatory non- protein amino acid identified in grass pea (Lathyrus sativus L.) and in ginseng (Panax spp). Its presence in the drought tolerant grass pea is thought to be responsible for the crippling disease neurolathyrism, while in the longevity promoting ginseng it is named dencichine and praised for its haemostatic properties. Some properties of beta-ODAP are put in perspective.

We tested the hypothesis that in the euphotic zone of oligotrophic oceanic waters, the growth-limiting nutrients for bacterioplankton vary over a diel cycle due to competition with phyto- plankton, leading to distinct diel patterns in bacterioplankton. During a cruise across the subtropical Atlantic Ocean, a distinct diel periodicity in bacterioplankton was detectable, with highest bacterial abundance in the early morning and declining over the day. A peak in the frequency of dividing cells (FDC, 20 to 25% of the total bacterial community) was recorded at midnight; FDC rates were consis- tently low during the day and cell volume increased over the day until the early night period. No diel variations in these parameters were detectable in the layers below the deep chlorophyll maximum (DCM). Bio-assay experiments with freshly collected, N+P amended water were started in the early morning. No significant bacterial growth was detectable in the early morning while, in the bio-assays that started at noon, significant bacterial growth was detectable in the N+P treatments. P amendment alone also caused an increase in bacterial abundance although to a lesser extent. Thus, we have evi- dence that the bacterioplankton in the euphotic layer of the subtropical Atlantic are not limited by the availability of N+P in the morning (most likely limited in C) but at noon when they are competing with phytoplankton for the inorganic N+P sources. Based on the concomitantly measured decline in inorganic N+P in the bio-assays, we estimated the C:N:P ratio of newly produced bacteria. The mean atomic C:N:P ratio for the bacteria in the N+P amended bio-assay experiments started in the early morning was C:N:P = 15:13:1 while the corresponding ratio for bacteria in the bio-assays started at noon was C:N:P = 118:11:1. This diel variation in the C:N:P ratio was caused by the 1 order of mag- nitude higher bacterial C production at noon as compared to the early morning. Thus, excess uptake of N+P occurs during the morning and luxury C uptake during noon. This plasticity in the C:N:P ratio of bacterioplankton might be a strategy to overcome, at least partly, periods of shortage in a specific nutrient species in the oligotrophic subtropical Atlantic.

Feta cheeses (five trials) of different sodium content were made, using ewes' milk, from split lots of curd by varying the salting procedure, i.e. dry salting with NaCl (control) or mixtures of NaCl/KCl(3:1 or 1:1, w/w, basis) and filling the cans with brine made with NaCl or the above NaCl/KCl mixtures, respectively. in order to study the influence of the partial substitution of NaCl by KCl on the proteolysis during cheese ripening. The extent and characteristics of proteolysis in the cheeses were monitored during aging by using Kjeldahl determination of soluble nitrogen fractions (water-soluble nitrogen, trichloroacetic acid-soluble nitrogen, phosphotungstic acid-soluble nitrogen), the cadmium-ninhydrin method for the determination of total free amino acids (FAA), urea-polyacrylamide gel electrophoresis of cheese proteins followed by densitometric analysis of the chi (s1)- and beta -casein fractions, reverse-phase HPLC analysis of the water-soluble extracts of cheeses, and ion-exchange HPLC analysis of FAA. The results showed that proteolysis was similar in control and experimental cheeses at all sampling ages, indicating that the partial substitution of NaCl by KCI in the manufacture of Feta cheese had no significant effect on the extent and characteristics of proteolysis during cheese aging. (C) 2001 Elsevier Science Ltd. All rights reserved. International Dairy Journal

Dry deposition of NH$and NOx(NO and NO#) can affect plant metabolism at the cellular and whole-plant level.Gaseous pollutants enter the plant mainly through the stomata, and once in the apoplast NH$dissolves to formNH%+, whereas NO#dissolves to form NO$−and NO#−. The latter compound can also be formed after exposureto NO. There is evidence that NH$-N and NOx-N can be reversibly stored in the apoplast. Temporary storagemight affect processes such as absorption rate, assimilation and re-emission. Once formed, NO$−and NO#−canbe reduced, and NH%+can be assimilated via the normal enzymatic pathways, nitrate reductase (NR), nitritereductase and the glutamine synthetase}glutamate synthase (GS}GOGAT) cycle. Fumigation with lowconcentrations of atmospheric NH$increasesin vitroglutamine synthetase activity, but whether this involves bothor only one of the GS isoforms is still an open question. There seems to be no correlation between fumigation withlow concentrations of NH$andin vitroGDH activity. The contribution of atmospheric NH$and NO#depositionto the N budget of the whole plant has been calculated for various atmospheric pollutant concentrations andrelative growth rates (rgrs). It is concluded that at current ambient atmospheric N concentrations the directimpact of gaseous N uptake by foliage on plant growth is generally small.Key words : Apoplastic storage, gaseous nitrogen deposition, glutamate dehydrogenase, glutamine synthetase,plant nitrogen demand, sulphur dioxide root}shoot interaction, ammonia, nitrogen oxides.

Dry deposition of NH3 and NOx (NO and NO2) can affect plant metabolism at the cellular and whole-plant level. Gaseous pollutants enter the plant mainly through the stomata, and once in the apoplast NH3 dissolves to form NH4+, whereas NO2 dissolves to form NO3- and NO2-. The latter compound can also be formed after exposure to NO. There is evidence that NH3-N and NOx-N can be reversibly stored in the apoplast. Temporary storage might affect processes such as absorption rate, assimilation and re-emission. Once formed, NO3- and NO2- can be reduced, and NH4+ can be assimilated via the normal enzymatic pathways, nitrate reductase (NR), nitrite reductase and the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle. Fumigation with ion; concentrations of atmospheric NH3 increases in vitro glutamine synthetase activity, but whether this involves both or only one of the GS isoforms is still an open question. There seems to be no correlation between fumigation with low concentrations of NH3 and in vitro GDH activity. The contribution of atmospheric NH3 and NO2 deposition to the N budget of the whole plant has been calculated for various atmospheric pollutant concentrations and relative growth rates (RGRs). It is concluded that at current ambient atmospheric N concentrations the direct impact of gaseous N uptake by foliage on plant growth is generally small.

Oligotrophy, or the inability of bacterial cells to propagate at elevated nutrient concentrations, is a controversial phenomenon in microbiology. The exact cause of the unculturability of many indigenous marine bacteria on standard laboratory media has still not been resolved. Unfortunately the physiology of such cells is difficult to investigate as long as high cell density cultures cannot be obtained. An extensive evaluation of experiments relating to oligotrophy and the cultivation of marine bacteria is presented in this review. When incorporating the findings of studies performed with molecular biological methods, the picture emerges that indigenous marine bacteria can be cultivated under certain conditions and that the 'oligotrophic way of life' is a transient characteristic. Although strong generalisations should not be made with respect to a biological system as diverse as the world's oceans, it should be anticipated that cells with unique physiological characteristics appear to exist in the oceanic system. When combining conventional physiological approaches with molecular biological techniques it is feasible to unveil the phenotypes that go with the encountered genotypes. In view of the enormous complexity of the oceanic system this will prove an ambitious, yet resourceful undertaking.

Oligotrophy, or the inability of bacterial cells to propagate at elevated nutrient concentrations, is a controversial phenomenon in microbiology. The exact cause of the unculturability of many indigenous marine bacteria on standard laboratory media has still not been resolved. Unfortunately the physiology of such cells is difficult to investigate as long as high cell density cultures cannot be obtained. An extensive evaluation of experiments relating to oligotrophy and the cultivation of marine bacteria is presented in this review. When incorporating the findings of studies performed with molecular biological methods, the picture emerges that indigenous marine bacteria can be cultivated under certain conditions and that the 'oligotrophic way of life' is a transient characteristic. Although strong generalisations should not be made with respect to a biological system as diverse as the world's oceans, it should be anticipated that cells with unique physiological characteristics appear to exist in the oceanic system. When combining conventional physiological approaches with molecular biological techniques it is feasible to unveil the phenotypes that go with the encountered genotypes. In view of the enormous complexity of the oceanic system this will prove an ambitious, yet resourceful undertaking.

Despite considerable progress in recent years, many questions regarding fish larval nutrition remain largely unanswered, and several research avenues remain open. A holistic understanding of the supply line of nutrients is important for developing diets for use in larval culture and for the adaptation of rearing conditions that meet the larval requirements for the optimal presentation of food organisms and/or microdiets. The aim of the present review is to revise the state of the art and to pinpoint the gaps in knowledge regarding larval nutritional requirements, the nutritional value of live feeds and challenges and opportunities in the development of formulated larval diets. © 2013 Wiley Publishing Asia Pty Ltd., This study benefited from participation in LARVANET – COST action FA0801 (EU RTD framework programme. K. Hamre received funding from the Norwegian Ministry of Fisheries and the Research Council of Norway (CODE-199482). M. Yúfera received financial support from the Spanish Ministry of Science and Innovation MICINN + FEDER/ERDF (projects AGL2007-64450-C02-01 and Consolider Ingenio 2010 Program-project Aquagenomics CSD2007-0002). L. Conceição participated in this review in the framework of project HYDRAA – PTDC/MAR/71685/2006, granted by Fundação para a Ciência e a Tecnologia (FCT), Portugal, with the support of FEDER. I. Rønnestad received funding from the Research Council of Norway (CODE- 199482 and GutFeeling- 190019) and the EC FP7 (LIFECYCLE- 222719).

Marine bacteria in Resurrection Bay near Seward, Alaska, and in the central North Sea off the Dutch coast were cultured in filtered autoclaved seawater following dilution to extinction. The populations present before dilution varied from 0.11 x 10(9) to 1.07 x 10(9) cells per liter. The mean cell volume varied between 0.042 and 0.074 mum3, and the mean apparent DNA content of the cells ranged from 2.5 to 4.7 fg of DNA per cell. All three parameters were determined by high-resolution flow cytometry. All 37 strains that were obtained from very high dilutions of Resurrection Bay and North Sea samples represented facultatively oligotrophic bacteria. However, 15 of these isolates were eventually obtained from dilution cultures that could initially be cultured only on very low-nutrient media and that could initially not form visible colonies on any of the agar media tested, indicating that these cultures contained obligately oligotrophic bacteria. It was concluded that the cells in these 15 dilution cultures had adapted to growth under laboratory conditions after several months of nutrient deprivation prior to isolation. From the North Sea experiment, it was concluded that the contribution of facultative oligotrophs and eutrophs to the total population was less than 1% and that while more than half of the population behaved as obligately oligotrophic bacteria upon first cultivation in the dilution culture media, around 50% could not be cultured at all. During one of the Resurrection Bay experiments, 53% of the dilution cultures obtained from samples diluted more than 2.5 x 10(5) times consisted of such obligate oligotrophs. These cultures invariably harbored a small rod-shaped bacterium with a mean cell volume of 0.05 to 0.06 mum3 and an apparent DNA content of 1 to 1.5 fg per cell. This cell type had the dimensions of ultramicrobacteria. Isolates of these ultramicrobacterial cultures that were eventually obtained on relatively high-nutrient agar plates were, with respect to cell volume and apparent DNA content, identical to the cells in the initially obligately oligotrophic bacterial dilution culture. Determination of kinetic parameters from one of these small rod-shaped strains revealed a high specific affinity for the uptake of mixed amino acids (a-degrees(A) 1,860 liters/g of cells per h), but not for glucose or alanine as the sole source of carbon and energy (a-degrees(A) +/- 200 liters/g of cells per h). The ultramicrobial strains obtained are potentially a very important part of picoplankton biomass in the areas investigated.

A rationale is given for a modelling approach to identify the mechanisms involved in the functioning and metabolic activity of tissues in the wall of the gastrointestinal tract. Maintenance and productive functions are discussed and related to the distinct compartments of the gastrointestinal tract and the metabolic costs involved. Functions identified are: tissue turnover; tissue proliferation; ion transport; nutrient transport; secretions of digestive enzymes, mucus and immunoglobulins; production of immune cells. The major nutrients involved include glucose, amino acids and volatile fatty acids.In vivomeasurements of net portal fluxes of these nutrients in pigs and ruminants are evaluated to illustrate the complexity of physiology and metabolic activity of the gastrointestinal tract. Experimental evidence indicates that high, but variable and specific, nutrient costs are involved in the functioning of the gastrointestinal tract.

A number of scientific studies have investigated aspects of soles ( Solea solea and S. senegalensis) ecology, population genetics and biology in their natural environment, and the species have been extensively studied in captivity during the last decade. Studies on the genetic population structure of sole indicate that several distinct breeding populations exist within its distributional range in European waters. Recent studies suggest a phylogenetic relatedness of S. solea and S. senegalensis, being found as closest sister lineages in most reconstructions. However, studies on molecular genetics and morphological traits give diagnostic differences that consistently lead to their taxonomic separation at the specific rank. Studies show that sole spawn readily in captivity, and the buoyant, fertilized eggs are easily collected. Stocking density during maturation should be 1-1.5 kg/m(2), and temperature should be kept above 16degreesC ( S. senegalensis) or between 8 and 12degreesC ( S. solea). In nature, the onset of spawning is related to a rise in temperature occurring during spring ( March-June). Salinity should be kept constant around 33-35& and the fish reared under simulated natural photoperiod ( LDN). In other cultured flatfish species, a change in the photoperiod is the key environmental signal used to manipulate and control maturation, but at present time there are no published work that verifies or contradicts this for either S. senegalensis or S. solea. Studies indicate that a mixture of inert and live food may increase the weaning success of sole fry, and this can be further enhanced by using attractants in the dry feed. Future experiments are needed to determine the ideal time to commence weaning and determine the minimum duration of this period. Studies on alternative feeding strategies are also required. The effect of temperature and photoperiod on juvenile growth has not been studied systematically in neither of the two species and the relative importance of a direct photoperiod effect on growth in sole therefore remains to be defined.

Marine bacteria in Resurrection Bay near Seward, Alaska, and in the central North Sea off the Dutch coast were cultured in filtered autoclaved seawater following dilution to extinction. The populations present before dilution varied from 0.11 x 10(9) to 1.07 x 10(9) cells per liter. The mean cell volume varied between 0.042 and 0.074 mum3, and the mean apparent DNA content of the cells ranged from 2.5 to 4.7 fg of DNA per cell. All three parameters were determined by high-resolution flow cytometry. All 37 strains that were obtained from very high dilutions of Resurrection Bay and North Sea samples represented facultatively oligotrophic bacteria. However, 15 of these isolates were eventually obtained from dilution cultures that could initially be cultured only on very low-nutrient media and that could initially not form visible colonies on any of the agar media tested, indicating that these cultures contained obligately oligotrophic bacteria. It was concluded that the cells in these 15 dilution cultures had adapted to growth under laboratory conditions after several months of nutrient deprivation prior to isolation. From the North Sea experiment, it was concluded that the contribution of facultative oligotrophs and eutrophs to the total population was less than 1% and that while more than half of the population behaved as obligately oligotrophic bacteria upon first cultivation in the dilution culture media, around 50% could not be cultured at all. During one of the Resurrection Bay experiments, 53% of the dilution cultures obtained from samples diluted more than 2.5 x 10(5) times consisted of such obligate oligotrophs. These cultures invariably harbored a small rod-shaped bacterium with a mean cell volume of 0.05 to 0.06 mum3 and an apparent DNA content of 1 to 1.5 fg per cell. This cell type had the dimensions of ultramicrobacteria. Isolates of these ultramicrobacterial cultures that were eventually obtained on relatively high-nutrient agar plates were, with respect to cell volume and apparent DNA content, identical to the cells in the initially obligately oligotrophic bacterial dilution culture. Determination of kinetic parameters from one of these small rod-shaped strains revealed a high specific affinity for the uptake of mixed amino acids (a-degrees(A) 1,860 liters/g of cells per h), but not for glucose or alanine as the sole source of carbon and energy (a-degrees(A) +/- 200 liters/g of cells per h). The ultramicrobial strains obtained are potentially a very important part of picoplankton biomass in the areas investigated.

In sheltered, eutrophicated estuaries, reduced nitrogen (NH x ), and pH levels in the water layer can be greatly enhanced. In laboratory experiments, we studied the interactive effects of NH x , pH, and shoot density on the physiology and survival of eelgrass (Zostera marina). We tested long-term tolerance to NH x at pH 8 in a 5-week experiment. Short-term tolerance was tested for two shoot densities at both pH 8 and 9 in a 5-day experiment. At pH 8, eelgrass accumulated nitrogen as free amino acids when exposed to high loads of NH x , but showed no signs of necrosis. Low shoot density treatments became necrotic within days when exposed to NH x at pH 9. Increased NH3 intrusion and carbon limitation seemed to be the cause of this, as intracellular NH x could no longer be assimilated. Remarkably, experiments with high shoot densities at pH 9 showed hardly any necrosis, as the plants seemed to be able to alleviate the toxic effects of high NH x loads through joint NH x uptake. Our results suggest that NH x toxicity can be important in worldwide observed seagrass mass mortalities. We argue that the mitigating effect of high seagrass biomass on NH x toxicity is a positive feedback mechanism, potentially leading to alternative stable states in field conditions.