STUDIES IN HISTOCHEMISTRY : LVII. Determination of the Total Dry Mass of Human Erythrocytes by Interference Microscopy and X-ray Microradiography

The total dry mass of human erythrocytes was determined by both interference microscopy and x-ray microradiography. The determination of mass per unit area, and calculation of total dry mass per cell were simplified by changing the shape of the cells to spheres which were then flattened to discs of constant thickness when smeared on glass slides for measurement of fixed cells by interferometry, and to oblate spheroids when smeared on parlodion-coated slides for measurement of fixed cells by x-ray absorption. From x-ray measurements of 100 smeared and alcohol-fixed cells a mean dry mass per cell of 33.7 x 10^-12 g was obtained. Interference measurements of 100 fresh cells suspended in isotonic saline gave a mean value of 32.4 x 10^-12 g while interference measurement of 100 smeared and alcohol-fixed cells gave a mean value of 30.8 x 10^-12 g. The first two values compare well with a mean corpuscular hemoglobin of 31.2 x 10^-12 g, obtained from determinations of erythrocyte count and hemoglobin, since 95 per cent of the dry mass of the cell is hemoglobin. The difference in interference values between the fixed and fresh cells is possibly due to a difference between the specific refractive increment of alcohol-denatured hemoglobin and that of the unmodified substance. The value for the latter was used since that of the former is unknown.     

Dry to wet weight biomass conversion constant for Tetrahymena elliotti (Ciliophora,Protozoa)

Summary The wet and dry weights of both axenic and monoxenic cultures of the ciliate Tetrahymena were determined directly. These estimates are dependent upon the method of volume determination. Assuming a prolate spheroidal shape for the ciliate, we calculate a mean wet weight of 0.4157±0.0713 pg m^-3 and a mean dry weight of 0.2793±0.0652 pg m^-3. Using electronic cell sizing, our estimates are 0.7869±0.1659 pg m^-3 and 0.5239±0.1101 pg m^-3, respectively. Independent of the method of volume determination, we estimate a mean biomass conversion ratio (dry weight/wet weight) of 0.59±0.08.     

Effect of zearalenone (F-2) on pea stem,maize root,and rat liver mitochondria

At 5 and 10 g ml^-1 concentration, zearalenone (F-2), a mycotoxin produced by a number of species of the genus Fusarium, causes an inhibition of the oxidative phosphorylation of isolated plant mitochondria, while at 20 and 40 g ml^-1 it causes uncoupling. However, when the mitochondria are pre-incubated for 20 min with F-2, the uncoupling appears to be the prevailing effect. F-2 is also able to inhibit the mitochondrial ATPase activity (Mg²⁺-dependent). Conversely, F-2 (40 g ml^-1) does not alter the ATP level of maize roots and only slightly affects the ATPase activity of pea stem and maize root microsomal fractions. In addition, F-2 (10–40 g ml^-1) inhibits ATP synthesis catalyzed by rat liver mitochondria. It is suggested that the phytotoxicity of F-2, also known for its ability to collapse the transmembrane electric potential of maize roots, may be mainly linked to its ability to increase the proton permeability of the cell, similar to the common uncouplers.Abbreviations F-2 zearalenone - DCCD N,N-dicyclohexylcarbodiimide - PCCP carbonyl cyanide, p-trifluoromethoxiphenylhydrazone - CBT Cerospora beticola toxin     

Rates of litterfall and organic matter turnover at three South Australian indigenous forest sites

A quantitative study of the litterfall rate, litter mass and the organic carbon content of litter and soil at three South Australian low open-forests was made monthly for 2 years. The soil and dominant vegetation at the sites were similar, but the mean annual rainfall differed; 635, 690 and 1050 mm respectively at the ‘dry’, ‘medium’ and ‘wet’ sites. Mean annual litterfall at the dry, medium and wet sites was 114, 205 and 233 g m^?2 respectively, and the mean mass of the litter layer was 885, 1153 and 858 g m^?2 Leafy material formed 82% of the litterfall at each site and maximum fall occurred in the summer. Leafy material formed 64–68% of the three litter layers and there was no general pattern of seasonal change of mass. The data obtained were used to calculate the half-life (T_1/2 of the litter, the values were 5.4, 3.9 and 2.6 years respectively at the dry, medium and wet sites. The values from the medium and wet sites fit within the range previously recorded for other open-forests and low open-forests, but indicate an extremely slow rate of decomposition at the dry site, which was located close to open-scrubland. The amounts of organic carbon in the 0–8 cm layer of soil at the dry, medium and wet sites were 777, 928and 950 g m^-2 respectively. However, without a detailed investigation of the source of soil carbon, these values could only be used to show that the organic carbon store to a depth of 8 cm is equivalent to 9–15 times the annual litterfall input.     

NUCLEAR GENE DOSAGE EFFECTS ON MITOCHONDRIAL MASS AND DNA

In order to assess the effect of nuclear gene dosage on the regulation of mitochondria we have studied serial sections of a set of isogenic haploid and diploid cells of Saccharomyces cerevisiae, growing exponentially in the absence of catabolite repression, and determined the amount of mitochondrial DNA per cell. Mitochondria accounted for 14% of the cytoplasmic and 12% of the total cellular volume in all cells examined regardless of their ploidy or their apparent stage in the cell cycle. The mean number of mitochondria per cell was 22 in the diploid and 10 in the haploids. The volume distribution appeared unimodal and identical in haploids and diploids. The mitochondrial DNA accounted for 12.6 ± 1.2% and 13.5 ± 1.3% of the total cellular DNA in the diploid and haploid populations, respectively. These values correspond to 3.6 x 10^-15 g, 2.2 x 10⁹ daltons, or 44 genomes (50 x 10⁶ daltons each) per haploid and twice that per diploid cell. On this basis, the average mitochondrion in these cells contains four mitochondrial genomes in both the haploid and the diploid.     

Morphological studies on the epiphyseal growth plate combined with biochemical and X-ray microprobe analyses

Summary The epiphyseal growth plate of the domestic pig was investigated topologically combining biochemical methods with electron microprobe microanalyses both correlated to histological controls. A lateral resolution of about 50 m was reached. Highest nuclease activity was found in the lower columnar cell zone, while alkaline phosphatase showed maximal activity in the hypertrophic area, connected with maximal values for extractable, organically bound phosphorus, and extractable Ca and Mg. Acid phosphatase activity reached maximal values in the zone of the lower primary spongiosa, while the extractable P_i had maximal values at the end of the zone of bone remodelling. Microprobe analyses have shown that the extracellular Ca content (per dry mass) remained relatively constant at 0.7% (about 58 mM/kg wet weight for 66% tissue fluid) in all zones of the plate increasing to 1% in the vicinity of the first foci of mineralization. The intracellular P content (per dry mass) was about 4.5%, the extracellular 0.1–0.2% (about 10–20 mM/kg wet weight) increasing also to about 1% in the vicinity of the first foci of mineralization. Thus the Ca x P product was much higher than the ion-product of 2 mM² which is necessary for an in vitro mineralization of connective tissue. The extracellular S content (per dry mass) as a probable indicator of sulfated proteoglycans was relatively constant at about 3.5% in the different zones but decreased to about 0.3% in the fully mineralized regions. This indicates a loss of sulfur containing substances with mineralization which is not so high since the concentrations per dry mass must be normalized to a unit volume of equal density of mass.     

Mass balance of nitrogen in constructed mangrove wetlands receiving ammonium-rich wastewater: Effects of tidal regime and carbon supply

A series of computer-controlled mangrove tide-tanks planted with Kandelia candel was constructed to investigate the removal and transformation of ammonium–nitrogen under two tidal regimes: (i) 12-h wet/12-h dry (long tidal regime) and (ii) 6-h wet/6-h dry/6-h wet/6-h dry daily (short tidal regime). All tanks were irrigated with NH₄Cl solution for nine water cycles (each cycle lasted for 5 weeks) at an amount of around 2.1 g NH₄Cl (equivalent to 0.52 g N) per tank per cycle. During the experiment, total Kjeldahl nitrogen (TKN), inorganic nitrogen (N) (NH₄⁺–, NO₂^?–, and NO₃^?–N) and carbon were completely removed by the mangrove system. The added NH₄⁺–N was not detected in tidal water or accumulated in sediment. The mass balance of nitrogen showed that the discharge of ammonium-rich wastewater to mangrove wetlands enhanced microbial nitrogen transformation, particularly nitrification and denitrification processes, with 15–30% of the total nitrogen inputs returned to atmosphere as N₂ gas. Growth of K. candel and macroalgae was stimulated by ammonium addition, and up to 3 and 7% of total N inputs were assimilated in plant and algal tissues, respectively. Constructed mangrove wetlands with short tidal regime had higher numbers of nitrifiers and significantly lower content of ammonium that those with long tidal regime. On the other hand, higher populations of denitrifiers and lower nitrate were found in mangroves with long tidal regime and with glucose addition.     

Body Mass Parameters,Lipid Profiles and Protein Contents of Zebrafish Embryos and Effects of 2,4-Dinitrophenol Exposure

Morphology and physiology of fish embryos undergo dramatic changes during their development until the onset of feeding, supplied only by endogenous yolk reserves. For obtaining an insight how these restructuring processes are reflected by body mass related parameters, dry weights (dw), contents of the elements carbon and nitrogen and lipid and protein levels were quantified in different stages within the first four days of embryo development of the zebrafish (Danio rerio). The data show age dependent changes in tissue composition. Dry weights decreased significantly from 79μgdw/egg at 0hours post fertilization (hpf) to 61 μgdw/egg after 96 hpf. The amounts of total carbon fluctuated between 460 mg g^-1 and 540 mg g^-1 dw, nitrogen was at about 100 mg g^-1 dw and total fatty acids were between 48–73 mg g^-1 dw. In contrast to these parameters that remained relatively constant, the protein content, which was 240 mg g^-1 at 0 hpf, showed an overall increase of about 40%. Comparisons of intact eggs and dechorionated embryos at stages prior to hatching (24, 30, 48 hpf) showed that the differences seen for dry weight and for carbon and nitrogen contents became smaller at more advanced stages, consistent with transition of material from the chorion to embryo tissue. Further, we determined the effect of 2,4-dinitrophenol at a subacutely toxic concentration (14 μM, LC10) as a model chemical challenge on the examined body mass related parameters. The compound caused significant decreases in phospholipid and glycolipid fatty acid contents along with a decrease in the phospholipid fatty acid unsaturation index. No major changes were observed for the other examined parameters. Lipidomic studies as performed here may thus be useful for determining subacute effects of lipophilic organic compounds on lipid metabolism and on cellular membranes of zebrafish embryos.     

Determination of the Biomasses of Small Bacteria at Low Concentrations in a Mixture of Species with Forward Light Scatter Measurements by Flow Cytometry

The forward light scatter intensity of bacteria analyzed by flow cytometry varied with their dry mass, in accordance with theory. A standard curve was formulated with Rayleigh-Gans theory to accommodate cell shape and alignment. It was calibrated with an extinction-culture isolate of the small marine organism Cycloclasticus oligotrophus, for which dry weight was determined by CHN analysis and ¹⁴C-acetate incorporation. Increased light scatter intensity due to formaldehyde accumulation in preserved cells was included in the standard curve. When differences in the refractive indices of culture media and interspecies differences in the effects of preservation were taken into account, there was agreement between cell mass obtained by flow cytometry for various bacterial species and cell mass computed from Coulter Counter volume and buoyant density. This agreement validated the standard curve and supported the assumption that cells were aligned in the flow stream. Several subpopulations were resolved in a mixture of three species analyzed according to forward light scatter and DNA-bound DAPI (4′,6-diamidino-2-phenylindole) fluorescence intensity. The total biomass of the mixture was 340 μg/liter. The lowest value for mean dry mass, 0.027 ± 0.008 pg/cell, was for the subpopulation of C. oligotrophus containing cells with a single chromosome. Calculations from measurements of dry mass, Coulter Counter volume, and buoyant density revealed that the dry weight of the isolate was 14 to 18% of its wet weight, compared to 30% for Escherichia coli. The method is suitable for cells with 0.005 to about 1.2 pg of dry weight at concentrations of as low as 10³ cells/ml and offers a unique capability for determining biomass distributions in mixed bacterial populations.     

CORRELATED MORPHOMETRIC AND BIOCHEMICAL STUDIES ON THE LIVER CELL : I. Morphometric Model, Stereologic Methods, and Normal Morphometric Data for Rat Liver

The basic morphological properties of liver cells are defined in the form of a morphometric model to permit integrated quantitative characterization of functionally important parameters. Stereologic methods which allow efficient and reliable quantitative evaluation of sectioned liver tissue are presented. Material, obtained by a rigorous three-stage sampling procedure from five normal rat livers, is systematically subjected to this analysis at four levels of magnification. This yields quantitative data which are expressed as "densities," i.e. content per 1 ml of tissue, as "specific dimensions" related to 100 g body weight, and as absolute dimensions per average "mononuclear" hepatocyte. Base line data relating to the normal rat liver are presented for the entire spectrum of parameters. As examples, 1 ml of liver tissue contains 169 x 10⁶ hepatocyte nuclei, some 90 x 10⁶ nuclei of other cells, and 280 x 10⁹ mitochondria. Hepatocyte cytoplasm accounts for 77% of liver volume, and the mitochondria for 18%. The surface area of endoplasmic reticulum membranes in 1 ml of liver tissue measures 11 m² of which are ⅔ of the rough form carrying some 2 x 10¹³ ribosomes. The surface area of mitochondrial cristae in the unit volume is estimated at 6 m². The validity and applicability of the method are discussed, and the data are compared with available information from other studies.     

Mass cultivation of the nitrogen-fixing cyanobacteriumGloeotrichia natans,indigenous to rice-fields

Gloeotrichia natans, a nitrogen fixing cyanobacterium common in rice fields in the Philippines, was used for studies to establish key features of its physiology and potential production in outdoor cultures. Under optimal growth conditions (38 °C, pH 8.0, no carbon enrichment) the specific growth rate of rice-field isolate was 0.076 h^–1. The pH of the medium (between 6.5 and 9.0) did not influence the growth rate, but it did affect phycobiliprotein content, as reflected by a change in colour. At pH 7.0 the culture was green-brown, with phycobiliproteins constituting up to 10% of the total protein, while at pH 9.0 the culture was brownish-black and the pigment content was as high as 28% of the total protein. In outdoor cultures the specific growth rate was related directly to cell density in the range of 0.7–1.5 g dry weight 1^–1 at a rate of stirring of 30 rpm, and inversely related to cell density at half this rate. At a stirring of 30 rpm, daily production of outdoor cultures harvested to maintain cell densities of 0.7, 1.15 andw 1.5 g 1^–1 were 14.7, 17.1 and 18.1 g m^–2 d^t, respectively. This rate of production was maintained for more than 45 days. Phycobiliprotein content in the culture kept at a density of 1.5 g 1^–1 reached 14% of the total biomass.     

Microbial Ecosystem in Sunderban Mangrove Forest Sediment,North-East Coast of Bay of Bengal,India

This is the first documentation of seasonal and spatial fluctuation of the culturable microbial population collected from different zones in the sediment of the Sunderban mangrove forest. The population of cellulose degrading bacteria, [mean value of CFU 6.189 ± 1.025 × 10⁶ (g dry weight of sediment)^?1] was found to be maximum during post monsoon in the deep forest region, whereas, the fungal population [mean value of CFU 3.424 ± 0.886 × 10⁶ (g dry weight of sediment)^?1] was found to be maximum during pre-monsoon in the rooted region. The abundances of microbes, in decreasing order, studied from different zones are nitrifying bacteria [mean value of CFU 1.125 ± 0.359 × 10⁶ (g dry weight of sediment)^?1], phosphorous solubilizing bacteria (PSB) [mean value of CFU 0.805 ± 0.322 × 10⁶ (g dry weight of sediment)^?1], free living nitrogen fixing bacteria [mean value of CFU 0.417 ± 0.120 × 10⁶ (g dry weight of sediment)^?1] and sulfur reducing bacteria (SRB) [mean value of CFU 0.356 ± 0.125 × 10⁶ (g dry weight of sediment)^?1]. The content of organic carbon in the soil decreased from the deep forest region to the rooted and unrooted region but a reverse profile was found for soil salinity and soil silicate concentration. The results from the present study indicate that the monsoon cycle has a pronounced effect on the microbially dominated biogeochemistry in the sediment and consequently on the ecology of the Sundarban mangrove forest.     

Effects of fire,mowing and nitrogen addition on root characteristics in tall-grass prairie

Abstract. Root harvests and root windows were used to study the influence of fire, mowing and nitrogen additions on root lengths, biomass, and nitrogen content in tall-grass prairie. Four years of nitrogen additions (10 g m² yr^?1) increased below-ground mass by 15 % and nitrogen concentration in that mass by 77 %. In general, live roots and rhizomes exhibited greater increases in nitrogen concentrations than detrital roots and rhizomes. After four years of treatment, live roots and rhizomes immobilized an additional 1.5 to 5 g/m² of nitrogen, depending upon specific treatment, while dead roots and rhizomes immobilized an additional 3 to 3.5 g/m². Average root growth parameters, as measured with root windows, were positively correlated with above-ground peak foliage biomass; however, the only significant correlation was between average new root growth and above-ground peak foliage biomass (r = 0.73, p ≤ 0.04). Root growth and decay, as measured by annual mean values for eight root windows over a four year interval, were insensitive to climatic and treatment effects.     

Identification of oxindole-3-acetic acid,and metabolic conversion of indole-3-acetic acid to oxindole-3-acetic acid in Pinus sylvestris seeds

Oxindole-3-acetic acid (OxIAA) has been identified in germinating seeds of Scots pine (Pinus sylvestris) using gas chromatography-mass spectrometry. Seeds germinated for 5 d contained 2.7 ng OxIAA·g^-1 (dry weight) whereas ungerminated seeds contained 0.2 ng·g^-1. Isotopically labelled OxIAA was formed in seeds incubated with [1-¹⁴C]-, [2-¹⁴C]- or [²H₅]indole-3-acetic acid.Abbreviations DDC sodium diethyldithiocarbamate - GC gas chromatography - HPLC high-performance liquid chromatography - IAA indole-3-acetic acid - MS mass spectrometry - OxIAA oxindole-3-acetic acid - PVP polyvinylpyrrolidone - TMS trimethylsilyl     

Fish as sources and sinks of nutrients in lakes

1. The release of total phosphorus (TP) and nitrogen (N in ammonium) was measured for the five most abundant fish species (>85% of biomass) in Mouse and Ranger Lakes, two biomanipulated, oligotrophic lakes in Ontario. 2. The specific release rate of both nutrients was significantly related to fish mass; log₁₀ TP release rate (μg h^?1) = 0.793 (±0.109) [log₁₀ wet mass (g)] + 0.7817 (±0.145), and log₁₀ N release rate (μg h^?1) = 0.6946 (±0.079) [log₁₀wet mass (g)] + 1.7481 (±0.108). 3. When fish nutrient release was standardized for abundance (all populations, 1993–95) and epilimnetic volume, fish were estimated to contribute 0.083 (±0.061) μg TP L^?1 day^?1, and 0.41 (±0.17) μg N L^?1 day^?1 in Mouse L., and 0.062 (±0.020) μg TP L^?1 day^?1 and 0.31 (±0.08) μg N L^?1 day^?1 in Ranger L. 4. In comparison, concurrent rates of total planktonic P regeneration were 1.02 (±0.45) μg L^?1 day^?1 (Mouse L.) and 0.85 (±0.19) μg L^?1 day^?1 (Ranger L.). Fish represented 8% of planktonic P release in Mouse L. and 7% in Ranger L. 5. Fish dry mass had mean elemental body compositions of 39.3% carbon, 10.9% nitrogen, and 4.0% phosphorus (all fish combined), with a mean molar C : N : P ratio of 27 : 6 : 1. This comprised about 55% and 23% of the total epilimnetic particulate P and N respectively. 6. Turnover times of P and N in fish were approximately 103 and 48 days respectively. In comparison, planktonic turnover times of particulate P in Mouse and Ranger Lakes were 4.3 and 4.4 days respectively. Given their high P content and low turnover rates, fish appear to be important P sinks in lakes.     

Contemporary and pre-industrial global reactive nitrogen budgets

Increases and expansion of anthropogenic emissions of both oxidized nitrogen compounds, NO_x, and a reduced nitrogen compound, NH₃, have driven an increase in nitrogen deposition. We estimate global NO_x and NH₃ emissions and use a model of the global troposphere, MOGUNTIA, to examine the pre-industrial and contemporary quantities and spatial patterns of wet and dry NO_y and NH_x deposition. Pre-industrial wet plus dry NO_x and NH_x deposition was greatest for tropical ecosystems, related to soil emissions, biomass burning and lightning emissions. Contemporary NO_y+NH_x wet and dry deposition onto Northern Hemisphere (NH) temperate ecosystems averages more than four times that of preindustrial N deposition and far exceeds contemporary tropical N deposition. All temperate and tropical biomes receive more N via deposition today than pre-industrially. Comparison of contemporary wet deposition model estimates to measurements of wet deposition reveal that modeled and measured wet deposition for both NO ₃ ^– and NH ₄ ⁺ were quite similar over the U.S. Over Western Europe, the model tended to underestimate wet deposition of NO ₃ ^– and NH ₄ ⁺ but bulk deposition measurements were comparable to modeled total deposition. For the U.S. and Western Europe, we also estimated N emission and deposition budgets. In the U.S., estimated emissions exceed interpolated total deposition by 3-6 Tg N, suggesting that substantial N is transported offshore and/or the remote and rural location of the sites may fail to capture the deposition of urban emissions. In Europe, by contrast, interpolated total N deposition balances estimated emissions within the uncertainty of each.Abbreviations EMEP European Monitoring and Evaluation Program - GEIA Global Emissions Inventory Activity - NADP/NTN National Atmospheric Deposition Program/National Trends Network in the US - NH Northern Hemisphere - NH_x=NH₃+NH ₊ ⁴ NO_x=NO+NO₂ NO_y total odd nitrogen=NO_x+HNO₃+HONO+HO₂NO₂+NO₃+radical (NO₃ ^.)+Peroxyacetyl nitrates+N₂O₅+organic nitrates - SH Southern Hemisphere - Gg 10⁹ g - Tg 10¹² g     

Buoyant Densities and Dry-Matter Contents of Microorganisms: Conversion of a Measured Biovolume into Biomass

Several isolates of bacteria and fungi from soil, together with cells released directly from soil, were studied with respect to buoyant density and dry weight. The specific volume (cubic centimeters per gram) of wet cells as measured in density gradients of colloidal silica was correlated with the percent dry weight of the cells and found to be in general agreement with calculations based on the partial specific volume of major cell components. The buoyant density of pure bacterial cultures ranged from 1.035 to 1.093 g/cm³, and their dry-matter content ranged from 12 to 33% (wt/wt). Average values proposed for the conversion of bacterial biovolume into biomass dry weight are 1.09 g/cm³ and 30% dry matter. Fungal hyphae had buoyant densities ranging from 1.08 to 1.11 g/cm³, and their dry-matter content ranged from 18 to 25% (wt/wt). Average values proposed for the conversion of hyphal biovolume into biomass dry weight are 1.09 g/cm³ and 21% dry matter. Three of the bacterial isolates were found to have cell capsules. The calculated buoyant density and percent dry weight of these capsules varied from 1.029 g/cm³ and 7% dry weight to 1.084 g/cm³ and 44% dry weight. The majority of the fungi were found to produce large amounts of extracellular material when grown in liquid cultures. This material was not produced when the fungi were grown on either sterile spruce needles or membrane filters on an agar surface. Fungal hyphae in litter were shown to be free from extracellular materials.     

Total Mercury in Sediments,Macrophytes, and Fish from a Shallow Steppe Lake in Eastern Austria

During summer 2011, samples of sediment, macrophytes, and fish tissues from the shallow, slightly alkaline Lake Neusiedl, Austria, were evaluated for their total Hg content. This is the first report of Hg levels from this lake. Sediments displayed Hg contents between 0.025 and 0.113 μg g^?1 dw (dry weight), significantly correlating with the proportion of organic components pointing to a small anthropogenic impact on the lake's Hg content. Hg Levels in plants and fish were unexpectedly high: both investigated submerged plant species, Potamogeton pectinatus and Myriophyllum spicatum, showed mean values of 0.245±0.152 and 0.298±0.115 μg g^?1 dw, respectively. Biomagnification was evident when comparing muscle samples of the planktivorous fish species rudd Scardinus erythrophthalmus (n=10, mean=0.084 μg g^?1 ww (wet weight)) with the piscivorous perch Perca fluviatilis (n=21, mean=0.184 μg g^?1 ww) or pike‐perch Sander lucioperca (n=9, mean=0.205 μg g^?1 ww). Significantly lower values were found in the muscle of the piscivorous pike Esox lucius (n=25, mean=0.135 μg g^?1 ww), pointing to a specific Hg metabolism of this fish, presumably under the particular physicochemical properties of the lake. Hg Concentrations in fish could pose a risk to piscivorous birds in this protected wetland system.     

DIRECT COUNTING AND SIZING OF MITOCHONDRIA IN SOLUTION

Resistive particle counting has been developed for the accurate sizing and counting of mitochondria in solution. The normal detection limit with a 30 µ aperture is 0.48 µ diameter, or 0.056 µ³ particle volume The mean volume of rat liver mitochondria was 0.42 µ³ or 0.93 µ in diameter. The average value for numbers of particles per milligram of mitochondrial protein was 4.3 x 10³, and per gram of rat liver was about 11 x 10¹⁰. These values compare satisfactorily with those derived by light microscopy and electron microscopy. The mean volume for mitochondria from rat heart was 0 60 µ³ and from rat kidney cortex, 0.23 µ³. These values agree within 15% of those determined by electron microscopy of whole tissue. Mitochondrial fragility and contaminating subcellular organelles were shown to have little influence on the experimentally determined size distributions The technique may be applied to rapid swelling studies, as well as to estimations of the number and size of mitochondria from animals under different conditions such as liver regeneration and hormonal, pathological, or drug-induced states Mitochondrial DNA, RNA, cytochrome c-oxidase, cytochrome (a ÷ a₃), and iron were nearly constant per particle over large differences in particle size. Such data may be particularly valuable for biogenesis studies and support the hypothesis that the net amount per particle of certain mitochondrial constituents remains constant during mitochondrial growth and enlargement     

Outdoor mass culture of Spirulina maxima in sea-water

Summary The results of a whole year experiment on the outdoor mass culture of Spirulina maxima strain 4Mx on fertilized sea-water are reported. Carbonate and phosphate precipitation in the sea-water media was prevented by maintaining a low concentration of phosphate and by controlling the pH in the range of 8.0–8.3. The mean annual yield of biomass on sea-water plus urea as nitrogen source was 7.35 g (dry weight) m^-2· day^-1, a value slightly lower than that obtained on the standard bicarbonate medium (8.14 g · m^-2 · day^-1). On sea-water plus nitrate the yield was only 5.2 g·m^-2·day^-1. The nitrogen content of the biomass was higher in summer and lower in winter. The seasonal effect was more evident when nitrate was the nitrogen source.