Occurrence of nitrifiers and diversity of ammonia-oxidizing bacteria in developing drinking water biofilms

We studied the population dynamics of nitrifying bacteria during the development of biofilms up to 233 or 280 days on polyvinylchloride pipes connected to two full-scale drinking water distribution networks supplying processed and chloraminated surface water. The numbers of nitrifiers in biofilms were enumerated at intervals of 10–64 days by the most probable number (MPN) method at waterworks and at several study sites in distribution network areas. The numbers of nitrifiers increased towards the distal sites. The highest detected MPN counts of ammonia-oxidizing bacteria (AOB) for study areas 1 and 7 were 500 MPN cm⁻² and 1.0×10⁶ MPN cm⁻², and those of nitrite-oxidizing bacteria (NOB) 96 MPN cm⁻² and 2.2×10³ MPN cm⁻², respectively. The diversity of AOB was determined by PCR amplifying, cloning and sequencing the partial ammonia monooxygenase (amoA) gene of selected biofilm samples presenting different biofilm ages. The PCR primers used, A189 and A682, also amplified a fragment of particulate methane monooxygenase (pmoA) gene of methane-oxidizing bacteria. The majority of biofilm clones (24 out of 30 studied) contained Nitrosomonas amoA-like sequences. There were only two pmoA-like sequences of Type I methanotrophs, and four sequences positioned in amoA/pmoA sequence groups of uncultured bacteria. From both study area very similar or even completely identical Nitrosomonas amoA-like sequences were obtained despite of high difference in AOB numbers. The results show that the conditions in newly formed biofilms in drinking water distribution systems favor the growth of Nitrosomonas-type AOB.     

A short-chain dehydrogenase gene from Pichia stipitis having D-arabinitol dehydrogenase activity

An NAD⁺-dependent D-arabinitol dehydrogenase (polyol dehydrogenase) gene was isolated from Pichia stipitis CBS 6054 and cloned in Saccharomyces cerevisiae. The gene was isolated by screening of a λ-cDNA library with a zymogram technique. D -Arabinitol, xylitol, D -glucitol and galactitol are substrates for the recominant protein. With D -arabinitol as substrate the reaction product is D -ribulose. The molecular weight of the native tetramer enzyme is 110 000 Da and the monomer is 30 000 Da. The amino acid sequence is homologous to the short-chain dehydrogenase family. It is 85·5% identical to a D -arabinitol dehydrogenase from Candida albicans. The gene in P. stipitis was induced by D -arabinitol and P. stipitis was able to grow on D -arabinitol. The physiological role of D -rabinitol metabolism is discussed.     

Gas Chromatographic headspace analysis of Chardonnay and Semillon wines after thermal processing

Volatile compounds of Chardonnay and Semillon wines were analysed using a dynamic headspace method based on selective capillary column trapping and gas chromatography with another column. The effect of thermal treatment on the volatile composition of the wines was studied. Some grape derived norisoprenoid compounds, important to the aroma of aged wine, were formed at 45°C by hydrolysis from the glycosidic precursors. The heat treatment increased the proportion of 1,1,6-Trimethy1-1,2-dihydronaphthalene (TDN) and vitispirane in Chardonnay and Semillon wines. Headspace volatiles of model wines, to which the glycosidic fraction of Chardonnay and Semillon juices was added, were also analysed.

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Gaschromatographische Kopfraum-Analyse der erhitzten Chardonnay- und Semillon-Weine

Zusammenfassung Flüchtige Verbindungen der Weine Chardonnay und Semillon wurden mittels der dynamischen Kopfraum-Methode untersucht. Die flüchtigen Substanzen wurden selektiv mit einer Capillarsäule abgefangen und gaschromatographisch mit einer anderen Säule behandelt. Der Effekt einer Hitzebehandlung auf die Zusammensetzung der flüchtigen Verbindungen der Weine wurde untersucht. Durch Hydrolyse bei 45 °C wurden aus glykosidischen Vorstufen einige Norisoprenoid-Verbindung gebildet, die für das Aroma länger gelagerter Weine wichtig sind. Durch die Hitzebehandlung wurden die Anteile von 1,1,6-Trimethyl-1,2-dihydronaphtalen (TDN) und Vitispirane der Chardonnay- und Semillon-Weine erhöht. Zusätzlich wurden die flüchtigen Verbindungen solcher Referenzweine analysiert, denen die glykosidische Fraktion der Chardonnay- und Semillon-Traubensäfte zugesetzt worden war.

     

Selenium in Finnish foods after beginning the use of selenate-supplemented fertilisers

In Finland all agricultural multinutrient fertilisers have been supplemented with sodium selenate since 1984 in order to increase the selenium (Se) content of domestic foods and to raise the population's low Se intake. In the present study the Se contents of 125 food items, most of them produced locally but some imported, have been analysed using electrothermal atomic absorption spectrophotometry. The quality of Se analyses was verified by accuracy and precision measures of certified references and non-certified controls. In all cases the Se-supplemented fertilisation increased the Se levels of domestic agricultural products. The mean Se values (mg kg^?1 dry weight) in different food groups varied as follows: cereals 0.05–0.35; vegetables and fruit < 0.01,–1.20; meat and meat products 0.16–4.90; fish 0.54–3.80; dairy products and eggs 0.05–1.30; and ready to eat foods 0.04–0.78. The highest Se contents were in kidney, liver and roe whereas the lowest concentrations were found in some fruits and berries. Generally, foods of high protein content, such as meat, fish and high protein dairy products, were abundant sources of Se. The present Se intake, calculated according to Finnish national food statistics and the present results, is about 0.11 mg day^?1 at an energy level of 10 MJ (2400 keal). Cereals, meat, dairy products, eggs and fish, respectively, contributed 26%, 29%, 20%, 10% and 9% of the total intake. The Se intake now very probably meets the recommendations. Only diets with a very exceptional composition provide less than 0.05 mg or more than 0.2 mg per 10 MJ.     

Bioavailability of sediment-associated PCDD/Fs and PCDEs: relative importance of contaminant and sediment characteristics and biological factors

Factors that determine accumulation of sediment-associated polychlorinated dibenzo-p-dioxins and furans and polychlorinated diphenyl ethers into semipermeable membrane devices (SPMDs) and benthic oligochaete worms (Lumbriculus variegatus) were examined. These factors included both physical-chemical and structural characteristics of the contaminants (water solubility, lipophilicity, dipole moment, molecular size, and conformation) and sediment characteristics (organic carbon content, particle size, aromaticity, and polarity of organic carbon). The results of partial least squares regression analysis indicated that lipophilicity alone is not a sufficient predictor for contaminant bioaccumulation potential, even though it is a significant contributor. It was shown that contaminant molecular size and conformation (specifically planarity/nonplanarity) as well as sediment characteristics also have a significant role. The studied factors contributed up to 63-88% of the variation in accumulation data for SPMDs and 50-65% for oligochaetes. Comparison of (bio)accumulation factors (BAF28d for oligochaetes and AF28d for SPMDs) revealed that accumulation of contaminants in oligochaetes is largely influenced by biological factors (e.g., feeding habits), while the physical-chemical nature of the process is emphasized for SPMDs.     

Occurrence of nitrifying bacteria and nitrification in Finnish drinking water distribution systems

Microbiological nitrification process may lead to chemical, microbiological and technical problems in drinking water distribution systems. Nitrification activity is regulated by several physical, and chemical, and operational factors. However, the factors affecting nitrification in the distribution systems in boreal region, having its specific environmental characteristics, are poorly known. We studied the occurrence and activity of nitrifying bacteria in 15 drinking water networks distributing water with very different origin and treatment practices. The waters included chloraminated surface water, chlorinated surface water, and non-disinfected groundwater. The networks were located in eight towns in different parts of Finland. Our results showed that nitrifying bacteria are common in boreal drinking water distribution systems despite their low temperature. Surprisingly high numbers and activities of nitrifiers were detected in pipeline sediment samples. The numbers of ammonia-oxidizing bacteria and their oxidation potentials were highest in chloraminated drinking water delivering networks, whereas the nitrite-oxidizing bacteria were present in the greatest numbers in those networks that used non-disinfected groundwater. The occurrence of nitrifying bacteria in drinking water samples correlated positively with the numbers of heterotrophic bacteria and turbidity, and negatively with the content of total chlorine. Although nitrifying bacteria grew well in drinking water distribution systems, the problems with nitrite accumulation are rare in Finland.     

Proteome analysis of recombinant xylose-fermenting Saccharomyces cerevisiae

Introduction of an active xylose utilization pathway into Saccharomyces cerevisiae, which does not naturally ferment pentose sugars, is likely to have a major impact on the overall cellular metabolism as the carbon introduced to the cells will now flow through the pentose phosphate pathway. The metabolic responses in the recombinant xylose-fermenting S. cerevisiae were studied at the proteome level by comparative two-dimensional gel electrophoresis of cellular proteins within a pH range of 3-10. Glucose-limited chemostat cultivations and corresponding chemostat cultivations performed in media containing xylose as the major carbon source were compared. The cultivations were studied in aerobic and anaerobic metabolic steady states and in addition at time points 5, 30 and 60 min after the switch-off of oxygen supply. We identified 22 proteins having a significant abundance difference on xylose compared to glucose, and 12 proteins that responded to change from aerobic to anaerobic conditions on both carbon sources. On xylose in all conditions studied, major changes were seen in the abundance of alcohol dehydrogenase 2 (Adh2p), acetaldehyde dehydrogenases 4 and 6 (Ald4p and Ald6p), and DL-glycerol 3-phosphatase (Gpp1p). Our results give indications of altered metabolic fluxes especially in the acetate and glycerol pathways in cells growing on xylose compared to glucose.     

Role of Polyamine-Induced Dimerization of Antizyme in Its Cellular Functions

The polyamines, spermine (Spm) and spermidine (Spd), are important for cell growth and function. Their homeostasis is strictly controlled, and a key downregulator of the polyamine pool is the polyamine-inducible protein, antizyme 1 (OAZ1). OAZ1 inhibits polyamine uptake and targets ornithine decarboxylase (ODC), the rate-limiting enzyme of polyamine biosynthesis, for proteasomal degradation. Here we report, for the first time, that polyamines induce dimerization of mouse recombinant full-length OAZ1, forming an (OAZ1)₂-Polyamine complex. Dimerization could be modulated by functionally active C-methylated spermidine mimetics (MeSpds) by changing the position of the methyl group along the Spd backbone—2-MeSpd was a poor inducer as opposed to 1-MeSpd, 3-MeSpd, and Spd, which were good inducers. Importantly, the ability of compounds to inhibit polyamine uptake correlated with the efficiency of the (OAZ1)₂-Polyamine complex formation. Thus, the (OAZ1)₂-Polyamine complex may be needed to inhibit polyamine uptake. The efficiency of polyamine-induced ribosomal +1 frameshifting of OAZ1 mRNA could also be differentially modulated by MeSpds—2-MeSpd was a poor inducer of OAZ1 biosynthesis and hence a poor downregulator of ODC activity unlike the other MeSpds. These findings offer new insight into the OAZ1-mediated regulation of polyamine homeostasis and provide the chemical tools to study it.     

Intraspecific Competition, Growth, Chemistry, and Susceptibility to Voles in Seedlings of Betula pendula

We studied the effects of the intensity of intraspecific competition, as indicated by seedling density, and competitive success within populations, as indicated by seedling size, on the secondary chemistry of the stems of silver birch seedlings and their palatability to field voles. We found that the size of seedlings and their total phenolic concentrations were inversely related to stand density. Voles, however, did not discriminate between seedlings grown at the densities studied. Variation in competitive success was associated with several phenomena. Seedling size was positively related to the concentrations of several secondary compounds and to vole preferences, but negatively related to nitrogen concentration. We suggest that the ecological consequences of variation in seedling size cannot be predicted without knowledge of the mechanism behind the variation.