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Climatic and agricultural factors in nutrient exports from two watersheds in Ohio.

Export of agricultural nutrients and sediment to lakes and oceans is of great environmental concern in many agricultural watersheds. Recent years have seen efforts to reduce loads through agricultural practices such as conservation tillage, efficient fertilization, and reservation of erodible areas. Monitoring the efficacy of such efforts is complicated by the fact they take place against a varying climatic and hydrologic background. In this study, statistical analysis was used to identify those climatic, hydrologic, and agricultural variables that best explained variations in nitrate, phosphorus, and total suspended solids over the period 1976-1995 in two large agricultural watersheds that feed Lake Erie, those of the Maumee and Sandusky Rivers. The dominant variable was stream discharge; after curvefits to remove its influence, the residual loads were tested via stepwise linear regression to reveal the most significant explanatory variables. Loads of nitrate, total suspended solids, and total phosphorus tended to decrease when previous months were wet, except in the summer, and to decrease when snow cover was extensive. It is speculated that stores of nitrate in the soil were lost during wet periods through increased crop uptake and/or leaching. Nitrogen fertilizer application in the Maumee watershed decreased following dry periods, but not enough to decrease stream loads. Soluble reactive phosphorus loads were negatively correlated to conservation tillage and reserves, and positively correlated to fertilizer and manure sources. Results for total phosphorus were similar to those for total suspended solids, on which most transported phosphorus is adsorbed.

Agriculture↗

Phenotypic characterization of copper-resistant mutants of Methylosinus trichosporium OB3b.

Cultures of Methylosinus trichosporium OB3b grown in the presence of very low concentrations of copper synthesize a soluble methane monooxygenase (sMMO) that efficiently catalyzes the oxidation of trichloroethylene and other organic pollutants. Recently, we isolated five M. trichosporium OB3b mutants that express sMMO activity when grown in the presence of elevated copper concentrations (P.A. Phelps, S. K. Agarwal, G. E. Speitel, Jr., and G. Georgiou, Appl. Environ. Microbiol. 58:3701-3708, 1992). Here we show that, in contrast to the results for the wild-type cells, the addition of copper to mutant cultures grown on methane and nitrate as the nitrogen source has no noticeable effect on the growth rate and sMMO expression. In vitro experiments indicated that the copper-resistant phenotype does not arise from an increased stability of sMMO to copper deactivation. Furthermore, the mutant cultures exhibit altered speciation of copper in the extracellular fluid and have substantially decreased levels of cell-associated copper. On the basis of these results, we propose that the mutant phenotype arises from defects in copper uptake and metabolism rather than from changes in sMMO expression or enzyme stability.

Biodegradation, Environmental↗

Performance of sulphonic cation exchangers in the recovery of ammonium from basic and slight acidic solutions.

Two sulphonated polystyrene-divinylbenzene cation exchangers (gel type Purolite SGC 100 x 10 MBH and macroporous Purolite C160 MBH) have been investigated for NH(4)(+) ions uptake from the ammonium-rich simulated solutions, corresponding to the caustic condensate of the nitrogen fertilizers production. One component (NH(3) or NH(4)NO(3)) solutions and the mixtures with varying molar ratio of these compounds have been used at the total concentration 0.214 mol/L. Batch and column experiments have been conducted to establish the influence of the matrix structure on the performance of the cation exchangers investigated during the sorption and the desorption. Batch sorption isotherms and breakthrough curves have shown the similar behaviour of the cation exchangers in the removal of NH(4)(+) and NH(3). On decreasing the influent pH from 11.4 to 5.74 a decrease in breakthrough capacity (BC) from 2.57 to 1.93 mol/L was observed. The distribution coefficients (K(d)), calculated from the batch sorption isotherms, are higher for the basic feed solution than for slightly acidic one. Both the degree of the cation exchanger regeneration (N/N(0)) and the efficiency of the NH(4)(+) ions recovery (N(R)), obtained using 0.7 bed volume (BV) of eluent (20% nitric acid) are lower for Purolite C 160 MBH than those for Purolite SGC 100 x 10 MBH. Using 5 BV of eluent the efficiency of the ammonium recovery amounted to 100% for both cation exchangers investigated. With respect to the efficiency of NH(4)(+) ions sorption and regeneration cation exchangers investigated are applicable for the recovery of ammonium ions from caustic condensate in the nitrogen fertilizers production.

Adsorption↗

Photoenhanced uptake of gaseous NO2 on solid organic compounds: a photochemical source of HONO?

In several recent field campaigns the existence of a strong daytime source of nitrous acid was demonstrated. The mechanism of this source remains unclear. Accordingly, in the present laboratory study, the effect of light (in the range 300-500 nm) on the uptake kinetics of NO2 on various surfaces taken as proxies for organic surfaces encountered in the troposphere (as organic aerosol but also ground surfaces) was investigated. In this collaborative study, the uptake kinetics and product formation rate were measured by different flow tube reactors in combination with a sensitive HONO instrument. Uptake on light absorbing aromatic compounds was significantly enhanced when irradiated with light of 300-420 nm, and HONO was formed with high yield when the gas was humidified. Especially organic substrates containing a combination of electron donors, such as phenols, and of compounds yielding excited triplet states, such as aromatic ketones, showed a high reactivity towards NO2. Based on the results reported a mechanism is suggested, in which photosensitised electron transfer is occurring. The results show that HONO can be efficiently formed during the day in the atmosphere at much longer wavelengths compared to the recently proposed nitrate photolysis.

Aerosols↗

Glutamine Transfer from Xylem to Phloem and Translocation to Developing Leaves of Populus deltoides.

The distribution of (14)C from xylem-borne [(14)C]glutamine, the major nitrogen compound moving in xylem sap of cottonwood (Populus deltoides Bartr. ex Marsh), was followed in rapidly growing shoots with a combination of autoradiographic, microautoradiographic, and radioassay techniques. Autoradiography and (14)C analyses of tissues showed that xylem-borne glutamine did not move with the transpiration stream into mature leaves. Instead, most of it was transferred from xylem to phloem in the upper stem and then translocated to young developing tissues. Microautoradiography showed that metaxylem parenchyma, secondary xylem parenchyma, and rays were the major areas of uptake from xylem vessels in the stem. Accumulation in phloem (high (14)C concentrations in sieve tubes) took place in internodes subtending recently mature leaves. Little (14)C from xylem-borne glutamine was found in phloem of mature leaves, which indicates restricted retransport of glutamine that did enter the leaf. In the primary tissues of the upper stem, most (14)C was found in the phloem. Cottonwood stems have an efficient uptake and transfer system that enhances glutamine movement to developing tissues of the upper stem.

Journal Article↗

Transport and fate of nitrate in headwater agricultural streams in Illinois.

Nitrogen inputs to the Gulf of Mexico have increased during recent decades and agricultural regions in the upper Midwest, such as those in Illinois, are a major source of N to the Mississippi River. How strongly denitrification affects the transport of nitrate (NO(3)-N) in Illinois streams has not been directly assessed. We used the nutrient spiraling model to assess the role of in-stream denitrification in affecting the concentration and downstream transport of NO(3)-N in five headwater streams in agricultural areas of east-central Illinois. Denitrification in stream sediments was measured approximately monthly from April 2001 through January 2002. Denitrification rates tended to be high (up to 15 mg N m(-2) h(-1)), but the concentration of NO(3)-N in the streams was also high (>7 mg N L(-1)). Uptake velocities for NO(3)-N (uptake rate/concentration) were lower than reported for undisturbed streams, indicating that denitrification was not an efficient N sink relative to the concentration of NO(3)-N in the water column. Denitrification uptake lengths (the average distance NO(3)-N travels before being denitrified) were long and indicated that denitrification in the streambed did not affect the transport of NO(3)-N. Loss rates for NO(3)-N in the streams were <5% d(-1) except during periods of low discharge and low NO(3)-N concentration, which occurred only in late summer and early autumn. Annually, most NO(3)-N in these headwater sites appeared to be exported to downstream water bodies rather than denitrified, suggesting previous estimates of N losses through in-stream denitrification may have been overestimated.

Agriculture↗

Some aspects of the ecophysiology of cyanobacteria.

Fresh waters rich in nutrients often show mass development of cyanobacteria. The kind of cyanobacteria to be found depends on the properties of the lake. In non-stratified shallow lakes, the most common species Oscillatoria agardhii. In stratified lakes, cyanobacteria can be found in restricted zones of the deeper part of the lakes, and always possess cells with very active gas vacuoles. The most common species are Microcystis aeruginosa, Oscillatoria agardhii var isothrix, Oscillatoria "var red" and different Anabaena species. In lakes with prolonged nitrogen limitation, genera such as Anabaena and Aphanizomenon are common. Two important factors determine the distribution of these organisms: the light climate and the availability of nutrients. We have limited our discussion in this paper to the influence of the light climate. This influence of the light climate on growth can be determined in different ways. The influence of light intensity on the growth rate of Oscillatoria agardhii can be described using a Monod-like relation with light inhibition at light intensities above 40 W m-2. Studies with other species of cyanobacteria like Microcystis aeruginosa and Aphanizomenon flos-aquae support the assumption that most cyanobacteria living in fresh water are extremely sensitive to high light intensities. The energy balance of phototrophic growth can be described by the equation mu = qE . c - mue where mu is the specific growth rate, mue the specific maintenance energy rate and qE the specific light energy uptake rate; c represents the growth efficiency factor with which light energy is converted into cell material. In this equation, there are two factors which determine the growth of the organisms: ue and c. It was found that the value of mue of Oscillatoria was extremely low (mue approximately 0.001 h-1) compared with a eukaryotic organism (Scenedesmus: mue 0.008 h-1). The value of c was found to be dependent on the growth rate, but did not greatly differ from values found in eukaryotic algal cells. Data obtained from cultures of Oscillatoria grown under light-dark cycles, so that total energy input over 24 h was growth-limiting, showed good regulation of both the carbohydrate synthesis rate and the growth rate. In comparing the data of light-limited growth of cyanobacteria, the conclusion can be drawn that these organisms are favoured at low light intensities by their low specific maintenance energy rate and their pigment composition. A number of competition experiments support the conclusion that the light climate is the most important steering factor for the distribution of cyanobacteria in fresh waters.

Cyanobacteria↗

Iron uptake from ferrioxamine and from ferrirhizoferrin by germinating spores of Rhizopus microsporus.

Mucormycosis caused by the fungus Rhizopus has been documented in iron overloaded patients and more particularly in dialysis patients, both when treated with desferrioxamine B (DFO). This iron and aluminium chelator is thought to play a role in the pathogenesis of this infection. We therefore investigated in vitro the cellular pharmacology of iron chelated by DFO in the fungus Rhizopus. In a medium, designed for fungal cultivation, Rhizopus microsporus var. rhizopodiformis takes up iron from ferric-DFO complex (55Fe.DFO) and from 55Fe.rhizoferrin, the siderophore synthesized and secreted by Rhizopus [Drechsel et al., Biol. Metals 4: 238-243, 1991]. In both cases, iron accumulation is partially saturable with the duration of exposure and the chelator concentration. Fe.DFO binds to Rhizopus; iron becomes trapped and remains associated with the fungus, whereas the iron-depleted siderophore is released in the extracellular medium. In a medium designed for mammalian cell cultivation and in the absence of human serum, the fungal iron accumulation both from 55Fe.DFO and from 55Fe.rhizoferrin is proportional to the chelator concentration. Human serum at 40% does not influence the iron accumulation from Fe.DFO but it significantly affects that from Fe.rhizoferrin which, in the presence of serum, only occurs at concentration > 5 microM. This difference finds its explanation in the iron transfer observed between Fe.rhizoferrin and seric apotransferrin, the latter making the metal unavailable to Rhizopus. By contrast, no iron transfer takes place between Fe.DFO and apotransferrin, allowing fungal iron utilization from this complex, even at very low concentrations. The iron uptake, being inhibited by NaN3 and KCN, is energy-dependent; being inhibited by bipyridyl, it requires prior reduction of ferric iron; being unaffected by the covalent linkage of Fe.DFO to albumin, it does not require the entry of Fe.DFO within the fungus. These in vitro results strongly suggest that, upon administration of DFO to iron overloaded or dialysis patients, the formed Fe.DFO is efficiently used as an iron source by Rhizopus, even in the presence of serum apotransferrin or rhizoferrin. The consequent promotion of the growth of Rhizopus helps explain the increased risk of mucormycosis in DFO-treated patients.

2,2'-Dipyridyl↗

Adsorption of hydrocarbons on mesoporous SBA-15 and PHTS materials.

Plugged hexagonal templated silica (PHTS) materials are synthesized using a high TEOS/EO(20)PO(70)EO(20) ratio in the SBA-15 synthesis. This generates internal microporous nanocapsules or plugs in part of the channels, which could be inferred from the two-step desorption branch. These materials exhibit a tunable amount of open and plugged pores and a very high micropore volume (up to 0.24 mL/g) and are more stable than the conventional micellar templated structures known so far. In this study the adsorption properties of PHTS are investigated and compared to those of its plug-free analogue SBA-15. For this purpose nitrogen, n-hexane, n-heptane, c-hexane, 3-methylpentane, 1-hexene, and water were adsorbed on SBA-15 and PHTSs with a different ratio of open and plugged mesopores. The adsorption of n-hexane, c-hexane, n-heptane, and 3-methylpentane on SBA-15 and PHTS-A demonstrated that the presence of the plugs had an effect on the uptake of adsorbate in the low relative pressure region, the position of the capillary condensation step, and the total adsorbed amount of adsorbate. The results showed that n-heptane and 3-methylpentane cannot access part of the micropore system of SBA-15 and PHTS-A. Adsorption of c-hexane and n-hexane on PHTS-A indicated that not only the kinetic diameter but also the shape of the molecule is an important factor for being able to be adsorbed into the micropores or past the plugs. Moreover, these two adsorbates were the most efficient in filling up the available pore volume. From the adsorption of n-hexane on PHTSs with a different ratio of open and plugged pores, it was concluded that the size of the plugs differed, which depends on the synthesis conditions. Water adsorption isotherms proved SBA-15 and PHTS-B to be more hydrophobic than PHTS-A. n-Hexane, 1-hexene, and toluene were adsorbed on SBA-15 and the PHTSs to investigate the influence of the polarity of the adsorbate. The isotherms showed higher uptakes for polar adsorbates on more hydrophobic materials and vice versa.

Journal Article↗

Enhanced photodegradation of 2,4,6-trichlorophenol over palladium phthalocyaninesulfonate modified organobentonite.

This article explores a supported photocatalyst of palladium(II) phthalocyaninesulfonate (PdPcS) onto organoclay for removal of 2,4,6-trichlorophenol (TCP) from water under visible light (lambda > or = 450 nm) irradiation. The composite clay was not only a good sorbent for uptake of TCP from water, but it also exhibited notable activity for TCP oxidation via singlet molecular oxygen, generated in situ from PdPcS photosensitization. Complete dechlorination of TCP could be achieved after TCP was totally oxidized. The initial rate of TCP degradation was observed to increase with the initial amount of TCP sorption, the kinetics following well the Langmuir-Hinshelwood equation. The resulting rate constant of TCP oxidation was found to increase with the alkyl chain length of the intercalated surfactant from dodecyltrimethylammonium to cetyltrimethylammonium and to octadecyltrimethylammonium. Such the trend in photoactivity was in agreement with the catalyst capacity for TCP sorption from aqueous medium. The result showed that enrichment of toxic TCP on the catalyst was indeed an efficient way for enhancement of the photosensitized degradation of target pollutant. However, PdPcS loading led to a notable decrease in TCP sorption, because of partial blocking of the sorptive sites. The optimal loading of PdPcS was about 1.0 wt %. The catalyst was characterized by nitrogen adsorption, X-ray diffraction, visible diffuse reflectance spectroscopy, and TCP sorption, which revealed that the incorporated PdPcS, mainly in the form of photoactive monomer, coexisted with the intercalated surfactant in the bentonite interlayers. Seven repeated experiments demonstrated that the composite clay was relatively stable and could be repeatedly used for sorption and degradation of TCP pollutant via molecular oxygen and visible light.

Journal Article↗

Locoregional administration of 5-fluoro-2'-deoxyuridine (FdUrd) in Novikoff hepatoma in the rat: effects of dose and infusion time on tumor growth and on FdUrd metabolite levels in tumor tissue as determined by 19F-NMR spectroscopy.

The influence of infusion time and dose on the anticancer efficacy of 5-fluoro-2'-deoxyuridine (FdUrd) was investigated using a locoregional therapy model: Novikoff hepatoma transplanted i.m. into the thigh of Wistar rats and FdUrd infusion via a catheter implanted in the femoral artery. In experiment A the FdUrd dose (five daily doses of 12, 19 and 30 mg/kg) and the duration of administration (bolus, 1 h, 5 h, and 24 h) were varied. The change in tumor volume following treatment and the number of rats showing regression vs progression served as indicators of therapy response. The results showed a clear dose dependence, and for each infusion time the 30 mg/kg dose was the most effective, without any signs of general toxicity. At this dose the longest infusion time (24 h) was less effective (regression in three of six rats) compared with 1-h or 5-h treatments (four of five in regression). In experiment B either one or five daily FdUrd doses (15, 30, 60 mg/kg) were administered i.a. for the same infusion times used in experiment A. After treatment, tumors were explanted ex vivo and approximately 1-g tissues samples were immediately frozen in liquid nitrogen for storage. 19F-NMR spectroscopy at 11.7 T was used to quantify FdUrd metabolites [5-fluorouracil (FUra), alpha-fluoro-beta-alanine (F beta Ala), 5-fluorouracil nucleosides and nucleotides (F-Nuc)] in the solid tumor tissue samples (maintained at 4 degrees C) with a detection threshold of about 5 nmol/g. The metabolite signal pattern indicated that FdUrd is first converted to FUra, followed by anabolism primarily to nucleotides in the oxy form (e.g. FUTP). The total amount of fluorine detected in tumor tissue increased with dose and decreased with infusion time. For all treatments FNuc could be detected, even after 24 h infusion, and their levels showed a good linear correlation with the total F. The major catabolite F beta Ala was present in tumor at low levels that correlated poorly with total F, indicating recirculation from other organs (e.g. liver) as the main source. Thus, the NMR method can provide detailed information regarding the efficiency of locoregional treatment (catheter function, drug uptake and metabolism). Initial results of non-invasive in vivo NMR experiments are also presented.

Animals↗

Climate change has only a minor impact on nutrient resorption parameters in a high-latitude peatland.

Nutrient resorption from senescing plant tissues is an important determinant of the fitness of plant populations in nutrient-poor ecosystems, because it makes plants less dependent on current nutrient uptake. Moreover, it can have significant "afterlife" effects through its impact on litter chemistry and litter decomposability. Little is known about the effects of climate change on nutrient resorption. We studied the effects of climate change treatments (including winter snow addition, and spring and/or summer warming) on nutrient resorption of four dominant species in a nutrient-poor subarctic peatland. These species were Betula nana (woody deciduous), Vaccinium uliginosum (woody deciduous), Calamagrostis lapponica (graminoid) and Rubus chamaemorus (forb). After five years of treatments both mature and senesced leaf N concentrations showed a small but significant overall reduction in response to the climate treatments. However, the effects were species-specific. For example, in the controls the N concentration in senesced leaves of Calamagrostis (3.0+/-0.2 mg N g(-1)) was about four times lower than for Rubus (11.2+/-0.2 mg N g(-1)). There were no significant treatment effects on N resorption efficiency (% of the N pool in mature leaves that is resorbed during senescence). The nitrogen resorption efficiency of Calamagrostis (about 80%) was higher than in the other three species (about 60%). Thus, climate change has only a minor impact on nutrient resorption parameters. However, given the substantial interspecific differences in these parameters, substantial changes in plant-soil feedbacks may be expected as a result of the observed changes in the species composition of high-latitude vegetation. These changes are species-specific and thus difficult to predict.

Analysis of Variance↗

Mechanisms of pulmonary NO2 absorption.

Although NO2-induced cytotoxic responses have been well characterized, the specific mechanisms responsible for initiating toxicity remain equivocal. The inhomogeneous distribution of epithelial injury suggests that differential interactions between NO2 and the lung surfaces may contribute to the extent of regional responses. Consequently, we have initiated studies to characterize the mechanisms which govern NO2 absorption and the initiation of the toxic cascade. Due to limitations in whole animal models, we have utilized numerous in vitro exposure models. Herein we examine our recent investigations. In brief synopsis: NO2 uptake is governed by reaction between inhaled NO2 and constituents of the pulmonary surface lining layer (SLL). The predominant reaction pathway involves hydrogen abstraction producing HNO2 and an organic radical. NO2 uptake is first-order with respect to NO2 ([NO2] < 10 ppm), is aqueous substrate-dependent, and is saturable. Conditions at the gas/liquid interface proper modulate the rate of transfer into the aqueous phase. Most likely, NO2 does not diffuse unreacted through the SLL. Absorption is proportional to inspired dose. The clearance efficiency may be modulated by ventilation frequency and the effective surface area of the exposed air space surfaces. We propose that the profile and concentration of SLL constituents mediate both the dosimetry and the extent of epithelial responses. Due to differential lining layer conditions, the relationship between absorbed dose and response may be complex and may exhibit anatomic and host variability.

Absorption↗

Effects of heavy metals on nitrogen uptake by Paxillus involutus and mycorrhizal birch seedlings.

The effects of the heavy metals Cu, Cd, Ni, Pb and Zn on [(14)C]methylamine and [(14)C]aminoisobutyric acid uptake were studied in the free-living fungus Paxillus involutus and in mycorrhizal and non-mycorrhizal birch roots. The uptake of both N sources by P. involutus was inhibited by the five metals tested. However, Cu(2+) and Pb(2+) had a greater inhibitory effect. Non-competitive inhibitions were determined between heavy metals and [(14)C]methylamine uptake. [(14)C]Methylamine uptake was reduced by one third by 2 µM Cd(2+) and Cu(2+) in non-mycorrhizal roots, whereas that of mycorrhizal roots was not affected. However, it was reduced by 30 to 80% by 200 µM Cd(2+) and Cu(2+) irrespective of the mycorrhizal status. [(14)C]Aminoisobutyric acid uptake in mycorrhizal roots was not significantly affected by Cd(2+) and Cu(2+), whereas that of non-mycorrhizal roots was decreased by 77% at 200 µM Cu(2+). [(14)C]Aminoisobutyric acid uptake was 4.5 to 6 fold higher in mycorrhizal roots, compared with non-mycorrhizal roots, even under metal exposure. The high efficiency of N acquisition by mycorrhizal birch seedlings under metal exposure might be regarded as a mechanism of stress avoidance.

Journal Article↗

Growth of Pseudomonas aeruginosa on nitrous oxide.

Three strains of Pseudomonas aeruginosa were grown anaerobically on exogenous N2O in a defined medium under conditions that assured the maintenance of highly anaerobic conditions for periods of 1 week or more. The bacteria were observed reproducibly to increase their cell density by factors of 3 to 9, but not more, depending on the initial amount of N2O. Growth on N2O was cleanly blocked by acetylene. Cell yields, CO2 production, and N2O uptake all increased with initial PN2O at PN2O less than or equal to 0.1 atm. Growth curves were atypical in the sense that growth rates decreased with time. This is the first observation of growth of P. aeruginosa on N2O as the sole oxidant. N2O was shown to be an obligatory, freely diffusible intermediate during growth of strains PAO1 and P1 on nitrate. All three strains used this endogenous N2O efficiently for growth. For strains PAO1 and P1, it was confirmed that exogenous N2O had little effect on the cell yields of cultures growing with nitrate; thus, for these strains exogenous N2O neither directly inhibited growth nor was used significantly for growth. On the other hand, strain P2 grew abundantly on exogenous N2O when small and growth-limiting concentrations of nitrate or nitrate (2 to 10 mM) were included in the medium. The dramatic effect of these N-anions was realized in large part even when the exogenous N2O was introduced immediately after the quantitative conversion of anion-nitrogen to N2. No evidence was found for a factor in filter-sterilized spent medium that stimulated fresh inocula to grow abundantly on N2O.(ABSTRACT TRUNCATED AT 250 WORDS)

Anaerobiosis↗

Culture treatments for enhancing post-thaw recovery of cryopreserved suspension cells of potato cv. Desiree.

An efficient and reproducible protocol has been developed for the cryopreservation of cell suspension cultures of the potato (Solanum tuberosum L.) cv. Desiree. An evaluation was made of the effectiveness of different pre-culture and post-thaw treatments on cell growth, as measured by changes in biomass. Cell suspensions were cultured in UM medium supplemented with 0.25, 0.5, 0.625, 0.75 or 1.0 M sucrose prior to cryopreservation. Sucrose-treated cells were harvested from suspension and 0.75 ml packed cell volumes placed in 2 ml capacity polypropylene vials with 0.5 ml of chilled cryoprotectant (glycerol 46.0 g 1(-1), dimethylsulphoxide 39.0 g 1(-1), sucrose 342.0 g 1(-1) proline 5.0 g 1(-1); pH 5.8). Cells were frozen at -0.5 degrees C min(-1) from 0 to -35 degrees C, held at -35 degrees C for 35 min and stored, for 10 days, in liquid nitrogen (-196 degrees C). The most effective pre-treatment, in terms of subsequent post-thaw cell viability as assessed by fluorescein diacetate uptake or triphenyltetrazolium chloride reduction, was culture with 0.75 M sucrose. For this treatment, the mean absorbance (490 nm) following triphenyltetrazolium chloride reduction was 88% greater (p < 0.05) than control and 59% greater (p < 0.05) for thawed cells also cultured on supporting filter paper discs.

Cell Survival↗

The Sinorhizobium meliloti ABC transporter Cho is highly specific for choline and expressed in bacteroids from Medicago sativa nodules.

In Sinorhizobium meliloti, choline is the direct precursor of phosphatidylcholine, a major lipid membrane component in the Rhizobiaceae family, and glycine betaine, an important osmoprotectant. Moreover, choline is an efficient energy source which supports growth. Using a PCR strategy, we identified three chromosomal genes (choXWV) which encode components of an ABC transporter: ChoX (binding protein), ChoW (permease), and ChoV (ATPase). Whereas the best homology scores were obtained with components of betaine ProU-like systems, Cho is not involved in betaine transport. Site-directed mutagenesis of choX strongly reduced (60 to 75%) the choline uptake activity, and purification of ChoX, together with analysis of the ligand-binding specificity, showed that ChoX binds choline with a high affinity (KD, 2.7 microM) and acetylcholine with a low affinity (KD, 145 microM) but binds none of the betaines. Uptake competition experiments also revealed that ectoine, various betaines, and choline derivatives were not effective competitors for Cho-mediated choline transport. Thus, Cho is a highly specific high-affinity choline transporter. Choline transport activity and ChoX expression were induced by choline but not by salt stress. Western blotting experiments with antibodies raised against ChoX demonstrated the presence of ChoX in bacteroids isolated from nitrogen-fixing nodules obtained from Medicago sativa roots. The choX mutation did not have an effect on growth under standard conditions, and neither Nod nor Fix phenotypes were impaired in the mutant, suggesting that the remaining choline uptake system(s) still present in the mutant strain can compensate for the lack of Cho transporter.

ATP-Binding Cassette Transporters↗

Investigation on the mechanism of H(2)S removal by biological activated carbon in a horizontal biotrickling filter.

The use of supporting media for the immobilization of microorganisms is widely known to provide a surface for microbial growth and a shelter that protects the microorganisms from inhibitory compounds. In our previous studies, activated carbon (AC) alone used as a support medium for H(2)S biological removal was proved prompt and efficient in a bench-scale biofilter and biotrickling filter. In this study, the mechanisms of H(2)S elimination using microbial immobilized activated carbon, i.e., biological activated carbon (BAC), are investigated. A series of BAC as supporting medium were taken from the inlet to outlet of a bench-scale horizontal biotrickling filter to examine the different effects of physical/chemical adsorption and microbial degradation on the overall removal of H(2)S. The surface properties of BAC together with virgin and exhausted carbon (after H(2)S breakthrough test, non-microbial immobilization) were characterized using the sorption of nitrogen (Braunner-Emmett-Teller test), scanning electron microscopy (SEM), surface pH, thermal, carbon-hydrogen-nitrogen-sulfur (CHNS) elemental and Fourier transform infrared (FTIR) analyses. Tests of porosity and surface area provide detailed information about the pore structure of BAC along the bed facilitating the understanding of potential pore blockages due to biofilm coating. A correlation between the available surface area and pore volume with the extent of microbial immobilization and H(2)S uptake is evidenced. SEM photographs show the direct carbon structure and biofilm coated on carbon surface. FTIR spectra, differential thermogravimetric curves and CHNS results indicate less diversity of H(2)S oxidation products on BAC than those previously observed on exhausted carbon from H(2)S adsorption only. The predominant oxidation product on BAC is sulfuric acid, and biofilm is believed to enhance the oxidation of H(2)S on carbon surface. The combination of biodegradation and physical adsorption of using BAC in removal of H(2)S could lead to a long-term (i.e., years) good performance of biotrickling filters and biofilters based on BAC compared to carbon adsorption only.

Bacteria↗