Superior residual fertiliser value in soil with phosphorus recycled from urine in layered double hydroxides.

Layered double hydroxides (LDHs) of magnesium (Mg) and aluminium (Al) are ion exchangers that can be used as slow release phosphorus (P) fertilisers. These LDHs can be used successfully to concentrate P from waste streams such as urine. This study was set up to test the fertiliser potential of P derived from urine and concentrated on LDHs. Ryegrass was grown in a pot trial using a P- and N-deficient soil where different urine derived fertilisers, i.e. LDH-P, stored urine and urine mixed with sludge as a source of P were compared to different mineral N and P doses in a full factorial design. Plants were grown for 75 days with four cuttings and did not exhibit salinity stress in stored urine treatments. Plant growth and P uptake responded to N, P doses in mineral fertilizer treatments with significant N-P interaction. The fertiliser use efficiency of urine fertilisers was lower than that of mineral fertilisers at equivalent total nutrient input for stored urine, due to lower N availability, and for urine mixed with sludge due to lower P availability. In contrast, the yield and P uptake of ryegrass grown on LDH loaded with P from urine (LDH-P) showed equal fertiliser P use as mineral fertiliser. Interestingly, the residual soil P after harvest, scored by the sum of isotopically exchangeable P in soil and the P uptake, was higher for LDH-P than for mineral P, confirming slow release properties of LDH that limit loss of P by fixation in soil.

Inefficacy of vancomycin and teicoplanin in eradicating and killing Staphylococcus epidermidis biofilms in vitro.

Biofilm-associated bacteria display a decreased susceptibility towards antibiotics. Routine assessment of antibiotic susceptibility of planktonic bacteria therefore offers an insufficient prediction of the biofilm response. In this study, in vitro biofilms of eight clinical Staphylococcus epidermidis strains were subjected to treatment with vancomycin, teicoplanin, oxacillin, rifampicin and gentamicin. In addition, the biofilms were subjected to combinations of an antibiotic with rifampicin. The effects on the biofilms were assessed by crystal violet staining to determine the total biofilm biomass, staining with XTT to determine bacterial cell viability, and microscopy. Combining these methods showed that treatment of S. epidermidis biofilms with glycopeptides increased the total biofilm biomass and that these antibiotics were not effective in killing bacteria embedded in biofilms. The decreased killing efficacy was more pronounced in biofilms produced by strains that were classified as 'strong' biofilm producers. Rifampicin, oxacillin and gentamicin effectively killed biofilm-associated bacteria of all tested strains. Combining antibiotics with rifampicin increased the killing efficacy without influencing the total biofilm biomass. When vancomycin or teicoplanin were combined with rifampicin, the increase in biofilm biomass was neutralised and also the killing efficacy was influenced in a positive way. We conclude that the combined methodology used in this study showed that glycopeptides were not effective in eradicating S. epidermidis biofilms but that combination with rifampicin improved the killing efficacy in vitro.

Labile synthetic cadmium complexes are not bioavailable to Pseudokirchneriella subcapitata in resin buffered solutions.

The Free Ion Activity Model (FIAM) predicts that cadmium (Cd) uptake by organisms is identical for solutions with the same free Cd(2+) concentration and inorganic composition. Clear exceptions to the FIAM have been shown for Cd uptake by plant roots, periphyton and human cells where labile Cd complexes increase bioavailability and which has been attributed to their role in enhancing Cd diffusion towards the uptake cells. Here, we assessed the role of labile Cd complexes on Cd uptake by algae, for which diffusion limitations should be less pronounced due to their smaller size. Long-term (3 days) Cd uptake by the green algae Pseudokirchneriella subcapitata was measured in resin buffered solutions with or without synthetic ligands and at three Cd(2+) ion activities (pCd 8.2-5.7). The free Cd(2+) activity was maintained during the test using a metal-selective resin located in the algal bottles. Total dissolved Cd increased up to 35-fold by adding the synthetic ligands at constant Cd(2+) activity. In contrast, Cd uptake by algae increased maximally 2.8 fold with increasing concentration of the synthetic ligands and the availability of the complexes were maximally 5.2% relative to Cd(2+) for NTA and CDTA complexes. It is concluded that labile Cd complexes do not greatly enhance Cd bioavailability to the unicellular algae and calculations suggest that Cd transport from solution to these small cells is not rate limiting.

A resin-buffered nutrient solution for controlling metal speciation in the algal bottle assay.

Metal speciation in solution is uncontrolled during algal growth in the traditional algal bottle assay. A resin-buffered nutrient solution was developed to overcome this problem and this was applied to test the effect of chloride (Cl⁻) on cadmium (Cd) uptake. Standard nutrient solution was enriched with 40 mM of either NaNO3 or NaCl, and was prepared to contain equal Cd²âº but varying dissolved Cd due to the presence of CdCl(n)(2-n) complexes. Both solutions were subsequently used in an algal assay in 100 mL beakers that contained only the solution (designated "-R") or contained the solution together with a cation exchange sulfonate resin (2 g L⁻¹, designated "+R") as a deposit on the bottom of the beaker. Pseudokirchneriella subcapitata was grown for 72 h (1.4 × 105-1.4 × 106 cells mL⁻¹) in stagnant solution and shaken three times a day. Growth was unaffected by the presence of the resin (p>0.05). The Cd concentrations in solution of the -R devices decreased with 50-58% of initial values due to Cd uptake. No such changes were found in the +R devices or in abiotic controls. Cd uptake was unaffected by either NaNO3 or NaCl treatment in the +R device, confirming that Cd²âº is the preferred Cd species in line with the general concept of metal bioavailability. In contrast, Cd uptake in the -R devices was two-fold larger in the NaCl treatment than in the NaNO3 treatment (p<0.001), suggesting that CdCl(n)(2-n) complexes are bioavailable in this traditional set-up. However this bioavailability is partially, but not completely, an apparent one, because of the considerable depletion of solution ¹°9Cd in this set-up. Resin-buffered solutions are advocated in the algal bottle assay to control trace metal supply and to better identify the role of metal complexes on bioavailability.

Long-term dynamics of the atrazine mineralization potential in surface and subsurface soil in an agricultural field as a response to atrazine applications.

The dynamics of the atrazine mineralization potential in agricultural soil was studied in two soil layers (topsoil and at 35-45 cm depth) in a 3 years field trial to examine the long term response of atrazine mineralizing soil populations to atrazine application and intermittent periods without atrazine and the effect of manure treatment on those processes. In topsoil samples, (14)C-atrazine mineralization lag times decreased after atrazine application and increased with increasing time after atrazine application, suggesting that atrazine application resulted into the proliferation of atrazine mineralizing microbial populations which decayed when atrazine application stopped. Decay rates appeared however much slower than growth rates. Atrazine application also resulted into the increase of the atrazine mineralization potential in deeper layers which was explained by the growth on leached atrazine as measured in soil leachates recovered from that depth. However, no decay was observed during intermittent periods without atrazine application in the deeper soil layer. atzA and trzN gene quantification confirmed partly the growth and decay of the atrazine degrading populations in the soil and suggested that especially trzN bearing populations are the dominant atrazine degrading populations in both topsoil and deeper soil. Manure treatment only improved the atrazine mineralization rate in deeper soil layers. Our results point to the importance of the atrazine application history on a field and suggests that the long term survival of atrazine degrading populations after atrazine application enables them to rapidly proliferate once atrazine is again applied.

Stable isotope analysis of dissolved organic carbon in soil solutions using a catalytic combustion total organic carbon analyzer-isotope ratio mass spectrometer with a cryofocusing interface.

Stable carbon isotopes are a powerful tool to assess the origin and dynamics of carbon in soils. However, direct analysis of the (13)C/(12)C ratio in the dissolved organic carbon (DOC) pool has proved to be difficult. Recently, several systems have been developed to measure isotope ratios in DOC by coupling a total organic carbon (TOC) analyzer with an isotope ratio mass spectrometer. However these systems were designed for the analysis of fresh and marine water and no results for soil solutions or (13)C-enriched samples have been reported. Because we mainly deal with soil solutions in which the difficult to oxidize humic and fulvic acids are the predominant carbon-containing components, we preferred to use thermal catalytic oxidation to convert DOC into CO(2). We therefore coupled a high-temperature combustion TOC analyzer with an isotope ratio mass spectrometer, by trapping and focusing the CO(2) cryogenically between the instruments. The analytical performance was tested by measuring solutions of compounds varying in the ease with which they can be oxidized. Samples with DOC concentrations between 1 and 100 mg C/L could be analyzed with good precision (standard deviation (SD) < or = 0.6 per thousand), acceptable accuracy, good linearity (overall SD = 1 per thousand) and without significant memory effects. In a (13)C-tracer experiment, we observed that mixing plant residues with soil caused a release of plant-derived DOC, which was degraded or sorbed during incubation. Based on these results, we are confident that this approach can become a relatively simple alternative method for the measurement of the (13)C/(12)C ratio of DOC in soil solutions.

The impact of agricultural soil erosion on the global carbon cycle.

Agricultural soil erosion is thought to perturb the global carbon cycle, but estimates of its effect range from a source of 1 petagram per year(-1) to a sink of the same magnitude. By using caesium-137 and carbon inventory measurements from a large-scale survey, we found consistent evidence for an erosion-induced sink of atmospheric carbon equivalent to approximately 26% of the carbon transported by erosion. Based on this relationship, we estimated a global carbon sink of 0.12 (range 0.06 to 0.27) petagrams of carbon per year(-1) resulting from erosion in the world's agricultural landscapes. Our analysis directly challenges the view that agricultural erosion represents an important source or sink for atmospheric CO2.

Zoonotic transmission of Cryptococcus neoformans from a magpie to an immunocompetent patient.

We report a case of cryptococcal meningitis in an immunocompetent female patient with exposure to a pet magpie (Pica pica). Genetically indistinguishable isolates were cultured from the cerebrospinal fluid of the patient and excreta of the bird. Our data strongly suggest zoonotic transmission of Cryptococcus neoformans var. grubii from a magpie to this patient.

Radiocaesium soil-to-wood transfer in commercial willow short rotation coppice on contaminated farm land.

The feasibility of willow short rotation coppice (SRC) for energy production as a revaluation tool for severely radiocaesium-contaminated land was studied. The effects of crop age, clone and soil type on the radiocaesium levels in the wood were assessed following sampling in 14 existing willow SRC fields, planted on radiocaesium-contaminated land in Sweden following Chernobyl deposition. There was only one plot where willow stands of different maturity (R6S2 and R5S4: R, root age and S, shoot age) and clone (Rapp and L78183 both of age category R5S4) were sampled and no significant differences were found. The soils differed among others in clay fraction (3-34%), radiocaesium interception potential (515-6884 meq kg(-1)), soil solution K (0.09-0.95 mM), exchangeable K (0.58-5.77 meq kg(-1)) and cation exchange capacity (31-250 meq kg(-1)). The soil-to-wood transfer factor (TF) of radiocaesium differed significantly between soil types. The TF recorded was generally small (0.00086-0.016 kg kg(-1)), except for willows established on sandy soil (0.19-0.46 kg kg(-1)). Apart from the weak yet significant exponential correlation between the Cs-TF and the solid/liquid distribution coefficient (R2 = 0.54) or the radiocaesium interception potential, RIP (R2 = 0.66), no single significant correlations between soil characteristics and TF were found. The wood-soil solution 137Cs concentration factor (CF) was significantly related to the potassium concentration in the soil solution. A different relation was, however, found between the sandy Trödje soils (CF = 1078.8 x m(K)(-1.83), R2 = 0.99) and the other soils (CF = 35.75 x m(K)(-0.61), R2 =0.61). Differences in the ageing rate of radiocaesium in the soil (hypothesised fraction of bioavailable caesium subjected to fast ageing for Trödje soils only 1% compared to other soils), exchangeable soil K (0.8-1.8 meq kg(-1) for Trödje soils and 1.5-5.8 meq kg(-1) for the other soils) and the ammonium concentration in the soil solution (0.09-0.31 mM NH4+ for the Trödje soils compared to 0.003-0.11 mM NH4+ for the other soils) are put forward as potential factors explaining the higher CF and TF observed for the Trödje soils. Though from the dataset available it was not possible to unequivocally predict the Cs-soil-to-wood-transfer, the generally low TFs observed point to the particular suitability for establishment of SRC on radiocaesium-contaminated land.

Reliability of the ica, aap and atlE genes in the discrimination between invasive, colonizing and contaminant Staphylococcus epidermidis isolates in the diagnosis of catheter-related infections.

OBJECTIVES: To evaluate the usefulness of detecting two genes involved in biofilm formation (icaA and aap) and one gene involved in initial adhesion (atlE) for discrimination between contaminant, colonizing and invasive Staphylococcus epidermidis isolates involved in catheter-related infections. PATIENTS: The first group contained 29 isolates that were isolated from the skin of healthy volunteers (contaminant isolates). The second group contained 16 isolates recovered from catheters (>1000 CFUs on quantitative catheter culture) from asymptomatic patients without bacteremia. These isolates were considered to be colonizing isolates. The third group contained 34 isolates grown in >or=2 different blood cultures from patients with a systemic inflammatory response. These isolates were considered to be invasive isolates. RESULTS: The prevalence of atlE did not differ between the three groups. The icaA and aap genes were significantly more prevalent in colonizing isolates (88% aap; 88% icaA) than in invasive isolates (68% aap, P = 0.179; 59% icaA, P = 0.055) and than in skin isolates (52% aap, P = 0.02; 38% icaA, P = 0.002). CONCLUSIONS: The high prevalence of aap and icaA in skin isolates and their higher prevalence in colonizing than in invasive isolates led to a low specificity when these genes were used to differentiate between contamination, colonization and invasive infection. We conclude that, although the prevalence of these genes differs in the three groups, their presence cannot be used for clinical decision-making.

Predictions of in situ solid/liquid distribution of radiocaesium in soils.

Previous work has demonstrated that plant uptake of radiocaesium (RCs) is related to the activity concentration of RCs in soil solution, which is linked to the soil/soil solution distribution coefficient, K(D). The solid-liquid distribution of RCs is generally studied in soil suspensions in the laboratory and there are few reported measurements for in situ soil solutions. From a data set of 53 different soils (contaminated with either 134CsCl or 137CsCl) used in pot trials to investigate grass uptake of RCs, we analysed the variation of in situ K(D) with measured soil properties. The soils differed widely in % clay (0.5-58%), organic matter content (1.9-96%) and pH (2.4-7.0, CaCl2). The K(D) varied between 29 and 375,000 L kg-' (median 1460 L kg(-1)). Stepwise multiple regression analysis showed a significant correlation between the log K(D) and pH (p < 0.001), log %clay (p < 0.01) and log exchangeable K (p < 0.001) (overall R2 = 0.70). The in situ K(D) values were further compared to K(D)S predicted using an existing model, which assumes that RCs sorption occurs on specific sites and regular ion-exchange sites on the soil solid phase. Sorption of RCs on specific sites was quantified from the radiocaesium interception potential (RIP) measured for each soil and the soil solution concentrations of K+ and NH4+. The in situ log K(D) correlated well with the predicted K(D) (R2 = 0.85 before plant growth, R2 = 0.83 after plant growth). However, the observations were fivefold to eightfold higher than the predictions, particularly for the mineral soils. We attribute the under-prediction to the long contact times (minimum 4 weeks) between the RCs tracers and our experimental soils relative to the short (24 h) contact times used in RIP measurements. We conclude that our data confirmed the model but that ageing of RCs in soil is a factor that needs to be considered to better predict in situ KD values.

Use of gDNA as internal standard for gene expression in staphylococci in vitro and in vivo.

An internal RNA standard proved less suitable in bacterial gene expression experiments. We therefore developed a method for simultaneous RNA and gDNA (genomic DNA) isolation from in vitro and in vivo samples containing staphylococci and combined it with quantitative PCR. The reliability of gDNA for bacterial quantification and for standardisation in gene expression experiments was evaluated. Quantitative PCR proves equivalent to quantitative culture for in vitro samples, and superior for in vivo samples. In gene expression experiments, gDNA permits a good standardisation for the initial amount of bacteria. The average interassay variability of the in vitro expression is 20.1%. The in vivo intersample variability was 73.3%. This higher variability can be attributed to the biological variation of gene expression in vivo. This method permits exact quantification of the number of bacteria and the expression of genes in staphylococci in vivo (e.g., in biofilms, evolution in time) and in vitro.

Potential nitrification rate as a tool for screening toxicity in metal-contaminated soils.

A potential nitrification rate test (PNR) was used to identify metal toxicity in field-contaminated soils. The test was applied to metal salt-spiked soils, to 27 uncontaminated soils, and to 15 soils that are contaminated by former metal smelting activities. Four agricultural soils (pH 4.5-6.6) were spiked with various rates of CdCl2 (0-200 mg Cd/kg dry wt) or ZnCl2 (0-3,000 mg Cd/kg dry wt) and were equilibrated more than nine months prior to testing. The soil Zn EC50s of the PNR were between 150 and 350 mg Zn/kg dry weight. No continuous decrease of the nitrification with increasing Cd application was observed. The nitrification rate was reduced by between 50 and 80% at the highest Cd application in all soils. The PNRs of 27 uncontaminated soils varied widely (0-21 mg N/kg/d), but most of this variability is explained by soil pH (R2 = 0.77). The PNRs of the 15 contaminated soils were 0 to 44% of the values predicted for an uncontaminated soil at corresponding pH. Significant toxicity in field-contaminated soils was identified if the PNR was outside the 95% prediction interval of the PNR for an uncontaminated soil at corresponding pH and was found in seven soils. These soils contain 160 to 34,000 mg Zn/kg dry weight and 5 to 104 mg Cd/kg dry weight and had a pH >5.7. No toxicity could be detected below pH 5.6, where even a zero PNR value is within the 95% prediction interval of uncontaminated soils. It is concluded that the nitrification is sensitive to metal stress but that its power as a soil bioassay is low because of the high variability of the endpoint between uncontaminated soils. The ecological significance of the assay is discussed.

Quantification of expression of Staphylococcus epidermidis housekeeping genes with Taqman quantitative PCR during in vitro growth and under different conditions.

The aims of the present study were (i) to develop and test a sensitive and reproducible method for the study of gene expression in staphylococci and (ii) to study the expression of five housekeeping genes which are involved in nucleic acid metabolism (gmk, guanylate kinase; the dihydrofolate reductase [DHFR] gene), glucose metabolism (tpi, triosephosphate isomerase), and protein metabolism (the 16S rRNA gene; hsp-60, heat-shock protein 60) during in vitro exponential and stationary growth. A modified method for instant mRNA isolation was combined with gene quantification via Taqman real-time quantitative PCR. The detection limit of our method was 10 copies of RNA. The average intersample variability was 16%. A 10-fold increase in the expression of the hsp-60 gene was induced by exposure to a 10 degrees C heat shock (37 to 47 degrees C) for 10 min. During in vitro growth, the expression of all five housekeeping genes showed rapid up-regulation after inoculation of the bacteria in brain heart infusion medum and started to decline during the mid-exponential-growth phase. Maximal gene expression was 110- to 300-fold higher than gene expression during stationary phase. This indicates that housekeeping metabolism is a very dynamic process that is extremely capable of adapting to different growth conditions. Expression of the 16S rRNA gene decreases significantly earlier than that of other housekeeping genes. This confirms earlier findings for Escherichia coli that a decline in bacterial ribosomal content (measured by 16S rRNA gene expression) precedes the decline in protein synthesis (measured by mRNA expression).

Factors controlling sediment and phosphorus export from two Belgian agricultural catchments.

Sediment and total phosphorus (TP) export vary through space and time. This study was conducted to determine the factors controlling sediment and TP export in two agricultural catchments situated in the Belgian Loess Belt. At the outlet of these catchments runoff discharge was continuously measured and suspended sediment samples were taken during rainfall events. Within the catchments vegetation type and cover, soil surface parameters, erosion features, sediment pathways, and rainfall characteristics were monitored. Total P content and sediment characteristics such as clay, organic carbon, and suspended sediment concentration were correlated. Total sediment and TP export differ significantly between the monitored catchments. Much of the difference is due to the occurrence of an extreme event in one catchment and the morphology and spatial organization of land use in the catchments. In one catchment, the direct connection between erosive areas and the catchment outlet by means of a road system contributed to a high sediment delivery ratio (SDR) at the outlet. In the other catchment, the presence of a wide valley in the center of the catchment caused sediment deposition. Vegetation also had an effect on sediment production and deposition. Thus, many factors control sediment and TP export from small agricultural catchments; some of these factors are related to the physical catchment characteristics such as morphology and landscape structure and are (semi)permanent, while others, such as vegetation cover and land use, are time dependent.

Functional characterization of colloidal phosphorus species in the soil solution of sandy soils.

The impact of mobile colloids on the transport of phosphorus in the subsurface environment is not well understood. We hypothesized that interactions between metals, organic matter, and P control the dynamics of mobile colloidal P species in excessively fertilized sandy soils. The effect of UV irradiation and additions of 32P, orthophosphate, Fe, Al, and NaF on the concentration of colloidal P was examined using gel filtration chromatography. In addition, molybdate unreactive P (MUP) was characterized using phosphomonoesterase assays. The high molecular mass reactive P (HMMRP) fraction did not react to orthophosphate additions, increased upon Al and Fe additions and decreased upon NaF addition and UV irradiation. These results support the hypothesis that HMMRP is present as organic matter-metal-orthophosphate complexes. The concentration of high molecular mass unreactive P (HMMUP) decreased upon UV irradiation. The MUP concentration slightly decreased upon incubation with phytase and acid phosphatase. These observations fitted well to the "protection" hypothesis, where hydrolyzable P bonds are protected from monoesterase attack through occlusion in colloidal material. Taken together, this study indicates the high potential for subsurface P loss by colloidal particles in soils excessively fertilized with animal manure.

Evaluation of the intrinsic mtbe biodegradation potential in MTBE-contaminated soils.

MTBE has only recently being used as an octane enhancer in gasoline in Europe and is considered as a more recent groundwater contaminant on this continent. In this study we examined if during the recent contamination history, European MTBE contaminated aquifers had developed MTBE degrading microbial communities. Different MTBE contaminated and non-contaminated aquifers and soils were tested for their intrinsic biodegradation potential. The role of the oxygen concentration, the availability of nutrients and the influence of the presence of a co-contaminant like benzene on the MTBE biodegradation capabilities of the indigenous microorganisms were examined. All studied soil samples showed degradation of benzene under all tested conditions. On the other hand only one aquifer showed the capacity to degrade MTBE as demonstrated by the disappearance of MTBE and the production of TBA, the main degradation product of MTBE.

Symbiotic performance of herbaceous legumes in tropical cover cropping systems.

Increasing use of herbaceous legumes such as mucuna ( Mucuna pruriens var. utilis [Wright] Bruck) and lablab ( Lablab purpureus [L.] Sweet) in the derived savannas of West Africa can be attributed to their potential to fix atmospheric nitrogen (N2). The effects of management practices on N2 fixation in mucuna and lablab were examined using 15N isotope dilution technique. Dry matter yield of both legumes at 12 weeks was two to five times more in in situ mulch (IM) than live mulch (LM) systems. Land Equivalent Ratios, however, showed 8 to 30% more efficient utilization of resources required for biomass production under LM than IM systems. Live mulching reduced nodule numbers in the legumes by one third compared to values in the IM systems. Similarly, nodule mass was reduced by 34 to 58% under LM compared to the IM systems. The proportion of fixed N2 in the legumes was 18% higher in LM than IM systems. Except for inoculated mucuna, the amounts of N fixed by both legumes were greater in IM than LM systems. Rhizobia inoculation of the legumes did not significantly increase N2 fixation compared to uninoculated plots. Application of N fertilizer reduced N2 fixed in the legumes by 36 to 51% compared to inoculated or uninoculated systems. The implications of cover cropping, N fertilization, and rhizobia inoculation on N contributions of legumes into tropical low-input systems were discussed.

acs1 of Haemophilus influenzae type a capsulation locus region II encodes a bifunctional ribulose 5-phosphate reductase- CDP-ribitol pyrophosphorylase.

The serotype-specific, 5.9-kb region II of the Haemophilus influenzae type a capsulation locus was sequenced and found to contain four open reading frames termed acs1 to acs4. Acs1 was 96% identical to H. influenzae type b Orf1, previously shown to have CDP-ribitol pyrophosphorylase activity (J. Van Eldere, L. Brophy, B. Loynds, P. Celis, I. Hancock, S. Carman, J. S. Kroll, and E. R. Moxon, Mol. Microbiol. 15:107-118, 1995). Low but significant homology to other pyrophosphorylases was only detected in the N-terminal part of Acs1, whereas the C-terminal part was homologous to several short-chain dehydrogenases/reductases, suggesting that Acs1 might be a bifunctional enzyme. To test this hypothesis, acs1 was cloned in an expression vector and overexpressed in Escherichia coli. Cells expressing this protein displayed both ribitol 5-phosphate dehydrogenase and CDP-ribitol pyrophosphorylase activities, whereas these activities were not detectable in control cells. Acs1 was purified to near homogeneity and found to copurify with ribitol 5-phosphate dehydrogenase and CDP-ribitol pyrophosphorylase activities. These had superimposable elution profiles from DEAE-Sepharose and Blue-Sepharose columns. The dehydrogenase activity was specific for ribulose 5-phosphate and NADPH in one direction and for ribitol 5-phosphate and NADP+ in the other direction and was markedly stimulated by CTP. The pyrophosphorylase showed activity with CTP and ribitol 5-phosphate or arabitol 5-phosphate. We conclude that acs1 encodes a bifunctional enzyme that converts ribulose 5-phosphate into ribitol 5-phosphate and further into CDP-ribitol, which is the activated precursor form for incorporation of ribitol 5-phosphate into the H. influenzae type a capsular polysaccharide.