thiourea and allylthiourea

Nitrification inhibitors (NIs) such as dicyandiamide (DCD), 3,4-dimethylpyrazole phosphate (DMPP), and allylthiourea (AT) are commonly used to suppress ammonia oxidization at different time scales varying from a few hours to several months. Although the responses of NIs to edaphic and temperature conditions have been studied, the influence of the aforementioned factors on their inhibitory effect remains unknown. In this study, laboratory-scale experiments were conducted to assess the short-term (24 h) influence of eight abiotic and biotic factors on the inhibitory effects of DCD, DMPP, and AT across six cropped and non-cropped soils at two temperature conditions with three covariates of soil texture. Simultaneously, the dominant contributions of ammonia-oxidizing archaea (AOA) and bacteria (AOB) to potential ammonia oxidization (PAO) were distinguished using the specific inhibitor 2 phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO). Our results revealed that AT demonstrated a considerably greater inhibitory effect (up to 94.9% for an application rate of 75 mg of NI/kg of dry soil) than DCD and DMPP. The inhibitory effect of AT was considerably affected by the relative proportions of silt, sand, and clay in the soil and total PAO. In contrast to previous studies, the inhibitory effects of all three NIs remained largely unaffected by the landcover type and temperature conditions for the incubation period of 24 h. Furthermore, the efficacy of all three tested NIs was not affected by the differential contributions of AOA and AOB to PAO. Collectively, our results suggested a limited influence of temperature on the inhibitory effects of all three NIs but a moderate dependence of AT on the soil texture and PAO. Our findings can enhance the estimation of the inhibitory effect in soil, and pure cultures targeting the AOA and AOB supported ammonia oxidization and, hence, nitrogen dynamics under NI applications.

Topics: Ammonia; Guanidines; Nitrification; Oxidation-Reduction; Phosphates; Pyrazoles; Soil; Soil Microbiology; Thiourea

Ammonia oxidation is mainly performed by ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Allylthiourea (ATU) has been found to specifically inhibit ammonia oxidation. However, the effect of ATU on AOA and AOB transcription has been infrequently studied. In the present study, we examined the responses of AOA and AOB activity and DNA/cDNA community structure to ATU exposure. The ammonia oxidation activity in the 100-mg/L ATU group was 4.3% of that in the control group after 7 days. When exposed to ATU, the gene abundance of AOA was favored compared with that of AOB, and there were no statistically significant differences in the abundance of AOB amoA in DNA and cDNA between the two groups. Compared with the control group, the gene abundance of AOA significantly increased by 5.23 times, while the transcription of AOA significantly decreased by 0.70 times. Moreover, the transcriptional ratio of AOA in the ATU group was only 0.05 times as high as that in the control group. ATU selectively affected AOB and completely inhibited Nitrosomonas europaea and Bacterium amoA.22.HaldeII.kultur at the genetic level. Under ATU exposure, all AOA clusters were transcribed, but three AOB clusters were not transcribed. Our results indicated that the ammonia oxidation potential of the soil of water level fluctuation areas, based on ATU inhibition, was associated mainly with AOA amoA gene abundance and AOB community shifts in DNA and cDNA.

Topics: Ammonia; Archaea; Bacteria; DNA, Complementary; Oxidation-Reduction; Phylogeny; Soil; Soil Microbiology; Thiourea; Water

Functional carbon materials have been developed and applied in various sewage treatment applications in recent years. This article reports the fabrication, characterization, and application of a new kind of poly (allylthiourea-co-acrylic acid) (PAT⁻PAC) hydrogel-based carbon monolith. The results indicated that the poly acrylic acid component can endow the PAT⁻PAC hydrogel with an increased swelling ratio and enhanced thermal stability. During the carbonization process, O⁻H, N⁻H, C=N, and ⁻COO⁻ groups, etc. were found to be partly decomposed, leading to the conjugated C=C double bonds produced and the clear red shift of C=O bonds. Particularly, it was found that this shift was accelerated under higher carbonization temperature, which ultimately resulted in the complex conjugated C=C network with oxygen, nitrogen, and sulfur atoms doped in-situ. The as-obtained carbon monoliths showed good removal capacity for Ni(II) ions, organic solvents, and dyes, respectively. Further analysis indicated that the Ni(II) ion adsorption process could be well described by pseudo-second-order and Freundlich models under our experimental conditions, respectively. The adsorption capacity for Ni(II) ions and paraffin oil was as high as 557 mg/g and 1.75 g/g, respectively. More importantly, the as-obtained carbon monoliths can be recycled and reused for Ni(II) ions, acetone, and paraffin oil removal. In conclusion, the proposed PAT⁻PAC-based carbonaceous monoliths are superior adsorbents for wastewater treatment.

Topics: Acrylates; Adsorption; Carbon; Polymers; Thiourea; Wastewater; Water Pollutants, Chemical; Water Purification

Topics: Biological Oxygen Demand Analysis; Biomass; Bioreactors; Heterotrophic Processes; Kinetics; Models, Theoretical; Nitrification; Oxygen; Sewage; Thiourea; Waste Disposal, Fluid

Allylthiourea (ATU) and chlorate (ClO

Topics: Ammonium Compounds; Chlorates; Chlorides; Filtration; Nitrification; Research; Thiourea; Water Purification

Topics: Ammonia; Ampicillin; Archaea; Betaproteobacteria; Gene Dosage; Geologic Sediments; Guanidines; High-Throughput Nucleotide Sequencing; Nitrosomonas; Oxidation-Reduction; Phylogeny; Rivers; Smell; Thiourea

Four novel Pd

Topics: Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Cell Survival; Coordination Complexes; DNA; HeLa Cells; Humans; Inhibitory Concentration 50; Palladium; Platinum; Thiourea; X-Ray Diffraction

Nitrification is a critical process for the balance of reduced and oxidized nitrogen pools in nature, linking mineralization to the nitrogen loss processes of denitrification and anammox. Recent studies indicate a significant contribution of ammonia-oxidizing archaea (AOA) to nitrification. However, quantification of the relative contributions of AOA and ammonia-oxidizing bacteria (AOB) to in situ ammonia oxidation remains challenging. We show here the production of nitric oxide (NO) by Nitrosopumilus maritimus SCM1. Activity of SCM1 was always associated with the release of NO with quasi-steady state concentrations between 0.05 and 0.08 μM. NO production and metabolic activity were inhibited by the nitrogen free radical scavenger 2-phenyl-4,4,5,5,-tetramethylimidazoline-1-oxyl-3-oxide (PTIO). Comparison of marine and terrestrial AOB strains with SCM1 and the recently isolated marine AOA strain HCA1 demonstrated a differential sensitivity of AOB and AOA to PTIO and allylthiourea (ATU). Similar to the investigated AOA strains, bulk water column nitrification at coastal and open ocean sites with sub-micromolar ammonia/ammonium concentrations was inhibited by PTIO and insensitive to ATU. These experiments support predictions from kinetic, molecular and biogeochemical studies, indicating that marine nitrification at low ammonia/ammonium concentrations is largely driven by archaea and suggest an important role of NO in the archaeal metabolism.

Topics: Ammonia; Ammonium Compounds; Aquatic Organisms; Archaea; Betaproteobacteria; Cyclic N-Oxides; Denitrification; Imidazoles; Nitric Oxide; Nitrification; Nitrogen; Oxidation-Reduction; Thiourea

The fate of the most commonly used brominated flame retardant, tetrabromobisphenol A (TBBPA), in wastewater treatment plants is obscure. Using a (14)C-tracer, we studied TBBPA transformation in nitrifying activated sludge (NAS). During the 31-day incubation, TBBPA transformation (half-life 10.3 days) was accompanied by mineralization (17% of initial TBBPA). Twelve metabolites, including those with single benzene ring, O-methyl TBBPA ether, and nitro compounds, were identified. When allylthiourea was added to the sludge to completely inhibit nitrification, TBBPA transformation was significantly reduced (half-life 28.9 days), formation of the polar and single-ring metabolites stopped, but O-methylation was not significantly affected. Abiotic experiments confirmed the generation of mono- and dinitro-brominated forms of bisphenol A in NAS by the abiotic nitration of TBBPA by nitrite, a product of ammonia-oxidizing microorganisms (AOMs). Three biotic (type II ipso-substitution, oxidative skeletal cleavage, and O-methylation) and one abiotic (nitro-debromination) pathways were proposed for TBBPA transformation in NAS. Apart from O-methylation, AOMs were involved in three other pathways. Our results are the first to provide information about the complex metabolism of TBBPA in NAS, and they are consistent with a determining role for nitrifiers in TBBPA degradation by initiating its cleavage into single-ring metabolites that are substrates for the growth of heterotrophic bacteria.

Topics: Ammonia; Bacteria; Benzhydryl Compounds; Flame Retardants; Half-Life; Halogenation; Methylation; Nitrification; Phenols; Polybrominated Biphenyls; Sewage; Thiourea; Wastewater

The first step of nitrification is carried out by ammonia-oxidizing bacteria (AOB) and archaea (AOA). It is largely unknown, by which mechanisms these microbes are capable of coexistence and how their respective contribution to ammonia oxidation may differ with varying soil characteristics. To determine how different levels of ammonium availability influence the extent of archaeal and bacterial contributions to ammonia oxidation, microcosm incubations with controlled ammonium levels were conducted. Net nitrification was monitored and ammonia-oxidizer communities were quantified. Additionally, the nitrification inhibitor allylthiourea (ATU) was applied to discriminate between archaeal and bacterial contributions to soil ammonia oxidation. Thaumarchaeota, which were the only ammonia oxidizers detectable at the start of the incubation, grew in all microcosms, but AOB later became detectable in ammonium amended microcosms. Low and high additions of ammonium increasingly stimulated AOB growth, while AOA were only stimulated by the low addition. Treatment with ATU had no effect on net nitrification and sizes of ammonia-oxidizing communities suggesting that the effective concentration of ATU to discriminate between archaeal and bacterial ammonia oxidation is not the same in different soils. Our results support the niche-differentiating potential of ammonium concentration for AOA and AOB, and we conclude that ammonium limitation can be a major reason for absence of detectable AOB in soil.

Topics: Ammonia; Ammonium Compounds; Archaea; Bacteria; Grassland; Nitrification; Oxidation-Reduction; Soil; Soil Microbiology; Thiourea

Source waters sampled from Perpetual Spouter hot spring (pH 7.03, 86.4°C), Yellowstone National Park, WY, have low concentrations of total ammonia, nitrite, and nitrate, suggesting nitrogen (N) limitation and/or tight coupling of N cycling processes. Dominant small-subunit rRNA sequences in Perpetual Spouter source sediments are closely affiliated with the ammonia-oxidizing archaeon "Candidatus Nitrosocaldus yellowstonii" and the putatively nitrogen-fixing (diazotrophic) bacterium Thermocrinis albus, respectively, suggesting that these populations may interact at the level of the bioavailable N pool, specifically, ammonia. This hypothesis was evaluated by using a combination of geochemical, physiological, and transcriptomic analyses of sediment microcosms. Amendment of microcosms with allylthiourea, an inhibitor of ammonia oxidation, decreased rates of acetylene reduction (a proxy for N2 fixation) and nitrite production (a proxy for ammonia oxidation) and decreased transcript levels of structural genes involved in both nitrogen fixation (nifH) and ammonia oxidation (amoA). In contrast, amendment of microcosms with ammonia stimulated nitrite production and increased amoA transcript levels while it suppressed rates of acetylene reduction and decreased nifH transcript levels. Sequencing of amplified nifH and amoA transcripts from native sediments, as well as microcosms, at 2 and 4 h postamendment, indicates that the dominant and responsive populations involved in ammonia oxidation and N2 fixation are closely affiliated with Ca. Nitrosocaldus yellowstonii and T. albus, respectively. Collectively, these results suggest that ammonia-oxidizing archaea, such as Ca. Nitrosocaldus yellowstonii, have an apparent affinity for ammonia that is higher than that of the diazotrophs present in this ecosystem. Depletion of the bioavailable N pool through the activity of ammonia-oxidizing archaea likely represents a strong selective pressure for the inclusion of organisms capable of nitrogen fixation in geothermal communities. These observations help to explain the strong pattern in the codistribution of ammonia-oxidizing archaea and diazotrophs in circumneutral-to-alkaline geothermal springs.

Topics: Acetylene; Ammonia; Archaea; Biodiversity; Geologic Sediments; Hot Springs; Molecular Sequence Data; Nitrites; Nitrogen Fixation; Oxidation-Reduction; Oxidoreductases; RNA, Ribosomal, 16S; Thiourea; Wyoming

Anaerobic ammonium-oxidizing (anammox) bacteria, aerobic ammonia-oxidizing archaea (AOA) and bacteria (AOB) are three groups of ammonia/ammonium-oxidizing prokaryotes (AOPs) involved in the biochemical nitrogen cycling. In this study, the effects of allylthiourea (ATU), pH, and salinity on these three groups from mangrove sediment were investigated through microcosm incubation in laboratory. ATU treatments (50, 100, and 500 mg L(-1)) obviously affected the community structure of anammox bacteria and AOB, but only slightly for AOA. ATU began to inhibit anammox bacteria growth slightly from day 10, but had an obvious inhibition on AOA growth from the starting of the study. At 100 mg L(-1) of ATU or higher, AOB growth was inhibited, but only lasted for 5 days. The pH treatments showed that acidic condition (pH 5) had a slight effect on the community structure of anammox bacteria and AOA, but an obvious effect on AOB. Acidic condition promoted the growth of all groups of AOPs in different extent, but alkaline condition (pH 9) had a weak effect on AOB community structure and a strong effect on both anammox bacteria and AOA. Alkaline condition obviously inhibited anammox bacteria growth, slightly promoted AOA, and slightly promoted AOB in the first 20 days, but inhibited afterward. Salinity treatment showed that higher salinity (20 and 40 ‰) resulted in higher anammox bacteria diversity, and both AOA and AOB might have species specificity to salinity. High salinity promoted the growth of both anammox bacteria and AOB, inhibited AOA between 5 and 10 days, but promoted afterward. The results help to understand the role of these microbial groups in biogeochemical nitrogen cycling and their responses to the changing environments.

Topics: Ammonia; Ammonium Compounds; Anaerobiosis; Archaea; Bacteria; Biota; DNA, Archaeal; DNA, Bacterial; Geologic Sediments; Hydrogen-Ion Concentration; Molecular Sequence Data; Oxidation-Reduction; Salinity; Seawater; Sequence Analysis, DNA; Thiourea

Nitrification inhibitors have been used for decades to improve nitrogen fertilizer utilization in farmland. However, their effect on ammonia-oxidizing Archaea (AOA) in soil is little explored. Here, we compared the impact of diverse inhibitors on nitrification activity of the soil archaeon Ca. Nitrososphaera viennensis EN76 and compared it to that of the ammonia-oxidizing bacterium (AOB) Nitrosospira multiformis. Allylthiourea, amidinothiourea, and dicyandiamide (DCD) inhibited ammonia oxidation in cultures of both N. multiformis and N. viennensis, but the effect on N. viennensis was markedly lower. In particular, the effective concentration 50 (EC50) of allylthiourea was 1000 times higher for the AOA culture. Among the tested nitrification inhibitors, DCD was the least potent against N. viennensis. Nitrapyrin had at the maximal soluble concentration only a very weak inhibitory effect on the AOB N. multiformis, but showed a moderate effect on the AOA. The antibiotic sulfathiazole inhibited the bacterium, but barely affected the archaeon. Only the NO-scavenger carboxy-PTIO had a strong inhibitory effect on the archaeon, but had little effect on the bacterium in the concentrations tested. Our results reflect the fundamental metabolic and cellular differences of AOA and AOB and will be useful for future applications of inhibitors aimed at distinguishing activities of AOA and AOB in soil environments.

Topics: Ammonia; Archaea; Benzoates; Fertilizers; Guanidines; Imidazoles; Nitrification; Nitrosomonadaceae; Oxidation-Reduction; Picolines; Sulfathiazole; Sulfathiazoles; Thiourea

A flow biosensor for the detection of toxicity in water using the ammonia-oxidizing bacterium (AOB) Nitrosomonas europaea as a bioreceptor and a polymeric membrane ammonium-selective electrode as a transducer is described. The system is based on the inhibition effects of toxicants on the activity of AOB, which can be evaluated by measuring the ammonium consumption rates with the ammonium-selective membrane electrode. The AOB cells are immobilized on polyethersulfone membranes packed in a holder, while the membrane electrode is placed downstream in the flow cell. Two specific inhibitors of the ammonia oxidation-allylthiourea and thioacetamide-have been tested. The IC50 values defined as the concentration of an inhibitor causing a 50% reduction in the ammonia oxidation activity have been measured as 0.17 μM and 0.46 μM for allylthiourea and thioacetamide, respectively. The proposed sensor offers advantages of simplicity, speed and high sensitivity for measuring toxicity in water.

Topics: Ammonia; Biosensing Techniques; Cells, Immobilized; Electrodes; Hydrogen-Ion Concentration; Nitrosomonas; Oxidation-Reduction; Polymers; Potentiometry; Sulfones; Temperature; Thioacetamide; Thiourea; Water

The long-term addition of N2H4 to completely autotrophic nitrogen removal over nitrite (CANON) sequencing batch reactors (SBRs) recovered and enhanced their autotrophic nitrogen removal capacity while simultaneously reducing their production of NO3(-). The total nitrogen (TN) removal rate and TN removal efficiency of the process increased from 0.202±0.011 to 0.370±0.016 kg N/m(3)/d and from 65.1±3.75% to 77.4±3.8%, respectively, and the molar ratio of NO3(-) production to NH4(+) removal (MRNN) decreased to 0.058. The most effective concentration of N2H4 addition was approximately 3.99 mg/L. N2H4 could increase the specific growth rate of anaerobic ammonium-oxidizing bacteria (AnAOB) and inhibit aerobic ammonia oxidation. The electrons released from the oxidation of additional N2H4 using hydrazine dehydrogenase (HDH), which substituted the electrons from NO2(-) oxidation to NO3(-), replenished the consumption of AnAOB anabolism and significantly reduced the consequent NO3(-) production.

Topics: Aerobiosis; Ammonia; Autotrophic Processes; Bacteria, Anaerobic; Bioreactors; Electrons; Equipment Design; Fermentation; Hydrazines; Nitrites; Nitrogen; Oxidoreductases; Oxygen; Sewage; Thiourea; Time Factors; Waste Disposal, Fluid; Wastewater; Water Pollutants, Chemical; Water Purification

A bulk polymerization method was used to easily and efficiently prepare homoveratric acid (3,4-dimethoxyphenylacetic acid)-imprinted polymers from eight basic monomers: 2-vinylpyridine, 4-vinylpyridine, 1-vinylimidazole, N-allylaniline, N-allylpiperazine, allylurea, allylthiourea, and allylamine, in the presence of homoveratric acid as a template in N,N-dimethylformamide as a porogen. The imprinted polymer prepared from allylamine had the highest affinity to the template, showing an imprinting factor of 3.43, and allylamine polymers MIP8/NIP8 were selected for further studies. Their binding properties were analyzed using the Scatchard method. The results showed that the imprinted polymers have two classes of heterogeneous binding sites characterized by two pairs of K(d), B(max) values: K(d)(1) = 0.060 μmol/mL, B(max)(1) = 0.093 μmol/mg for the higher affinity binding sites, and K(d)(2) = 0.455 μmol/mL, B(max)(2) = 0.248 μmol/mg for the lower affinity binding sites. Non-imprinted polymer has only one class of binding site, with K(d) = 0.417 μmol/mL and B(max) = 0.184 μmol/mg. A computational analysis of the energies of the prepolymerization complexes was in agreement with the experimental results. It showed that the selective binding interactions arose from cooperative three point interactions between the carboxylic acid and the two methoxy groups in the template and amino groups in the polymer cavities. Those results were confirmed by the recognition studies performed with the set of structurally related compounds. Allylamine polymer MIP8 had no affinity towards biogenic amines. The obtained imprinted polymer could be used for selective separation of homoveratric acid.

Topics: Allylamine; Imidazoles; Models, Molecular; Molecular Imprinting; Piperazine; Piperazines; Polymers; Pyridines; Thiourea; Urea; Vanillic Acid

Soluble methane monooxygenase (sMMO) can degrade many chlorinated and aromatic pollutants. It is produced by certain methanotrophs such as Methylosinus trichosporium when grown on methane under copper limitation but, due to its low aqueous solubility, methane cannot support dense biomass growth. Since it is water soluble, methanol may be a more attractive growth substrate, but it is widely believed that sMMO is not produced on methanol. In this study, when the growth-limiting substrate was switched from methane to methanol, in the presence of the particulate MMO inhibitor, allylthiourea, growth of M. trichosporium OB3b continued unabated and sMMO activity was completely retained. When allylthiourea was then removed, sMMO activity was maintained for an additional 24 generations, albeit at a slightly lower level due to the presence of 0.70 microM of Cu(2+) in the feed medium. While a biomass density of only 2 g l(-1) could be obtained on methane, 7.4 g l(-1) was achieved by feeding methanol exponentially, and 29 g l(-1) was obtained using a modified feeding strategy employing on-line carbon dioxide production measurement. It was concluded that methanol can be employed to produce large amounts of M. trichosporium biomass containing sMMO.

Topics: Algorithms; Biomass; Carbon Dioxide; Copper; Formaldehyde; Methane; Methanol; Methylosinus trichosporium; Oxygenases; Solubility; Thiourea

Tyrosinase (TYR) was thought to play a critical role during trematode egg production. In this study, we analyzed two genes (SjTYR1 and SjTYR2), derived from Schistosoma japonicum genome databases, which encode proteins with significant homologies to mammalian TYR. They exhibited the typical TYR topology, including two copper-binding domains and a highly conserved cysteine-rich domain. Semi-quantitative reverse transcription polymerase chain reaction showed that two SjTYR genes were mainly expressed in the female adult worm. A complementary DNA coding the putative common copper center domain of each SjTYR was cloned and inserted into a pET-28a-c(+) prokaryotic expression vector. After purification, the recombinant proteins expressed in Escherichia coli were used to produce their specific antibodies. The native active SjTYRs enzyme appeared to function as a homodimer, the subunits of which were linked to each other via covalent disulfide bonds. Both female and male worms possessed monophenol oxidase and diphenol oxidase activities of TYR. The relative enzymatic activities were 0.165 min(-1) mg(-1) and 0.0805 min(-1) mg(-1), which were inhibited by a copper-chelating agent (allyl thiourea) and correlated with disruption of female egg production. Our results revealed that SjTYRs might play a significant role during eggshell formation.

Topics: Amino Acid Sequence; Animals; Catechol Oxidase; Cloning, Molecular; Dimerization; Enzyme Inhibitors; Escherichia coli; Gene Expression; Gene Expression Profiling; Helminth Proteins; Male; Molecular Sequence Data; Monophenol Monooxygenase; Protein Structure, Tertiary; Schistosoma japonicum; Sequence Alignment; Sequence Homology; Thiourea

Methanotrophs expressing soluble methane monooxygenase (sMMO) may find use in a variety of industrial applications. However, sMMO expression is strongly inhibited by copper, and the growth rate may be limited by the aqueous solubility of methane. In this study, addition of allylthiourea decreased intracellular copper in Methylosinus trichosporium OB3b, allowing sMMO production at Cu/biomass ratios normally not permitting sMMO synthesis. The presence of about 1.5 micromoles intracellular Cu g(-1) dry biomass resulted in sMMO activity of about 250 micromoles 1-napthol formed per hour gram dry biomass whether this intracellular Cu concentration was achieved by Cu limitation or by allylthiourea addition. No loss of sMMO activity occurred when the growth substrate was switched from methane to methanol when allylthiourea had been added to growth medium containing copper. Addition of copper to medium that was almost copper-free increased the yield of dry biomass from methanol from 0.20 to 0.36 g g(-1), demonstrating that some copper was necessary for good growth. This study demonstrated a method by which sMMO can be produced by M. trichosporium OB3b while growing on methanol in copper-containing medium.

Topics: Bacterial Proteins; Biomass; Copper; Methane; Methanol; Methylosinus trichosporium; Oxygenases; Solubility; Thiourea

The oxidizing ability of a nitrifying consortium exposed to p-cresol (25 mg CL(-1)) was evaluated in batch cultures. Biotransformation of the phenolic compound was investigated by identifying the different intermediates formed. p-Cresol inhibited the ammonia-oxidizing process with a decrease of 83% in the specific rate of ammonium consumption. After 48 h, ammonium consumption efficiency was 96+/-9% while nitrate yield reached 0.95+/-0.06 g NO(3)(-)-Ng(-1)NH(4)(+)-N consumed. High value for nitrate production yield showed that the nitrifying metabolic pathway was only affected at the specific rate level being nitrate the main end product. The consortium was able to totally oxidize p-cresol at a specific rate of 0.17+/-0.06 mg p-cresol-Cmg(-1) microbial protein h(-1). p-Cresol was first transformed to p-hydroxybenzaldehyde and p-hydroxybenzoate, which were later completely mineralized. In the presence of allylthiourea, a specific inhibitor of ammonia monooxygenase (AMO), p-cresol was oxidized to the same intermediates and in a similar pattern as obtained without the AMO inhibitor. AMO seemed not to be involved in the p-cresol oxidation process. When p-hydroxybenzaldehyde was added (25 mg CL(-1)), the nitrifying process was inhibited in the same way as observed with p-cresol, indicating that p-hydroxybenzaldehyde could be the main compound responsible for nitrification inhibition. p-Hydroxybenzaldehyde was accumulated during 15 h before complete consumption at a specific rate value eight times lower than the p-cresol consumption rate. Results showed that p-hydroxybenzaldehyde oxidation was the limiting step in p-cresol mineralization by the nitrifying consortium.

Topics: Benzaldehydes; Bioreactors; Biotransformation; Cresols; Nitrates; Oxidoreductases; Sewage; Thiourea; Waste Disposal, Fluid

The effects of three metabolic inhibitors (acetylene, methanol, and allylthiourea [ATU]) on the pathways of N2 production were investigated by using short anoxic incubations of marine sediment with a 15N isotope technique. Acetylene inhibited ammonium oxidation through the anammox pathway as the oxidation rate decreased exponentially with increasing acetylene concentration; the rate decay constant was 0.10+/-0.02 microM-1, and there was 95% inhibition at approximately 30 microM. Nitrous oxide reduction, the final step of denitrification, was not sensitive to acetylene concentrations below 10 microM. However, nitrous oxide reduction was inhibited by higher concentrations, and the sensitivity was approximately one-half the sensitivity of anammox (decay constant, 0.049+/-0.004 microM-1; 95% inhibition at approximately 70 microM). Methanol specifically inhibited anammox with a decay constant of 0.79+/-0.12 mM-1, and thus 3 to 4 mM methanol was required for nearly complete inhibition. This level of methanol stimulated denitrification by approximately 50%. ATU did not have marked effects on the rates of anammox and denitrification. The profile of inhibitor effects on anammox agreed with the results of studies of the process in wastewater bioreactors, which confirmed the similarity between the anammox bacteria in bioreactors and natural environments. Acetylene and methanol can be used to separate anammox and denitrification, but the effects of these compounds on nitrification limits their use in studies of these processes in systems where nitrification is an important source of nitrate. The observed differential effects of acetylene and methanol on anammox and denitrification support our current understanding of the two main pathways of N2 production in marine sediments and the use of 15N isotope methods for their quantification.

Topics: Acetylene; Anaerobiosis; Bacteria; Enzyme Inhibitors; Geologic Sediments; Methanol; Nitrites; Nitrogen; Nitrogen Isotopes; Nitrous Oxide; Oxidation-Reduction; Quaternary Ammonium Compounds; Thiourea

In activated sludge, protozoa feed on free-swimming bacteria and suspended particles, inducing flocculation and increasing the turnover rate of nutrients. In this study, the effect of protozoan grazing on nitrification rates under various conditions in municipal activated sludge batch reactors was examined, as was the spatial distribution of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) within the activated sludge. The reactors were monitored for ammonia, nitrite, nitrate, and total nitrogen concentrations, and bacterial numbers in the presence and absence of cycloheximide (a protozoan inhibitor), allylthiourea (an inhibitor of ammonia oxidation), and EDTA (a deflocculating agent). The accumulations of nitrate, nitrite, and ammonia were lower in batches without than with protozoa grazing. Inhibition of ammonia oxidation also decreased the amount of nitrite and nitrate accumulation. Inhibiting protozoan grazing along with ammonia oxidation further decreased the amounts of nitrite and nitrate accumulated. Induction of deflocculation led to high nitrate accumulation, indicating high levels of nitrification; this effect was lessened in the absence of protozoan grazing. Using fluorescent in situ hybridization and confocal laser scanning microscopy, AOB and NOB were found clustered within the floc, and inhibiting the protozoa, inhibiting ammonia oxidation, or inducing flocculation did not appear to lower the number of AOB and NOB present or affect their position within the floc. These results suggest that the AOB and NOB are present but less active in the absence of protozoa.

Topics: Ammonia; Animals; Bacteria; Cycloheximide; Eukaryota; In Situ Hybridization, Fluorescence; Microscopy, Confocal; Nitrites; Predatory Behavior; Sewage; Thiourea; Waste Disposal, Fluid

Halogenated fumigants have been used extensively in production agriculture to control soilborne pests. These types of pesticides are highly volatile and are prone to affect air quality and imperil public health. In the present study, a chemical tarp approach, termed a reactive surface barrier (RSB), was developed to reduce the emission of fumigant 1,3-dichloropropene (1,3-D) from the soil surface. The agrochemicals thiourea and allylthiourea were tested as active reagents for the construction of a RSB, where these soil amendments react with 1,3-D to form nonvolatile isothiuronium ions at the soil surface and, thereby, impede fumigant emission into the atmosphere. The feasibility of the method largely depends on the reactivity of 1,3-D and the RSB agrochemicals in soil as well as on the mobility, persistence, and toxicity of the transformation products. Therefore, the reaction kinetics and transformation mechanism of 1,3-D by thiourea and allylthiourea were studied comprehensively in aqueous solution and soil. A catalytic process occurring at the surface of soil colloids facilitated the reaction between 1,3-D and thiourea in amended soils. The rate of 1,3-D transformation in thiourea-amended soil increased with decreasing soil moisture or increasing thiourea amendment level. In a field trial, a thiourea RSB reduced cumulative 1,3-D emissions by more than 80% relative to that in bare soil surface. The present results clearly indicate that this chemical remediation technology has great potential to control the emissions of volatile halogenated organic contaminants and to mitigate atmospheric pollution.

Topics: Allyl Compounds; Chromatography, Gas; Gas Chromatography-Mass Spectrometry; Hydrocarbons, Chlorinated; Insecticides; Soil; Surface Properties; Thiourea

Diphenyliodonium (DPI) is known to irreversibly inactivate flavoproteins. We have found that DPI inhibits both membrane-bound methane monooxygenase (pMMO) from Methylococcus capsulatus and ammonia monooxygenase (AMO) of Nitrosomonas europaea. The effect of DPI on NADH-dependent pMMO activity in vitro is ascribed to inactivation of NDH-2, a flavoprotein which we proposed catalyzes reduction of the quinone pool by NADH. DPI is a potent inhibitor of type 2 NADH:quinone oxidoreductase (NDH-2), with 50% inhibition occurring at approximately 5 micro M. Inhibition of NDH-2 is irreversible and requires NADH. Inhibition of NADH-dependent pMMO activity by DPI in vitro is concomitant with inhibition of NDH-2, consistent with our proposal that NDH-2 mediates reduction of pMMO. Unexpectedly, DPI also inhibits pMMO activity driven by exogenous hydroquinols, but with approximately 100 micro M DPI required to achieve 50% inhibition. Similar concentrations of DPI are required to inhibit formate-, formaldehyde-, and hydroquinol-driven pMMO activities in whole cells. The pMMO activity in DPI-treated cells greatly exceeds the activity of NDH-2 or pMMO in membranes isolated from those cells, suggesting that electron transfer from formate to pMMO in vivo can occur independent of NADH and NDH-2. AMO activity, which is known to be independent of NADH, is affected by DPI in a manner analogous to pMMO in vivo: approximately 100 micro M is required for 50% inhibition regardless of the nature of the reducing agent. DPI does not affect hydroxylamine oxidoreductase activity and does not require AMO turnover to exert its inhibitory effect. Implications of these data for the electron transfer pathway from the quinone pool to pMMO and AMO are discussed.

Topics: Biphenyl Compounds; Electron Transport; Enzyme Inhibitors; Methylococcus capsulatus; Nitrosomonas; Onium Compounds; Oxidoreductases; Oxygenases; Quinone Reductases; Thiourea

Topics: Animals; Disease Models, Animal; Liver; Mice; Mice, Inbred BALB C; Schistosomiasis japonica; Thiourea

Topics: Anti-Infective Agents, Local; Homeopathy; Humans; Philosophy, Medical; Placebo Effect; Professional Competence; Skin Diseases; Thiourea

To observe the effect of acryl thiourea, an inhibitor of phenol oxidase, on pathological changes in the liver of mice infected with Schistosoma japonicum.. From day 22 to day 42 postinfection with cercariae, the mice of the acryl thiourea group were each injected i.p. with acryl thiourea at a dose of 300 mg/kg every other day. The mice were killed on the 42nd day postinfection to observe the pathological changes in the liver.. Compared to the infected control group, the liver tissue of the acryl thiourea group showed scattered foci of inflammatory cell infiltration, the mean diameter and area of the foci were significantly reduced (P < 0.01), and there were no eggs in the center of the foci except for some granules.. After i.p. injections of acryl thiourea, no typical egg granuloma was found in the liver of infected mice. This was possibly due to the inhibition of schistosome phenol oxidase activity and so the female adult schistosomes could not produce normal eggs.

Topics: Animals; Female; Liver; Mice; Monophenol Monooxygenase; Parasite Egg Count; Schistosoma japonicum; Schistosomiasis japonica; Thiourea

A screening method for nitrification-inhibition determination has been evaluated at two laboratories, one in Sweden and one in Spain. Allyl-thiourea (ATU), methanol, chromium and zinc were used as reference toxicants in combination with different kinds of full-scale sludge and sludge produced in the laboratory. Different results were obtained with different combinations of activated sludge and toxicants. It was found that activated sludge often exposed to small amounts of a variety of toxic substances may build up a general resistance to toxic substances. Specific adaptation to certain substances was also observed. Domestic activated sludge responded in the same way to ATU regardless of whether it originated from Sweden or Spain. Synthetic sludge and domestic sludge exhibited the same inhibition of nitrification when tested with ATU as a toxicant. Synthetic activated sludge from two laboratory-scale plants run in parallel did not exhibit exactly the same sensitivity to certain toxicants. The variation in repeated tests with the synthetic sludges was about the same as for repeated tests with full-scale sludge and for other biological toxicity tests. Limits of detection were calculated to be about 5% for a single sample with three blanks as references at both the laboratories.

Topics: Chromium; Methanol; Nitrogen; Reproducibility of Results; Schistosomicides; Sewage; Thiourea; Waste Disposal, Fluid; Zinc

Anaerobic nitrate consumption was accompanied by the accumulation of nitrite in the range of 3-52 micromoll(-1) in laboratory sediment cores continuously perfused with nitrate from below. Highest interstitial nitrite concentrations were observed in organically poor sediment cores with a low extractable protein content. Supplemented with an additional source of fixed nitrogen (ammonium), the nitrate-perfused sediments exhibited considerably lower nitrite concentrations than the respective control treatments. The simultaneous addition of ammonium and chloramphenicol, however, resulted in high nitrite concentrations. This suggests that in the absence of chloramphenicol an ammonium-fuelled bacterial protein de novo synthesis prevented the accumulation of nitrite in the course of anaerobic nitrate consumption. Additional DOC (glucose, methanol) generally increased the anaerobic nitrate consumption rate and the interstitial nitrite concentrations. The different scales of these increases were probably related to the degradability of the respective DOC compound and to the taxonomic composition of the microbial community evolving under different feeding regimes.

Topics: Anaerobiosis; Anti-Bacterial Agents; Bacterial Proteins; Biodegradation, Environmental; Chloramphenicol; Fresh Water; Geologic Sediments; Glucose; Methanol; Nitrates; Nitrites; Porosity; Quaternary Ammonium Compounds; Schistosomicides; Thiourea; Water Pollutants, Chemical; Water Pollution

Topics: Anti-Infective Agents, Local; Attitude of Health Personnel; Greece; Humans; Materia Medica; Professional Practice; Skin Diseases; Thiourea; United Kingdom

Cloned luciferase-encoding operons were transferred by conjugation to a natural isolate of the ammonia-oxidizing bacterial strain Nitrosomonas sp. RST41-3, thereby establishing conjugation as a tool for gene transfer into Nitrosomonas strains. Luminescence was dependent on the pH of the medium and the concentration of the substrate ammonium chloride. Moreover, the luminescence of the transconjugants was reduced immediately by micromolar concentrations of nitrapyrin and allylthiourea, which are specific inhibitors of nitrification. Our results indicate that luminescent Nitrosomonas strains may be useful as a probe to detect nitrification conditions in the natural environment as well as in sewage plants.

Topics: Ammonium Chloride; Biodegradation, Environmental; Culture Media; Gene Transfer Techniques; Genes, Bacterial; Hydrogen-Ion Concentration; Luciferases; Luminescent Measurements; Nitrites; Nitrosomonas; Picolines; Thiourea; Transfection

An expression vector for the luxAB genes, derived from Vibrio harveyi, was introduced into Nitrosomonas europaea. Although the recombinant strain produced bioluminescence due to the expression of the luxAB genes under normal growing conditions, the intensity of the light emission decreased immediately, in a time-and dose-dependent manner, with the addition of ammonia monooxygenase inhibitors, such as allylthiourea, phenol, and nitrapyrin. When whole cells were challenged with several nitrification inhibitors and toxic compounds, a close relationship was found between the change in the intensity of the light emission and the level of ammonia-oxidizing activity. The response of bioluminescence to the addition of allylthiourea was considerably faster than the change in the ammonia-oxidizing rate, measured as both the O2 uptake and NO2- production rates. The bioluminescence of cells inactivated by ammonia monooxygenase inhibitor was recovered rapidly by the addition of certain substrates for hydroxylamine oxidoreductase. These results suggested that the inhibition of bioluminescence was caused by the immediate decrease of reducing power in the cell due to the inactivation of ammonia monooxygenase, as well as by the destruction of other cellular metabolic pathways. We conclude that the assay system using luminous Nitrosomonas can be applied as a rapid and sensitive detection test for nitrification inhibitors, and it will be used to monitor the nitrification process in wastewater treatment plants.

Topics: Enzyme Inhibitors; Genes, Bacterial; Kinetics; Luciferases; Luminescence; Nitrites; Nitrosomonas; Oxidoreductases; Oxygen Consumption; Phenol; Picolines; Plasmids; Restriction Mapping; Thiourea; Transformation, Bacterial; Vibrio

It has been found that heterotrophic nitrification by Thiosphaera pantotropha can be inhibited by thiosulphate in batch and chemostat cultures. Allythiourea and nitrapyrin, both classically considered to be specific inhibitors of autotrophic nitrification, inhibited nitrification by Tsa. pantotropha in short-term experiments with resting cell suspensions. Hydroxylamine inhibited ammonia oxidation in chemostat cultures, but was itself fully oxidized. Thus the total nitrification rate for the culture remained the same. Heterotrophic nitrification by another organism, a strain of "Pseudomonas denitrificans" has also been shown to be inhibited by thiosulphate in short term experiments and in the chemostat. During these experiments it became evident that this strain is able to grow mixotrophically (with acetate) and autotrophically in a chemostat with thiosulphate as the energy source.

Topics: Acetates; Ammonia; Culture Media; Hydroxylamines; Nitrogen; Oxygen; Picolines; Pseudomonas; Thiobacillus; Thiosulfates; Thiourea

Topics: Bisphenol A-Glycidyl Methacrylate; Calorimetry; Chromatography, High Pressure Liquid; Composite Resins; Dental Bonding; Light; Methacrylates; Methylmethacrylate; Methylmethacrylates; Thiourea

To elucidate the nature of negative and positive phases of background delta-waves (BDW), their interaction with evoked potentials was studied in the rabbit visual cortex. BDW appeared after injection of drugs synchronizing ECoG: amyzyl, aminozin, barbiturates, and during asphyxia. The flashes evoked slow positive responses lasting about 0.5 to 1.0 s. These responses blocked the negative BDW. The results of the study and published data suggest that background delta-activity consists of segments which reflect the desynchronized state and correspond to positive BDW, and of segments which reflect active inhibition in the cerebral cortex and correspond to negative BDW.

Topics: Animals; Barbiturates; Delta Rhythm; Electroencephalography; Evoked Potentials, Visual; Female; Male; Photic Stimulation; Rabbits; Thiourea; Visual Cortex

It has been found that treatment of mice infected with Schistosoma mansoni with thiosinamine for five days had a significant effect on the formation of normal egg-shells within the ootype of female worms. The protein material, not organized into complete egg-shells, was nevertheless tanned and the surface of these amorphous masses was formed into microspines. Normal egg-shell formation was restored following drug withdrawal. The process of egg-shell formation consists of the integration of the physical moulding of egg-shell precursors derived from the vitelline cells, associated with the chemical process of protein tanning. It is suggested that thiosinamine treatment in vivo results in disruption of egg-shell formation by causing a breakdown in the moulding process and not by the inhibition of protein tanning involving the enzyme polyphenoloxidase. Treatment of worms with the drug under in vitro conditions resulted in a more enhanced effect of egg-shell formation.

Topics: Animals; Female; Mice; Microscopy, Electron; Ovum; Schistosoma mansoni; Schistosomiasis; Schistosomicides; Thiourea

Topics: Animals; Body Weight; Body Weights and Measures; Iodides; Iodine; Rats; Thiourea; Thyroid Gland; Thyroxine; Viscera

Topics: Adolescent; Adrenal Cortex Hormones; Adult; Anti-Bacterial Agents; Drug Therapy, Combination; Glucocorticoids; Humans; Hydrocortisone; Infant; Lacrimal Apparatus Diseases; Lacrimal Duct Obstruction; Male; Middle Aged; Nasolacrimal Duct; Sodium Salicylate; Thiourea; Young Adult

Topics: Iodides; Iodine; Thiourea; Thyroid Gland; Thyroxine; Viscera

Topics: Fibroma; Humans; Keloid; Thiourea

Topics: Amino Acids; Conjunctiva; Cornea; Humans; Injections; Lipotropic Agents; Thiourea; Trachoma

Topics: Anti-Bacterial Agents; Thiourea; X-Rays

Topics: Cicatrix; Humans; Thiourea; Tissue Adhesions

Topics: Humans; Scleroderma, Localized; Scleroderma, Systemic; Thiourea

Topics: Arthritis; Arthritis, Rheumatoid; Humans; Rheumatic Diseases; Sodium Benzoate; Thiourea

Topics: Goiter; Neoplasms; Thiourea; Thyroid Gland; Thyroid Neoplasms

Topics: Humans; Raynaud Disease; Sympathectomy; Thiourea

Topics: Dupuytren Contracture; Thiourea

Topics: Animals; Rats; Thiourea; Thyroid Gland; Thyroidectomy