Enzyme activity in terrestrial soil in relation to exploration of the Martian surface

Urease activity in soil is persistent for long periods under low water, low temperature, and sterile regimes, and it was suggested that some form of enzyme-protective mechanism exists in soil. Dublin soil was extracted by sonication in water followed by adding a mixture of salts. Urease activity is associated with the organo-mineral complex thus obtained and is resistant to the activities of proteolytic enzymes. Clay free soil organic matter prepared subsequently by filtration also exhibits urease activity which is resistant to proteolysis. Models consisting of enzymes with bentonite and lignin were found to mimic this resistance to proteolysis. A model system is presented which suggests both the origin and location of soil ureases and a reason for their persistence in nature

Effects of Commercial Diazinon and Imidacloprid on Microbial Urease Activity in Soil and Sod

Diazinon [O,O-diethyl O-2-isopropyl-6-methyl(pyrimidine-4-yl) phosphorothioate] and imidacloprid [1-(1-[6-chloro-3-pyridinyl]methyl)-N-nitro-2-imidazolidinimine] are applied to lawns for insect control simultaneously with nitrogenous fertilizers such as urea, but their potential effect on urease activity and nitrogen availability in turfgrass management has not been evaluated. Urease activity in enzyme assays, washed cell assays, and soil slurries was examined as a function of insecticide concentration. Intact cores from field sites were used to assess the effect of insecticide application on urease activity in creeping bentgrass (Agrostis palustris Huds.) and bluegrass (Poa pratensis L.) sod. Bacterial urease fromBacillus pasteurii and plant urease from jack bean [Canavalia ensiformis (L.) DC.] were unaffected by the insecticides. Both insecticides inhibited the growth of Proteus vulgaris, a urease-producing bacterium, but only diazinon significantly reduced urease activity in washed cells; neither insecticide inhibited urease activity in sonicated cells. Neither diazinon nor imidacloprid inhibited urease activity in Woolper soil (fine, mixed, mesic Typic Argiudoll) slurries, but diazinon slightly inhibited urease activity in Maury soil (fine, mixed, semiactive, mesic Typic Paleudalf) slurries. Imidacloprid had no effect on urease activity in creeping bentgrass or bluegrass sod at up to 10 times the commercial application rate. Diazinon briefly, but significantly, reduced urease activity in bluegrass sod. Co-application of imidacloprid and urea appears to be benign with respect to urease activity in soil and sod. Diazinon, in contrast, appears to have a significant, short-term, inhibitory effect on the microbial urease-producing community, but that effect depends on soil type

Effects of Commercial Diazinon and Imidacloprid on Microbial Urease Activity in Soil and Sod

Diazinon [O,O-diethyl O-2-isopropyl-6-methyl(pyrimidine-4-yl) phosphorothioate] and imidacloprid [1-(1-[6-chloro-3-pyridinyl]methyl)-N-nitro-2-imidazolidinimine] are applied to lawns for insect control simultaneously with nitrogenous fertilizers such as urea, but their potential effect on urease activity and nitrogen availability in turfgrass management has not been evaluated. Urease activity in enzyme assays, washed cell assays, and soil slurries was examined as a function of insecticide concentration. Intact cores from field sites were used to assess the effect of insecticide application on urease activity in creeping bentgrass (Agrostis palustris Huds.) and bluegrass (Poa pratensis L.) sod. Bacterial urease fromBacillus pasteurii and plant urease from jack bean [Canavalia ensiformis (L.) DC.] were unaffected by the insecticides. Both insecticides inhibited the growth of Proteus vulgaris, a urease-producing bacterium, but only diazinon significantly reduced urease activity in washed cells; neither insecticide inhibited urease activity in sonicated cells. Neither diazinon nor imidacloprid inhibited urease activity in Woolper soil (fine, mixed, mesic Typic Argiudoll) slurries, but diazinon slightly inhibited urease activity in Maury soil (fine, mixed, semiactive, mesic Typic Paleudalf) slurries. Imidacloprid had no effect on urease activity in creeping bentgrass or bluegrass sod at up to 10 times the commercial application rate. Diazinon briefly, but significantly, reduced urease activity in bluegrass sod. Co-application of imidacloprid and urea appears to be benign with respect to urease activity in soil and sod. Diazinon, in contrast, appears to have a significant, short-term, inhibitory effect on the microbial urease-producing community, but that effect depends on soil type

Urease Activity in a Kentucky Bluegrass Turf

The components of a turfgrass ecosystem, including plants, an intervening layer of thatch and the underlying soil, influence the fate of topically applied urea fertilizer. The loss of urea N by ammonia volatilization may be governed by the rate of urea hydrolysis. The main objective of this study was to determine the extent of urease activity associated with turfgrass plant tissue, thatch, and the underlying soil. This information may help elucidate the mechanism of ammonia loss following urea application. Because a turfgrass stand frequently possesses an extensive thatch layer that may serve as the primary plant growth medium, additional objectives included: i) determining the effects of air drying and seasonal variation on the activity of urease in thatch; ii) determining the variability in thatch urease activity by analyzing multiple field samples; and iii) determining the variation of urease activity within a thatch profile. Turfgrass clippings, thatch, and underlying Flanagan silt loam soil (Aquic Argiudoll) samples were taken from a field-grown Kentucky bluegrass (Poa pratensis L.) turf in either September 1980 or March 1981. On a dry weight basis, urease activity was 18 to 30 times higher from turfgrass clippings and thatch than from soil. Air drying thatch increased urease activity by 20 % over moist samples while air drying soil samples had no apparent effect. Greenhouse incubation of winter-dormant thatch samples increased urease activity 40 %, presumably in response to the duration of increased temperature. Thatch urease activity varied between sampling sites but still remained extremely high compared to soil activity. Within each thatch sample (1 X 1 X 2 cm), urease activity was highest in the upper 1.0 cm of the profile. It was concluded that thatch urease activity was variable in nature depending upon seasonal conditions which contrasts sharply with extremely stable soil urease activities. These findings suggest that, because of the high level of urease in thatch, ammonia volatilization will occur from most urea-treated turfgrass stands, regardless of the type of underlying soil unless the urea is thoroughly washed into the soil

Using pH and Cell Growth to Measure the Urease Activity of Sporosarcina pasteurii in Stuart’s Urea Broth with Bromothymol Blue

Honorable Mention Winner Sporosarcina pasteurii can perform microbially-induced calcite precipitation (MICP)—when bacteria hydrolyze urea and precipitate calcium carbonate crystals. This has potential applications in biocementation, though there are many barriers to implementation. One way to overcome these barriers is to measure cell growth and urease activity of S. pasteurii. Because urea hydrolysis increases the surrounding pH, urease activity could be measured using Stuart’s Urea Broth with the pH indicator bromothymol blue. Additionally, a standard curve was generated to quantify pH change, and cell growth of S. pasteurii was measured before and after urea hydrolysis. It was found that that higher concentrations of bacteria resulted in higher pH values and a faster pH increase, indicating higher urease activity. It was also found that cell growth declines during urea hydrolysis, and the growth media seems to influence this decline. These findings suggest that higher cell concentrations should be used in MICP applications to produce the highest urease activity and that the cell growth of S. pasteurii may not increase with urease activity

Metabolic activity, urease production, antibiotic resistance and virulence in dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus

In this paper, the metabolic activity in single and dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus isolates was investigated. Our results demonstrated that there was less metabolic activity in dual species biofilms compared to S. aureus biofilms. However, this was not observed if S. aureus and S. epidermidis were obtained from the same sample. The largest effect on metabolic activity was observed in biofilms of S. aureus Mu50 and S. epidermidis ET-024. A transcriptomic analysis of these dual species biofilms showed that urease genes and genes encoding proteins involved in metabolism were downregulated in comparison to monospecies biofilms. These results were subsequently confirmed by phenotypic assays. As metabolic activity is related to acid production, the pH in dual species biofilms was slightly higher compared to S. aureus Mu50 biofilms. Our results showed that S. epidermidis ET-024 in dual species biofilms inhibits metabolic activity of S. aureus Mu50, leading to less acid production. As a consequence, less urease activity is required to compensate for low pH. Importantly, this effect was biofilm-specific. Also S. aureus Mu50 genes encoding virulence-associated proteins (Spa, SpIF and Dps) were upregulated in dual species biofilms compared to monospecies biofilms and using Caenorhabditis elegans infection assays, we demonstrated that more nematodes survived when co-infected with S. epidermidis ET-024 and S. aureus mutants lacking functional spa, spIF or dps genes, compared to nematodes infected with S. epidermidis ET-024 and wild type S. aureus. Finally, S. epidermidis ET-024 genes encoding resistance to oxacillin, erythromycin and tobramycin were upregulated in dual species biofilms and increased resistance was subsequently confirmed. Our data indicate that both species in dual species biofilms of S. epidermidis and S. aureus influence each other's behavior, but additional studies are required necessary to elucidate the exact mechanism(s) involved

Studying the Relationship Between Indigenous Microbial Communities, Urease Activity, and Calcite Precipitation in Artificial Mixes of Clay and Sand

Microbial-induced calcium carbonate precipitation (MICP) is evolving as a new method of improving the mechanical properties of soil. This environmentally friendly technique is a bio-geo-chemical process where microbes play a key role in increasing soil strength through precipitating calcium carbonate. Past studies at Boise State University have indicated that MICP via bio-stimulation could be a viable alternative for expansive clayey soil treatments. However, these studies raised a new question about the relationship between soil composition, urease activity, and calcite precipitation. To answer this question, batch studies were conducted using autoclaved-sterilized sand mixed with different percentages of non-sterile natural clay and tested for urease activity. Moreover, to investigate the difference in urease activity between sand and clay bacterial communities, experiments were repeated on samples with different amounts of non-sterile sand and autoclaved-sterilized clay. MICP-treated clay/sand mixes were then evaluated for calcite precipitation. Our results showed that soil mixes with higher clay content have more urease activity and higher levels of calcite precipitation for both sand-autoclaved and clay-autoclaved soil mixes. Test results indicate that urease activity could potentially be used as an indicator of MICP performance in different soil compositions

Plant growth regulators induced urease activity in Cucurbita pepo L. cotyledons

This study is aimed to investigate the activity of urease (EC 3.5.1.5, urea amidohydrolase) that catalyzes the hydrolysis of urea in 5-day-old Cucurbita pepo cotyledons subjected to various concentrations of different growth regulators. The treatment of C. pepo cotyledons with different concentrations (100–600 μmol) of different auxins [indole-3-acetic acid (IAA), indole butyric acid (IBA), indole propionic acid (IPA) and naphthalene acetic acid (NAA)]; or with different concentrations (100–300 μmol) of different cytokinins [kinetin, zeatin and benzyladenine (6-BA)] resulted in a significant increase of urease activity, compared to control. The optimal effects were recorded for each of 500 μmol of IAA and 300 μmol of zeatin treatments. A gradual increase in urease activity was detected in cotyledons treated with various concentrations (0.2–1.0 mM) of 28-homobrassinolide (HBL), in relative to control. A substantial increase in urease activity was observed in cotyledons subjected to different concentrations of triazole (10–60 mg L–1), containing either triadimefon (TDM) or hexaconazole (HEX), compared to control. The combination of 300 μmol zeatin with any of protein inhibitors, namely 5-fluorouridine (FUrd), cordycepin and α-amanitin, resulted in the alleviation of their inhibitory effect on the urease activity