Effect of soil structure on temporal and spatial dynamics of bacteria

Soil is a complex heterogeneous system comprising of highly variable and dynamic micro-habitats that have significant impacts on the growth and activity of resident microbiota. A question addressed in this research is how soil structure affects the temporal dynamics and spatial distribution of bacteria. Using repacked microcosms, the effect of bulk-density, aggregate sizes and water content on growth and distribution of introduced Pseudomonas fluorescens and Bacillus subtilis bacteria was determined. Soil bulk-density and aggregate sizes were altered to manipulate the characteristics of the pore volume where bacteria reside and through which distribution of solutes and nutrients is controlled. X-ray CT was used to characterise the pore geometry of repacked soil microcosms. Soil porosity, connectivity and soil-pore interface area declined with increasing bulk-density. In samples that differ in pore geometry, its effect on growth and extent of spread of introduced bacteria was investigated. The growth rate of bacteria reduced with increasing bulk-density, consistent with a significant difference in pore geometry. To measure the ability of bacteria to spread thorough soil, placement experiments were developed. Bacteria were capable of spreading several cm’s through soil. The extent of spread of bacteria was faster and further in soil with larger and better connected pore volumes. To study the spatial distribution in detail, a methodology was developed where a combination of X-ray microtopography, to characterize the soil structure, and fluorescence microscopy, to visualize and quantify bacteria in soil sections was used. The influence of pore characteristics on distribution of bacteria was analysed at macro- and microscales. Soil porosity, connectivity and soil-pore interface influenced bacterial distribution only at the macroscale. The method developed was applied to investigate the effect of soil pore characteristics on the extent of spread of bacteria introduced locally towards a C source in soil. Soil-pore interface influenced spread of bacteria and colonization, therefore higher bacterial densities were found in soil with higher pore volumes. Therefore the results in this showed that pore geometry affects the growth and spread of bacteria in soil. The method developed showed showed how thin sectioning technique can be combined with 3D X-ray CT to visualize bacterial colonization of a 3D pore volume. This novel combination of methods is a significant step towards a full mechanistic understanding of microbial dynamics in structured soils.

Cadmium phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.) in three contrasting agroclimatic conditions of Chile

The phytoextraction process implies the use of plants to promote the elimination of metal contaminants in the soil. In fact, metal-accumulating plants are planted or transplanted in metal-contaminated soil and cultivated in accordance with established agricultural practices. The objective of the present study was to evaluate the productivity and Cd phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.), as well as the effect on residual Cd concentration in the soil. Both species of lupines were grown at three CdCl2 rates (0, 1, and 2 mg kg-1), under three agroclimatic conditions in Chile in 2013. In the arid zone (Pan de Azúcar, 73 mm precipitation), narrow-leafed lupine production was significantly (P < 0.05) higher than white lupine (4.55 vs. 3.26 Mg DM ha-1, respectively). In locations with higher precipitation (Santa Rosa, 670 mm; Carillanca, 880 mm), narrow-leafed lupine DM production was slightly higher than in Pan de Azúcar, but white lupine was approximately three times higher. Total plant Cd concentrations in white and narrow-leafed lupine increased as Cd rates increased in the three environments, but they were much higher in narrow-leafed lupine than white lupine; 150%, 58%, and 344% higher in Pan de Azúcar, Santa Rosa, and Carillanca, respectively. Cadmium uptake (g Cd ha-1) and apparent recovery were also higher (P < 0.05) in narrow-leafed lupine in two environments (Pan de Azúcar and Carillanca). These results suggest that narrow-leafed lupine present higher potential as phytoremediation species than white lupine.

An Investigation of Presumptive Synergism of Oil Palm Bunch Ash and Sawdust Amendments in Remediation of crude oil Spiked Soil

Presumed synergistic effect of combined amendment of crude oil spiked soil with oil palm bunch ash and sawdust was carried out in a laboratory experiment. Two kilogram (2 kg) of sandy soil was placed in each of five plastic vessels labeled TA, TB, TC, TD and TE. TA was left in its natural state while the others were each polluted with 6.7% v/w of crude oil. TB was not given any remediation amendment. TC and TD were each amended with 13.3% of oil palm bunch ash and sawdust respectively while TE was amended with 13.3% each of oil palm bunch ash and sawdust. The setups were replicated five times and watered twice weekly. Results showed that soil pH increased from 8.7±0.04 to 10.5±0.06, 5.3±0.01 to 8.5±0.04 and 5.6±0.18 to 11.5±0.15 for TC, TD and TE respectively. Percentage total petroleum hydrocarbon contents reduced by 65% for TC, TD and 52% for TE. Total organic carbon increased from 7.6±0.7 to 8.5±0.5%%, reduced from 4.0±0.1% to 3.7±0.3% and from 4.1±0.1% to 2.2±1.0% TC, TD and TE respectively. Total nitrogen increased from 0.66±0.1 to 0.69±0.0% for TC, remained nearly the same for TD and reduced from 0.4±0.0 to 0.2±0.0% for TE while average phosphorus increased from 0.4±0.0 to 23.0±4.2 mg/kg, 0.3±0.0 to 1.8±0.4 mg/kg and from 0.2±1.0 mg/kg to 52.6±4.6 mg/kg for TC, TD and TE respectively. Conclusively, combined amendment with oil palm bunch ash and sawdust did not induce synergism in soil total petroleum hydrocarbon content reduction.

MICROBIOLOGICAL ANALYSIS OF TRADITIONALLY FERMENTED MILK SOLD IN KINIGI SECTOR OF MUSANZE DISTRICT IN RWANDA

A research work entitled: “Microbiological analysis of traditionally fermented milk (Ikivuguto) sold in Kinigi Sector of Musanze District,” was carried out at Higher Learning Institution of Applied Sciences (INES-Ruhengeri) Laboratory of Microbiology located near Volcanoes in the Northern Province of Rwanda. The main objective of this work was to determine the microbiological quality of traditionally fermented milk, which is consumed by Kinigi Center local people. The hypothesis was to analyze if traditionally fermented milk commercialized in Kinigi restaurants contained pathogenic bacteria such as fecal coliforms and Escherichia coli , in addition to staphylococci and yeasts. Milk samples were collected from Kinigi sector and examined in the microbiology laboratory in order to assess the microbiological quality and safety of traditionally fermented milk in rural areas. The samples were analyzed qualitatively and quantitatively for the microbes found in fermented milk sold in Kinigi Center, and the results were as follows: 7.21x107 CFU/ml for total counts; 3.89x107 CFU/ml for Lactobacillus ; 2.77x107 CFU/ml for yeasts; 1.196x105 CFU/ml for total coliforms; 9.63x104 CFU/ml for fecal coliforms and 8.92x103 CFU/ml for staphylococci. Biochemical tests were carried out and the results showed that identified pathogens were E. coli, Providencia alcalifaciens , and the staphylococci group. It was found that fermented milk contained genera and species of Staphylococcus haemolyticus , Staphylococcus aureus , Staphylococcus intermedius , Staphylococcus xylosus and Staphylococcus saprophyticus . Findings showed that the commercial milk samples were cross-contaminated by different pathogens from environment. These contaminations could have been due to improper handling, presence of flies, soil erosion, dust from atmosphere, as well as contaminated milk vessels or pots, stirrers and unpasteurized water. It was concluded that local farmers and milk retailers did not adhere to required hygienic conditions for milk safety. In this regard, the sold traditional fermented milk does not meet health and safety standards because people did not respect good manufacturing practices. The hypothesis and main objective were confirmed, because traditionally fermented milk of Kinigi was cross-contaminated before consumption. Thus, it would be better to train farmers in the areas of product hygiene, sanitation and safety during milking, processing and marketing.

The influence of organic amendments on soil aggregate stability from semiarid sites

Restoring the native vegetation is the most effective way to regenerate soil health. Under these conditions, vegetation cover in areas having degraded soils may be better sustained if the soil is amended with an external source of organic matter. The addition of organic materials to soils also increases infiltration rates and reduces erosion rates; these factors contribute to an available water increment and a successful and sustainable land management. The goal of this study was to analyze the effect of various organic amendments on the aggregate stability of soils in afforested plots. An experimental paired-plot layout was established in southern of Spain (homogeneous slope gradient: 7.5%; aspect: N170). Five amendments were applied in an experimental set of plots: straw mulching; mulch with chipped branches of Aleppo Pine (Pinus halepensis L.); TerraCotten hydroabsobent polymers; sewage sludge; sheep manure and control. Plots were afforested following the same spatial pattern, and amendments were mixed with the soil at the rate 10 Mg ha-1. The vegetation was planted in a grid pattern with 0.5 m between plants in each plot. During the afforestation process the soil was tilled to 25 cm depth from the surface. Soil from the afforested plots was sampled in: i) 6 months post-afforestation; ii) 12 months post-afforestation; iii) 18 months post-afforestation; and iv) 24 months post-afforestation. The sampling strategy for each plot involved collection of 4 disturbed soil samples taken from the surface (0–10 cm depth). The stability of aggregates was measured by wet-sieving. Regarding to soil aggregate stability, the percentage of stable aggregates has increased slightly in all the treatments in relation to control. Specifically, the differences were recorded in the fraction of macroaggregates (≥ 0.250 mm). The largest increases have been associated with straw mulch, pinus mulch and sludge. Similar results have been registered for the soil organic carbon content. Independent of the soil management, after six months, no significant differences in microaggregates were found regarding to the control plots. These results showed an increase in the stability of the macroaggregates when soil is amended with sludge, pinus mulch and straw much. This fact has been due to an increase in the number cementing agents due to: (i) the application of pinus, straw and sludge had resulted in the release of carbohydrates to the soil; and thus (ii) it has favored the development of a protective vegetation cover, which has increased the number of roots in the soil and the organic contribution to it.

Impact of HydroPolymers on the soil biological components in mediterranean drylands

Soil degradation affects more than 52 million ha of land in counties of the European Union. This problem is particularly serious in Mediterranean areas, where the effects of anthropogenic activities (tillage on slopes, deforestation, and pasture production) add to problems caused by prolonged periods of drought and intense and irregular rainfall. Soil microbiota can be used as an indicator of the soil healthy in degraded areas. This is because soil microbiota participates in the cycle elements and in the organic matter decomposition. All this helps to the young plants establishment and in long term protect the soils against the erosion. During dry periods in the Mediterranean areas, the lack of water entering the soil matrix leads to a loss of soil microbiological activity and it turns into a lower soil production capabilities. Under these conditions, the aim of this study was to evaluate the positive effect on soil biological components produced by an hydro absorbent polymer (Terracottem). The aim of the experiment was to evaluate the impact assessment of an hydropolymer (Terracottem) on the soil biological components. An experimental flowerpot layout was established in June 2015 and 12 variants with different amount of Terracottem were applied as follow: i) 3.0 kg.m3 ; ii) 1.5 kg.m3 and; iii) 0 kg.m3. In all the variants were tested the further additives: a) 1% of glucose, b) 50 kg N.ha-1 of Mineral nitrogen, c) 1% of Glucose + 50 kg N.ha-1 of Mineral nitrogen d) control (no additive). According to natural conditions, humidity have been kept at 15% in all the variants. During four weeks, mineral nitrogen leaching and soil respiration have been measured in each flowerplot. Respiration has been quantified four times every time while moistening containers and alkaline soda lime has been used as a sorbent. The amount of CO 2 increase has been measured with the sorbent. Leaching of mineral nitrogen has been quantified by ion exchange resins (IER). IER pouches have been placed on the bottom of each container, and after completion of the experiment mineral nitrogen leaching has been evaluated by distillation and titration method. Results from respiration have shown statistically significant differences between the variants. According to control, soil with polymers have shown significant difference when comparing respiration with independence of the additive used. CO 2 production in the first week has exceeded the sum of the outputs of the following weeks. Mineral nitrogen leaching measurement has shown statistically significant differences. The lowest leaching has been occurred in control variant, while the highest in variant containing only the addition of mineral nitrogen. Research results may conclude that the biological part of the test soil is not limited by a lack of components, the only thing that suppresses its activity is the lack of moisture. After moistening it leads to a rapid growth of soil activity, without causing the nutrients loss. Besides, Terracottem has affected soil activity neither positively nor negatively, but it considers being a suitable tool for reducing the drought impact in arid and semi-arid areas.

SAMPLING BIAS IN EVALUATING THE PROBABILITY OF SEISMICALLY INDUCED SOIL LIQUEFACTION WITH SPT & CPT CASE HISTORIES

Several deterministic and probabilistic methods are used to evaluate the probability of seismically induced liquefaction of a soil. The probabilistic models usually possess some uncertainty in that model and uncertainties in the parameters used to develop that model. These model uncertainties vary from one statistical model to another. Most of the model uncertainties are epistemic, and can be addressed through appropriate knowledge of the statistical model. One such epistemic model uncertainty in evaluating liquefaction potential using a probabilistic model such as logistic regression is sampling bias. Sampling bias is the difference between the class distribution in the sample used for developing the statistical model and the true population distribution of liquefaction and non-liquefaction instances. Recent studies have shown that sampling bias can significantly affect the predicted probability using a statistical model. To address this epistemic uncertainty, a new approach was developed for evaluating the probability of seismically-induced soil liquefaction, in which a logistic regression model in combination with Hosmer-Lemeshow statistic was used. This approach was used to estimate the population (true) distribution of liquefaction to non-liquefaction instances of standard penetration test (SPT) and cone penetration test (CPT) based most updated case histories. Apart from this, other model uncertainties such as distribution of explanatory variables and significance of explanatory variables were also addressed using KS test and Wald statistic respectively. Moreover, based on estimated population distribution, logistic regression equations were proposed to calculate the probability of liquefaction for both SPT and CPT based case history. Additionally, the proposed probability curves were compared with existing probability curves based on SPT and CPT case histories.

An Analysis of the Potential Risk Exposure to Lead (Pb) through Urban Community Gardens

Community gardening in cities is increasing, driven by social interaction and food security. City soils are sinks for heavy metals; including neurotoxic lead (Pb). Exposure routes are primarily through inhalation/ingestion of soil, or second by ingestion of plants that have accumulated Pb. This research evaluates soil at three Liberty City, Florida sites estimating risk of Pb exposure through primary and secondary pathways. Soil cores were collected from Liberty City, and red Malabar spinach (Basella rubra) was grown in Pb soil treatments in a greenhouse. Total soil Pb levels and plant tissues were measured after acid digestion, by ICP-OES. In Liberty City, two sites had hotspots with areas of elevated soil Pb levels. Plants grown on Pb contaminated soil all accumulated statistically significant Pb concentrations. Therefore, there is a potential risk of Pb exposure to residents in Liberty City by exposure in hotspot sites through both the primary and secondary pathways.

Influence of Soil Biogeochemical Properties on the Invasiveness of Old World Climbing Fern (Lygodium microphyllum)

The state of Florida has one of the most severe exotic species invasion problems in the United States, but little is known about their influence on soil biogeochemistry. My dissertation research includes a cross-continental field study in Australia, Florida, and greenhouse and growth chamber experiments, focused on the soil-plant interactions of one of the most problematic weeds introduced in south Florida, Lygodium microphyllum (Old World climbing fern). Analysis of field samples from the ferns introduced and their native range indicate that L microphyllum is highly dependent on arbuscular mycorrhizal fungi (AMF) for phosphorus uptake and biomass accumulation. Relationship with AMF is stronger in relatively dry conditions, which are commonly found in some Florida sites, compared to more common wet sites where the fern is found in its native Australia. In the field, L. microphyllum is found to thrive in a wide range of soil pH, texture, and nutrient conditions, with strongly acidic soils in Australia and slightly acidic soils in Florida. Soils with pH 5.5 - 6.5 provide the most optimal growth conditions for L. microphyllum, and the growth declines significantly at soil pH 8.0, indicating that further reduction could happen in more alkaline soils. Comparison of invaded and uninvaded soil characteristics demonstrates that L. microphyllum can change the belowground soil environment, with more conspicuous impact on nutrient-poor sandy soils, to its own benefit by enhancing the soil nutrient status. Additionally, the nitrogen concentration in the leaves, which has a significant influence in the relative growth rate and photosynthesis, was significantly higher in Florida plants compared to Australian plants. Given that L. microphyllum allocates up to 40% of the total biomass to rhizomes, which aid in rapid regeneration after burning, cutting or chemical spray, hence management techniques targeting the rhizomes look promising. Over all, my results reveal for the first time that soil pH, texture, and AMF are major factors facilitating the invasive success of L. mcirophyllum. Finally, herbicide treatments targeting rhizomes will most likely become the widely used technique to control invasiveness of L. microphyllum in the future. However, a complete understanding of the soil ecosystem is necessary before adding any chemicals to the soil to achieve a successful long-term invasive species management strategy.

Initial growth of sunflower in soils with high concentrations of boron and heavy metals.

Phytoremediation studies have been conducted in an area contaminated by heavy metals, located in Piracicaba - SP, Brazil. This area was contaminated accidentally by the addition of auto scrap shredding to the soil and was limed later to reduce heavy metal mobility in the environment. Previous characterization showed that it also presents high concentration of boron, which has limited the initial plant development of some species. As sunflower plants require a high boron supply and the literature describes its use in the phytoremediation of soils contaminated with heavy metals under some conditions, the aim of this work was to evaluate its potential for the remediation of this area. In the present study, the results of preliminary tests are presented, aiming at the evaluation of sunflower plant germination and its initial development when cultivated in the contaminated soil described. Two sunflower hybrids were sown in soils treated with different rates of boron and in the soil from the contaminated area in study. The results showed that sunflower plants had a normal initial development, even in the soil from the contaminated area. Therefore, sunflower is a promising crop and further studies will be developed to evaluate the sunflower efficiency in phytoextraction or phytostabilization of heavy metals in areas where boron contamination also occurs, as is the case in the study area.

Annual crop rotation of tropical pastures with no-till soil as affectedby lime surface application.

Soil acidity and low natural fertility are the main limiting factors for grain production in tropical regionssuch as the Brazilian Cerrado. The application of lime to the surface of no-till soil can improve plant nutrition, dry matter production, crop yields and revenue. The present study, conducted at the Lageado Experimental Farm in Botucatu, State of São Paulo, Brazil, is part of an ongoing research project initi-ated in 2002 to evaluate the long-term effects of the surface application of lime on the soil?s chemical attributes, nutrition and kernel/grain yield of peanut (Arachis hypogaea), white oat (Avena sativa L.) and maize (Zea mays L.) inter cropped with palisade grass (Urochloa brizantha cv. Marandu), as well as the forage dry matter yield of palisade grass in winter/spring, its crude protein concentration, estimated meat production, and revenue in a tropical region with a dry winter during four growing seasons. The experiment was designed in randomized blocks with four replications. The treatments consisted of four rates of lime application (0, 1000, 2000 and 4000 kg ha−1), performed in November 2004. The surface application of limestone to the studied tropical no-till soil was efficient in reducing soil acidity from the surface down to a depth of 0.60 m and resulted in greater availability of P and K at the soil surface. Ca and Mg availability in the soil also increased with the lime application rate, up to a depth of 0.60 m. Nutrient absorption was enhanced with liming, especially regarding the nutrient uptake of K, Ca and Mg by plants.Significant increases in the yield components and kernel/grain yields of peanut, white oat and maize were obtained through the surface application of limestone. The lime rates estimated to achieve the maximum grain yield, especially in white oat and maize, were very close to the rates necessary to increase the base saturation of a soil sample collected at a depth of 0?0.20 m to 70%, indicating that the surface liming of 2000 kg ha−1is effective for the studied tropical no-till soil. This lime rate also increases the forage dry matter yield, crude protein concentration and estimated meat production during winter/spring in the maize-palisade grass inter cropping, provides the highest total and mean net profit during the four growing seasons, and can improve the long-term sustainability of tropical agriculture in the Brazilian Cerrado.

Carbon Stocks in Compartments of Soil Organic Matter 31 Years after Substitution of Native Cerrado Vegetation by Agroecosystems.

ABSTRACT: Changes in carbon stocks in different compartments of soil organic matter of a clayey Latossolo Vermelho Distrófico (Typic Haplustox), caused by the substitution of native savanna vegetation (cerrado sensu stricto) by agroecosystems, were assessed after 31 years of cultivation. Under native vegetation, a stock of 164.5 Mg ha-1 C was estimated in the 0.00-1.00 m layer. After 31 years of cultivation, these changes in soil C stocks were detected to a depth of 0.60 m. In the case of substitution of cerrado sensu stricto by no-tillage soybean-corn rotation, a reduction of at least 11 % of the soil C pools was observed. However, the adoption of no-tillage as an alternative to tillage with a moldboard plow (conventional system) reduced CO2 emissions by up to 12 %.

Effectiveness of a new toothbrush design versus a conventional tongue scraper in improving breath odor and reducing tongue microbiota

For centuries, specific instruments or regular toothbrushes have routinely been used to remove tongue biofilm and improve breath odor. Toothbrushes with a tongue scraper on the back of their head have recently been introduced to the market. The present study compared the effectiveness of a manual toothbrush with this new design, i.e., possessing a tongue scraper, and a commercial tongue scraper in improving breath odor and reducing the aerobic and anaerobic microbiota of tongue surface. The evaluations occurred at 4 moments, when the participants (n=30) had their halitosis quantified with a halimeter and scored according to a 4-point scoring system corresponding to different levels of intensity. Saliva was collected for counts of aerobic and anaerobic microorganisms. Data were analyzed statistically by Friedman's test (p<0.05). When differences were detected, the Wilcoxon test adjusted for Bonferroni correction was used for multiple comparisons (group to group). The results confirmed the importance of mechanical cleaning of the tongue, since this procedure provided an improvement in halitosis and reduction of aerobe and anaerobe counts. Regarding the evaluated methods, the toothbrush's tongue scraper and conventional tongue scraper had a similar performance in terms of breath improvement and reduction of tongue microbiota, and may be indicated as effective methods for tongue cleaning.

Effect of different cleansers on the surface of removable partial denture

Removable partial dentures (RPD) demand specific hygienic cleaning and the combination of brushing with immersion in chemical solutions has been the most recommended method for control of biofilm. However, the effect of the cleansers on metallic components has not been widely investigated. This study evaluated the effect of different cleansers on the surface of RPD. Five disc specimens (12 mm x 3 mm metallic disc centered in a 38 x 18 x 4 mm mould filled with resin) were obtained for each experimental situation: 6 solutions [Periogard (PE), Cepacol (CE), Corega Tabs (CT), Medical Interporous (MI), Polident (PO), 0.05% sodium hypochlorite (NaOCl), and distilled water (DW) control] and 2 Co-Cr alloys [DeguDent (DD) and VeraPDI (VPDI)] were used for each experimental situation. A 180-day immersion was simulated and the measurements of roughness (Ra, µm) of metal and resin were analyzed using 2-way ANOVA and Tukey’s test. The surface changes and tarnishes were examined with a scanning electronic microscopy (SEM). In addition, energy dispersive x-ray spectrometry (EDS) analysis was carried out at representative areas. Visually, NaOCl and MI specimens presented surface tarnishes. The roughness of materials was not affected by the solutions (p>0.05). SEM images showed that NaOCl and MI provided surface changes. EDS analysis revealed the presence of oxygen for specimens in contact with both MI and NaOCl solutions, which might suggest that the two solutions promoted the oxidation of the surfaces, thus leading to spot corrosion. Within the limitations of this study, it may be concluded that the NaOCl and MI may not be suitable for cleaning of RPD.

Validation of the Deardorff model for estimating energy balance components for a sugarcane crop

The quantification of the available energy in the environment is important because it determines photosynthesis, evapotranspiration and, therefore, the final yield of crops. Instruments for measuring the energy balance are costly and indirect estimation alternatives are desirable. This study assessed the Deardorff's model performance during a cycle of a sugarcane crop in Piracicaba, State of São Paulo, Brazil, in comparison to the aerodynamic method. This mechanistic model simulates the energy fluxes (sensible, latent heat and net radiation) at three levels (atmosphere, canopy and soil) using only air temperature, relative humidity and wind speed measured at a reference level above the canopy, crop leaf area index, and some pre-calibrated parameters (canopy albedo, soil emissivity, atmospheric transmissivity and hydrological characteristics of the soil). The analysis was made for different time scales, insolation conditions and seasons (spring, summer and autumn). Analyzing all data of 15 minute intervals, the model presented good performance for net radiation simulation in different insolations and seasons. The latent heat flux in the atmosphere and the sensible heat flux in the atmosphere did not present differences in comparison to data from the aerodynamic method during the autumn. The sensible heat flux in the soil was poorly simulated by the model due to the poor performance of the soil water balance method. The Deardorff's model improved in general the flux simulations in comparison to the aerodynamic method when more insolation was available in the environment.