Spatial heterogeneity of a microbial community in a sandy soil ecosystem

A fieldwork was carried out in Caesarea sand dunes, Israel, to determine the influence of fine-scale landscape-patch abiotic-factor heterogeneity on microbial activity in a Mediterranean region. Soil organisms in terrestrial systems are unevenly distributed in time and space, and are often aggregated. Spatio-temporal patchiness in the soil environment is thought to be crucial for the maintenance of soil biodiversity, providing diverse microhabitats that are tightly interwoven with resource partitioning. Determination of a ‘scale unit’ to help understand ecological processes has become one of the important and most debatable problems in recent years. To better understand the distribution of soil microbial communities at multiple spatial scales, a survey was conducted to examine the spatial organization of the community structure in two sandy soil ecosystems. One-hundred forty-four soil samples were collected from two patches 4000 m apart from each other. Basal respiration (CO₂ evolution without the addition of any external substrate), microbial biomass, functional diversity, and community-level physiological profile (CLPP) in soil were measured with a MicroResp? system. Soil abiotic analysis was performed by soil standard analytical methods. The results demonstrated that bacterial distributions can be highly structured, even within a habitat that appears to be relatively homogeneous at the plot and field scale. Different subsets of the microbial community were distributed differently across the plot. This is due to spatial heterogeneity associated with soil physical, chemical, and biological properties. Although spatial variability in the distribution of soil microorganisms is generally regarded as random, this variability often has a predictable spatial structure. This study provided evidence that a spatially explicit approach to soil ecology can enable the identification of factors that drive the spatial heterogeneity of populations and activities of soil organisms, at scales ranging from meters to hundreds of meters. Furthermore, there is increasing evidence that spatial soil ecology can yield new insights into the factors that maintain and regulate soil biodiversity, as well as on how the spatial distribution of soil organisms influences plant growth and plant community structure.     

Role of perennial plants in determining the activity of the microbial community in the Negev Desert ecosystem

Perennial plants are known to be one of the most influential parameters in desert ecosystems affecting microbial activity. In this study, we examined the importance of these perennial shrubs and attempted to determine the most influential factor that contributes the most to the ecosystem by separating the physical part and the organic contribution of perennial plants. The study site is located in the northern Negev Desert, Israel, where 50 Hammada scoparia shrubs and 50 artificial plants were randomly marked to be used as a tool for the above objectives. Soil samples were collected monthly in the vicinity of the canopies of both shrubs while control samples were collected from the open areas between the shrubs. All samples were collected from the upper (0–10 cm) and the deeper (10–20 cm) soil layers. The contribution to microbial activity was measured by evaluation of the microbial community functions in soil. The results of the research showed a trend of a strong influence of the perennial H. scoparia shrubs on microbial community function. The functional aspects of the microbial community that were measured were CO₂ evolution, microbial biomass, microbial functional diversity, and the physiological profile of the community. High values for all parameters were observed under the vicinity of the H. scoparia shrubs, while the artificial plants were found to have a weaker effect on the community according to soil depth. The upper-soil layer at both locations (artificial plant canopy and control samples) showed higher values of the functioning parameters than that of the deeper soil layer. The results indicate the importance of the organic contribution (plant litter) in comparison to the physical part of desert shrubs.     

Incorporation of S into soil organic matter in the field as determined by the natural abundance of stable S isotopes

In agricultural systems with low S inputs, soil organic matter is a major source of S and the transformations between organic and inorganic S pools are important for the supply of S to plants. This study was conducted to determine the effect of S fertilizer on the size and activity of organic S pools. For 5 years S fertilizer with a known composition of stable S isotopes was applied to a rotation on a loamy soil and a coarse sandy soil at rates higher than the plant demand. Total organic S in soil organic matter was not affected by sulphur application, but a small increase occurred in the sulphate ester fractions (P<0.05). Inorganic sulphate concentrations in the soil reflected the S application in the year of sampling, whereas S applied in earlier years was not recognized. Organic matter below the plough layer in both soils was enriched with S, possibly as a result or organic matter leaching or an increased clay content in the subsoils. At 0–20 cm, the C:S ratio in organic matter was ca. 100 for both soils, decreasing to 73 and 46 at 60–80 cm for the coarse sandy soil and the loamy soils, respectively. In both soils, isotope data showed that ca. 30% of organic-bonded S at 0–20 cm originated from fertilizer S applied during the last 5 years, irrespective of the S application rate. At 20–40 cm the rate of incorporations was lower and at 40–60 cm no incorporation of fertilizer S into organic matter was recognized. The fertilizer application did not induce net changes in the total organic S fraction, but isotope data indicated that a considerable part of the organic S pool was involved in S cycling in the field.     

Metabolism of the food-associated carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by human intestinal microbiota

2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine is a putative human carcinogenic heterocyclic aromatic amine formed from meat and fish during cooking. Although the formation of hazardous PhIP metabolites by mammalian enzymes is well-documented, nothing is known about the PhIP transformation potency of human intestinal bacteria. In this study, the in vitro metabolism of PhIP by human fecal samples was investigated. Following anaerobic incubation of PhIP with stools freshly collected from six healthy volunteers, we found that PhIP was extensively transformed by the human intestinal bacteria. HPLC analysis showed that the six human fecal microbiota transformed PhIP with efficiencies from 47 to 95% after 72 h incubation, resulting in one major derivative. ESI-MS/MS, HRMS, 1D (1H, 13C, DEPT) and 2D (gCOSY, gTOCSY, gHMBC, gHSQC) NMR, and IC analysis elucidated the complete chemical identity of the microbial PhIP metabolite as 7-hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3',2':4,5]imidazo[1,2-a]pyrimidin-5-ium chloride. At present, no information is available about the biological activity of this newly discovered bacterial PhIP metabolite. Our findings however suggest that bacteria derived from the human intestine play a key role in the activation or detoxification of PhIP, a digestive fate ignored so far in risk assessments. Moreover, the variation in transformation efficiency between the human microbiota indicates interindividual differences in the ability to convert PhIP. This may predict individual susceptibility to carcinogenic risk from this suspected dietary carcinogen.     

Carbon flow into microbial and fungal biomass as a basis for the belowground food web of agroecosystems

The origin and quantity of plant inputs to soil are primary factors controlling the size and structure of the soil microbial community. The present study aimed to elucidate and quantify the carbon (C) flow from both root and shoot litter residues into soil organic, extractable, microbial and fungal C pools. Using the shift in C stable isotope values associated with replacing C3 by C4 plants we followed root- vs. shoot litter-derived C resources into different soil C pools. We established the following treatments: Corn Maize (CM), Fodder Maize (FM), Wheat + maize Litter (WL) and Wheat (W) as reference. The Corn Maize treatment provided root- as well as shoot litter-derived C (without corn cobs) whereas Fodder Maize (FM) provided only root-derived C (aboveground shoot material was removed). Maize shoot litter was applied on the Wheat + maize Litter (WL) plots to trace the incorporation of C4 litter C into soil microorganisms. Soil samples were taken three times per year (summer, autumn, winter) over two growing seasons. Maize-derived C signal was detectable after three to six months in the following pools: soil organic C (C_org), extractable organic C (EOC), microbial biomass (C_mic) and fungal biomass (ergosterol). In spite of the lower amounts of root- than of shoot litter-derived C inputs, similar amounts were incorporated into each of the C pools in the FM and WL treatments, indicating greater importance of the root- than shoot litter-derived resources for the soil microorganisms as a basis for the belowground food web. In the CM plots twice as much maize-derived C was incorporated into the pools. After two years, maize-derived C in the CM treatment contributed 14.1, 24.7, 46.6 and 76.2% to C_org, EOC, C_mic and ergosterol pools, respectively. Fungi incorporated maize-derived C to a greater extent than did total soil microbial biomass.     

Ants as indicators of soil-based ecosystem services in agroecosystems of the Colombian Llanos

Ants represent a widespread and functionally diverse taxonomic group that are both sensitive to land management and serve as important regulators of key soil processes. Building upon this idea, we sought to understand the impacts of agricultural management on ant communities in the Orinoco River Basin of eastern Colombia and to identify species that could be used as indicators of soil-based ecosystem services. Ants were collected and identified from the soil and litter layer within 75 fields (nine TSBF sub-samples along a transect in each field) divided among five common agricultural land uses in the region: 1) annual crops (maize, soy and rice), 2) rubber plantations, 3) oil palm plantations, 4) improved pastures (based on Brachiaria spp.), and 5) semi-natural savannas. As expected, land management was found to greatly influence ant communities. Improved pastures showed the highest species richness (6.9 species per transect) and semi-natural savanna the greatest abundance of ants (145 individuals per transect). Within each of these fields a suite of soil and agroecosystem characteristics were measured and combined into synthetic indicators of five soil-based ecosystem services: 1) nutrient provision, 2) water storage and regulation, 3) maintenance of soil structure, 4) climate regulation services and 5) soil biodiversity and biological activity. Ant species were then associated with these synthetic indicators using the IndVal method to identify indicator species for each of the five consolidated ecosystem services measured. In total, 14 indicator species were identified and found to be significantly associated with either the high or low provision of each of the five services. The development of such bioindicators offers a rapid and relatively inexpensive tool to facilitate land management and policy decisions in this biologically diverse and rapidly changing region of Colombia.     

How the development of barren land into orchards affects soil ecosystem in Tibet, China

Fruit production is an important strategy for alleviating poverty on the Tibetan Plateau and leads to the conversion of natural barren land into orchards. This study aimed to understand how the conversion of barren land to peach (Prunus persica) orchards affects soil nutrients, heavy metals, and fungal communities in the 0-40 cm profile (at 20 cm intervals) in an experiment including three treatments, barren land (BL), peach orchards planting for 4 years (Y4), and peach orchards planting for 10 years (Y10). Results of the experiment showed that compared with BL, Y4 reduced the availability of some macronutrients (N and K) and micronutrients (Fe and Mn) due to the exclusive application of chemical fertilizer at the seedling stage. Conversely, Y10, which included six years of green cultivation management, using a combination of sheep manure and chemical fertilizer as well as alfalfa (Medicago sativa Linn) intercropping, effectively improved soil macronutrients, but did not enhance the availability of Fe and Mn. Although the investigated heavy metals (As, Hg, Pb, Cr, and Cd) in both the Y4 and Y10 soils were found to pose a low risk to food safety and soil environment, Hg, Cr, and As tended to accumulate in the subsoil (20-40 cm). Furthermore, the variations in the fungal community composition and functional groups were mainly driven by the interaction effects of macronutrients, micronutrients, and heavy metals, but their independent contribution to specific key functional groups cannot be overlooked. For example, Y4 and Y10 decreased the relative abundance of soil saprotrophic and lichenized fungi, mainly due to the loss of micronutrients (Fe and Mn). However, as a result of macronutrient input and dung saprotrophic fungi enrichment, orchard soils promoted the growth of pathogens that play critical roles in fungal co-occurrence networks. These findings indicate that supplementation with N or K fertilizer or manure at the seedling stage and fertilizers rich in Fe and Mn throughout the growth period would be beneficial to the balance of soil nutrients and provide insights into linking the variations in soil nutrients and heavy metals to the function of the fungal community during the conversion of barren land to orchards in alpine soil ecosystems. The risks posed by heavy metal accumulation and fungal pathogen enrichment should be actively prevented.     

Pine sawdust as stimulator of the microbial community in post-arable afforested soil

Sawdust and wood residues are considered to be stimulators of microbiological change in soil and could be used in post-arable sites designed for afforestation. In this study, we discuss changes in bacteria numbers within selected groups in relation to soil type (arable and forest) and method of sawdust application. Sawdust was spread along planting rows and mixed with the soil or put directly under roots of planted seedlings. Applying sawdust under the roots increased the number of cellulolytic bacteria in all experimental treatments. On post-arable soil, adding sawdust in rows increased the number of copiotrophic bacteria and reduced the number of fluorescent bacteria and actinomycetes (ACT). In control forest soils, the number of copiotrophic, oligotrophic and spore-forming bacteria decreased simultaneously with increased ACT in all treatments. The number of fluorescent bacteria increased when sawdust was applied on forest sites under planted trees and decreased when sawdust was spread in planting rows. Oligotrophic bacteria also decreased in the latter case. We present a list of bacteria identified by DNA – 16S rRNA gene sequences. Adding sawdust to arable soils significantly changed the quantitative and qualitative composition of microbial communities.     

Hydrolase activity as an indicator of the state of microbial communities in vermicomposts

The hydrolase activity was studied using the fluorescein diacetate (FDA) hydrolysis assay in order to assess the microbial activity in composts and vermicomposts obtained with participation of Eisenia fetida andrei, Aporrectodea caliginosa, and A. rosea earthworms from cattle manure, leaves, and peat. The hydrolase activity (the Michaelis—Menten constant, K _m ) was found to be lower in the vermicomposts than in the composts and correlated with a corresponding decrease in the length of fungal hyphae. The determination of the hydrolase activity according to the reaction of FDA hydrolysis may be recommended to control the process of vermicomposting and the quality of vermicomposts. Thus, the process of vermicomposting may be characterized with the use of the functional (hydrolase activity) and structural (microbial biomass, length of fungal hyphae) approaches.     

两种母岩发育森林土壤微生物生物量碳代谢的差异性

针对不同母岩发育土壤的微生物生物量碳代谢特征及驱动因子不明确的科学问题,以石灰岩和碎屑岩两种母岩发育的森林土壤为研究对象,利用18O-H2O标记测定微生物生长速率、微生物呼吸速率、微生物生物量碳利用效率（CUE）以及微生物周转时间,并结合土壤理化性质、土壤有机质矿物保护特性和土壤酶活性以及微生物生物量和群落组成,明确岩性对森林土壤微生物生物量碳代谢的影响机制。结果表明：石灰岩发育土壤的pH和0.05mm～0.002 mm粒径含量高于碎屑岩发育土壤,而有机碳（SOC）、全氮（TN）、可溶性碳（DOC）、C:P和N:P却低于碎屑岩发育土壤（P<0.05）;石灰岩发育土壤交换性钙镁（(Ca+Mg)）和游离态铁铝（(Fe+Al)d）含量高于比碎屑岩发育土壤,但非晶态铁铝（(Fe+Al)o）含量则低于碎屑岩发育土壤;石灰岩发育土壤碳氮磷循环、相关酶活性均显著低于碎屑岩发育土壤（P<0.05）;石灰岩发育土壤微生物生物量磷（MBP）高于碎屑岩发育土壤,但微生物生物量碳（MBC）、真菌细菌比（F:B）和革兰氏阳性菌阴性菌比（G+:G-）则显著低于碎屑岩发育土壤（P < 0.05）;石灰岩发育土壤微生物生长速率和周转速率显著高于碎屑岩发育土壤（P < 0.05）,但微生物呼吸速率和CUE在两种土壤之间差异并不显著。土壤微生物生长速率和微生物周转速率均与土壤pH、(Ca+Mg):(Fe+Al)o、(Ca+Mg):SOC、(Fe+Al)d:SOC和革兰氏阴性细菌呈显著正相关（P<0.05）,而与DOC、铁铝结合态有机碳、酶活性、MBC:MBN、F:B和G+:G-比呈显著负相关关系（P < 0.05）。此外,土壤CUE与MBC和MBC:MBN呈显著负相关（P < 0.05）;微生物呼吸速率仅与酚氧化酶活性呈显著负相关（P<0.05）。两种岩石发育的森林土壤微生物生物量碳代谢受生物和非生物因素的控制,这一研究结果为解释不同母岩发育森林土壤有机碳库的差异提供参考。     

Regional pattern of the mobile water fraction in soils as determined by disc infiltrometer experiments

In order to characterize the partitioning of solutes into the mobile and the immobile fraction of pore water at the soil surface, field measurements of infiltration in unsaturated loamy sands were carried out using a disc infiltrometer in a region of about 10 km² at the Lüneburger Heide, Germany. The aims of these measurements were to obtain the mobile water fraction and the hydraulic properties of the soil, to relate them to other measured soil properties like the soil texture, and to assess their spatial variability in the region. From the infiltration results we obtained the hydraulic conductivity K₀ and the sorptivity S₀ using a three-dimensional infiltration analysis and the two-term infiltration equation, respectively. The mobile water fraction θ_m/θ of the soil was estimated from the infiltration results of a second disc infiltrometer filled with bromide solution. The estimated values of K₀ and S₀ were in good agreement with those reported in the literature as typical for loamy sands. The average value of θ_m/θ was calculated to be 71 % under the experimental conditions. Neither the horizontal spatial distribution of water and bromide concentration beneath the disc infiltrometer nor the change of the infiltrometer during the experiment had a notable influence on the experimental results. The spatial variability of the data was analyzed by means of variograms. All variograms were spatially structured and indicated a high small scale variation of the variables. In conclusion, the technique applied is suitable for the estimation of the hydraulic and solute transport properties of soils, especially at larger scales.     

Variation in the flavonol glycoside composition of almond seedcoats as determined by maldi-tof mass spectrometry

Seedcoats of 16 almond varieties were screened for flavonol glycosides by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Flavonol glycosides were extracted by a simple methanolic extraction followed by a quick cleanup procedure with a Sep-Pak C(18) cartridge. Each of the 16 seedcoat samples exhibited a unique composition. Four flavonol glycosides, isorhamnetin rutinoside, isorhamnetin glucoside, kaempferol rutinoside, and kaempferol glucoside, were detected and quantified with use of rutin as an internal standard. Individual peak ratios were very consistent across triplicate analyses of all samples; the average standard deviation was 9%. In all almond varieties, isorhamnetin rutinoside was the most abundant flavonol glycoside, and the total content ranged from 75 to 250 microg/g.     

Determination of microbial phenolic acids in human faeces by UPLC-ESI-TQ MS

The aim of the present work was to develop a reproducible, sensitive, and rapid UPLC-ESI-TQ MS analytical method for determination of microbial phenolic acids and other related compounds in faeces. A total of 47 phenolic compounds including hydroxyphenylpropionic, hydroxyphenylacetic, hydroxycinnamic, hydroxybenzoic, and hydroxymandelic acids and simple phenols were considered. To prepare an optimum pool standard solution, analytes were classified in 5 different groups with different starting concentrations according to their MS response. The developed UPLC method allowed a high resolution of the pool standard solution within an 18 min injection run time. The LOD of phenolic compounds ranged from 0.001 to 0.107 μg/mL and LOQ from 0.003 to 0.233 μg/mL. The method precision met acceptance criteria (<15% RSD) for all analytes, and accuracy was >80%. The method was applied to faecal samples collected before and after the intake of a flavan-3-ol supplement by a healthy volunteer. Both external and internal calibration methods were considered for quantification purposes, using 4-hydroxybenzoic-2,3,4,5-d4 acid as internal standard. For most analytes and samples, the level of microbial phenolic acids did not differ by using one or another calibration method. The results revealed an increase in protocatechuic, syringic, benzoic, p-coumaric, phenylpropionic, 3-hydroxyphenylacetic, and 3-hydroxyphenylpropionic acids, although differences due to the intake were only significant for the latter compound. In conclusion, the UPLC-DAD-ESI-TQ MS method developed is suitable for targeted analysis of microbial-derived phenolic metabolites in faecal samples from human intervention or in vitro fermentation studies, which requires high sensitivity and throughput.     

Limitations of soil microbial biomass carbon as an indicator of soil pollution in the field

The size of the soil microbial biomass carbon (SMBC) has been proposed as a sensitive indicator for measuring the adverse effects of contaminants on the soil microbial community. In this study of Australian agricultural systems, we demonstrated that field variability of SMBC measured using the fumigation-extraction procedure limited its use as a robust ecotoxicological endpoint. The SMBC varied up to 4-fold across control samples collected from a single field site, due to small-scale spatial heterogeneity in the soil physicochemical environment. Power analysis revealed that large numbers of replicates (3-93) were required to identify 20% or 50% decreases in the size of the SMBC of contaminated soil samples relative to their uncontaminated control samples at the 0.05% level of statistical significance. We question the value of the routine measurement of SMBC as an ecotoxicological endpoint at the field scale, and suggest more robust and predictive microbiological indicators.     

Arylsulfatase activity of microbial biomass in soils as affected by cropping systems

 The impacts of crop rotations and N fertilization on different pools of arylsulfatase activity (total, intracellular, and extracellular) were studied in soils of two long-term field experiments in Iowa to assess the contibution of the microbial biomass to the activity of this enzyme. Surface-soil samples were taken in 1996 and 1997 in corn, soybeans, oats, or meadow (alfalfa) plots that received 0 or 180 kg N ha^–1 before corn, and an annual application of 20 kg P ha^–1 and 56 kg K ha^–1. The arylsulfatase activity in the soils was assayed at optimal pH (acetate buffer, pH 5.8) before and after chloroform fumigation; microbial biomass C (C_mic) and N (N_mic) were determined by chloroform-fumigation methods. All pools of arylsulfatase activity in soils were significantly affected by crop rotation and plant cover at sampling time, but not by N fertilization. Generally, the highest total, intracellular, and extracellular arylsulfatase activities were obtained in soils under cereal-meadow rotations, taken under oats or meadow, and the lowest under continuous cropping systems.Total, intracellular, and extracellular arylsulfatase activities were significantly correlated with C_mic (r>0.41, P<0.01) and N_mic (r>0.38, P<0.01) in soils. The averages of specific activity values, i.e., of arylsulfatase activity of the microbial biomass, expressed per milligram C_mic, ranged from 315 to 407 μg p-nitrophenol h^–1. The total arylsulfatase activity was significantly correlated with the intracellular activity, with r values >0.79 (P<0.001). In general, about 45% of the total arylsulfatase activity was extracellular, and 55% was associated with the microbial biomass in soils, indicating the importance of the microflora as an enzyme source in soils. Received: 23 April 1998     

Metabolism and lack of DNA reactivity of the mycotoxin ochratoxin a in cultured rat and human primary hepatocytes

It is still unclear whether the carcinogenic mycotoxin ochratoxin A (OTA) is bioactivated to DNA-binding metabolites in rodents and humans. Therefore, we have incubated cultured rat and human primary hepatocytes with noncytotoxic concentrations of (3)H-OTA ranging from 10(-7) to 10(-5) M for 8 h and determined its metabolism and covalent DNA binding. In rat hepatocytes, OTA was metabolized to small amounts of three products, which were further studied by electrospray ionization (ESI)-MS/MS techniques. In addition to 4-hydroxy-OTA, which is a known product of OTA biotransformation, two novel metabolites were detected and tentatively identified as hexose and pentose conjugates of OTA. The in vitro induction with 3-methylcholanthrene (3MC) increased the formation of 4-hydroxy-OTA but did not alter the formation of the conjugated metabolites. No covalent binding of (3)H-OTA or its metabolites to DNA was observed in rat hepatocytes with or without 3MC induction with a limit of detection of 2 adducts per 10(9) nucleotides. However, the cellular ratio of reduced glutathione to oxidized glutathione was significantly decreased by treatment with OTA. In cultured human hepatocytes, (3)H-OTA was only very poorly metabolized, and no covalent DNA binding was observed. In conclusion, the results of this in vitro study do not support the notion that OTA has the potential to undergo metabolic activation and form covalent DNA adducts in rodents and humans.     

Characterization of metabolites of hydroxycinnamates in the in vitro model of human small intestinal epithelium caco-2 cells

Hydroxycinnamic acids are antioxidant phenolic compounds which are widespread in plant foods, contribute significantly to total polyphenol intakes, and are absorbed by humans. The extent of their putative health benefit in vivo depends largely on their bioavailability. However, the mechanisms of absorption and metabolism of these phenolic compounds have not been described. In this study, we used the in vitro Caco-2 model of human small intestinal epithelium to investigate the metabolism of the major dietary hydroxycinnamates (ferulate, sinapate, p-coumarate, and caffeate) and of diferulates. The appearance of metabolites in the medium versus time was monitored, and the various conjugates and derivatives produced were identified by HPLC-DAD, LC/MS, and enzyme treatment with beta-glucuronidase or sulfatase. Enterocyte-like differentiated Caco-2 cells have extra- and intracellular esterases able to de-esterify hydroxycinnamate and diferulate esters. In addition, intracellular UDP-glucuronosyltransferases and sulfotransferases existing in Caco-2 cells are able to form the sulfate and the glucuronide conjugates of methyl ferulate, methyl sinapate, methyl caffeate, and methyl p-coumarate. However, only the sulfate conjugates of the free acids, ferulic acid, sinapic acid, and p-coumaric acid, were detected after 24 h. The O-methylated derivatives, ferulic and isoferulic acid, were the only metabolites detected following incubation of Caco-2 cells with caffeic acid. These results show that the in vitro model system differentiated Caco-2 cells have the capacity to metabolize dietary hydroxycinnamates, including various phase I (de-esterification) and phase II (glucuronidation, sulfation, and O-methylation) reactions, and suggests that the human small intestinal epithelium plays a role in the metabolism and bioavailability of these phenolic compounds.