Interactions of bacteria,protozoa and plants leading to mineralization of soil nitrogen

The capacity of bacteria and protozoa to mineralize soil nitrogen was studied in microcosms with sterilized soil with or without wheat plants. The effect of small additions of glucose or ammonium nitrate or both, twice a week was also tested. Plant dry weight and N-content, number of microorganisms and biomass plus inorganic N were determined after 6 weeks.The introduction of plants profoundly influenced the N transformations. In the presence of root-derived carbon, much more N was mineralized from the organic matter and immobilized mainly in plant biomass. “Total observable change in biomass N plus inorganic N” was negative in the unvegetated soils without additions, while a mineralization of 1.7 mg N microcosm^?1 was observed in microcosms with wheat plants grown with bacteria only. When protozoa were included, the N taken up by plants increased by 75%. Sugar additions resulted in an 18% increase of total N in the shoots when protozoa were present, but had no significant effect in the absence of grazers. Plants with the same root weight were more efficient in their uptake of inorganic N when protozoa were present. Plants grown with protozoa also had a lower R/S ratio, indicating a less stressed N availability situation. The lowest ratio was found with N additions in the presence of protozoa.The results indicate that, with energy supplied by plant roots or with external glucose additions, soil bacteria can mineralize N from the soil organic matter to support their own growth. Grazing of the bacteria is necessary to make bacterial biomass N available for plant uptake.     

The impact of protozoa on the availability of bacterial nitrogen to plants

Summary Microbial N from ¹⁵N-labelled bacterial biomass was investigated in a microcosm experiment, in order to determine its availability to wheat plants. Sterilized soil was inoculated with either bacteria (Pseudomonas aeruginosa alone or with a suspension of a natural bacterial population from the soil) or bacteria and protozoa to examine the impact of protozoa. Plant biomass, plant N, soil inorganic N and bacterial and protozoan numbers were determined after 14 and 35 days of incubation. The protozoa reduced bacterial numbers in soil by a factor of 8, and higher contents of soil inorganic N were found in their presence. Plant uptake of N increased by 20010 in the presence of protozoa. Even though the total plant biomass production was not affected, the shoot: root ratios increased in the presence of protozoa, which is considered to indicate an improved plant nutrient supply. The presence of protozoa resulted in a 65010 increase in mineralization and uptake of bacterial ¹⁵N by plants. This effect was more pronounced than the protozoan effect on N derived from soil organic matter. It is concluded that grazing by protozoa strongly stimulates the mineralization and turnover of bacterial N. The mineralization of soil organic N was also shown to be promoted by protozoa.Communication No. 9 of the Dutch Programme on Soil Ecology of Arable Farming Systems     

Grazing of protozoa on rhizosphere bacteria alters growth and reproduction of Arabidopsis thaliana

Plant roots are densely colonized by bacteria which form the basis of the rhizosphere bacterial food web with protozoa as most effective predators. We established a well defined laboratory system with Arabidopsis thaliana as model plant allowing to investigate in detail the effect of rhizosphere interactions on plant performance. We used this system to analyse separate and combined effects of natural rhizobacteria and the protozoa Acanthamoeba castellanii on plants.Protozoa and bacteria increased plant growth with the effect of protozoa markedly exceeding that of bacteria only. Arabidopsis immediately responded to the presence of protozoa by increasing carbon but not nitrogen uptake. Later protozoa enhanced plant uptake of nitrogen from organic material and prolonged vegetative growth of Arabidopsis resulting in strongly increased seed production. It is concluded that the immediate plant response was based on changes in rhizosphere signalling inducing increased plant carbon fixation rather than on protozoa-mediated increase in nitrogen availability. The subsequently increased plant nitrogen uptake presumably originated from nitrogen fixed in bacterial biomass made available by protozoan grazing, i.e. the microbial loop in soil. The results suggest that Arabidopsis prepared for the upcoming mobilization of nitrogen by increasing carbon fixation and root carbon allocation which paid-off later by increased nutrient capture and strongly increased plant reproduction.     

Protozoan predation and the turnover of soil organic carbon and nitrogen in the presence of plants

Summary The impact of protozoan grazing on the dynamics and mineralization of ¹⁴C- and ¹⁵N-labelled soil organic material was investigated in a microcosm experiment. Sterilized soil was planted with wheat and either inoculated with bacteria alone or with bacteria and protozoa or with bacteria and a 1:10 diluted protozoan inoculum. ¹⁴C–CO₂ formation was continuously monitored. It served as an indicator of microbial activity and the respiration of soil organic C. The activity of protozoa increased the turnover of ¹⁴C-labelled substrates compared to soil without protozoa. The accumulated ¹⁴C–CO₂ evolved from the soils with protozoa was 36% and 53% higher for a 1:10 and for a 1:1 protozoan inoculum, respectively. Protozoa reduced the number of bacteria by a factor of 2. In the presence of protozoa, N uptake by plants increased by 9% and 17% for a 1:10 and a 1:1 protozoan inoculum, respectively. Both plant dry matter production and shoot: root ratios were higher in the presence of protozoa. The constant ratio of ¹⁵N: ¹⁴⁺¹⁵N in the plants for all treatments indicated that in the presence of protozoa more soil organic matter was mineralized. Bacteria and protozoa responded very rapidly to the addition of water to the microcosms. The rewetting response in terms of the ¹⁴C–CO₂ respiration rate was significantly higher for 1 day in the absence and for 2 days in the presence of protozoa after the microcosms had been watered. It was concluded that protozoa improved the mineralization of N from soil organic matter by stimulating the turnover of bacterial biomass. Pulsed events like the addition of water seem to have a significant impact on the dynamics of food-chain reactions in soil in terms of C and N mineralization.Communication No. 19 of the Dutch Programme on Soil Ecology of Arable Farming Systems     

Grazing of a common species of soil protozoa (Acanthamoeba castellanii) affects rhizosphere bacterial community composition and root architecture of rice (Oryza sativa L.)

We performed a controlled experiment with rice seedlings (Oryza sativa L.) growing in Petri dishes on homogeneous nutrient agar containing a simple rhizosphere food web consisting of a diverse bacterial community and a common soil protozoa, Acanthamoeba castellanii, as bacterial grazer. Presence of amoebae increased bacterial activity and significantly changed the community composition and spatial distribution of bacteria in the rhizosphere. In particular, Betaproteobacteria did benefit from protozoan grazing. We hypothesize that the changes in bacterial community composition affected the root architecture of rice plants. These effects on root architecture affect a fundamental aspect of plant productivity. Root systems in presence of protozoa were characterized by high numbers of elongated (L-type) laterals, those laterals that are a prerequisite for the construction of branched root systems. This was in sharp contrast to root system development in absence of protozoa, where high numbers of lateral root primordia and short (S-type) laterals occurred which did not grow out of the rhizosphere region of the axile root. As a consequence of nutrient release from grazed bacteria and changes in root architecture, the nitrogen content of rice shoots increased by 45% in presence of protozoa. Our study illustrates that interactions over three trophic levels, i.e. between plants, bacteria and protozoa significantly modify root architecture and nutrient uptake by plants.     

Early decomposer assemblages of soil organisms in litterbags with vetch and rye roots

The assemblages of microbial (bacteria and fungi), microfaunal (protozoa and nematodes) and mesofaunal (microarthropods) populations were studied in decomposing root residues from hairy vetch (Vicia villosa Roth) and rye (Secale cereale L.) in a litterbag field experiment. Litterbags containing vetch or rye root residues were buried in soil at the same day as either vetch or rye winter catch crops were incorporated into the field soil from which the materials were gathered. The litterbags were sampled after 6 weeks in the field. In vetch, bacterial and fungal biomasses were similar whereas fungi dominated microbial biomass in rye. The biomass of the bacterial consuming fauna dominated by nematodes and microarthropods was similar to the biomass of bacteria in vetch as opposed to in rye where bacterivore biomass was lower than bacterial biomass. This suggests a much higher bacterial production in vetch compared to rye. Furthermore, in vetch dauer larvae of bacteria feeding nematodes prevailed, which is also a sign of high bacterial production followed by food shortage for the bacterivores. Bacterivorous and predatory nematodes with capability of consuming protozoa showed an inverse relationship to flagellated protozoa. This suggests that these nematodes controlled the protozoan biomass constituting a lower fraction of the bacterivore biomass in vetch compared to in rye. Such intraguild predator-prey relationship is therefore indicated for microbivorous organisms among bacterivorous and predatory nematodes (the intraguild predator) protozoa (the intraguild prey) and bacteria (the common prey). The much higher fungal biomass in rye than in vetch litterbags was not reflected in the biomass of the fungal feeders. Due to the generally lower intrinsic rate of increase of the fungivores, as well as of the omnivores and predators, in comparison with the bacterial feeders, they were not able to generate dense populations at this early stage of decomposition.     

Influence of texture on habitable pore space and bacterial-protozoan populations in soil

Summary Soil texture affects pore space, and bacterial and protozoan populations in soil. In the present study we tested the hypothesis that bacteria are more protected from protozoan predation in fine-textured soils than in coarse-textured soils because they have a larger volume of protected pore space available to them. The experiment consisted of three sterilized Orthic Black Chernozemic soils (silty clay, clay loam, and sandy loam) inoculated with bacteria, two treatments (with and without protozoa), and five sampling dates. The soils were amended with glucose and mineral N on day 0. On day 4 bacterial numbers in all three soils were approximately 3×10⁹ g^–1 soil. The greatest reduction in bacteria due to protozoan grazing occurred between day 4 and day 7. Compared to the treatment without protozoa, bacteria in the treatment with protozoa were reduced by 68, 50, and 75% in the silty clay, clay loam, and sandy loam, respectively. On day 4, 2 days after the protozoan inoculation, all protozoa were active. The numbers were 10330, 4760, and 15 380 g^–1 soil for the silty clay, clay loam, and sandy loam, respectively. Between day 4 and day 7, the period of greatest bacterial decline, total protozoa increased greatly to 150480, 96160, and 192100 g^–1 soil for the three soils, respectively. Most protozoa encysted by day 7. In all soils the addition of protozoa significantly increased CO₂–C evolution per g soil relative to the treatment without protozoa. Our results support the hypothesis that bacteria are more protected from protozoan predation in fine-textured soils than in coarse-textured soils.     

Oscillating dynamics of bacterial populations and their predators in response to fresh organic matter added to soil: The simulation model ‘BACWAVE-WEB’

Recently, regular oscillations in bacterial populations and growth rates of bacterial feeding nematodes (BFN) were shown to occur after addition of fresh organic matter to soil. This paper presents a model developed to investigate potential mechanisms of those oscillations, and whether they were initiated by bacteria-substrate interactions or by predatory regulation of bacteria. The model was also used to investigate mineral nitrogen release during short-term organic matter decomposition. Experimental data originated from several microcosm experiments with a sandy soil amended with clover-grass mixture. Numbers of bacteria and BFN, and nitrate and ammonium content in soil were measured daily during about a month, whereas protozoa were counted occasionally. A substrate-based food web model was constructed with 3 plant residue and 5 soil organic matter compartments, 3 trophic groups of bacteria (copiotrophic, oligotrophic and hydrolytic), and two predatory groups (BFN and protozoa). Both carbon and nitrogen flows between these compartments were modelled. Fluctuations in microbial populations in soil after plant residue incorporation could be reproduced with and without participation of predators. The first two peaks in bacterial numbers were mainly related to bacteria-substrate interactions, while predators (particularly protozoa) influenced bacterial dynamics during later stages of bacterial community development. Oligotrophic bacteria had a stabilizing effect on fluctuations of other trophic groups, and were the main source of nutrients for predators. A peak in soil ammonium occurred within 1 week after residue incorporation. Nitrate increased sigmoidally after a short lag phase. The final nitrate concentration was primarily determined by bacterial dynamics, and to a lesser extent by protozoa and nematodes. This model indicates the importance of substrate-consumer relations for regulation of populations at different trophic levels and nutrient release from fresh organic matter added to soil.     

Prey-predator dynamics in communities of culturable soil bacteria and protozoa: differential effects of mercury

We investigated whether the prey-predator dynamics of bacteria and protozoa were affected by inorganic mercury at concentrations of 0, 3.5 and 15 mg Hg(II) kg soil⁻¹. The amount of bioavailable Hg was estimated using a biosensor-assay based on the mer-lux gene fusion. The numbers of bacterial CFUs on the general medium 1/100 tryptic soy agar (TSA) were significantly decreased when the soil had been amended with Hg. In contrast, no effect was seen on the number of CFUs on the Pseudomonas-specific medium Gould's S1 agar. Protozoan numbers estimated by the most probable number (MPN) method with 1/100 TSB as growth medium were also negatively affected by Hg. The different fractions of protozoa were affected to different degrees suggesting that amoebae were less sensitive than slow-growing flagellates, which again were less sensitive than the fast-growing flagellates. In contrast, Hg did not induce any detectable changes in the diversity of flagellate morphotypes. In the treatment with 15 mg Hg kg⁻¹ a transiently increased number of bacteria was seen at day 6 probably concomitant with a decrease in the numbers of protozoa. This might indicate that Hg affected the prey-predator dynamics in communities of culturable bacteria and protozoa in soil. Furthermore, we showed that the number of Pseudomonas spp. was not affected by Hg whereas the number of bacteria growing on a general medium was.     

Protease and deaminase activities in wheat rhizosphere and their relation to bacterial and protozoan populations

Protease and deaminase activities and population dynamics of bacteria and protozoa were measured in the rhizosphere of wheat to study their interactions with the mineralization of nitrogen. The experimental design allowed the separation of roots and soil material by means of a gauze. The most pronounced rhizosphere effect was detected for all the measured variables in the soil closest to the gauze. The number of bacteria was significantly higher in the presence than in the absence of plants up to 4 mm away from the soil-root interface and the closer to this interface the higher the number. Protozoan and bacterial population dynamics were positively correlated; generally, populations of flagellates and amoebae were comparable and their sum accounted for the population of total protozoa. For both enzyme activities the rhizosphere effect extended up to 2 mm away from the soil-root interface. The histidinase activity was of bacterial origin, while it is likely that bacteria, protozoa and root hair all contributed to the overall caseinase activity. Decomposition of root exudates and native organic matter in the rhizosphere, reflected by a growing microbial population, is associated with nitrogen mineralization through increases in caseinhydrolysing and L-histidine-deaminating activities. The adopted soil-plant microcosm is suitable for the study of the rhizosphere effect over time of incubation and distance gradient from the soil-root interface.     

土壤原生动物 ——研究方法及其在土传病害防控中的作用

原生动物是原生生物的一种,是土壤食物网中的消费者,能捕食细菌和真菌等其他微生物.除了对土壤微生物群落和物质循环产生重要影响外,根际原生动物与细菌、真菌等土壤微生物共同组成生物网络屏障,并在抵御土传病原菌入侵作物根系的过程中发挥着重要作用.然而,相对于根际有益细菌和有益真菌,国内外关于原生动物防控土传病害的效果及作用机制...     

Evidence for gluconic acid production by Enterobacter intermedium as an efficient strategy to avoid protozoan grazing

The effect of pure gluconic acid and of gluconic-acid-producing bacteria on the activity of three protozoan species, Colpoda steinii (a ciliate), Vahlkampfia sp. (an amoeba) and Neobodo designis (a flagellate), was determined in vitro and in soil microcosms. Pure gluconic acid was shown to mediate disappearance of active cells, due to encystment and/or death of protozoa, at 0.15 mM in saline medium. Similarly, the presence of gluconic acid inhibited excystment of the three protozoa tested. Enterobacter intermedium 60-2G (Wt), a gluconic acid-producing rhizobacterium, elicited the same effects on protozoa when co-cultured in the presence of 5 g L⁻¹ glucose. However, the effect was not observed when glucose was omitted from the medium. Similarly, a pqqA isogenic mutant strain, unable to produce gluconic acid from glucose, exhibited a reduced effect on protozoan activity. Rhizosphere-microcosm studies performed with wheat (Triticum aestivum L.) confirmed the reduced ability of the pqqA mutant to limit protozoa reproduction compared to the Wt strain. Since the sodium salt of gluconic acid did not cause any significant stress to protozoa and considering that addition of 50 mM Tris-Cl (pH 7.2) abolished the deleterious effect of gluconic acid, acidification of the medium appeared as the key factor that induced encystment/death of protozoa. We propose that production and excretion of gluconic acid should be considered an efficient mechanism evolved by bacteria to escape, tolerate or defend themselves against protozoan grazing in rhizosphere environments.     

Populations of predatory protozoa in field soils after 5 years of elemental s fertilizer application

The effect of 5 yr of repeated application of elemental S (S°) fertilizer on predatory protozoa in soil was investigated. Protozoa that feed on the bacteria Arthrobacter globiformis and Enterobacter aerogenes or the fungi Fusarium solani and Neurospora crassa were enumerated by most probable number (MPN) methods. The application of S° fertilizer reduced the microbial biomass and its activity in soil. Soils treated with 44kg S° ha⁻¹ yr⁻¹ for 5 yr exhibited a 30–71% decline in MPN of protozoa feeding on bacteria and more than a 84% decline in the population of mycophagous amoebae. This decline in protozoa populations parallelled changes in microbial biomass, especially in the case of mycophagous amoebae and fungal biomass. The adverse effect of repeated S° applications on microbial biomass and predatory protozoa was long lasting. Since nutrient transformations (e.g. mineralization) in soil are influenced by microbial interactions, our results suggest reduced nutrient turnover via microbial predation in S° treated soils.     

Bacteria and protozoa in soil microhabitats as affected by earthworms

The effects of incorporation of elm leaves (Ulmus glabra) into an agricultural sandy loam soil by earthworms (Lumbricus festivus) on the bacterial and protozoan populations were investigated. Three model systems consisting of soil, soil with leaves, and soil with leaves and earthworms, respectively, were compared. The total, viable, and culturable number of bacteria, the metabolic potentials of bacterial populations, and the number of protozoa and nematodes were determined in soil size fractions. Significant differences between soil fractions were shown by all assays. The highest number of microorganisms was found in microaggregates of 2–53 μm and the lowest in the <0.2μm fraction. A major part of the bacteria in the latter fraction was viable, but non-culturable, while a relatively higher number of culturable bacteria was found in the macroaggregates. The number of colony-forming units and 5-cyano-2,3-ditolyl tetrazolim chloride (CTC)-reducing bacteria explained a major part of the variation in the number of protozoa. High protozoan activity and predation thus coincided with high bacterial activity. In soil with elm leaves, fungal growth is assumed to inhibit bacterial and protozoan activity. In soil with elm leaves and earthworms, earthworm activity led to increased culturability of bacteria, activity of protozoa, number of nematodes, changed metabolic potentials of the bacteria, and decreased differences in metabolic potentials between bacterial populations in the soil fractions. The effects of earthworms can be mediated by mechanical mixing of the soil constituents and incorporation of organic matter into the soil, but as the earthworms have only consumed a minor part of the soil, priming effects are believed partly to explain the increased microbial activity. Received: 7 January 1996     

A new method for assessing soil microorganism diversity and evidence of vitamin deficiency in low diversity communities

Summary A proposed new method for assessing the diversity of a soil microbial community is based on the species-typical ester-linked phospholipid fatty acids in the membranes of living cells. Soils that support only a few dominant species (bacteria, fungi, protozoa or algae) are expected to show few dominant fatty acids and vice versa. The phospholipid fatty-acid diversity in nine soils from Central Switzerland was calculated using Shannon's formula. By means of a respiration test, it was further established that the low-diversity soils responded significantly and positively (respiration increase) to small additions of a vitamin mixture containing thiamin, pyridoxin, calpan, folic acid, and biotin. The results indicate a connection between microbial diversity and a yet unspecified vitamin deficiency within the population. Whether the vitamin deficiency is the cause or the effect of the reduced diversity remains to be established.     

The mycorrhizal fungus (Glomus intraradices) affects microbial activity in the rhizosphere of pea plants (Pisum sativum)

Pea plants were grown in γ-irradiated soil in pots with and without addition of the AM fungus Glomus intraradices at sufficient N and limiting P. Depending on the growth phase of the plant presence of AM had negative or positive effect on rhizosphere activity. Before flowering during nutrient acquisition AM decreased rhizosphere respiration and number of protozoa but did not affect bacterial number suggesting top-down regulation of bacterial number by protozoan grazing. In contrast, during flowering and pod formation AM stimulated rhizosphere respiration and the negative effect on protozoa decreased. AM also affected the composition of the rhizosphere bacterial community as revealed from DNA analysis (DGGE). With or without mycorrhiza, rhizosphere respiration was P-limited on very young roots, not nutrient limited at more mature roots and C-limited at withering. This suggests changes in the rhizosphere community during plant growth also supported by changes in the bacteria (DGGE).     

Trophic interactions between rhizosphere bacteria and bacterial feeders influenced by phosphate and aphids in barley

The aim was to study the effects of P fertilization and leaf aphid attack on the trophic interactions of bacteria and bacterial feeders in the rhizospheres of barley plants. The density of protozoa peaked in the rhizospheres of plants fertilized with N and P, whereas nematodes peaked in the rhizospheres of plants to which only N had been added. Fingerprinting of bacterial communities by length heterogeneity polymerase chain reaction revealed differences in community structure between NP rhizospheres and N rhizospheres as well as aphid-related differences within N rhizospheres. Specifically, α-proteobacteria increased with P addition. To evaluate if differences in bacteria in terms of their quality as food could partly explain the observed differences in protozoan and nematode abundances, growth of the flagellate Cercomonas sp. was assessed with 935 bacteria isolated from the different treatments. This assay indicated that bacterial isolates were of higher food quality to Cercomonas sp. in NP than in N rhizospheres when plants were subjected to aphid attack. Bacteria of high and low food quality for Cercomonas sp., respectively, were fed to the nematode Caenorhabditis elegans and larval production examined. α-Proteobacteria supported the growth of Cercomonas sp. well, whereas Actinobacteria did not. In contrast, C. elegans reproduced poorly on most α-proteobacteria but were able to reproduce well on some Actinobacteria. These results suggest that the different response of protozoa and nematodes to P addition could be mediated through a food quality-related change in community composition of bacteria and that leaf aphid attack may interfere with nutrient effects on bacterial assemblages of rhizospheres.     

The “soil microbial loop” is not always needed to explain protozoan stimulation of plants

Protozoa stimulate plant growth, but we do not completely understand the underlying mechanisms, and different hypotheses seek to explain this phenomenon. To test these hypotheses, we grew the grass Yorkshire fog (Holcus lanatus) in pots with soil, which contained either (1) no organisms but bacteria – or (2) bacteria and protozoa. Half of the pots received a glucose treatment so as to mimic an additional root exudation. We measured plant growth and plant nitrogen uptake, along with various microbial pools and processes that support plant growth. Protozoan presence significantly enhanced soil nitrogen mineralization, plant nitrogen uptake from organic nitrogen sources, plant nitrogen content, and plant growth. By contrast, we found no evidence that glucose addition, mimicking root exudation, increased soil nitrogen availability and plant nitrogen uptake. Moreover, although protozoan presence affected bacterial community structure, it did not affect the proportion of IAA-producing bacteria in the community or plant root morphology. These results refute the “soil microbial loop” hypotheses, which suggest that protozoan stimulation of plant growth results from complex interactions between plant roots, bacteria and protozoa. Our experiment thus favours the simple explanation that increased nitrogen availability is the key factor behind the positive protozoan effect on plant growth. To exploit natural resources in an efficient and environmentally friendly way, we need to understand in detail the functioning of ecosystems. This study stresses that to achieve this, it is still urgent, besides investigating intricate food-web and signal compound interactions, also to focus on the basic stoichiometric and energetic aspects of organisms.     

Humic preparations and the assessment of their biological activity for certification purposes

The diversity of responses of test organisms (higher plants, microalgae, protozoa, crustaceans, bacteria, and mammal cell cultures in vitro) to the action of humates produced industrially from different raw materials was discussed in relation to the urgent problem of the certification of structural and functional properties of commercial humates. It was proposed to include higher plants (for assessing the effect of stimulation) and a set of biotests using test organisms of different trophic levels (for determining toxic concentrations) in the program of assessing the biological activity of humic preparations by laboratory testing methods.     

Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.