Population dynamics of the mycoparasite, sporidesmium sclerotivorum, and its host, sclerotinia minor, in soil

The infection and survival of sclerotia of Sclerotinia minor and the production ofmacroconidia of the mycoparasite, Sporidesmium sclerotivorum, were studied in vitro when each fungus was added to soil at various initial inoculum densities. The rate at which S. sclerotivorum invaded host sclerotia and caused their decay varied with the amount of the mycoparasite added to soil. The results suggest that approximately 5 macroconidia of the mycoparasite g^?1 of soil are needed to successfully infect sclerotia and bring about their decay, when soils are sampled and mixed every 2 weeks. The rate at which S. sclerotivorum infects sclerotia of S. minor and causes their decay is also dependent on the initial inoculum density of the host. Each infected sclerotium supports the production of about 15,000 new macroconidia in soil regardless of the initial inoculum density of the host. It is concluded that successful biological control by S. sclerotivorum is dependent on the soil population of both the host and the mycoparasite.     

In vitro experiments simulating the possible fates of aphid honeydew sugars in soil

In vitro intact soil cores were treated with sugar amendments at a rate equivalent to that of honeydew produced by a population of tree dwelling aphids. The priming action induced by such amendments was dependent upon the soil type, nature of sugar added and degree of repetition of amendment. A hypothesis has been proposed to explain the presence of a positive priming effect in a woodland soil and a negative priming effect in a grassland soil after equivalent sucrose additions.     

In vitro effects of Fusarium blight-controlling fungicides on pathogens of Poa pratensis

An experimental iprodione fungicide,3-(3,5dichlorophenyl)-N-(1-methylethyl)-2,4-dioxo-1-imidazolidinecarboxamide, controls Fusarium blight of Kentucky bluegrass (Poa pratensis L.), but it also amplifies the proportion of crowns colonized by Fusarium and the number of its propagules in soils. In contrast, the disease, the proportion of infected crowns, and the numbers of propagules in soil are generally suppressed by benomyl, methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate. Triadimefon, 1-(4-chlorophenoxy)-3,3-dimethyl-1-(1H-1,2,4-triazol-1-yl)-2-butanone, also controls the disease but is not stimulatory or inhibitory of fusaria. Iprodione and benomyl were studied for their effects on growth and sporulation of Fusarium acuminatum isolated from diseased crowns; iprodione had no or slightly stimulatory effects, and benomyl greatly suppressed these processes, except in a benomyl-tolerant strain.Toxicities of iprodione and benomyl to 1555 identified Fusarium isolates from Kentucky bluegrass turf were determined, as were the toxicities of iprodione to 23 turfgrass pathogens. Of the Fusarium spp, only F. solani was significantly inhibited by iprodione, whereas all were inhibited by benomyl. Iprodione-sensitive fungi included species of Bipolaris, Corticium, Curvularia, Drechslera, Rhizoctonia, Sclerotinia, and Typhula. Insensitive fungi included Colletotrichum, Fusarium, Gaeumannomyces, and Pythium.Investigations with selective fungicides indicate that the primary causal agent of Fusarium blight is not among the fusaria, and that re-interpretation of the disease and its etiology is necessary.     

Ultrastructure of developing chlamydospores of Fusarium solani f. Cucurbitae in vitro

Propagules from macroconidial cultures of Fusarium solani f. cucurbitae in vitro were examined at intervals during chlamydospore formation by transmission electron microscopy. There was a gradual breakdown of the original macroconidial cell wall. Simultaneously, synthesis of new cell wall material occurred. The significance of autolytic cell wall degradation as it affects the morphology of chlamydospores formed from macroconidial cells in soil is discussed.     

Energy efficiency and soil conservation in conventional,minimum tillage and no-tillage

The objective of this research was to determine the capacity of a soil tillage system in soil conservation, in productivity and in energy efficiency. The minimum tillage and no-tillage systems represent good alternatives to the conventional (plough) system of soil tillage, due to their conservation effects on soil and to the good production of crops (Maize, 96%-98% of conventional tillage for minimum tillage, and 99.8% of conventional tillage for no till; Soybeans, 103%-112% of conventional tillage for minimum tillage and 117% of conventional tillage for no till; Wheat, 93%-97% of conventional tillage for minimum tillage and 117% of conventional tillage for no till. The choice of the right soil tillage system for crops in rotation help reduce energy consumption, thus for maize: 97%-98% energy consumption of conventional tillage when using minimum tillage and 91% when using no-tillage; for soybeans: 98% energy consumption of conventional tillage when using minimum tillage and 93 when using no-tillage; for wheat: 97%-98% energy consumption of conventional tillage when using minimum tillage and 92% when using no-tillage. Energy efficiency is in relation to reductions in energy use, but also might include the efficiency and impact of the tillage system on the cultivated plant. For all crops in rotation, energy efficiency (energy produced from 1 MJ consumed) was the best in no-tillage — 10.44 MJ ha^− 1 for maize, 6.49 MJ ha^− 1 for soybean, and 5.66 MJ ha^− 1 for wheat. An analysis of energy-efficiency in agricultural systems includes the energy consumed-energy produced-energy yield comparisons, but must be supplemented by soil energy efficiency, based on the conservative effect of the agricultural system. Only then will the agricultural system be sustainable, durable in agronomic, economic and ecological terms. The implementation of minimum and no-tillage soil systems has increased the organic matter content from 2% to 7.6% and water stable aggregate content from 5.6% to 9.6%, at 0–30 cm depth, as compared to the conventional system. Accumulated water supply was higher (with 12.4%-15%) for all minimum and no-tillage systems and increased bulk density values by 0.01%-0.03% (no significant difference) While the soil fertility and the wet aggregate stability have initially been low, the effect of conservation practices on the soil characteristics led to a positive impact on the water permeability in the soil. Availability of soil moisture during the crop growth period led to a better plant watering condition. Subsequent release of conserved soil water regulated the plant water condition and soil structure.     

Effects of nutrients and acidity on phialospore germination of Trichoderma in vitro

Phialospores of Trichoderma species required an external source of nutrients and CO₂ for germination on agar discs. Both unwashed agar and unwashed spores contributed sufficient quantities of nutrients to support germination.In a nutrient-depleted system germination was much greater under acid conditions than under neutral conditions. The addition of malt extract to the agar largely eliminated the acidity effect with T. viride Pers. ex S.F. Gray, T. harzianum Rifai and T. koningii Oud. The germination of T. polysporum (Link ex Pers.) Rifai and T. saturnisporum Hammill was strongly inhibited by neutral conditions and this inhibition was not completely overcome by nutrient additions. As spores of most species aged, they required more nutrients and became more sensitive to neutral conditions.Dextrose was less effective than malt extract in promoting germination and amino acids were generally inhibitory. A high level of Al in solution inhibited germination and the degree of inhibition varied from one species to another.     

Interaction between the soil yeast Rhodotorula mucilaginosa and the arbuscular mycorrhizal fungi Glomus mosseae and Gigaspora rosea

The effect of the soil yeast, Rhodotorula mucilaginosa LBA, on Glomus mosseae (BEG n°12) and Gigaspora rosea (BEG n°9) was studied in vitro and in greenhouse trials. Hyphal length of G. mosseae and G. rosea spores increased significantly in the presence of R. mucilaginosa. Exudates from R. mucilaginosa stimulated hyphal growth of G. mosseae and G. rosea spores. Increase in hyphal length of G. mosseae coincided with an increase in R. mucilaginosa exudates. No stimulation of G. rosea hyphal growth was detected when 0.3 and 0.5 ml per petri dish of yeast exudates was applied. Percentage root length colonization by G. mosseae in soybean (Glycine max L. Merill) and by G. rosea in red clover (Trifolium pratense L. cv. Huia) was increased only when the soil yeast was inoculated before G. mosseae or G. rosea was introduced. Beneficial effects of R. mucilaginosa on arbuscular mycorrhizal (AM) colonization were found when the soil yeast was inoculated either as a thin agar slice or as a volume of 5 and 10 ml of an aqueous solution. R. mucilaginosa exudates (20 ml per pots) applied to soil increased significantly the percentage of AM colonization of soybean and red clover.     

Potential gene exchange between Bacillus thuringiensis subsp. kurstaki and Bacillus spp. in soil in situ

The possible transfer of genes from Bacillus thuringiensis subsp. kurstaki (Btk) to indigenous Bacillus spp. was investigated in soil samples from stands of cork oak in Orotelli (Sardinia, Italy) collected 5 years after spraying of the stands with a commercial insecticidal preparation (FORAY 48B) of Btk. Two colonies with a morphology different from that of Btk were isolated and identified as Bacillus mycoides by morphological and physiological characteristics and by 16S rDNA analysis. Amplification by the polymerase chain reaction (PCR) of the DNA of the two isolated B. mycoides colonies with primers used for the identification of the Btk cry genes showed the presence of a fragment of 238 bp of the cry1Ab9 gene that had a similarity of 100% with the sequence of the cry1Ab9 gene present in GenBank, indicating that the isolates of B. mycoides acquired part of the sequence of this gene from Btk. No cells of Btk or B. mycoides carrying the 238-bp fragment of the cry1Ab9 gene were isolated from samples of unsprayed control soil. However, the isolates of B. mycoides were not able to express the partial Cry1Ab protein. Hybridization with probes for IS231 and the cry1Ab9 gene suggested that the inverted repeated sequence, IS231, was probably involved in the transfer of the 238-bp fragment from Btk to B. mycoides. These results indicate that transfer of genes between introduced Btk and indigenous Bacillus spp. can occur in soil under field conditions.     

Transgenic Bt plants decompose less in soil than non-Bt plants

Bt plants are plants that have been genetically modified to express the insecticidal proteins (e.g. Cry1Ab, Cry1Ac, Cry3A) from subspecies of the bacterium, Bacillus thuringiensis (Bt), to kill lepidopteran pests that feed on corn, rice, tobacco, canola, and cotton and coleopteran pests that feed on potato. The biomass of these transgenic Bt plants (Bt+) was decomposed less in soil than the biomass of their near-isogenic non-Bt plant counterparts (Bt−). Soil was amended with 0.5, 1, or 2% (wt wt⁻¹) ground, dried (50 °C) leaves or stems of Bt corn plants; with 0.5% (wt wt⁻¹) ground, dried biomass of Bt rice, tobacco, canola, cotton, and potato plants; with biomass of the near-isogenic plants without the respective cry genes; or not amended. The gross metabolic activity of the soil was determined by CO₂ evolution. The amounts of C evolved as CO₂ were significantly lower from soil microcosms amended with biomass of Bt plants than of non-Bt plants. This difference occurred with stems and leaves from two hybrids of Bt corn, one of which had a higher C:N ratio than its near-isogenic non-Bt counterpart and the other which had essentially the same C:N ratio, even when glucose, nitrogen (NH₄NO₃), or glucose plus nitrogen were added with the biomass. The C:N ratios of the other Bt plants (including two other hybrids of Bt corn) and their near-isogenic non-Bt counterparts were also not related to their relative biodegradation. Bt corn had a significantly higher lignin content than near-isogenic non-Bt corn. However, the lignin content of the other Bt plants, which was significantly lower than that of both Bt and non-Bt corn, was generally not statistically significantly different, although 10-66% higher, from that of their respective non-Bt near-isolines. The numbers of culturable bacteria and fungi and the activity of representative enzymes involved in the degradation of plant biomass were not significantly different between soil amended with biomass of Bt or non-Bt corn. The degradation of the biomass of all Bt plants in the absence of soil but inoculated with a microbial suspension from the same soil was also significantly less than that of their respective inoculated non-Bt plants. The addition of streptomycin, cycloheximide, or both to the soil suspension did not alter the relative degradation of Bt+ and Bt− biomass, suggesting that differences in the soil microbiota were not responsible for the differential decomposition of Bt+ and Bt− biomass. All samples of soil amended with biomass of Bt plants were immunologically positive for the respective Cry proteins and toxic to the larvae of the tobacco hornworm (Manduca sexta), which was used as a representative lepidopteran in insect bioassays (no insecticidal assay was done for the Cry3A protein from potato). The ecological and environmental relevance of these findings is not clear.     

Crop rotation and soil temperature influence the community structure of Aspergillus flavus in soil

Aspergillus flavus, the most important cause of aflatoxin contamination, has two major morphotypes commonly termed ‘S’ and ‘L’ strains. Strain S isolates, on average, produce more aflatoxins than the strain L isolates. The S strain has been implicated as the primary causal agent of several contamination events in both North America and Africa. Strain S incidence and A. flavus propagules were quantified periodically in 11 agricultural fields in South Texas from spring 2001 through spring 2003. Both A. flavus populations and S strain incidence varied significantly among seasons, with warm seasons having higher average quantities of A. flavus (718 CFU g⁻¹) and higher incidences of the S strain (32.3%) than cold seasons (403 CFU g⁻¹ and 16.9% incidence). Previous crop influenced both the quantity of A. flavus and S strains incidence. Corn favors higher soil populations of A. flavus (1628 CFU g⁻¹) compared to cotton (374 CFU g⁻¹) and sorghum (237 CFU g⁻¹). In the agroecosystem of South Texas, both cotton (23.7%) and sorghum (23.5%) favored greater S strain incidence compared to corn (14.0%). Soil surface temperature greatly influenced fungal communities with propagule density decreasing when daily average soil temperature was either below 18 °C or above 30 °C, and the proportion of A. flavus belonging to the S strain increasing as soil temperature increased. The results suggest it may be possible to manipulate crop rotations in order to reduce aflatoxin severity, and that periods of increased soil temperature drive selection of the highly toxigenic S strain of A. flavus in warm climates.     

Field inoculation of Medicago with V-A mycorrhiza and Rhizobium in phosphate-fixing agricultural soil

Field inoculation of Medicago sativa with its symbiotic partners Rhizobium meliloti and the endomycorrhizal fungus Glomus mosseae was assayed under standard agricultural conditions in untreated arable phosphate-fixing soil. Glomus mosseae was successfully introduced and efficiently stimulated plant growth, N incorporation and P uptake. In contrast with a previous filed experiment, Rhizobium meliloti was also effective when inoculated alone. The dual inoculation of Rhizobium + Glomus more than doubled yield compared to an uninoculated control.     

垄覆膜集雨对苜蓿草地土壤水分动态及利用效率的影响

我国西北黄土高原干旱半干旱区年降雨稀少和土壤水分周期性亏缺, 导致人工草地生产力低下、水分利用效率低。本试验在旱作条件下, 将垄覆膜集雨措施应用于紫花苜蓿种植, 研究了沟垄宽比和覆膜方式对2 年龄苜蓿草地土壤水分动态及水分利用效率的影响。结果表明: 垄覆膜集雨在集雨前期(4 月中旬至6 月上旬)显著提高0~20 cm 土层土壤含水量, 在集雨中后期(6 月中旬至9 月下旬)显著提高0~120 cm 土层平均含水量,越冬期增加土壤水分入渗能力, 提高20~120 cm 土层平均含水量; 且垄覆膜处理的集雨效率高于土垄处理。随着生育时期的延伸, 垄覆膜处理0~120 cm 土壤平均贮水量呈先降后升的趋势, 土垄处理0~120 cm 土壤平均贮水量呈由高到低的趋势; 在苜蓿生长后期垄覆膜处理的蓄墒能力显著高于土垄处理。垄覆膜处理的平均水分利用效率为34.91 kg·mm^-1·hm^-2, 为对照(平作, CK)的2.25 倍, 土垄处理的平均水分利用效率为28.47kg·mm^-1·hm^-2, 为CK 的1.83 倍, 垄覆膜相对土垄处理平均水分利用效率提高22.62%; 垄覆膜处理以沟垄宽比为60 cm∶60 cm 和60 cm∶75 cm 的水分利用效率最高。     

Effect of soil moisture and temperature on the survival of phytophthora cinnamomi rands in soil

Soil moisture and temperature affect hyphal lysis and sporangial formation in Phytophthora cinnamomi. Hyphal lysis was most rapid in soils at, or below, field capacity when incubated at 25–27° C. Sporangia were not formed in soils below field capacity and only very slowly in waterlogged soil. Sporangia were not formed at temperatures below 15°C.     

Effects of Lumbricus terrestris, Allolobophora chlorotica and Eisenia fetida on microbial community dynamics in oil-contaminated soil

Oil spills are one of the most common types of soil pollution. Bioremediation has become an attractive alternative to physicochemical methods of remediation, where feasible. Earthworms have been shown to stimulate the degradation of petroleum hydrocarbons in soil, and it was hypothesized that the role of earthworms in remediation lies in the enhancement of an oil degrading microbial community. The aim of this study was to characterize microbial activity and community dynamics in oil-contaminated soil incubated with or without earthworms. Three earthworm species (Eisenia fetida, Allolobophora chlorotica and Lumbricus terrestris) were incubated in crude oil polluted soil (ca. 10,000 mg/kg total petroleum hydrocarbons (TPH)) and a reference soil for 28 d. Control treatments with manual mixing and/or cattle dung amendment were also included. In the oil-contaminated soil, respiration and concentration of microbial biomass was significantly enhanced by earthworm amendment, and TPH concentrations decreased significantly. These effects were less evident in treatments with A. chlorotica, possibly due to a difference in behavior, since individuals of this endogeic species were found in a state of inactivity (aestivation). Microbial community dynamics were described by phospholipid fatty acid (PLFA) analyses. After 28 d, similar shifts in the soil PLFA composition were observed in the oil-contaminated soil irrespective of worm species. Fungal:bacterial ratios were increased in the presence of worms, but also by addition of dung as a food source, indicating a non-specific effect of metabolizable substrates. In contrast, the fatty acids 17:1ω8 (=Δ9-heptadecenoic acid) and 20:4ω6c (arachidonic acid) were specifically stimulated by the presence of earthworms in the oil-contaminated soil. The results showed that earthworms can contribute positively to bioremediation of oil-contaminated soil, but that the effect may be species-dependent.     

Changes in soil aggregation and glomalin-related soil protein content as affected by the arbuscular mycorrhizal fungal species Glomus mosseae and Glomus intraradices

Arbuscular mycorrhizal (AM) fungi are key organisms of the soil/plant system, influencing soil fertility and plant nutrition, and contributing to soil aggregation and soil structure stability by the combined action of extraradical hyphae and of an insoluble, hydrophobic proteinaceous substance named glomalin-related soil protein (GRSP). Since the GRSP extraction procedures have recently revealed problems related to co-extracting substances, the relationship between GRSP and AM fungi still remains to be verified. In this work the hypothesis that GRSP concentration is positively correlated with the occurrence of AM fungi was tested by using Medicago sativa plants inoculated with different isolates of Glomus mosseae and Glomus intraradices in a microcosm experiment. Our results show that (i) mycorrhizal establishment produced an increase in GRSP concentration - compared to initial values - in contrast with non-mycorrhizal plants, which did not produce any change; (ii) aggregate stability, evaluated as mean weight diameter (MWD) of macroaggregates of 1-2 mm diameter, was significantly higher in mycorrhizal soils compared to non-mycorrhizal soil; (iii) GRSP concentration and soil aggregate stability were positively correlated with mycorrhizal root volume and weakly correlated with total root volume; (iv) MWD values of soil aggregates were positively correlated with values of total hyphal length and hyphal density of the AM fungi utilized.The different ability of AM fungal isolates to affect GRSP concentration and to form extensive and dense mycelial networks, which may directly affect soil aggregates stability by hyphal enmeshment of soil particles, suggests the possibility of selecting the most efficient isolates to be utilized for soil quality improvement and land restoration programs.     

Survival of Coniothyrium minitans associated with sclerotia of Sclerotinia sclerotiorum in soil

The development and survival of the mycoparasite Coniothyrium minitans associated with sclerotia of the plant pathogen Sclerotinia sclerotiorum was studied in pasteurised and non-sterile (untreated) soil. Using scanning electron microscopy, developing pycnidia were first seen within the sclerotial medulla at 7 days post-inoculation with the mycoparasite in pasteurised soil. However, by 14 days post-inoculation, pycnidia had developed fully in both pasteurised and non-pasteurised treatments, and conidial droplets were exuded onto the outer surface of the infected sclerotia. Thirty days post-inoculation, irrespective of soil treatment, the majority of the sclerotial medulla had been converted to pycnidia, with the sclerotial rind remaining largely intact. The pycnidia and dried intact droplets were still observed 6 months post-inoculation with C. minitans, although the conidia on the outer surface of the dried droplets had largely collapsed by this stage. Germinability studies at 10 months post-inoculation showed that approximately 13% of the conidia in dried droplets were still viable. This work shows the potential for infected sclerotia of S. sclerotiorum to provide a unique reservoir for the survival of C. minitans.     

Metabolism of fluoroacetate by a soil Pseudomonas SP. and Fusarium solani

Enzymes (haloacetate halidohydrolases) capable of cleaving the C-F bond of fluoroacetate and some other organofluorine compounds have been isolated and partially purified from a soil pseudomonad and from the common soil fungus Fusarium solani. Both enzymes readily released F^? from monofluoroacetate and fluoroacetamide but were without effect on a wide range of other organic F-compounds. The enzymes also cleaved the C-Cl and C-Br bonds in mono-chloroacetate and monobromoacetate. Inorganic F^? acted as a competitive inhibitor of the enzymes. The molecular weights of both enzymes were about 62,000. Thus the properties of the halidohydrolases from both organisms were similar in many respects but the bacterial enzyme was more stable at 55°C and exhibited an unusual difference in temperature coefficient (Q₁₀ value) over its higher (30–55°C) and lower (15–30°C) temperature ranges.     

Temporal shifts in diversity and quantity of nirS and nirK in a rice paddy field soil

Denitrification is an important part of the nitrogen cycle in the environment, and diverse bacteria, archaea, and fungi are known to have denitrifying ability. Rice paddy field soils have been known to have strong denitrifying activity, but the microbes responsible for denitrification in rice paddy field soils are not well known. Present study analyzed the diversity and quantity of the nitrite reductase genes (nirS and nirK) in a rice paddy field soil, sampled four times in one rice-growing season. Clone library analyses suggested that the denitrifier community composition varied over sampling time. Although many clones were distantly related to the known NirS or NirK, some clones were related to the NirS from Burkholderiales and Rhodocyclales bacteria, and some were related to the NirK from Rhizobiales bacteria. These denitrifiers may play an important role in denitrification in the rice paddy field soil. The quantitative PCR results showed that nirK was more abundant than nirS in all soil samples, but the nirK/nirS ratio decreased after water logging. These results suggest that both diversity and quantity changed over time in the rice paddy field soil, in response to the soil condition.     

Survival of sclerotia of sclerotinia sclerotiorum in soil

Factors affecting longevity of sclerotia in sandy clay loam (s.c.l.) and sandy loam (s.l.) were examined, using sclerotia from a laboratory culture of S. sclerotiorum and from natural infestations on beans and lettuce.Survival of sclerotia from culture and lettuce was compared in s.l. Recovery and viability were less, and incidence of Fusarium, Mucor and Trichoderma spp. greater, in sclerotia from lettuce than from culture. Rinds of sclerotia from lettuce were more perforated than those from culture.Burial of sclerotia at 4 cm for 35 weeks reduced recovery of sclerotia to zero in s.c.l. and by 50% in s.l. At the soil surface recovery was reduced by 55% in s.c.l. and by 10% in s.l. Less than 50% of sclerotia recovered were viable. Neither a chloropicrin-methyl bromide fumigant nor a tomato compost treatment affected recovery or viability. Fumigation increased incidence of Trichoderma spp. and decreased incidence of Fusurium and Mucor spp. isolated from sclerotia.Apothecia were produced over 6 weeks in s.c.l. and over 20 weeks in s.l. Production was increased by the low rate of fumigant in s.c.l. and by tomato compost in s.l.     

Feeding activity of the earthworm Eisenia andrei in artificial soil

Quantitative information on the feeding activity of earthworms is scarce but this information is valuable in many eco(toxico)logical studies. In this study, the feeding activity of the compost worm Eisenia andrei is examined in artificial soil (OECD medium), with and without a high-quality food source (cow manure), and at two temperatures (10 and 20 °C). Methods are provided to estimate the most important parameters: gut load, selection of organic matter (OM), digestion efficiency, compaction, gut retention time, and fraction of manure in the diet. Lanthanides (Lu and Tm) were successfully used as inert markers in soil and manure, and we applied Bayesian statistics to analyse the data and fully capture the compounded uncertainty in the parameter estimates. Results show that the compost worm does not feed on soil indiscriminately but is able to select an OM-enriched diet from apparently homogeneous OECD medium. When manure is present on the soil surface, approximately three-quarters of the diet still consists of soil particles. The gut load of the worms was approximately 10% (dwt gut/wwt empty worm), varying little with the treatments. Unfortunately, the digestion efficiency could only be reliably estimated at 20 °C, and was approximately 40%. Temperature clearly affected feeding as a 10° temperature decrease nearly doubled the gut retention time (from 2.9 to 5.5 h), which corresponds to a two-fold decrease in feeding rate. The present data may be used to interpret toxicity and accumulation studies with E. andrei in OECD medium. However, care must be taken, as it seems possible that feeding is influenced by the size of the worm and subtle differences in experimental set-up.