Spatial distribution of spiders and epedaphic Collembola in an environmentally heterogeneous forest floor habitat

Describing the biotic and abiotic processes that are responsible for the formation of spatial patterns in predators and their prey is crucial for improving our understanding of food–web interactions. We studied the spatial distribution of four abundant spider species and three common groups of epedaphic Collembola prey in a beech-dominated (Fagus sylvatica) forest floor habitat and related the observed patterns to environmental heterogeneity, overall predator activity (all ground beetles and spiders) and prey availability (all Collembola) at the local scale. Spiders and epedaphic Collembola were sampled over 392 days in a spatially explicit design based on a regular grid of 25 pitfall traps (inter-trap distance 100 m). Environmental heterogeneity was characterized by cover of moss and litter as well as the amount of dead wood at each trap location. We first used the index of dispersion to characterize the spatial distribution of spider species and Collembola and then related the observed patterns to environmental heterogeneity, predator and prey availability while testing for spatial autocorrelation within the same models. All taxa were significantly more aggregated than expected from the assumption of random distribution. The distribution of spider species was positively (Coelotes terrestris) or negatively (Tenuiphantes zimmermanni and Tapinocyba insecta) related to the cover of moss and negatively related to litter cover (C. terrestris) or the local availability of prey (T. insecta). The distribution of Collembola was negatively related to local litter cover (Lepidocyrtus spp.) and positively related to the amount of medium deadwood pieces (all other Entomobryidae). Our study suggests that none of the spider species preferred areas of low overall predator activity density. Moreover, it does not indicate association of spider species to prey-rich areas at the analyzed scale of 100 m. It further highlights the importance of environmental heterogeneity, as different habitat properties differentially affected the local activity density of spiders and Collembola and thus considerably contributed to the understanding of distribution patterns.     

Cereal aphid control by generalist predators in presence of belowground alternative prey: Complementary predation as affected by prey density

Generalist predators are important antagonists of pest species in agroecosystems. Increasing populations of alternative prey through detrital subsidies is one way to foster these predators. However, alternative prey may also distract generalist predators from their prey thereby diminishing the efficiency of biological control. To develop reliable predictions for biological control, it is essential to evaluate the relative importance of generalist predators, pests, alternative prey and their respective interactions. We investigated the effects of an assemblage of generalist predators on the grain aphid Sitobion avenae in winter wheat. Treatments with 10, 100 and 1000 aphids were established inside 2 m² sized caged plots with control and reduced density of predators. Three weeks after the experiment started, samples were taken to estimate the size of aphid populations and those of alternative prey. Three prey taxa were significantly reduced by generalist predators: the grain aphid S. avenae (Aphidina), the click beetle Adrastus pallens (Elateridae) and the springtail Isotoma viridis (Collembola). Springtails were decreased by generalist predators independent of aphid densities, indicating complementary predation of springtails and aphids. At high aphid densities, grain aphid population peaks were decreased to the threshold level of economic damage, demonstrating efficient aphid suppression by the predator community. Click beetle numbers declined only at low and medium aphid densities. The results suggest that generalist predators preferentially fed on click beetles at low and medium aphid densities and switched to aphids at high aphid densities. Early-season predators likely had the greatest influence on aphid suppression. Our results indicate that alternative prey from the belowground system forms a substantial food resource for generalist predators, suggesting that the belowground subsystem modulates predator–prey interactions above the ground.     

Earthworm primers for DNA-based gut content analysis and their cross-reactivity in a multi-species system

Specific primers can be used in polymerase chain reactions (PCR) to amplify prey DNA from the gut content of generalist predators at high specificity and sensitivity. A prerequisite for applying this approach to field studies, however, is to confirm that primers are actually targeting specific prey species or prey groups and do not produce false positive results by amplifying DNA either from predator species or from the wide range of potential alternative prey found under natural conditions. Here, we report on a new group-specific primer pair for earthworms designed from cytochrome oxidase c subunit 1 (COI) sequences of 11 earthworm species found in Central Europe that can be used to detect consumption of earthworms by invertebrate predators. Besides inter-specific also considerable intra-specific variation was found for COI sequences among most of the earthworm species. We, therefore, combined a universal forward primer with an earthworm-specific reverse primer which amplified a 523 bp product from all 11 species tested. Earthworm DNA amplification was also successful in the presence of excess DNA of a predator species. The primer pair was tested against 82 non-target invertebrate species commonly found in the same habitats, including potential prey for generalist predators and predators themselves. The earthworm primer was highly specific: only one of the non-target species showed a product of similar length as the earthworms, whereas PCR with 12 non-target species produced amplicons whose length differed from that of earthworms. We conclude that the new primer will be a useful tool to investigate the role earthworms play as a food resource in soil food-webs. Moreover, we suggest that future studies utilizing DNA-based approaches for prey detection should select non-target species for cross-reactivity tests according to their abundance and importance rather than choosing representatives of taxonomic units; this will help validate the results achieved using species- or group-specific primers and guarantee their meaningful ecological interpretation.     

Prey detection through olfaction by the soil-dwelling larvae of the carabid predator Pterostichus melanarius

Immunological and behavioural studies have shown that adult carabids such as Pterostichus melanarius (Illiger) are predators of slugs. However, little is known about the trophic interactions and spatial ecological of carabid larvae in the soil. Having demonstrated that the larvae will readily consume slugs in the laboratory and can reduce slug numbers in semi-field soil microcosms, experiments were designed to test the hypothesis that the larvae respond to chemical cues from their prey. Electroantennograms (EAG) showed that P. melanarius larvae were able to detect the presence of a range of volatiles using olfactory cues. There were significant EAG responses to both live and dead slugs (Derocerus reticulatum) and Diptera larvae (Calliphora sp.). Dead prey did not induce a response that was significantly different from that to live prey. Behavioural bioassays demonstrated attraction to odours from both live and dead slugs beneath the soil, but when offered a choice there was (after 6 h) a significantly greater attraction to live slugs. These results demonstrate for the first time that P. melanarius larvae appear to use olfactory cues to locate their subterranean prey.     

Amplification facilitators and multiplex PCR: Tools to overcome PCR-inhibition in DNA-gut-content analysis of soil-living invertebrates

Polymerase chain reaction (PCR) can be used to detect prey within the gut contents of predators and allows specific trophic interactions to be studied among soil-dwelling invertebrates which cannot be examined by other approaches. PCR-inhibitory substances, however, are commonly found in DNA prepared from soil organisms or from biological material contaminated with soil. This can lead to false-negative results and the risk of not detecting trophic connections or of underestimating predation rates in field studies. In the present study, we developed mitochondrial DNA markers to detect Amphimallon solstitiale (Coleoptera: Scarabaeidae) in the gut contents of invertebrate predators. Larvae of A. solstitiale can cause serious damage in grasslands, field crops, and forests by feeding on roots. Adequate methodologies to study predation on these pests are lacking, and their invertebrate predator guild is, therefore, barely known. To test the new molecular markers for prey detection, larvae and eggs of A. solstitiale were fed to Poecilus versicolor larvae (Coleoptera: Carabidae), which are abundant below-ground predators in grassland ecosystems. Unfortunately, even when specific DNA extraction and purification methods were used, DNA extracts from predators were of poor quality and not amplifiable by PCR; this yielded false-negative results and a dramatically lower prey-detection rate. We overcame PCR-inhibition by applying ?1.28 μg μl⁻¹ bovine serum albumin to the PCR reaction mix. This enabled us to detect A. solstitiale DNA within fed carabid larvae up to 48 and 40 h post-feeding for 127 and 463 bp sized DNA fragments, respectively. When single A. solstitiale eggs were consumed by the carabid larvae, predation could be verified in 100% of the predators within the first 8 h of digestion; some carabid larvae even tested positive 32 h after feeding. Moreover, by multiplexing primers targeting both prey and predator, we were able to simultaneously screen for prey consumption and check for co-purified PCR inhibitors. Sensitivity in prey detection was not reduced compared to singleplex PCR. We recommend the multiplex approach because it considerably reduces time and costs compared to singleplex assays. We also show that multiplex PCR not only detects specific prey, but also can identify the predator itself. This allows the identification of taxa which are difficult or not identifiable based on morphological characters, such as soil-dwelling predatory beetle larvae.     

Biogeophysical factors influencing soil respiration and mineral nitrogen content in an old field soil

Microbivorous grazers are thought to enhance nutrient mineralization. The predicted effect of microbivory on nutrient cycling depends on the pore habitat model used. We evaluated CO₂ evolution and mineral N content of an old field soil to test two alternative habitat hypotheses. The exclusion hypothesis predicts that nematodes are separated from their microbial food resources in water-filled pores when soils dry, resulting in slower rates of biogeochemical transformations. The enclosure hypothesis predicts that nematode densities increase relative to their forage in smaller, isolated water volumes when soils dry, accelerating rates of biogeochemical transformations. We investigated the effect of soil moisture on the relationship between microbial biomass, microbivorous and predaceous nematodes, soil respiration and mineral N concentrations in an old field five times during the course of a year.We could evaluate the validity of the two habitat hypotheses for the entire field only in August 1997 because that was the only sampling date when maximum water-filled pore diameters were smaller than microbivorous nematode body diameters in all sampled field locations. The mean microbivorous and predaceous nematode abundances for the field in August were greater than 6300 kg⁻¹ and 80,000 kg⁻¹, respectively. Accordingly, the exclusion hypothesis was rejected. Predaceous nematode abundance was markedly higher in August than at any other sampling date. The high abundance of predators present suggests that detrital resources were not limiting productivity and that predators and microbivores were in enclosures, allowing predators to efficiently access their prey. Spatial maps, in agreement with linear correlation analyses, suggest that under our driest sampling conditions, soil respiration and mineral N content were controlled by microbivory and predation.     

Molecular detection of nematode predation and scavenging in oribatid mites: Laboratory and field experiments

Recent stable isotope analyses indicate that a number of putative detritivorous soil microarthropods is not typical detritivores but rather live as predators or scavengers. Using molecular gut content analyses the present study investigates if nematodes indeed form part of the diet of oribatid mites. First, in a no-choice laboratory feeding experiment two nematode species (Phasmarhabditis hermaphrodita and Steinernema feltiae) were offered to eight species of oribatid mites and one gamasid mite. Second, after feeding for 4 and 48 h on each nematode species the detection time of prey DNA in the oribatid mite species Steganacarus magnus was investigated. Third, in a field experiment nematode prey (P. hermaphrodita and S. feltiae) in the diet of microarthropods was investigated distinguishing between scavenging and predation. In the no-choice laboratory experiment not only the gamasid mite but also several of the studied oribatid mite species consumed nematodes. After feeding on nematodes for 4 h prey DNA was detectable in S. magnus for only 4 h, but after feeding for 48 h prey DNA was detectable for 128 h, indicating that the duration of feeding on prey is an important determinant for prey DNA detection. The field experiment confirmed that oribatid mite species including Liacarus subterraneus, Platynothrus peltifer and S. magnus intensively prey on nematodes. Interestingly, DNA of dead P. hermaphrodita was detectable to a similar degree as that of living individuals indicating that scavenging is of significant importance in decomposer food webs. Results of our study indicate that predation and scavenging on nematodes by “detritivorous” microarthropods in soil food webs need to be reconsidered.     

Numerical and behavioral responses of waterfowl to the invasive American mink: A conservation paradox

A behavioral response is usually the first adaptation of animals to environmental changes such as the introduction of an alien invasive species, and it seems to be the most important factor in reducing the negative impact of new predators on naive prey. This study examined changes in the abundance and behavior of breeding coots (Fulica atra) and great crested grebes (Podiceps cristatus) in response to invasion of the Mazurian Lakeland (NE Poland) by American mink (Neovison vison), first recorded here in 1984. Feral mink densities varied from 1.0 to 14.6 individuals per 10 km of shoreline. Mink were responsible for the losses of 17–58% of experimental clutches (artificial nests containing chicken eggs) that were deployed. Mink destroyed more experimental clutches than other predators; the effects of mink predation were similar in reeds and on the bank. Radio-tracking showed that mink avoided lake banks near human settlements. Mink have markedly affected the breeding distribution of both species and caused a greater than 20-fold decline in coot numbers. For coots and grebes breeding distribution and nesting success were positively associated with built-up areas along lake shores and also with whether the birds were breeding in colonies. Increased brood survival of coots and grebes near human settlements is a paradox in the sense that a man-made habitat change has provided refuge from an alien predator.     

Temporal variations of soil arthropods community structure in El Pedregal de San Ángel Ecological Reserve,Mexico City,Mexico

Community structure of soil arthropods associated to Pittocaulon (Senecio) praecox was investigated in two sites with different vegetal cover in El Pedregal de San Ángel Ecological Reserve, South of Mexico City during thirteen-month period. The relationship of composition and abundance with soil variables was studied. A total of 101 483 specimens belonging to 28 taxa and 191 morphospecies were collected in both sites (Open site: Abundance: 60,369, 27 taxa and 181 morphospecies; Closed site: 41 114, 26 taxa and 179 morphospecies), the value of Shannon's diversity index was marginally higher in Closed site (H′ = 3.67) than in Open site (H′ = 3.59), according to a modified t-test for comparing indexes (t₉₁₄₁₈ = 9.6946; p < 0.001). Most abundant groups in both sites were Cryptostigmata and Prostigmata, with variations during the study period. The highest amount of specimens was recovered during the rainy season (May–October). The most abundant guilds recorded in both sites were predators, fungivores, detritivores and herbivores. The composition and trophic guilds of edaphic arthropods in the sites were affected by the microhabitat characteristics due to soil characteristics. According to a two-way ANOVA test, a significant effect due to site (F_1,286 = 17.85, p < 0.001) and month (F_12,286 = 22.68, p < 0.001) on the density of edaphic arthropods was recorded, but no significant differences in abundance were found when the interaction between both variables (F_12,286 = 0.77, p > 0.05) was considered. The RDA shows that the soil variables explain 23.5% of the variation observed in the distribution of species, and that variation is related mainly to the presence of K, P, Na and Mg, and groups of predator arthropods are more related to the aforementioned variables, while detritivorous groups are more related to MO, N contents, relative humidity and pH. The microenvironmental characteristics recorded in both sites can explain differences in composition and abundance of soil arthropods.     

Habitat use and activity patterns of larval and adult Cantharis beetles in arable land

Generalist predators play a key role in agriculturally and environmentally sustainable systems of pest control. A detailed knowledge on their ecology, however, is needed to improve management practices to maximize their service of pest control. The present study examines the habitat use and activity patterns of larval and adult Cantharis beetles that are abundant predators in arable land. Laboratory experiments revealed that sixth instar larvae of Cantharis fusca and Cantharis livida significantly preferred high relative humidity levels of 85–90% to lower ones. This can explain their preference for meadows over fields due to the more favorable microclimatic conditions in the former habitats. Surface activity of sixth instar Cantharis larvae during autumn, winter and early spring occurred at soil temperatures above 0 °C. However, no correlation between surface activity and soil temperature, air temperature or relative humidity was found above 0 °C. Catches of sixth instar Cantharis larvae within fenced pitfall traps were higher in a meadow (Mean ± S.D.; 13.8 ± 7.63 individuals m⁻²) than in a field (4.60 ± 2.89 individuals m⁻²). Mark-recapture density estimations for sixth instar larvae indicated mean densities of 25.9 ± 5.63 (field) and 42.8 ± 16.0 individuals m⁻² (meadow). The same pattern was found for adult emergence rates in the field (0.17 ± 0.39 adults m⁻²) and meadow (1.83 ± 1.17 adults m⁻²) as well as for adult densities in the vegetation (field 4.89 ± 3.62 adults 60 m⁻²; meadow 12.5 ± 11.2 adults 60 m⁻²). It is concluded that especially in winter elements that provide plant cover should be incorporated in arable fields to enhance larval cantharid population densities and to attract them from their prime grassland habitats into arable sites.     

Quantifying potato pathogen DNA in soil

An improved method for the direct extraction of DNA from soil involving processing of a relatively large sample (60 g) was developed. The accurate and reliable detection and quantification of the soil-borne potato pathogens Colletotrichum coccodes (black dot), Rhizoctonia solani (black scurf) and Spongospora subterranea (powdery scab) following inoculation of soils was demonstrated. With this method, low levels of target DNA (30–40 pg DNA/g soil) could be detected in field soils. DNA recovery was proportionate across a wide range of inoculum (R² > 0.86) and there was no effect of soil type on the recovery of C. coccodes. The method was used to assess levels of naturally occurring pathogen DNA in 122 soil samples obtained from commercial potato fields.     

The effect of organic matter on the structure of soils of different land uses

In this study, the soil structure of two soils (Haplic Chernozem and Eutric Fluvisol) of different land uses (forest, meadow, urban and agro-ecosystem – consisted of four crop rotations) in Slovakia was compared. The soil aggregate stability was determined with a dependence on the chemical composition of plant residues. The quantity and quality of the organic matter was assessed through the parameters of the C and N in size fractions of dry-sieved and water-resistant aggregates. The soil structure of the forest ecosystem was evaluated as the best of all of forms of land use. Differences in the soil structure under the grass vegetation of a meadow (natural conditions) and urban ecosystem were also recorded. The agro-ecosystem was characterised by a higher portion (55.95%) of the most valuable (agronomically) water-resistant aggregate size fraction of 0.5–3 mm. Values of the carbon management index showed that the larger water-resistant aggregates were, the greater were the changes in the organic matter (r = ?0.680, P < 0.05). In addition, a smaller content of dry-sieved aggregates of the 3–5 mm size fraction was observed with higher contents of soil organic carbon (SOC) (r = ?0.728, P < 0.05) and labile carbon (C_L) (r = ?0.760, P < 0.05); there were also greater changes in the soil organic matter and vice versa, higher contents of SOC (r = 0.744, P < 0.05) and C_L (r = 0.806, P < 0.05) greater contents of dry-sieved aggregates of size fraction 0.5–1 mm. The soil structure of agro-ecosystem was superior at a higher content of cellulose (r = ?0.712, P < 0.05) in the plant residues. The higher content of cellulose and hemicellulose in the plant residue of a previous crop was reflected in a smaller C_L content in the water-resistant aggregates (r = ?0.984, P < 0.05). A correlation was observed between a high content of lignin in the plant residue and a smaller SOC content in the water-resistant aggregates (r = ?0.967, P < 0.05). Lastly, a higher content of proteins in the plant residues (r = 0.744, P < 0.05) supported a greater content of dry-sieved aggregates of the 0.5–1 mm size fraction.     

Are enchytraeid worms (Oligochaeta) sensitive indicators of ammonia-N impacts on an ombrotrophic bog?

Enchytraeid worms (Oligochaeta) are the dominant mesofauna in wet acidic habitats. They have key roles in biogeochemical cycling, and can be used as biological indicators. Here we report the response of these worms to in situ ammonia-N (NH₃-N) deposition on an ombrotrophic bog. Three years of NH₃-N fumigation from an automated release system has created a gradient of NH₃-N concentrations downwind of the release pipe ranging from 83 μg m⁻³ (near source) to 4.5 μg m⁻³ NH₃-N (60 m from release pipe); the ambient NH₃-N concentration is 0.56 μg m⁻³ NH₃-N. Peat pH and mineral N content have increased near the ammonia release pipe. We hypothesised that enhanced N deposition at the site would have improved litter quality and thus, enchytraeid distribution would be increased along the transect compared to ambient. However, neither litter quality nor enchytraeid abundance and diversity were affected by NH₃-N despite increases in peat pH and mineral N. This suggests that three years of ammonia fumigation was not enough time for plant matter exposed to ammonia to become incorporated into the peat litter layer. Enchytraeids appear not to be sensitive indicators of NH₃ fumigation because there was no effect below-ground of peat chemistry on litter quality.     

Stable isotope labelling of earthworms can help deciphering belowground–aboveground interactions involving earthworms,mycorrhizal fungi,plants and aphids

Functional relationships between belowground detritivores and/or symbionts and aboveground primary producers and their herbivores are not well studied. In a factorial greenhouse experiment we studied interactions between earthworms (addition/no addition of Lumbricus terrestris; Clitellata: Lumbricidae) and arbuscular-mycorrhizal fungi (AMF; with/without inoculation of Glomus mosseae; Glomerales: Glomeraceae) on the leguminous herb Trifolium repens (Fabales: Fabaceae) and associated plant aphids (Aphis gossypii, A. craccivora; Hemiptera: Aphidoidea). In order to be able to trace organismic interactions, earthworms were dual-labelled with stable isotopes (¹⁵N-ammonium nitrate and ¹³C-glucose). We specifically wanted to investigate whether (i) isotopic signals can be traced from the labelled earthworms via surface castings, plant roots and leaves to plant aphids and (ii) these compartments differ in their incorporation of stable isotopes. Our results show that the tested organismic compartments differed significantly in their ¹⁵N isotope enrichments measured seven days after the introduction of earthworms. ¹⁵N isotope incorporation was highest in casts followed by earthworm tissue, roots and leaves, with lowest ¹⁵N signature in aphids. The ¹³C signal in roots, leaves and aphids was similar across all treatments and is for this reason not recommendable for tracing short-term interactions over multitrophic levels. AMF symbiosis affected stable isotope incorporation differently in different subsystems: the ¹⁵N isotope signature was higher below ground (in roots) but lower above ground (leaves and aphids) in AMF-inoculated mesocosms compared to AMF-free mesocosms (significant subsystem × AMF interaction). Aphid infestation was unaffected by AMF and/or earthworms. Generally, these results demonstrate that plants utilize nutrients excreted by earthworms and incorporate these nutrients into their roots, leaf tissue and phloem sap from where aphids suck. Hence, these results show that earthworms and plant aphids are functionally interlinked. Further, ¹⁵N-labelling earthworms may represent a promising tool to investigate nutrient uptake by plants and consequences for belowground-aboveground multitrophic interactions.     

Beetles community structures under different reclamation practices

Differences in beetle community structures between not re-cultivated over 60 years old forest and 10 years old re-cultivated meadow and shrubs were compared. On each locality there were arranged three replicates of 10 pitfall trap rows. Mean abundance of beetles was significantly greater on old forest stand than re-cultivated meadow. However no significant differences between localities in species richness and non parametric index of diversity (Shannon H′) were recorded. Rank–abundance curves for newly established communities best fits to geometric model of distribution (r = 0.98, P < 0.001) which is characteristic for species poor communities, where a single environmental resource is extremely important, meanwhile old stand, without any reclamation practice is best fitted to lognormal distribution model (r = 0.99, P < 0.001) which is typical for more stable and well developed communities. Principal component analysis for log (N + 1) transformed matrix of abundances described well 60% of the total variance. Two clear groups of species transects were derived: one concentrated assemblages of both reclaimed and newly established communities and the second one old non reclaimed forest stands. Significant feature of trophic guild structure in all three communities is the lowest abundance of decomposers meanwhile predators predominate on non re-cultivated old sites and herbivores on reclaimed young stands.     

Nitrogen mineralization in the ruderal sub-alpine communities in Mount Uludağ, Turkey

Nitrogen mineralization and nitrification in the soil of sub-alpine ruderal community of Mount Uludağ, Bursa, Turkey was measured for 1 year, under field conditions with Verbascum olympicum and Rumex olympicus being the dominant pioneer species under dry and wet sites, respectively. Seasonal fluctuations were observed in N mineralization and nitrification. The net N mineralization and nitrification were high in early summer and winter, due to high moisture. The annual net N mineralization rate (for the 0–15 cm soil layer) was higher under R. olympicus (188 kg N ha⁻¹ yr⁻¹) than under V. olympicum (96 kg N ha⁻¹ yr⁻¹). A significant positive correlation between net N mineralization and soil organic C (r² = 0.166), total N (r² = 0.141) and water content (r² = 0.211) was found. Our results indicate that N mineralization rate is high in soils of ruderal communities on disturbed sites and varies with dominant species and, a difference in net N mineralization rate can be attributed to organic C, total N and moisture content of soils.     

Detection and quantification of the nematophagous fungus Hirsutella minnesotensis in soil with real-time PCR

A real-time PCR assay was developed to quantify in soil the fungus Hirsutella minnesotensis, an important parasite of secondary-stage juvenile (J2) of the soybean cyst nematode. A primer pair 5′-GGGAGGCCCGGTGGA-3′ and 5′-TGATCCGAGGTCAACTTCTGAA-3′ and a TaqMan probe 5′-CGTCCGCCGTAAAACGCCCAAC-3′ were designed based on the sequence of the ITS region of the rRNA gene. The primers were highly species-specific. The PCR reaction system was very sensitive and able to detect as few as 4 conidia g^?1 soil. Regression analysis showed similar slopes and efficiency on DNA from pure culture (y = ?3.587x + 41.017, R² = 0.9971, E = 0.9055) and from Log conidia g^?1 soil (y = ?3.855x + 37.669, R² = 0.9139, E = 0.8172), indicating that the real-time PCR protocol can reliably quantify H. minnesotensis in the soil. The real-time PCR assay was applied to 20 soil samples from soybean fields, and compared with a parasitism assay. The real-time PCR assay detected H. minnesotensis in six of the soils, whereas the parasitism assay detected H. minnesotensis in the same six soils and three additional soils. The real-time PCR assay was weakly correlated (R² = 0.49) with the percentage of parasitized J2 in the six soils, indicating that different types of soil may interfere the efficiency of the real-time PCR assay, possibly due to the effect of soil types on efficacy of DNA extraction. The parasitism assay appeared to be more sensitive than real-time PCR in detecting presence of H. minnesotensis, but real-time PCR was much faster and less costly and provided a direct assessment of fungal biomass. Using the two assays in combination can obtain more complete information about the fungus in soil than either assay alone. Hirsutella parasitism was widespread and detected in 13 of the 20 field soils, indicating that these fungi may contribute to suppressiveness of soybean cyst nematode in nature and likely have high biological control potential for the nematode.     

Role of soil water content in the carbon and nitrogen dynamics of Lumbricus terrestris L. burrow soil

We evaluated the role of soil water content in controlling C and N dynamics within the drilosphere created by the anecic earthworm Lumbricus terrestris (L.). Mesocosms (volume = 3.1 l) were each amended with corn litter and three earthworms. Control treatments received no earthworms and no other earthworm species were present in the soil. WET and DRY treatments received a total of 9.25 cm and 3.25 cm of water, respectively. Water was added on weeks 1, 3, 7, and 10 at a rate of 2.0 cm per mesocosm for WET treatments and 0.5 cm per mesocosm for DRY treatments. Mesocosms were sampled destructively after incubation at 18–20 °C for 0, 3, 7, and 13 weeks. The water content of WET burrow soil ranged from 0.12 g g⁻¹ to 0.18 g g⁻¹ and was significantly higher than in the DRY treatment throughout the incubation period. The live weight of earthworms was significantly higher in the WET treatment only on week 13, whereas litter consumption was significantly lower in the DRY treatment for week 13. Carbon mineralization, measured as CO₂ evolved after a 24-h incubation, was consistently higher in WET than in DRY burrow soil. Effects of differences in soil water content were also apparent for biomass C and metabolic quotient. Soil water content did no affect the total C concentration of burrow soil. DRY burrow soil had consistently lower levels of nitrate than WET soil throughout the experiment. Lower levels of ammonium and inorganic N were observed for WET burrow soil on weeks 3 and 7. Water content did not have a significant effect on burrow soil total N. We concluded that the water content of the drilosphere affects both C and N dynamics and can affect the speciation of inorganic N; yet, the effects of soil water content do not appear to result from differences in the feeding activities of anecic earthworms.     

An outline of soil nematode succession on abandoned fields in South Bohemia

Secondary succession of nematodes was studied in 1–48-year-old abandoned fields on cambisols in South Bohemia, Czech Republic, and compared with cultivated field and sub-climax oak forests. Bacterivores were the predominant group in the cultivated field whereas in forests root-fungal feeders (mainly Filenchus) were almost as abundant as bacterivores. The total abundance of nematodes in the cultivated field averaged 868 × 10³ ind m⁻². During the first three years of succession the abundance practically did not change (775 × 10³ ind m⁻²), the fauna was still similar to that in cultivated field but the biomass increased mainly due to Aporcelaimellus. Then the abundance increased up to 3731 × 10³ ind m⁻² in 7–8-year-old abandoned fields, plant parasites (Helicotylenchus) dominated and the fungal-based decomposition channel was activated. Later the abundance stabilised at between 1086 and 1478 × 10³ ind m⁻² in 13–25-year-old successional meadow stages with high population densities of omnivores and predators. The total abundance of nematodes was low in the 12–13-year-old willow shrub stage (594 × 10³ ind m⁻²), increased in the 35–48-year-old birch shrub stage (1760 × 10³ ind m⁻²) and the nematode fauna developed towards a forest community. The diversity and maturity of nematode communities generally increased with the age of abandoned fields but the highest values were in meadow stages (81–113 species, 57–68 genera, MI 2.73–3.30). The development of meadow arrested succession towards forests or diverted succession towards a waterlogged ecosystem. The succession of nematodes was influenced by the method of field abandonment (bare soil vs. legume cover, mowing) that affected the formation of either a shrub or meadow stage, and by the soil water status. The composition of the nematode fauna indicated that the soil food web could recover faster from agricultural disturbance under successive meadows than under shrubs.     

Physiological studies and comparative analysis of rock phosphate solubilization abilities of Actinomycetales originating from Moroccan phosphate mines and of Streptomyces lividans

Actinomycete strains originating from Moroccan phosphate mines (MPM) were selected for their ability to use the insoluble ground hydroxyapatite called rock phosphate (RP), present in their biotope, as sole phosphate (P) source. Physiological studies carried out with these strains and with the reference strains, Streptomyces lividans and Streptomyces griseus, demonstrated that all strains were able to grown in a synthetic minimal medium (SMM) containing either soluble (SP) or insoluble (RP) phosphate as sole P source. The MPM strains and S. griseus took up glucose much more actively and exhaustively than S. lividans, constituting more abundant glycogen reserves than the latter. All strains took up soluble P at comparable rates, storing it as polyphosphates. In SMM + RP, a sharp increase in the concentration of soluble P was detected in the culture broths of all MPM strains and S. griseus, at stationary phase, but not in that of S. lividans. The P peak detected in the supernatant of these strains correlated with the successive appearance of two compounds absorbing at 320 nm and 430 nm, respectively. These compounds are thought to be strong ion chelators involved in the destruction of the hydroxyapatite structure leading to soluble P release. The good growth of S. lividans in SMM + RP indicated that this strain was also able to release P from RP but consummed it as soon as it was released, unlike the other strains. Our study is expected to lead to the development of a novel type of slow release bio-phosphate fertilizer constituted by the association of the MPM strains and ground RP. This novel product would precisely supply plant needs and thus limit the pollution of the environment.