Comparison of some novel polyculture and traditional monoculture vermicomposting reactors to decompose organic wastes

Efforts were made to evaluate the decomposition potentials of traditional monoculture and some novel polyculture vermireactors. Three earthworm species, i.e. Eisenia fetida (E. f.), Perionyx excavatus (P. ex.) and Lampito mauritii (L. m.), representing two different ecological categories: epigeic (E. fetida and P. excavatus) and anecic (L. mauritii), were used to design seven different vermireactors, i.e. Mono-(E. f.), Mono-(P. ex.), Mono-(L. m.), Poly-(E. f. + P. ex.), Poly-(P. ex. + L. m.), Poly-(E. f. + L. m.) and Poly-(E. f. + P. ex. + L. m.). The microbial load of vermireactors was evaluated through measuring dehydrogenises activities (DH-ase) and microbial biomass-N, while mineralization rate was measured in respect to changed level of some important nutrients in vermicomposted substrate. The vermicomposting caused decrease in pH (67.0–15.0%), organic C (46.1–28.4%) and C:N ratio (72.2–57.1%) and increase in total N (137.7–67.8%) as well as available P (107.9–16.9%) contents, at the end. The carbon and nitrogen mineralization rate showed the order: Poly-(E. f. + P. ex. + L. m.) > Poly-(E. f. + L. m.) > Poly-(P. ex. + L. m.) > Poly-(E. f. + P. ex.) > Mono-(E. f.) > Mono-(P. ex.) > Mono-(L. m.) for this study. The Poly-(E. f. + P. ex. + L. m.) vermireactor showed the maximum level of DH-ase activity 1926 ± 245 μg g⁻¹ substrate 24 h as well as microbial biomass-N 3059.1 ± 242.3 mg N g⁻¹ substrate, during experimentation. This study clearly suggests that burrowing earthworms in vermireactor not only promote the microbial colonization, but at the same time also accelerate the mineralization rate in decomposing waste. The polyculture vermicomposting, using burrowing earthworms with epigeics, could be more efficient than traditional monoculture vermireactors to decompose organic waste resources.     

Bioconversion of filter mud using vermicomposting employing two exotic and one local earthworm species

Three different earthworm species Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus in individual (Monocultures) and combinations (Polycultures) were utilized to compare the suitability of worm species for vermicomposting of filter mud as well as the quality of the end product. The filter mud blended with saw dust can be directly converted into good quality fertilizer (vermicompost). Eight different reactors including three monocultures and four polycultures of E. fetida, E. eugeniae and P. excavatus and one control were used for the experiment. Vermicomposting resulted in significant reduction in C/N ratio, pH, total organic matter (TOC) but increase in electrical conductivity (EC), total nitrogen (TN), total phosphorus (TP) and macronutrients (K, Ca and Na). Oxygen uptake rate (OUR) dropped up to 1.64–1.95 mg/g (volatile solids) VS/day for monoculture reactors and 1.45–1.78 mg/g VS/day for polycultures reactors, respectively, after 45 days of vermicomposting. Cocoon production and the earthworm biomass increased as vermicomposting progressed. On an overall the mono as well as polyculture reactors produced high quality stable compost free from pathogens and no specific differentiation could be inferred between the reactors.     

Plant community structure, soil properties and microbial characteristics in revegetated quarries

Quarries are an important type of degraded land in southern China requiring ecological improvement and rehabilitation. In this study, plant community structure, soil properties, and microbial biomass and community function were examined at different rehabilitated phases in three quarries, namely Turret Hill Quarry, Lam Tei Quarry and Shek O Quarry, in Hong Kong. Results show that plant species richness and the percentage of native species increased with rehabilitated ages in the three quarries. The highest coverage of woody species was found at older phases, while the lowest woody coverage occurred at younger phases. Soils were strongly to moderately acidic in reaction, and more acidic soils were found in the older than in the younger sites. Organic C as well as total N and P accumulated in soil along with secondary succession in the three quarries, which were positively correlated with woody species richness. Older phases had higher total microbial biomass C and N which were positively correlated with soil organic C, total N and extractable NO₃-N, as well as woody species coverage and native species richness as shown by the biplot of redundancy analysis. Diversity of utilized carbons suggested that metabolic abilities developed gradually with rehabilitation ages in Shek O Quarry, but Turret Hill Quarry and Lam Tei Quarry had similar patterns of carbon source utilization. Principal component analysis further revealed consistent differences in metabolic diversity. Woody coverage and native species richness were significantly correlated with carbon source utilization and functional diversity.     

Plants and fertilisers as drivers of change in microbial community structure and function in soils

Here we report on a range of studies designed to understand the link between diversity and function in soils and in particular how plants and fertilisers might interact with microbial community dynamics in soils. The data presented indicate that although plants and fertilisers do impact on microbial community structure, the relationship between diversity, community structure and function remains complex and difficult to interpret using currently available chemical and molecular fingerprinting techniques. The paper assesses plants and management practices as drivers of change in soil and argues that whilst understanding diversity per se is unlikely to contribute to our understanding of function, an appreciation of what causes communities to change and also the relative importance of such drivers, could lead to new insights into the sustainable management and conservation of soils and natural resources.     

Crystal structure of earthworm fibrinolytic enzyme component a: revealing the structural determinants of its dual fibrinolytic activity

Earthworm fibrinolytic enzyme component A (EFEa) from Eisenia fetida is a strong fibrinolytic enzyme that not only directly degrades fibrin, but also activates plasminogen. Proteolytic assays further revealed that it cleaved behind various P1 residue types. The crystal structure of EFEa was determined using the MIR method and refined to 2.3A resolution. The enzyme, showing the overall polypeptide fold of chymotrypsin-like serine proteases, possesses essential S1 specificity determinants characteristic of elastase. However, the beta strand at the west rim of the S1 specificity pocket is significantly elongated by a unique four-residue insertion (Ser-Ser-Gly-Leu) after Val217, which not only provides additional substrate hydrogen binding sites for distal P residues, but also causes extension of the S1 pocket at the south rim. The S2 subsite of the enzyme was partially occluded by the bulky side-chain of residue Tyr99. Structure-based inhibitor modeling demonstrated that EFEa's S1 specificity pocket was preferable for elastase-specific small hydrophobic P1 residues, while its accommodation of long and/or bulky P1 residues was also feasible if enhanced binding of the substrate and induced fit of the S1 pocket were achieved. EFEa is thereby endowed with relatively broad substrate specificity, including the dual fibrinolysis. The presence of Tyr99 at the S2 subsite indicates a preference for P2-Gly, while an induced fit of Tyr99 was also suggested for accommodation of bigger P2 residues. This structure is the first reported for an earthworm fibrinolytic enzyme component and serine protease originating from annelid worms.     

Vermicomposting--An effective tool for the management of invasive weed Parthenium hysterophorus

This study reports the results of vermicomposting with Eisenia fetida of Parthenium hysterophorus mixed with cow dung in different ratios (25%, 50% and 75%) in a 18 weeks experiment. In all the treatments, a decrease in pH, OC_total and C:N ratio, but increase in EC, N_total, P_aval, Ca_total, K_total and heavy metals was recorded. The cocoons production and growth rate (biomass gain worm⁻¹ day⁻¹) were maximum in 100% cow dung. The results indicated that parthenium can be a raw material for vermicomposting if mix with cow dung in appropriate quantity.     

Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment

This study focused on the diversity of bacterial communities from two series of two-stage constructed wetlands (CWs) treating tannery wastewater, under different hydraulic conditions. Series were separately planted with Typha latifolia and Phragmites australis in expanded clay aggregates and operated for 31 months. The effect of plant species, hydraulic loading and unit stage on bacterial communities was addressed through bacterial enumeration and denaturating gradient gel electrophoresis (DGGE). Diverse and distinct bacterial communities were found in each system unit, which was related in part to the type of plant and stage position (first or second unit in the series). Numerical analysis of DGGE profiles showed high diversity in each unit with an even distribution of species. No clear relation was established between the sample collection time, hydraulic loading applied and the bacterial diversity.     

Dynamics of a pig slurry microbial community during anaerobic storage and management

The microbial community of a pig slurry on a farm was monitored for 6 months using both molecular and cultural approaches. Sampling was carried out at all the different stages of effluent handling, from the rearing build-up to slurry spreading. Total DNA of each sample was extracted and analyzed by PCR-single-strand conformation polymorphism (SSCP) analysis using primers targeting the 16S rRNA genes from the archaeal and bacterial domains and also the Eubacterium-Clostridium, Bacillus-Streptococcus-Lactobacillus, and Bacteroides-Prevotella groups. A comparison of the SSCP profiles showed that there were rapid changes in the dominant bacterial community during the first 2 weeks of anaerobic storage and that the community was relatively stable thereafter. Several bacterial populations, identified as populations closely related to uncultured Clostridium and Porphyromonas and to Lactobacillus and Streptococcus cultured species commonly isolated from pig feces, remained present and dominant from the rearing build-up to the time of spreading. Enumeration of fecal indicators (enterococci and Escherichia coli) performed in parallel using cultural methods revealed the same trends. On the other hand, the archaeal community adapted slowly during pig slurry storage, and its diversity increased. A shift between two hydrogenotrophic methanogenic Methanobrevibacter populations from the storage pit to the pond was observed. Microorganisms present in pig slurry at the time of spreading could not be detected in soil after spreading by either molecular or cultural techniques, probably because of the detection limit inherent in the two techniques.     

Phoresy of the entomopathogenic nematode Steinernema feltiae by the earthworm Eisenia fetida

The free-living stage of entomopathogenic nematodes occurs in soil, and is an environmental-friendly alternative for biological control. However, their dispersal capability is limited. Earthworms improve soil characteristics, changing soil structure and influencing many edaphic organisms. Thus, earthworms could be used as vectors to introduce/disperse beneficial organisms. Nevertheless this interaction has not been studied in detail. This study presents the infectivity results of Steinernema feltiae after passing through the Eisenia fetida gut. Although entomopathogenic nematodes have no deleterious effects on earthworms, their passage through E. fetida gut seriously affected their mobility and virulence.     

Vermicomposting of tannery sludge mixed with cattle dung into valuable manure using earthworm Eisenia fetida (Savigny)

The present study revealed the role of earthworm in converting tannery sludge into a valuable product. Tannery sludge was toxic to earthworm, therefore it was mixed with cattle dung in different proportions viz. 0:100 (T₀), 10:90 (T₁₀), 25:75 (T₂₅), 50:50 (T₅₀) and 75:25 (T₇₅) on dry weight basis. The minimum mortality and highest population buildup of worms was in T₀ mixture. Nitrogen, sodium, phosphorus and pH increased from initial in the range of 7.3-66.6%, 16.90-70.58%, 8.57-44.8% and 2.8-13.65%, respectively. On the other hand potassium, organic carbon and electrical conductivity decreased in the range of 4.34-28.5%, 7.54-22.35% and 32.35-53.12%, respectively. C:N ratio decreased from 20.53% to 47.36% in the final products. Transition metals increased significantly from the initial value and within the permissible limit. The result indicated that vermicomposting with Eisenia fetida is better for changing this sludge into nutrient rich manure in a short period of time.     

Vermistabilization of primary sewage sludge

An integrated composting-vermicomposting process has been developed for utilization of primary sewage sludge (PSS). Matured vermicompost was used as bulking material and a source of active microbial culture during aerobic activated composting (AAC). AAC resulted in sufficient enrichment of bulking material with organic matter after 20 cycles of recycling and mixing with PSS and produced materials acceptable for vermicomposting. Vermicomposting caused significant reduction in pH, volatile solids (VS), specific oxygen uptake rate (SOUR), total organic carbon (TOC), C/N ratio and pathogens and substantial increase in electrical conductivity (EC), total nitrogen (TN) and total phosphorous (TP) as compared to compost. Environmental conditions and stocking density have profound effects on vermicomposting. Temperature of 20 °C with high humidity is favorable environmental condition for vermicomposting employing Eisenia fetida. Favorable stocking density range for vermiculture is 0.5-2.0 kg m⁻² (optimum: 0.5 kg m⁻²) and for vermicomposting is 2.0-4.0 kg m⁻² (optimum: 3.0 kg m⁻²), respectively.     

Earthworm effects on gaseous emissions during vermifiltration of pig fresh slurry

Treatment of liquid manure can result in the production of ammonia, nitrous oxide and methane. Earthworms mix and transform nitrogen and carbon without consuming additional energy. The objective of this paper is to analyse whether earthworms modify the emissions of NH₃, N₂O, CH₄ and CO₂ during vermifiltration of pig slurry.The experiment used mesocosms of around 50 L, made from a vermifilter treating the diluted manure of a swine house. Three levels of slurry were added to the mesocosms, with or without earthworms, during one month, in triplicate. Earthworm abundance and gas emissions were measured three and five times, respectively.There was a decrease in emissions of ammonia and nitrous oxide and a sink of methane in treatments with earthworms. We suggest that earthworm abundance can be used as a bioindicator of low energy input, and low greenhouse gas and ammonia output in systems using fresh slurry with water recycling.     

Evaluation of three composting systems for the management of spent coffee grounds

This study was conducted to evaluate the optimum composting approach for the management of spent coffee grounds from the restaurant and ready-to-serve coffee industry. Three composting systems were assessed, including in-vessel composting, vermicomposting bins, and aerated static pile bin composting, over study periods ranging from 47 to 98 days. Total carbon content was reduced by 5-7% in the spent coffee ground treatments across the three composting systems. Nitrogen and other mineral nutrient contents were conserved or enhanced from the initial to the final composts in all the composting systems assessed. Earthworm growth and survival (15-80%) was reduced in all the treatments but mortality rates were lower in coffee treatments with cardboard additions. A decline in earthworm mortality with cardboard additions was the result of reduced exposure to organic compounds and chemicals released through the decomposition of spent coffee grounds.     

From Metchnikoff to Monsanto and beyond: the path of microbial control

In 125 years since Metchnikoff proposed the use of Metarhizium anisopliae to control the wheat cockchafer and brought about the first field trials, microbial control has progressed from the application of naturalists' observations to biotechnology and precision delivery. This review highlights major milestones in its evolution and presents a perspective on its current direction. Fungal pathogens, the most eye-catching agents, dominated the early period, but major mycological control efforts for chinch bugs and citrus pests in the US had questionable success, and interest waned. The discoveries of Bacillus popilliae and Bacillus thuringiensis began the era of practical and commercially viable microbial control. A program to control the Japanese beetle in the US led to the discovery of both B. popilliae and Steinernema glaseri, the first nematode used as a microbial control agent. Viral insect control became practical in the latter half of the 20th century, and the first registration was obtained with the Heliothis nuclear polyhedrosis virus in 1975. Now strategies are shifting for microbial control. While Bt transgenic crops are now planted on millions of hectares, the successes of more narrowly defined microbial control are mainly in small niches. Commercial enthusiasm for traditional microbial control agents has been unsteady in recent years. The prospects of microbial insecticide use on vast areas of major crops are now viewed more realistically. Regulatory constraints, activist resistance, benign and efficacious chemicals, and limited research funding all drive changes in focus. Emphasis is shifting to monitoring, conservation, integration with chemical pesticides, and selection of favorable venues such as organic agriculture and countries that have low costs, mild regulatory climates, modest chemical inputs, and small scale farming.     

Afforestation alters community structure of soil fungi

Relatively little is known about the effect of afforestation on soil fungal communities. This study demonstrated that afforestation altered fungal community structure and that changes were correlated to pools of soil C. Pasture at three locations on the same soil type was afforested with Eucalyptus globulus or Pinus pinaster. The structure of fungal communities under the three land uses was measured after 13y using automated ribosomal intergenic spacer analysis (ARISA). Afforestation significantly altered the structure of fungal communities. The effect of location on the structure of fungal communities was limited to pasture soils; although these contained the same plant species, the relative composition of each species varied between locations. Differences in the structure of fungal communities between pasture, E. globulus and P. pinaster were significantly correlated with changes in the amount of total organic C and microbial biomass-C in soil. Afforestation of patches of agricultural land may contribute to conserving soil fungi in agricultural landscapes by supporting fungal communities with different composition to agricultural soils.     

Enzyme Activities and Chemical Changes in Wet Olive Cake after Treatment with Pleurotus ostreatus or Eisenia fetida

A laboratory experiment was conducted to evaluate the enzyme activities and chemical changes recorded in a recalcitrant phenolic-rich waste after treatment with Pleurotus ostreatus or Eisenia fetida. The waste used was wet olive cake (alperujo in Spanish), a waste produced in huge amounts by the olive oil industry. Both P. ostreatus and E. fetida were very effective in removing phenolic compounds, the initial concentration in the wet olive cake being reduced in both cases by around 90%. Laccase and manganese peroxidase activities were measured in the growth medium of P. ostreatus, and catechol 2,3 dioxygenase activity was only detected in the waste treated with Eisenia; these could be the main factors responsible for the oxidation of phenolic compounds. Increases of dehydrogenase and β-glucosidase activities were detected in the degraded wet olive cake by fungi or earthworms. In comparison with the natural wet olive cake, the degraded products had lower total organic carbon and humic acid contents but were rich in nitrogen and other nutrients, having lower C:N ratios. In addition, the toxicity of the wet olive cake against the seeds of Lepidium sativum significantly decreased after degradation. The low toxicity as well as moderate stability and maturity recorded in the wet olive cake treated with P. ostreatus or E. fetida imply that these products could be used as soil amendments.     

Influences of biological interactions on community structure within seagrass beds and sandprawn-dominated sandflats

The faunal composition of high-shore beds of eelgrass Zostera capensis versus unvegetated sandflats dominated by the sandprawn Callianassa kraussi was investigated by experimentally transplanting plots of eelgrass into sandflats where the sandprawn was either left unmanipulated or eliminated by prior faunal suffocation. After 12 months, multidimensional scaling defined five faunal clusters that were dictated primarily by the presence or absence of eelgrass rather than sandprawns. By 18 months, eelgrass and sandprawns played equal roles in defining faunal clusters. After 30 months, sandprawns had displaced the eelgrass transplants and faunal composition converged on that of sandflat control plots.Most species were either ‘eelgrass-associated’ or ‘sandflat-associated’, but three had unique responses to the treatments: two high-shore species (Assiminea globulus and Hydrobia sp.) that were abundant in eelgrass beds never appeared in the eelgrass transplants, whereas another eelgrass-occupant, Siphonaria compressa, became superabundant on the transplants for 18 months.Unexpectedly, the eelgrass fauna was never more rich or diverse than that of unvegetated sandflats, although abundance was greater in the eelgrass. As hypothesised, the sandflats were disproportionally populated by burrowers and eelgrass by non-burrowers, but a second hypothesis that these habitats should support mainly hard-bodied and soft-bodied animals respectively was rejected.The distinctively different faunas of eelgrass and unvegetated sandflats are seemingly respectively maintained by sediment stabilisation by eelgrass versus bioturbation by sandprawns. The transplants of eelgrass into sandflats developed an eelgrass-associated fauna provided sandprawns were excluded, but ultimately reverted to a sandflat fauna once sandprawns reinvaded.     

Changes in lignin content of leaf litters during mulching

Alkaline nitrobenzene oxidation, ozonation and methoxyl content determinations were applied to decomposing leaf litter of Ginkgo biloba L., Cinnamomum camphora sieb., Zelkova serrata Makino and Firmiana simplex W. F. Wight, respectively, during mulching to investigate the properties and estimate changes in lignin composition and content. Since the Klason lignin residue originated from components highly resistant to degradation by acid, the methoxyl content of Klason residue was used to estimate the lignin content of leaf litter. Quantitative analysis of presumed lignin-derived fragments, by use of alkaline nitrobenzene oxidation and ozonation methods, suggested that the estimated lignin content approximates that of the real lignin content of leaves, which is greatly overestimated by the Klason procedure. The estimated lignin contents ranged from 3.9 to 10.0% while the Klason lignan residue varied from 37.1 to 46.7% in un-mulched leaf litter. The absolute amounts of the measured lignin somewhat decreased during mulching, while the structure of lignin remaining in leaf litters after mulching was considered not to be very different from its original structure.     

Molecular and phenotypic characterisation of Bacillus thuringiensis isolated during epizootics in Cydia pomonella L

Twelve Bacillus thuringiensis strains were isolated from intestinal tracts of Cydia pomonella larvae during epizootics in different laboratory insect culture lines. Phenotypic and genetic similarity of these isolates, together with two cultured from Leucoma salicis larvae and 14 reference B. thuringiensis strains were determined. The epizootic bacteria did not form a single group based on numerical analysis of biochemical properties. Simple RAPD method with only one primer does not allow estimating the genetic similarity of B. thuringiensis strains. We propose a novel strategy based on combining several DNA patterns obtained by RAPD technique with different primers for B. thuringiensis typing. Majority of infections in the C. pomonella culture lines were caused by bacteria with different genotypes. However, two isolates cultured from infected insects at different time (one strain was isolated in 1990 and the other in 1992) had identical DNA fingerprint that suggested a possibility of these bacteria to survive in the laboratory and to cause infections in different years. The results of SDS-PAGE of whole-cell proteins revealed a possibility to apply protein profile analysis in epidemiological investigations of infections caused by B. thuringiensis. Strains with identical DNA patterns had very similar whole-cell protein profiles.     

Temporal change of composition and potential activity of the thermophilic archaeal community during the composting of organic material

To date, composting has been regarded as an aerobic process but it has been shown that composting piles are often sources of atmospheric methane. In order to gain a more comprehensive view on the diversity of methanogenic Archaea in compost, gas chromatographical methods and molecular cloning were used to study relationships of thermophilic archaeal communities and changes in methane production potential during compost maturation. According to the thermophilic methane production potential, wide differences could be detected between differently aged compost materials. In material derived from 3- and 4-week-old piles, low and no thermophilic methane production potential, respectively, was observed at 50 degrees C. Material from a 6-week-old pile showed the maximum methane production. With compost maturation, the production slowly decreased again with 6 weeks, 8 weeks, and mature compost showing an optimum methane production potential at 60 degrees C. At 70 degrees C, only 6-week-old material showed a comparable high production of methane. The 16S rRNA-based phylogenetic surveys revealed an increase of archaeal diversity with compost maturation. In the 6-week-old material, 86% of the sequences in the archaeal 16S rRNA library had the highest sequence similarities to Methanothermobacter spp. and the remaining 14% of the clones were related to Methanosarcina thermophila. Quantification of methanogens in 6-week-old material, on the basis of the methane production rate, resulted in values of about 2x10(7) cells per gram fresh weight. In 8-week-old and mature compost material, the proportion of sequences similar to Methanothermobacter spp. decreased to 34% and 0%, respectively. The mature compost material showed the highest variation in identified sequences, although 33% could be assigned to as yet uncultured Archaea (e.g. Rice cluster I, III, and IV). Our results indicate that compost harbours a diverse community of thermophilic methanogens, with changing composition during the maturation process, presumably due to altered pile conditions. Likewise, compost may act as a potential carrier for thermophilic methanogens in temperate soils because it is widely used as a soil amendment.