Publicacións BABS

I Villar, D Alves and S Mato

Product quality and microbial dynamics during vermicomposting and maturation of compost from pig manure.

Waste Management, 69: 498-507, 2017.


This research evaluates, through microbial dynamics, the use of earthworms Eisenia andrei for maturation of pre-composted pig manure in comparison with maturation under static conditions and with vermicomposting of fresh pig manure. Therefore, two substrates were used (fresh and pre-composted pig manure) and four treatments were developed: fresh manure vermicomposting, control of fresh manure without earthworms, pre-composting followed by vermicomposting and static maturation of pre-composted manure. In order to determine the microbial dynamics, the enzymatic activities and profiles of phospholipid fatty acids (PLFAs) were evaluated over a 112-days period. Physicochemical and biological parameters of the obtained products were also analyzed. The presence of earthworms significantly reduced (p < 0.05) microbial biomass and all the microbial groups (Gram + bacteria, Gram − bacteria, and fungi) in both substrates. The enzymatic activities (cellulase, β-glucosidase and acid phosphatase) behaved in a significantly distinctive manner (p < 0.05) depending on the treatment. Microbial communities had significant correlations (p < 0.05) with hydrolytic activities during static maturation of pre-composted manure. This indicates a direct effect of microbiota evolution on the degradative processes; however, complex earthworm-microbiota interactions were established in the presence of E. andrei. After earthworms’ removal from vermicompost of fresh substrate at 70 day, an increase in Gram + (4.4 times), Gram − (3.8 times) and fungi (2.8 times) were observed and, although the vermicompost achieved quality values, it is necessary to optimize the vermicompost aging phase period to improve the stability. Static maturation presented stability on microbial dynamics that indicated a slow degradation of organic compounds so that, maturation of pre-composted manure through vermicomposting is better option. © 2017 Elsevier Ltd

I Villar, D Alves, D Pérez-Díaz and S Mato

Changes in microbial dynamics during vermicomposting of fresh and composted sewage sludge.

Waste Management, 48: 409-417, 2016.


Municipal sewage sludge is a waste with high organic load generated in large quantities that can be treated by biodegradation techniques to reduce its risk to the environment. This research studies vermicomposting and vermicomposting after composting of sewage sludge with the earthworm specie Eisenia andrei. In order to determine the effect that earthworms cause on the microbial dynamics depending on the treatment, the structure and activity of the microbial community was assessed using phospholipid fatty acid analysis and enzyme activities, during 112 days of vermicomposting of fresh and composted sewage sludge, with and without earthworms. The presence of earthworms significantly reduced microbial biomass and all microbial groups (Gram+ bacteria, Gram- bacteria and fungi), as well as cellulase and alkaline phosphatase activities. Combined composting-vermicomposting treatment showed a lesser development of earthworms, higher bacterial and fungal biomass than vermicomposting treatment and greater differences, compared with the control without earthworms, in cellulase, β-glucosidase, alkaline and acid phosphatase. Both treatments were suitable for the stabilization of municipal sewage sludge and the combined composting-vermicomposting treatment can be a viable process for maturation of fresh compost. © 2015 Elsevier Ltd.

I Villar, D Alves, J Garrido and S Mato

Evolution of microbial dynamics during the maturation phase of the composting of different types of waste.

Waste Management, 54: 83-92, 2016.


During composting, facilities usually exert greater control over the bio-oxidative phase of the process, which uses a specific technology and generally has a fixed duration. After this phase, the material is deposited to mature, with less monitoring during the maturation phase. While there has been considerable study of biological parameters during the thermophilic phase, there is less research on the stabilization and maturation phase. This study evaluates the effects of the type of starting material on the evolution of microbial dynamics during the maturation phase of composting. Three waste types were used: sludge from the fish processing industry, municipal sewage sludge and pig manure, each independently mixed with shredded pine wood as bulking agent. The composting system for each waste type comprised a static reactor with capacity of 600 L for the bio-oxidative phase followed by stabilization and maturation phase in triplicate 200 L boxes for 112 days. Phospholipid fatty acids, enzyme activities and physico-chemical parameters were measured throughout the maturation phase. The evolution of the total microbial biomass, Gram + bacteria, Gram – bacteria, fungi and enzymatic activities (β-glucosidase, cellulase, protease, acid and alkaline phosphatase) depended significantly on the waste type (p < 0.001). The predominant microbial community for each waste type remained present throughout the maturation process, indicating that the waste type determines the microorganisms that are able to develop at this stage. While fungi predominated during fish sludge maturation, manure and municipal sludge were characterized by a greater proportion of bacteria. Both the structure of the microbial community and enzymatic activities provided important information for monitoring the composting process. More attention should be paid to the maturation phase in order to optimize composting. © 2016 The Authors.

I Villar, D Alves and S Mato

Seafood-processing sludge composting: Changes to microbial communities and physico-chemical parameters of static treatment versus for turning during the maturation stage.

PLoS ONE, 11 (12), 2016.


In general, in composting facilities the active, or intensive, stage of the process is done separately from the maturation stage, using a specific technology and time. The pre-composted material to be matured can contain enough biodegradable substrates to cause microbial proliferation, which in turn can cause temperatures to increase. Therefore, not controlling the maturation period during waste management at an industrial level can result in undesired outcomes. The main hypothesis of this study is that controlling the maturation stage through turning provides one with an optimized process when compared to the static approach. The waste used was sludge from a seafood-processing plant, mixed with shredded wood (1:2, v/v). The composting system consists of an intensive stage in a 600L static reactor, followed by maturation in triplicate in 200L boxes for 112 days. Two tests were carried out with the same process in reactor and different treatments in boxes: static maturation and turning during maturation when the temperature went above 55ºC. PLFAs, organic matter, pH, electrical conductivity, forms of nitrogen and carbon, hydrolytic enzymes and respiratory activity were periodically measured. Turning significantly increased the duration of the thermophilic phase and consequently increased the organic-matter degradation. PCA differentiated significantly the two treatments in function of tracking parameters, especially pH, total carbon, forms of nitrogen and C/N ratio. So, stability and maturity optimum values for compost were achieved in less time with turnings. Whereas turning resulted in microbialgroup stabilization and a low mono/sat ratio, static treatment produced greater variability in microbial groups and a high mono/sat ratio, the presence of more degradable substrates causes changes in microbial communities and their study during maturation gives an approach of the state of organic-matter degradation. Obtaining quality compost and optimizing the composting process requires using turning as a control mechanism during maturation. © 2016 Villar et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

M Pérez-Losada, J Eiroa, S Mato and J Domínguez

Phylogenetic species delimitation of the earthworms Eisenia fetida (Savigny, 1826) and Eisenia andrei Bouché, 1972 (Oligochaeta, Lumbricidae) based on mitochondrial and nuclear DNA sequences.

Pedobiologia, 49 (4): 317-324, 2005.


Three samples (11 individuals) of Eisenia fetida (Savigny, 1826) and four samples (20 individuals) of Eisenia andrei Bouché, 1972 (focal species), and two samples (six individuals) of Eisenia eiseni (Levinsen, 1884) (nonfocal species; outgroup) were tested for species delimitation using the phylogeny-based method of Wiens and Penkrot (2002). Thirty-seven and 17 DNA sequences of the nuclear 28S (2031 bp) and mitochondrial COI (658 bp) genes, respectively, were generated. Phylogenetic hypotheses for each gene and both combined were built using maximum parsimony, maximum likelihood, and Bayesian approaches. Fully resolved and well-supported trees (>90% bootstrap proportions and 1.0 posterior probabilities) showed both E. fetida and E. andrei samples as monophyletic. Application of Wiens and Penkrot’s protocol to those trees resulted in the recognition of these two taxa as different phylogenetic species. This result complements previous studies of crossability and allozyme genetics addressing their different biological status (biological species concept). Finally, our analyses also show that E. fetida is composed of reproductively isolated populations and suggest the presence of a hitherto unrecognized species or subspecies within this taxon. © 2005 Elsevier GmbH. All rights reserved.

P Ondina, J Hermida, A Outeiro and S Mato

Relationships between terrestrial gastropod distribution and soil properties in Galicia (NW Spain).

Applied Soil Ecology, 26 (1): 1-9, 2004.


This study investigated the influence of edaphic factors on the distribution of 17 terrestrial gastropod species over a large area of the northwest Iberian Peninsula. A total of 498 gastropod/soil samples were obtained, and a total of 19 edaphic variables determined. The resulting data matrix was analysed by detrended canonical correspondence analysis (DCCA). Our results indicate that the gastropods of the study area can be grouped on two types of criteria: first, chemical criteria (notably pH, cation exchange capacity, and Al content), and secondly physical criteria (notably texture and moisture content). In view of distribution with respect to these factors, two well-defined groups can be identified: one comprising Acanthinula aculeata, Euconulus fulvus, Punctum pygmaeum, Columella aspera and Oxychilus alliarius preferring coarse-textured acid soils, the other comprising Cochlicopa lubrica, Vertigo pygmaea, Zonitoides excavatus, Carychium tridentatum, Deroceras reticulatum and Deroceras lombricoides preferring wetter, finer-textured, less acid soils. Arion intermedius and Ponentina subvirescens were in general indifferent to the edaphic factors considered. © 2003 Elsevier B.V. All rights reserved.

M Aira, F Monroy, J Domínguez and S Mato

How earthworm density affects microbial biomas and activity in pig manure.

European Journal of Soil Biology, 38 (1): 7-10, 2002.


We studied the influence of the earthworm, Eisenia fetida, on the microbial populations during the vermicomposting of pig manure. Fresh pig manure was placed in replicated boxes with (two densities, 25 and 50) and without earthworms for a period of 16 d. Samples were destructively collected periodically and analyzed for microbial biomass nitrogen, microbial respiration, substrate-induced respiration (SIR) and substrate dehydrogenase activity. Microbial biomass N, microbial respiration, SIR and substrate dehydrogenase activity were significantly lower in the earthworm treatments after 8 d. © 2002 Éditions scientifiques et médicales Elsevier SAS. All rights reserved.

P Ondina and S Mato

Influence of vegetation type on the constitution of terrestrial gastropod communities in northwest Spain.

Veliger, 44 (1): 8-19, 2001.


We investigated the influence of three different vegetation types on the establishment of terrestrial gastropod communities in the northwest Iberian Peninsula, using both an ordination technique (Detrended Correspondence Analysis) and a classification technique (Two-Way Indicator Species Analysis) applied to a 498 X 47 site-by-species abundance matrix (total number of individuals 17,902). The results of these analyses indicate that meadow sites are characterized by a group of species comprising Cionella lubrica (Müller, 1774), Nesovitrea hammonis (Ström, 1765), and Zonitoides excavatus (Alder, 1830). Woodland sites are characterized by a group comprising Columella aspera Waldén, 1966, Oxychilus alliarius (Miller, 1822), Acanthinula aculeata (Müller, 1774), and Punctum pygmaeum (Draparnaud, 1801). Vitrea contracta (Westerlund, 1871), Discus rotundatus, and Aegopinella nitidula (Draparnaud, 1805) form a group of companion species to this latter community.

M G García, F Mariño and S Mato

Effect of the diet on growth and reproduction of Eisenia andrei (Oligochaeta, Lumbricidae) reared in individual cultures.

Pedobiologia, 43 (3): 267-275, 1999.


The effect of some vegetable bulking agents (oak leaves, pine bark and pine needles) mixed with activated sewage sludge on the growth of Eisenia andrei, Bouche 1972, was studied in cultures comprising of isolated individuals. Four diets were prepared: diet 1-sewage sludge + oak leaves (3:1 proportion, fresh weight); diet 2-sewage sludge + oak leaves (4:1); diet 3-sewage sludge + pine bark (2:1) and diet 4-sewage sludge + pine needles (3:1). Twenty juvenile worms (10-20 mg, wet mass) were maintained individually in 0.5 L replicates of a given test diet. Earthworms developed to full sexual maturity on each diet. Cocoon production was monitored in the four dietary mixtures; hatchling production was monitored in cocoons produced in diets 1, 2 and 3. Growth (mean weight) and reproduction appeared to be inversely related according to the identity of the sludge:bulking agent used for maintenance. Thus, maximum growth and the least number of cocoons (all non-viable) were recorded in the mixture of activated sewage sludge and pine needles. In contrast, the maximum reproduction rate and highest cocoon production was recorded in the 3:1 sludge:oak leaves mixture.

A Merino-Trigo, L Sampedro, F J Rodríguez-Berrocal, S Mato and M Páez De La Cadena

Activity and partial characterisation of xylanolytic enzymes in the earthworm Eisenia andrei fed on organic wastes.

Soil Biology and Biochemistry, 31 (12): 1735-1740, 1999.


This study demonstrates the occurrence of active xylanolytic enzymes in extracts from Eisenia andrei, Bouche 1972. Xylanase was the most active enzyme (2.06 U g-1), followed by acetyl xylan esterase (0.55 U g-1). Lower activities showed β-D- glucuronidase (0.17 U g-61), α-L-arabinosidase (0.04 U g-1) and β-xylosidase (0.03 U g-1). All of th xylanolytic enzymes displayed higher total activity when the earthworms were fed on a xylan-containing medium. Xylanase, α-L-arabinosidase and acetyl xylan esterase showed maximal activity at neutral pH. In contrast, β-xylosidase and β-glucuronidase displayed maximal activity at pH 3.0 and 4.0, respectively. All enzymes displayed a high activity within the physiological temperature range 35 to 40°C. K(m) and V(max) values of xylanolytic enzymes were also estimated. Kinetic parameters for xylanase were determined in extracts of earthworms fed on standard medium (K(m) = 6.91 ±1.120 mg ml-1; V(max) = 0.05 ± 0.002 U: mg-1), and in extracts of earthworms fed on a medium containing xylan (K(m) = 5.60±1.070 mg ml-1; V(max) = 0.12 ± 0.006 U mg-1).

P Ondina, S Mato, J Hermida and A Outeiro

Importance of soil exchangeable cations and aluminium content on land snail distribution.

Applied Soil Ecology, 9 (1-3): 229-232, 1998.


The influence of soil exchangeable cations and aluminium content on the distribution of various terrestrial gastropods was investigated. Calcium was the most important factor, though aluminium and magnesium content also had some effect. Potassium and sodium had no significant influence on distribution. The species most sensitive to the factors analysed were Cochlicopa lubrica, Vertigo pygmaea and Carychium tridentatum. © 1998 Elsevier Science B.V.

C Elvira, J Domínguez and S Mato

The growth and reproduction of Lumbricus rubellus and Dendrobaena rubida in cow manure Mixed cultures with Eisenia andrei.

Applied Soil Ecology, 5 (1): 97-103, 1997.


The growth and reproduction of the epigeic species Lumbricus rubellus and Dendrobaena rubida in cow manure and the possible interactions of these species with Eisenia andrei are studied. The mean growth rate of D. rubida was 3.84 mg day-1, reaching sexual maturity at 54 days and producing an average of 1.45 cocoons per week. After collection, 85% of the cocoons of this species were viable, incubation took an average of 21.7 days and an average of 1.67 worms emerged from each cocoon. The mean growth rate of L. rubellus was 8.02 mg day-1, maturing at 74 days and with a mean weekly production of 0.54 cocoons. After an incubation period of 36.5 days, 64% of the cocoons hatched, one worm emerging from each. The mixed cultures tested did not present any advantage over pure cultures. E. andrei showed higher growth rates in mixed cultures, while the growth rate of L. rubellus and D. rubida decreased slightly in mixed cultures as compared to pure cultures.

J Dominguez, M J I Briones and S Mato

Effect of the diet on growth and reproduction of Eisenia andrei (Oligochaeta, lumbricidae).

Pedobiologia, 41 (6): 566-576, 1997.


The effect of some vegetable bulking agents (straw, pine needles, pine bark, oak leaves and fern fronds) in mixtures with pig slurry (1:1 dry weight) on the growth and reproduction of Eisenia andrei, Bouche 1972 was studied in cultures with either 1 or 8 individuals. Unlike individual cultures, with no food limitation and no competition, the cultures with 8 individuals represent a more real situation, with food competition and mating processes. The maximum growth and reproduction rates were achieved, both in the individual and in the group cultures, in the mixtures with strew and pine needles. The earthworms showed low growth rates and very low reproductive rates in the oak leaves and fern mixtures, both in cultures with one individual or 8 individuals. The high growth rates obtained here are in agreement with others in the literature indicating the high potential of pig wastes for vermicomposting. Our results also confirm that there is a direct relationship between E. andrei biomass and clitellum development so that the minimum weight for maturation is approximately 0.4 g. In addition, the remarkably high reproduction rate and cocoon production in the cultures with one individual suggests that mating is not an obligatory requirement for cocoon production.

C Elvira, L Sampedro, J Dominguez and S Mato

Vermicomposting of wastewater sludge from paper-pulp industry withnitrogen rich materials.

Soil Biology and Biochemistry, 29 (3-4): 759-762, 1997.


The vermicomposting of pulp mill sludge mixed with sewage sludge, pig slurry and poultry slurry at different ratios was studied. Eisenia andrei (Bouche, 1972) showed high growth rates and high mortalities in all the mixtures considered.

C Elvira, M Goicoechea, L Sampedro, S Mato and R Nogales

Bioconversion of solid paper-pulp mill sludge by earthworms.

Bioresource Technology, 57 (2): 173-177, 1996.


Bioconversion of solid paper-pulp mill sludges and primary sewage sludge for 40 days at ratio of 3:1 dw:dw was studied in containers with and without earthworms (Eisenia andrei). This mixture was a suitable medium for optimum growth and reproduction of the earthworms. Regardless of the presence of earthworms, degradation occurred during the bioconversion period, but the presence of earthworms accelerated the mineralization of organic matter, favored the breakdown of structural polysaccharides and increased the humification rate. Consequently, the C/N ratio and the degree of extractability of heavy metals were lower in the worm-worked end product.

C Elvira, S Mato and R Nogales

Changes in heavy metal extractability and organic matter fractions after vermicomposting of sludges from a paper mill industry and wastewater treatment plant.

Fresenius Environmental Bulletin, 4 (8): 503-507, 1995.

M J Briones, R Mascato and S Mato

Autecological study of some earthworm species (Oligochaeta) by means of ecological profiles.

Pedobiologia, 39 (2): 97-106, 1995.

S Mato, M Garcia and D Otero

Composting of < 100 mm fraction of municipal solid waste.

Waste Management & Research, 12 (4): 315-325, 1994.


A low cost solution for management of municipal solid waste in small municipalities was investigated; composting experiments were done using domestic waste. Particles > 100 mm were removed, the screened substrate ( < 100 mm) was used for composting experiments on static piles. The results have shown that the size of waste does not affect composting. Though the inert material left greatly adds to the porosity, rainfall does not markedly affect the process; in fact excessive dryness gave the least stable end product, and total nitrogen, ammoniacal nitrogen and phosphorus content were the best pointers to the instability. © 1994, Sage Publications. All rights reserved.

M J Briones, R Mascato and S Mato

Biogeographical considerations in Asturias, Leon, Zamora and Salamanca (Spain) in relation to their earthworm fauna.

European Journal of Soil Biology, 30 (4): 149-155, 1994.

M J I Briones, R Mascato and S Mato

Relationships of earthworms with environmental factors studied by means of detrended canonical correspondence analysis.

Acta Oecologica, 13 (5): 617-626, 1992.


The relationship between 13 species and subspecies of earthworms from Spain and 20 soil factors were analysed. Two groups were distinguished: one is made up of Dendrobaena madeirensis, D. octaedra, D. pygmaea, Eisenia eiseni and Allolobophora oliveira, which prefer organic acid soils with low Ca and Mg content and high Al content. A second group is formed by other species which exhibit opposite preferences. In the latter group, A. caliginosa caliginosa, D. mammalis, Lumbricus rubellus, Eiseniella tetraedra and D. rubida could be separated from A. c. trapezoides, A. rosea and A. chlorotica by organic matter, texture and moisture content. -from Authors

D Trigo, S Mato, B F Souto and D J Diaz Cosin

Earthworms of continental portugal. Relationships with soil factors.

Bolletino di zoologia, 56 (4): 327-331, 1989.


The relationships between the 13 most common species of Portuguese earthworms and 20 soil characteristics have been investigated by analysis of ecological profiles, correspondence analysis and canonical correlation. Two groups emerged: one comprising basically the epigeic species Allolobophora oliveirae, Dendrobaena madeiren- sis, Dendrobaena octaedra and Eisenia eiseni, which tend to prefer organic acid soils with little calcium or magnesium and large quantities of aluminium in the exchange complex, and the other made up of the endogeic species Allolobophora caliginosa, Allolobophora mol- leri, Allolobophora rosea and Allolobophora chlorotica, which exhibit the opposite tendencies. © 1989 Taylor & Francis Group, LLC.

S Mato, R Mascato, D Trigo and D J Diaz Cosin

Vertical distribution in soil of earthworms in Sierra del Caurel. 1. Species and vegetation types.

Pedobiologia, 32 (3-4): 193-200, 1988.


Surface species predominated in beechwood Fagus sylvatica, and Quercus pyrenaica wood had proportionally the largest deep-dwelling populations. At the other sites adults exhibited a slight preference for the 5-15 cm layer and immatures for the 0-5 cm layer. -from Authors

D Trigo, R Mascato, S Mato and D J D Cosin

Biogeographical divisions of continental portugal as regards earthworm fauna.

Bolletino di zoologia, 55 (1-4): 85-92, 1988.


Biogeographical provinces have been defined for the earthworm fauna of continental Portugal by subjecting data obtained at 100 sites distributed among 23 zones to correspondence analysis and dendritic clustering so as to detect faunistically similar groups of zones. Various measures of species importance and interspecific association were calculated in order to characterize the provinces so detected. On the basis of our findings and the existing literature, ve propose that Galicia and Northern Portugal be included in a lalaico-Lusitanian Province characterized by the joint dominance f A. caliginosa and the three typical species D. madeirensis, Dotaedra and A. oliveirae, which are accompanied by other, more videly distributed species; and that continental Portugal south of he River Mondego be considered an Atlantic Province character-zed by the dominance of A. caliginosa, A. rosea and the «A. molleriii tomplex species, and by the absence of the three typical Northern species and all species of the genus Lumbricus. © 1988 Taylor & Francis Group, LLC.

R Mascato, S Mato, D Trigo, F Marino and D J Diaz Cosin

Soil factors and earthworm distribution in two areas of Galicia: comparison of some statistical methods [Factores del suelo y distribucion de las Lombrices de tierra en dos zonas de Galicia: Comparacion de diferentes metodos estadisticos].

Revue d’Ecologie et de Biologie du Sol, 24 (2): 111-135, 1987.


Distinguishes 2 groups 1) associated with high loam and Ca contents, high pH in water and in potassium chloride, and low coarse sand, C, N and Al contents; and 2) with the opposite tendencies. -from English summary