Advanced techniques for characterization of organic matter from anaerobically digested grapemarc...

Environ Monit Assess (2012) 184:2079–2089DOI 10.1007/s10661-011-2101-z

Advanced techniques for characterization of organicmatter from anaerobically digested grapemarc distilleryeffluents and amended soils

Gennaro Brunetti · Karam Farrag · Cesar Plaza ·Nicola Senesi

Received: 4 June 2010 / Accepted: 26 April 2011 / Published online: 15 May 2011© Springer Science+Business Media B.V. 2011

Abstract The effects of grapemarc distilleryeffluents on the quality of soil organic matter is ex-tremely important to ensure the environmentally-safe and agronomically efficient use of thesematerials as organic amendment. In this work,the effects of the application of untreated (UG)and anaerobically digested grapemarc distilleryeffluents, either added with (AGM) or withoutmycorrhiza (AG), on soil humic acid (HA) wereinvestigated in field plot experiments in compar-ison to HAs from a control soil and an inor-ganic fertilized soil. The humic acid-like fractions(HALs) isolated from UG, AG and soils werecharacterized for compositional, structural and

G. Brunetti · N. SenesiDipartimento di Biologia e Chimica Agroforestaleed Ambientale, Università di Bari, Via Amendola165/A, 70126 Bari, Italy

K. Farrag (B)Central Lab for Environmental Quality Monitoring(CLEQM), National Water Research Center(NWRC), Cairo, Egypte-mail: [email protected],[email protected]

C. PlazaCentro de Ciencias Medioambientales,Consejo Superior de Investigaciones Cientìficas,Serrano 115 dpdo., 28006 Madrid, Spain

functional properties by the use of elemental andfunctional group analysis, and ultraviolet/visible,Fourier transform infrared and fluorescence spec-troscopies. Results obtained indicated that anaer-obic digestion of effluents produced an extendedmineralization with loss of organic C and stabi-lization of residual organic matter by increasingthe content of HALs in the effluent. With re-spect to control soil HA, HALs isolated fromUG and AG were characterized by smaller acidicfunctional group contents, a prevalent aliphaticcharacter and smaller aromatic polycondensa-tion and humification degrees. The chemical andspectroscopic characteristics of native soil HAwere not substantially modified by applicationof UG, AG and AGM to soil, which suggeststhe occurred incorporation of the effluent HALinto native soil HA. In conclusion, these resultsshowed the possibility of a beneficial and saferecycling of grapemarc distillery effluents as soilamendment.

Keywords Grapemarc distillery effluents ·Anaerobic digestion · Humic acids ·Humification parameters · Soil amendment

Introduction

Effluents and by-products of food processingand agro-based industries are among the most

2080 Environ Monit Assess (2012) 184:2079–2089

common and abundant organic waste materialsresulting from agriculture and food industry. Re-cycling of these organic materials requires a pri-ority attention for avoiding their dispersion inthe environment. On the other hand, it is wellknown the vital role of soil organic matter (OM)on global soil fertility and agricultural production,and its key role in soil protection from degra-dation and contamination. Thus, a goal of majorimportance is to develop agricultural and environ-mental strategies for conservation of soil OM andsupport its reintegration in soils depleted by inten-sive modern agriculture practices. Consequently,the appropriate agriculture recycling of these or-ganic waste materials represents an optimal andnatural solution for achieving the recent wastepolicies in the EU (CEC 2005) which promotethe use of “wastes” as “resources” to be recy-cled instead of disposal. In this respect, recyclingof by-products of agro-food industries, includingwastes produced by wineries and distilleries, suchas grapemarc, appear to be the most appropriateecological approach to reduce their environmen-tal load and restore soil OM.

Grapemarc, including peel, stalks and seedsis the primary solid waste generated from wineproduction which, if not treated correctly, maycause various environmental hazards such aswater pollution, foul odours, and spread of dis-eases (Arvanitoyannis et al. 2006; Mcmanus 2008).Grapemarc is a valuable resource as a soil fertil-izer (Flavel et al. 2005) and could be recycled as asoil conditioner in view of its organic and nutrientcontents (Diaz et al. 2002).

Results of many chemical, biological and agro-nomical studies indicated that soil application ofany waste materials of biosynthetic origin, with-out being subjected to any chemical and/or bio-logical treatment, is not agronomically efficient,and may represent a serious risk to the en-vironment (Brunetti et al. 1995; Senesi et al.1996; Senesi and Brunetti 1996). In particular,Bustamante et al. (2008) reported that regard-less the richness of grapemarc residues in OMand macronutrients such as K, the presence ofpolyphenols that may cause phytotoxic and an-timicrobial effects makes it necessary to subject

these materials to appropriate treatments beforetheir use in agriculture.

The most common treatments of organic wastesinclude composting, vermicomposting, anaero-bic and aerobic digestion, thermophilic anaer-obic digestion, sequencing batch reactor, elec-trodialysis, wet oxidation, pyrolysis, incinera-tion, solid-state fermentation and ozonation (DeBertoldi et al. 1996; Edwards 2004; Williams2005; Arvanitoyannis et al. 2006). However,not enough data are currently available on anaer-obic digestion of winery and distillery wastes andtheir attitude for agricultural use. Anaerobic di-gestion is a complex biochemical process carriedout in a number of steps by several types of mi-croorganisms that decompose OM in the absenceof oxygen at a proper temperature and acidity.Anaerobic digestion of large quantities of munic-ipal, industrial and agricultural solid wastes canprovide biogas as well as other benefits such as re-duction in waste volume and production of biofer-tilisers and soil conditioners (Edelmann et al.2000; Grommen and Verstraete 2002; Lema andOmil 2001; Lettinga 2004).

Stability and maturity are the main aspects usu-ally used to assess the quality of OM in treatedwastes. During the treatment of organic wastes,mineralization and humification processes trans-form the raw OM in wastes into a more stabilizedmaterial that contains the so-called humic-likesubstances (HLS) that are suggested to representreliable indicators for stability and maturity dur-ing and after the treatment (Senesi et al. 1996;Paré et al. 1998).

Soil HS, of which humic acids (HAs) consti-tute the major fraction, contribute essential func-tions to soil global fertility and health (Stevenson1994). As a consequence, the knowledge of theamount and molecular, structural, functional andreactive properties of HLS, and especially theHA-like (HAL) fraction, contained in a givenorganic material is necessary to assess its qualityas soil amendment. Further, the effects of or-ganic amendment application on the status, qual-ity, chemistry and functions of native soil HAneed to be assessed for a thorough evaluationof the agronomic effectiveness and environmental

Environ Monit Assess (2012) 184:2079–2089 2081

impact of this practice (Senesi et al. 1996; Senesiand Plaza 2007).

A number of criteria and parameters includingthe so-called empirical humification parameters(humification index, degree of humification andhumification ratio) have been proposed to assessthe quality of OM present in organic materials(Senesi and Brunetti 1996; Senesi et al. 1996).However, because of the extreme diversity inthe origin and processing of organic substratesthese parameters like all other empirically basedmethods, including physical, chemical, microbi-ological tests and plant bioassays, are not suit-able for general application (Senesi et al. 2007).As a consequence, the most appropriate andreliable criteria to evaluate maturity of anyorganic amendment should be based on thechemical and physicochemical analyses of themolecular, structural and functional identity andproperties of HLS components, i.e., the HAL andfulvic acid-like fractions in the organic amend-ment. These data can be obtained by the useof spectroscopic methods, such as fluorescenceor/and infrared spectroscopy (Smidt et al. 2005;Senesi et al. 2007; Miano et al. 1988; Geyer et al.1998).

Fourier transform infrared spectroscopy (FT-IR) is a well-established technique for thediscrimination of functional groups and princi-pal structures in OM, including carbohydrates,lignins, cellulose, fats and/or lipids and proteina-ceous compounds (Senesi and Brunetti 1996;Brunetti et al. 2007; Artz et al. 2008). Fluores-cence spectroscopy for OM characterization hasadvanced by the use of excitation–emission ma-trix (EEM) spectroscopy that provides a three-dimensional picture of fluorescence intensity as afunction of excitation and emission wavelengths.The fluorescence regional integration techniquehas been used as a tool to analyze quantitativelyall the wavelength-dependent fluorescence inten-sity data from EEM spectra (Senesi et al. 1991,1996; Senesi and Brunetti 1996; Parlanti et al.2000; Provenzano et al. 2001; Sierra et al. 2005;Chen 2003). Without excluding the use of appro-priate empirical parameters, the current researchaims to identify and use a group of intrinsic pa-

rameters that allow a direct quality evolution ofthese OM materials.

The overall objective of the study is to evaluatethe anaerobic digestion process of a wine grape-marc so as to improve its quality as a soil organicamendment in short-term field and laboratory ex-periments. To accomplish this work, the specificobjectives were: (a) to evaluate the main chemicalcharacteristics and fertilizer value of grapemarcsubjected to anaerobic digestion by determiningthe molecular, structural and functional character-istics of the HALs isolated from grapemarc com-pared to those of HA from a native control soil;and (b) to determine the possible modifications ofHAs of soils amended with untreated grapemarcand anaerobically digested grapemarc, either in-oculated or without mycorrhiza, compared to theHA of the control soil and the same soil treatedwith mineral nitrogen fertilizer.

Materials and methods

Grapemarc substrates

Both field and laboratory experiments wereperformed using two grapemarc samples fromthe Bonollo distillery located in countryside ofAnagni (Frosinone, Lazio). Grapemarc (UG) isthe mixture of effluents resulting from the winedistillation, lees and vinasse, without any treat-ment. Stabilized grapemarc (AG) is the sameeffluent subjected to anaerobic digestion process,followed by concentration by press filtration orcentrifugation. Grapemarc added with mycor-rhizae is AG inoculated with mycorrhiza beforeapplication to soil.

Soils

Field experiments were conducted in the exper-imental farm of the Istituto Sperimentale Agro-nomico of Bari (MiPA) located in Foggia. Thesoil is classified as a fine, mixed, thermic TypicChromoxerert (Soil Survey Staff 1998), havingclay-loamy texture, moderately alkaline pH (8.3),an organic C content of 21.5 g kg−1 and a total N


content of 1.6 g kg−1. Soil samples were col-lected at a depth of 0–20 cm from plots of 16 m2

1 year after addition of untreated grapemarc(SUG), anaerobically digested grapemarc (SAG)and mycorrhiza inoculated grapemarc (SAGM),each equivalent to 140 U of N ha−1. Soil sampleswere also collected from untreated, control plot(S) and a plot added with a mineral fertilizer(SMF) at 140 U of N ha−1. Soil analyses werecarried out according to internationally recom-mended procedures and Italian official methods(Italy 1999).

Analyses of grapemarc substrates and soils

The grapemarc samples were characterized fortheir main chemical and physicochemical proper-ties according to the sludge analysis method ofCNR-IRSA (1985). The degree of humification(DH), humification rate (HR) and humificationindex (HI) were also determined according to theofficial methods of the Ministry for AgriculturalPolicies (Italy 1999).

Isolation of humic-like and humic acids

The HALs and HAs were isolated from grape-marcs and soils according to conventional proce-dures based on alkaline extraction followed byacid precipitation (Schnitzer 1982). Briefly, eachsample was treated with a solution 0.5 M NaOH +0.1 M Na4P2O7 using an extractant to sample ratioof 5:1 for soil samples and 10:1 for grapemarc.The mixtures were shaken mechanically in a N2

gas atmosphere for 24 h at room temperature.The alkaline supernatant solutions were then sep-arated from the solid phase by centrifugation at9,000×g for 15 min. This process was repeateduntil an exhaustive extraction was obtained. Thedifferent fractions collected were mixed together,then treated with HCl to pH∼1 and allowed tostand for 24 h to precipitate of the HA and HAL.The precipitates were separated by centrifuga-tion at 9,000×g for 15 min and then purified byrepeating two times dissolution in the alkalineextractant and precipitation with 6 M HCl toa pH∼1. The centrifuged HAs and HALs wererecovered with distilled water and then dialyzedagainst distilled water using a membrane having a

molecular weight cutoff of 8 kDa, until the dialysiswater gave a negative Cl− ion test with AgNO3.Finally, the dialyzed products were freeze-driedand stored at RT in plastic vials placed in a des-iccator containing P2O5.

Analyses of humic-like and humic acids

The determination of moisture and ash was car-ried out using calibrated platinum crucibles placedin an oven at 105◦C for several hours. About50 mg of sample were weighed accurately, main-tained (overnight) at 105◦C to constant weight,cooled in a desiccator and then weighed to calcu-late moisture. The same sample then was placedin muffle furnace at 550◦C until constant weight(overnight) and then transferred to a desiccator,cooled and weighed for ash calculation. The deter-mination of C, H, N and S was performed using aC, H, N, S analyzer Fisons, model EA 1108. Oxy-gen content was calculated by difference: O% =100 − (C + H + N + S)%. All obtained data werecorrected for moisture and ash contents. Theanalysis was performed in duplicate and repeatedagain when significant differences were found.The atomic ratios C/N, C/H and O/C were calcu-lated from the percentages of C, H, N, S and Ocorrected for moisture and ash.

Total acidity and carboxylic group contentswere determined according to conventional meth-ods (Schnitzer 1982), while the content of phenolicOH was calculated by difference between totalacidity and COOH groups. Each determinationwas performed twice. The E4/E6 ratio was cal-culated as the ratio of absorbances at 465 and665 nm measured by a Spectrophotometer PerkinElmer model Lambda 15 UV–Vis on solutions of3.0 mg of each sample dissolved in 10 ml of 0.05 MNaHCO3, with the pH adjusted to 8.3 with NaOH.

The FT-IR spectra were recorded over therange 4,000 to 400 cm−1 on pellets obtained bypressing under reduced pressure a mixture of 1 mgof sample and 400 mg of dried KBr, spectrometrygrade. A Nicolet 5PC FT-IR spectrophotometeroperating with a peak resolution of 2 cm−1, andOmnic 1.2 software were used to obtain the spec-tra. Three dimensional fluorescence spectra wereobtained in the form of EEM plots (contour maps)over an emission wavelength range from 300 to


600 nm by increasing sequentially the excitationwavelength from 250 to 550 nm. All spectra wererecorded using a luminescence spectrophotometerHitachi F-4500 Mod after overnight equilibrationat room temperature on aqueous solutions of sam-ples at a concentration of 100 mg L−1 and adjustedto pH 8 with 0.05 M NaOH.

Results and discussion

Chemical properties and humification parametersof grapemarc substrates

Results of analyses carried out on UG and AGsamples (Table 1) indicate that some significantmodifications occur after anaerobic digestion. Inparticular, with respect to the UG sample, the AGsample shows a higher pH value, probably due tomineralization of proteinaceous materials yieldingalkaline ammonia and/or loss of volatile acids. Theslight increase of total N content in AG may beattributed to the occurrence of nitrification anddenitrification processes. Both nitrification anddenitrification have been used to explain the oc-currence of fixed N removal in oxic environments(Hartnett and Devol 2003). They can also be usedto explain constant or even increasing N concen-trations in regions where denitrification is thoughtto be occurring (Codispoti and Christensen 1985;Laursen and Seitzinger 2002). The levels of P andK were also slightly greater in AG. Further, AGshows a loss of OM amounting to 27.7% of thatUG, which suggests the occurrence of extendedOM mineralization. The decrease of C/N ratio

measured in AG mainly reflects the amount ofC loss by mineralization. With respect to UG,both DH and HR of AG markedly increase,while HI decreases. Similar results have been ob-tained by Romero et al. (2007) after vermicom-posting of three different substrates from wineryand distillery wastes. These results suggest thatanaerobic digestion of grapemarc would promotethe humification process of OM, thus increasingits stability and the agronomical efficiently ofAG (Zucconi and De Bertoldi 1987; Senesi 1989;Senesi et al. 2007).

Chemical and structural characteristicsof humic-like and humic acids

Elemental and functional group compositionand E4/E6 ratio

The elemental and functional group compositionand E4/E6 ratio of the two grapemarc HALs ex-amined are presented in Table 2. With respect toHAL-UG, HAL-AG shows a significant increaseof C, N, H, S, total acidity and phenolic OH con-tents and a significant decrease of O content, C/Nratio and E4/E6. However, C/H and O/C ratiosand COOH content remain almost unchanged inthe two HAL samples. These results almost agreewith results of Romero et al. (2007), which foundan increase of O, S and acidic functional groupcontents, and a decrease of C and H contentsof HAL isolated from three different vermicom-posted grapemarcs.

The low E4/E6 ratios measured for HAL-AGwould suggest an increased level of molecular

Table 1 Chemicalproperties andhumification parametersof untreated grapemarc(UG) and stabilizedanaerobically digestedgrapemarc (AG)substrates

Parameters UG AG

Humidity (105◦C) (%) 73.6 74pH (1:10) – 7.1 8.1Potassium (K2O % d.w.) 1.68 1.72Total Nitrogen (% d.w.) 3.0 3.3Total phosphorus (P2O5% d.w.) 0.36 0.40Total organic carbon (g kg−1 d.w.) 417.2 301.4C/N ratio – 13.90 9.12Humic acid-like + Fulvic acid-like (g kg−1 d.w.) 150 176Degree of humification (DH) (%) 56.5 84.1Humification rate (HR) (%) 36.0 58.4Humification index (HI) – 0.77 0.19


Table 2 Compositionaland functionalcharacteristics of HALsisolated from untreatedgrapemarc (UG) andstabilized anaerobicallydigested grapemarc(AG), in comparison withHA of the control soil (S)

Parameters S UG AG

C (g kg−1) 563 ± 7 504 ± 7 556 ± 4H (g kg−1) 44 ± 1 73 ± 1 77 ± 0N (g kg−1) 41 ± 1 77 ± 1 99 ± 1S (g kg−1) 4 ± 1 3 ± 1 6 ± 0O (g kg−1) 348 ± 9 343 ± 6 262 ± 3C/N (atomic ratio) 15.9 7.7 6.6C/H (atomic ratio) 1.1 0.6 0.6O/C (atomic ratio) 0.5 0.5 0.4Total acidity (cmol kg−1) 760 330 440COOH (cmol kg−1) 480 200 210Phinolic OH (cmol kg−1) 270 130 230E4/E6 – 5.5 9.3 6.1

association and humification, whereas the largeE4/E6 ratios of HAL-UG would suggest thepresence of simple structural components of smallmolecular weight and low humification degree.These results are in agreement with Chen et al.(1977) who reported that the E4/E6 ratio is relatednegatively to the particle size, molecular weightand humification degree of HAs.

In general the properties of HAL-AG are rel-atively similar to those of S-HA, which confirmsthe partial humification process of OM occurredduring grapemarc anaerobic digestion.

FT-IR spectra

In general, the FT-IR spectra of HAL-UGand HAL-AG resemble each other and differmarkedly from that of HA-S (Fig. 1). The maindifferences with respect to HA-S are the greateraliphatic character, the presence of strong amideand polysaccharide components, and the low pres-ence of COOH groups. The main features ofthese spectra and their corresponding assignments(Bellamy 1975; Mac-Carthy and Rice 1985;Stevenson 1994) are the following: (a) a large

Fig. 1 Fourier transforminfrared spectra of HALsisolated from untreatedgrapemarc (UG) andstabilized anaerobicallydigested grapemarc(AG), in comparison withHA of the control soil (S)

Tra

nsm

ittan

ce

5001000150020002500300035004000

wavenumber (cm-1)

UG

S

AG

28522923

3384

1226

16241717

1419

2851

2922

3389

1078

1533

1241

1652

1518

2851

2922

3394

10711232

1654

1455

1452


band centered at about 3,400 cm−1, generallyattributed to O–H stretching and, secondarily,N–H stretching of various functional groups; (b)two peaks at about 2,920 and 2,850 cm−1 at-tributed to aliphatic C–H stretching, whose rel-ative intensity in HALs is significantly greaterthan in HA-S; (c) a peak at 1,717 cm−1, generallyattributed to C=O stretching of carbonyl groups,in particular COOH, which is more intense in HA-S and a weak shoulder in HAL-UG and HAL-AG; (d) an intense and broad band between 1,660and 1,620 cm−1, attributed mainly due to C=Caromatic structural vibrations, symmetric stretch-ing of COO− groups, C=O groups of H-bondedconjugated ketones and amide (amide I band); (e)a peak at about 1,525 cm−1, due to N–H defor-mation and C=N stretching of amides (amide IIband), present only in HALs; (f) a medium inten-sity peak at about 1,450 cm−1 due to aliphatic C–H groups stretching, whose presence is detectedonly in HALs; (g) a weak peak between 1,420–1,380 cm−1, assigned to antisymmetric stretchingof COO− groups; (h) a peak at about 1,230–40 cm−1, generally attributed to C-O stretchingand O–H deformation of COOH groups, whoseintensity is much greater in HA-S than in HALs;and, finally (i) a broad shoulder at 1070–80 cm−1,generally attributed to C-O stretching of polysac-charide components, which are more intense inHALs than in HA-S.

Fluorescence spectra

The 3D fluorescence EEM spectra (Fig. 2)of HAL-UG and HAL-AG feature twofluorophores centered at excitation/emissionwavelengths pairs (EEWP) of 505/530 and280/350 nm, respectively. Differently, theEEM spectrum of HA-S is characterized bya unique fluorophore at an EEWP of 480/534 nm.Further, the relative fluorescence intensity (RFI),expressed as arbitrary units (a.u.), of the peakat longer wavelength of HAL-AG is smaller(104 a.u.) and that of the peak at short wavelengthis larger (156 a.u.) than the corresponding valuesof HAL-UG (320 and 61 a.u., respectively).The intermediate wavelength unique peak ofHA-S features an RFI value of 164 a.u. Thus,

UG

AG

550

250

EX (nm)

550

250

EX (nm)

600300 EM (nm)

S 550

250

EX (nm)

Fig. 2 Fluorescence excitation–emission matrix spectra (ascontour maps) of HALs isolated from untreated grape-marc (UG) and stabilized anaerobically digested grape-marc (GS), in comparison with HA of the control soil (S)

the different RFI values of the two HAL peaksconfirm the structural and functional changesoccurring during the treatment. Further, theseresults suggest a higher humification degree ofHAL-AG with respect to HAL-UG.

Chemical and structural characteristicsof amended soil humic acids

The elemental and functional group compositionand E4/E6 ratio of HAs isolated from varioustreated soils examined (Table 3) appear similar.


Table 3 Compositionaland functionalcharacteristics of HAsisolated from SUG, SAGand SAGM and soilfertilized with mineralfertilizer (SMF), incomparison with HA ofthe control soil (S)

Parameters S SUG SAG SAGM SMF

C (g kg−1) 563 ± 7 598 ± 4 597 ± 8 589 ± 4 587 ± 7H (g kg−1) 44 ± 1 44 ± 0 43 ± 1 43 ± 1 45 ± 1N (g kg−1) 41 ± 1 41 ± 1 44 ± 0 41 ± 0 44 ± 1S (g kg−1) 4 ± 1 1 ± 0 1 ± 0 1 ± 1 2 ± 1O (g kg−1) 348 ± 9 317 ± 3 314 ± 6 327 ± 2 322 ± 9C/N (atomic ratio) 15.9 17.1 15.9 16.8 15.7C/H (atomic ratio) 1.1 1.1 1.1 1.2 1.1O/C (atomic ratio) 0.5 0.4 0.4 0.4 0.4Total acidity (cmol kg) 760 600 610 550 680COOH (cmol kg) 480 460 480 450 480Phenolic OH (cmol kg) 270 140 130 100 200E4/E6 – 5.5 5.3 5.5 5.3 5.5

However, regardless of the amendment type, theC content of amended soil HAs slightly increases,whereas the content of O, S, total acidity andphenolic OH slightly decrease. Further, the FT IRand fluorescence spectra of HAs from amendedand fertilized soils (Figs. 3 and 4, respectively) aresubstantially similar to each other and resemblethose typical of HA-S.

These results suggest that one year after grape-marc application, the elemental and acidic func-

tional group composition, the E4/E6 ratio, and FTIR and fluorescence spectra of amended soil HAstend to approach the corresponding values typicalof native soil HA. This result may be attributedto mineralization processes of simple and low-humified components of grapemarc OM, whichoccur in soil after their application. Accordingto Senesi et al. (2007), the greater the amountof HAL in an organic amendment, the more itscompositional and structural properties approach

Fig. 3 Fourier transforminfrared spectra of HAsisolated from SUG, SAG,SAGM and soil fertilizedwith mineral fertilizers(SMF), in comparisonwith HA of the controlsoil (S)

Tra

nsm

ittan

ce

500 1000 1500 20002500 3000 3500 4000

wavenumber (cm-1)

SUG

SAG

S

SMF

SAGM

28522923

3384

1241

1380

16241717

28512923

3406

1042

1379

1419

1624

1241

28512920

3415

1037

1419

1622

1042

28512921

3401

1041

1239

1238

1624

28512921

3407

1038

1241

1376

16241716


SMF

SAGM

S

SUG

550

250

EX (nm)

550

250

EX (nm)

550

250

EX (nm)

550

250

EX (nm)

550

250

EX (nm)

600300 EM (nm)

SAG

Fig. 4 Fluorescence excitation–emission matrix spectra (ascontour maps) of HAs isolated from SUG, SAG, SAGMand soil fertilized with mineral fertilizers (SMF), in com-parison with HA of the control soil (S)

those of soil HA, and the more agronomicallyefficient and environmentally safe can be consid-ered the organic amendment. Thus, it is apparentthat soil application of grapemarc is not traumaticbut even beneficial, at least for native organiccomponents of soil.

Conclusions

The increase of pH and content of P and K,and a decrease of TOC content and C/N ratioafter anaerobic digestion of grapemarc appearsto confer to the stabilized product characteristicsadequate for its use as a soil amendment. Fur-ther, the stabilized grapemarc features larger DHand HR and smaller HI than the untreated one.These results indicate that anaerobic digestion ofgrapemarc facilitates the humification process ofOM thus increasing its stability and agronomicefficiency.

In particular, the HAL fractions isolated fromanaerobic digested grapemarc are characterizedby C, N, H, S, total acidity and phenolic OHcontents larger and O content, C/N ratio andE4/E6 smaller than those of the untreated grape-marc, thus approaching the values typical of soilHA. These effects imply an increased degree ofmolecular association and the occurrence of par-tial humification processes of OM during grape-marc digestion. Apparently, anaerobic digestionof grapemarc distillery effluents can produce anextended mineralization with loss of organic Cand stabilization of the residual OM by increasingthe content of the HAL fraction in the effluent.

With respect to soil HA, the HAL fractionsare characterized by a smaller acidic functionalgroup content, a prevalent aliphatic characterand smaller aromatic polycondensation and hu-mification degrees. Soil application of grapemarc,especially if stabilized, appear not to produce sig-nificant changes of native soil HA. Thus, a partialincorporation of HAL into HA-S is expected tooccur. Further, the low humified portion of grape-marc OM appears to be rapidly (within one year)mineralized with benefit for plant nutrition result-ing from the immediate availability of releasednutrients.


Acknowledgements This research was conducted withinthe project Parsifal of MiPAF. Thanks are due to theBonollo distillery in Anagni (Fr) for providing grapemarcsamples. The authors are grateful to Dr. Donato Ferri,Istituto Sperimentale Agronomico (MiPA, Bari) and hiscolleagues for their kind collaboration during the fieldexperiments and for providing soil samples.

References

Artz, R. R. E., Chapman, S. J., Robertson, A. H. J., Potts,J. M., De’farge, L. F., Gogo, S., et al. (2008). FTIRspectroscopy can be used as a screening tool for or-ganic matter quality in regenerating cutover peatlands.Soil Biology & Biochemistry, 40, 515–527.

Arvanitoyannis, I. S., Ladas, D., & Mavromatis, A. (2006).Review: Wine waste treatment methodology. Inter-national Journal of Food Science & Technology, 41,1117–1151.

Bellamy, L. J. (1975). The infrared spectra of complex mole-cules. London: Chapman and Hall.

Brunetti, G., Senesi, N., Miano, T. M., & Benedetti, G.(1995). Trattamenti catalitici ed enzimatici di acquedi vegetazione di frantoi oleari ai fini del loro ricicloin agricoltura come ammendanti organico-umici per ilterreno. Scienza e Tecnica Agraria, 35(13), 3–25.

Brunetti, G., Plaza, C., Clapp, C. E., & Senesi, N. (2007).Compositional and functional features of humic acidsfrom organic amendments and amended soils inMinnesota, USA. Soil Biology & Biochemistry, 39,1355–1365.

Bustamante, M. A., Moral, R., Paredes, C., Pérez-Espinosa, A., Moreno-Caselles, J., & Pérez-Murcia,M. D. (2008). Agrochemical characterisation of thesolid by-products and residues from the winery anddistillery industry. Waste Management, 28, 372–380.

Chen, Y. (2003). Nuclear magnetic resonance, infra-redand pyrolysis: Application of spectroscopic method-ologies to maturity determination of composts. Com-post Science & Utilization, 11, 152–168.

Chen, Y., Senesi, N., & Schnitzer, M. (1977). Informationprovided on humic substances by E4/E6 ratios. SoilScience Society of America Journal, 41, 352–358.

CNR-IRSA (1985). Metodi analitici per l’analisi dei fanghi.Quaderni CNR-IRSA, n. 64.

Codispoti, L. A., & Christensen, J. P. (1985). Nitrification,denitrification and nitrous oxide cycling in the easterntropical South Pacific Ocean. Marine Chemistry, 16,277–300.

Commission of the European Communities (2005). Tack-ing sustainable use of resources foreword: A The-matic Strategy on the preventing and recycling of waste(p. 32). Brussels: COM.

De Bertoldi, M., Sequi, P., Lemmes, B., & Papi, T. (1996).The science of composting. London: Chapman & Hall.

Diaz, M., Madejon, E., Lopez, F., Lopez, R., & Cabrera,F. (2002). Optimization of the rate vinasse/grape marc

for co-composting process. Process Biochemistry, 37,1143–1150.

Edelmann, W., Schleiss, K., & Joss, A. (2000) Ecological,energetic and economic comparison of anaerobic di-gestion with different competing technologies to treatbiogenic wastes. Water Science and Technology, 41,263–273.

Edwards, C. A. (2004). Earthworm ecology. Boca Raton:CRC.

Flavel, T. C., Murphy, D. V., Lalor, B. M., & Fillery, I. R. P.(2005). Gross N mineralization rates after applicationof composted grape marc to soil. Soil Biology & Bio-chemistry, 37, 1397–1400.

Geyer, W., Brûggemann, L., & Hanschmann, G. (1998).Prediction of properties of soil humic substancesfrom FTIR spectra using partial least squares regres-sion. International Journal of Environmental Analyti-cal Chemistry, 718(2), 181–193.

Grommen, R., & Verstraete, W. (2002). Environmentalbiotechnology: The ongoing quest. Journal of Biotech-nology, 98, 113–123.

Hartnett, H. E., & Devol, A. H. (2003). Role of a strongoxygen deficient zone in the preservation and degra-dation of organic matter: A carbon budget for thecontinental margins of NW Mexico and WashingtonState. Geochimica et Cosmochimica Acta, 67, 247–264.

Italy (1999). Ministero per le Politiche Agricole, Metodiufficiali di analisi chimica del suolo. Decreto Ministe-riale del 13 Settembre. Gazzetta Ufficciale n 248 del21.10.1999.

Laursen, A. E., & Seitzinger, S. P. (2002). The role ofdenitrification in nitrogen removal and carbon miner-alization in Mid-Atlantic Bight sediments. ContinentalShelf Research, 22, 1397–1416.

Lema, J. M., & Omil, F. (2001). Anaerobic treatment: Akey technology for sustainable management of wastesin Europe. Water Science and Technology, 44, 133–140.

Lettinga, G. (2004). With anaerobic treatment approachtowards a more sustainable and robust environmentalprotection. In 10th international conference on anaer-obic digestion (Vol. 1, pp. 2–12). Montreal: Canada.29th August–3rd September 2004.

MacCarthy, P., & Rice, J. A. (1985). Spectroscopic meth-ods (other than NMR) for determining functionalityin humic substances. In J. R. Aiken, D. M. McKnight,R. L. Wershaw, & P. MacCarthy (Eds.), Humic sub-stances in soil, sediment and water. Geochemistry, iso-lation, and characterization (pp. 527–559). New York:Wiley.

Mcmanus, P. (2008). Mines, wines and thoroughbreds: To-wards regional sustainability in the Upper Hunter,Australia. Regional Studies, 42(9), 1275–1290.

Miano, T. M., Sposito, G., & Martin, J. P. (1988). Fluores-cence spectroscopy of humic substances. Soil ScienceSociety of America Journal, 52, 1016–1019.

Paré, T., Dinel, H., Schnitzer, M., & Dumontet, S. (1998).Transformations of carbon and nitrogen during com-posting of animal manure and shredded paper. Biol-ogy and Fertility of Soils, 26, 173–178.

Parlanti, E., Wörz, K., Geoffroy, M., & Lamotte, L. (2000).Dissolved organic matter fluorescence spectroscopy as


a tool to estimate biological activity in a coastal zonesubmitted to anthropogenic inputs. Organic Geochem-istry, 31, 1765–1781.

Provenzano, M. R., de Oliveira, S. C., Santiago Silva, M. R.,& Senesi, N. (2001). Assessment of maturity degree ofcomposts from domestic solid wastes by fluorescenceand Fourier transform infrared spectroscopies. Jour-nal of Agricultural and Food Chemistry, 49, 5874–5879.

Romero, E., Plaza, C., Senesi, N., Nogales, R., & Polo,A. (2007). Humic acid-like fractions in raw and ver-micomposted winery and distillery wastes. Geoderma,139, 397–406.

Schnitzer, M. (1982). Organic matter characterization. InA. L. Page, R. H. Miller, & D. R. Keeney (Eds.),Methods of soil analysis, Part 2. Chemical and micro-biological properties (2nd ed., pp. 581–594). Madison:American Society of Agronomy, Soil Science Societyof America.

Senesi, N. (1989). Composted materials as organic fertil-izers. Science of The Total Environment, 81/82, 521–542.

Senesi, N., & Brunetti, G. (1996). Chemical and physico-chemical parameters for quality evaluation of humicsubstances produced during composting. In M. DeBertoldi, P. Sequi, B. Lemmes, & T. Papi (Eds.),The science of composting (pp. 195–212). London:Blackie.

Senesi, N., & Plaza, C. (2007). Role of humificationprocesses in recycling organic wastes of various natureand sources as soil amendments. Clean, 35(1), 26–41.

Senesi, N., Miano, T. M., & Brunetti, G. (1996). Humic-like substances in organic amendments and effects onnative soil humic substances. In A. Piccolo (Ed.), Hu-

mic substances in terrestrial ecosystems (pp. 531–593).Amsterdam: Elsevier.

Senesi, N., Miano, T. M., Provenzano, M. R., & Brunetti,G. (1991). Characterization, differentiation, and clas-sification of humic substances by fluorescence spec-troscopy. Soil Science, 152, 259–271.

Senesi, N., Plaza, C., Brunetti, G., & Polo, A. (2007). Acomparative survey of recent results on humic-likefractions in organic amendment and effects on nativesoil humic substances. Soil Biology & Biochemistry,39, 1244–1262.

Sierra, M. M. D., Giovanela, M., Parlanti, E., & Soriano-Sierra, E. J. (2005). Fluorescence fingerprint of ful-vic and humic acids from varied origins as viewed bysingle-scan and excitation/emission matrix techniques.Chemosphere, 58, 715–733.

Smidt, E., Eckhardt, K., Lechner, P., Schulten, H., &Leinweber, P. (2005). Characterization of differentdecomposition stages of biowaste using FT-IR spec-troscopy and pyrolysis-field ionization mass spectrom-etry. Biodegradation, 16, 67–79.

Soil Survey Staff (1998). Keys to soil taxonomy (8th ed.).Washington: Natural Resources Conservation Service,USDA.

Stevenson, F. J. (1994). Humus chemistry: Genesis, compo-sition, reactions. New York: Wiley.

Williams, P. T. (2005). Waste treatment and disposal.Chichester: Wiley.

Zucconi, F., & De Bertoldi, M. (1987). Compostspecifications for the production and characterizationof compost from municipal solid waste. In M. DeBertoldi, P. Ferranti, P. L’Hermite, & F. Zucconi(Eds.), Compost: Production, quality and use (pp. 20–29). London: Elsevier.

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