IAMAW Conference

International Association
of Mediterranean Agro-Industrial Wastes
IAMAW Conference:
Eco-innovative Solutions
for Mediterranean Wastes and Wastewaters
Ecomondo2014, 8th November 2014, Rimini (Italy)
IAMAW website: www.iamawaste.org
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WELCOME
The International Association of Mediterranean Agro-industrial Wastes (IAMAW) is proud to
welcome you to the “IAMAW 4th International Conference 2014” that has been organized
in collaboration with CTS Ecomondo in the context of the biggest environmental fair in Italy,
“ Ecomondo 2014”.
Specific focus of the conference, perfectly in line with the missions of the Association and
the Ecomondo fair, is on “Eco-innovative Solutions for Mediterranean Wastes and
Wastewaters”.
The conference aims to be an opportunity to update, discuss and compare at industrial
research level, new management models, experiences and case studies that are in
demonstration stage, in scaling-up or already applied at industrial scale.
IAMAW President
Maurizio PETRUCCIOLI
IAMAW BOARD
ADMINISTRATION BOARD:
President
Maurizio Petruccioli
Vice-president
Rafael Borja
Vice-president
Milva Pepi
Secretary
Treasurer
Francesca Santori
Ermanno Federici
DIBAF – University of
Tuscia, Viterbo, Italy
Instituto de la Grasa –
CSIC, Sevilla, Spain
Stazione Zoologica Anton
Dohrn, Naples
ISRIM, Terni, Italy
Università degli Studi di
Perugia, Italy
NATIONAL MEMBERS:
Montserrat Sarrà
Universitat Autònoma de Barcelona,
Spain
Nicolas Kalogerakis
EE-TU-Crete, Chania, Greece
Erdinc Ikizoglu
Ege University, Izmir, Turkey
Maria Pilar Bernal
CEBAS-CSIC, Espinardo-Murcia,
Spain
José Duarte
LNEG-Unidade de Bioenergia,
Lisbon, Portugal
Lorenzo Bertin
DICAM – University of Bologna, Italy
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[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
[email protected]
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PROGRAM
Eco-innovative Solutions for Mediterranean Wastes and Wastewaters
Saturday, November 8, 2014. 9.30-14.00 Sala Diotallevi 2 Hall Sud
In the context of Green Economy, all sustainable innovations in monitoring, recycling and
treatment of agro-industrial wastes and wastewaters aimed to recover and produce
nutrients, biomolecules, energy and every added-value component have a great potential to
help businesses survive the recession, representing an opportunity of a competitive agroindustrial development. In this context, specific focus will be held to Mediterranean wastes
and wastewaters where the term Mediterranean refers to climatic condition and not
necessarily to geographic location. The conference aims to be an opportunity to update,
discuss and compare at industrial research level, new management models, experiences
and case studies that are in demonstration stage, in scaling-up or already applied at
industrial scale.
Session Chairs
Maurizio Petruccioli, President of IAMAW
Josè Cardoso Duarte, Past President of IAMAW
Program
9.30 - Introduction by the Presidents and brief report on the poster presentations
9.45 - Why a Mediterranean agro-industrial wastes cluster? Thoughts and Insights
Josè Cardoso Duarte (IAMAW, Lisbon, Portugal)
9.55 - Evaluation of Mediterranean wastes: a case study for Aegean region agro-industrial
wastes
Sayit Sargın, Fazilet Vardar Sukan (Ege University, Bornova-Izmir, Turkey)
10.20 - Renewable energy from thermophilic anaerobic digestion of winery residues: batch
and CSTR lab-scale study
David Bolzonella, Franco Cecchi (University of Verona; Interuniversity National Consortium
"Chemistry for the Environment"), Cinzia Da Ros, Cristina Cavinato, Paolo Pavan P.
(University Ca’Foscari, Venice)
10.40 - Experiences of birth business from the recovery of waste in the urban periphery
Graziano Bertogli (MAN.SE.F. Onlus, Milan)
Selected oral presentations
11.00 - Measure of biochemical methane potential (BMP) of by-products, residues and
alternative crops typical of the Mediterranean area.
Mariangela Soldano, Lorella Rossi, Nicola Labartino, Claudio Fabbri, Sergio Piccinini
(CRPA Lab, Sezione Ambiente ed Energia, Reggio Emilia, Italy)
11.15 - Effect of ultrasound and microwave pretreatments on two-phase olive mill solid
waste anaerobic digestibility
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Barbara Rincón, M. González de Canales, L. Bujalance and Rafael Borja (Instituto de la
Grasa – CSIC, Sevilla, Spain)
11.30 - The new role of the Sewage Treatment Works: the LIFE WW-SIP project experience
Francesca Santori (IAMAW, Terni, Italy)
11.45 - CarbGrowth: maximisation of greenhouse horticulture production with low quality
irrigation waters
Francisco M. del Amor Saavedra (IMIDA, La Alberca, Murcia, Spain)
12.00 - Fungal laccases production using agro-food wastes: a factorial design approach
Federica Spina, Annalisa Nanni, Alice Romagnolo, Giovanna Cristina Varese (University of
Turin, Italy), Marcello Fidaleo (University of Tuscia, Viterbo, Italy)
12.15 - An agro-industrial waste valorization: biopolymer production from dephenolized and
fermented grape pomace
Gonzalo A. Martinez, Joana B. Domingos, Stefano Rebecchi, Lorenzo Bertin, Fabio Fava
(DICAM, University of Bologna, Italy)
12.30 - Thermal application for agro industrial wastes utilization
Guillermo Rodríguez-Gutiérrez, Juan Fernández-Bolaños, Ana Jimenez-Araujo, Rocio
Rodríguez-Arcos, Aranzazu García-Borrego, Fátima Rubio-Senent, Antonio Lama-Muñoz,
Abdessalem Mrabet, Rafael Guillén-Bejarano (Instituto de la Grasa – CSIC, Sevilla, Spain)
12.45 - Valorization of corn-silage anaerobic digestate through the cultivation of the whiterot fungus Pleurotus ostreatus.
Guglielmo Santi, Simona Proietti, Stefano Moscatello, Alberto Battistelli (IBAF-CNR,
Porano, Italy), Valerio G. Muzzini, Emanuela Galli (IBAF-CNR, Monterotondo, Rome, Italy)
13.00 - Effects of dilution ratio, inoculum of yeasts and concentration of ammonium sulphate
on the bioremediation of olive mill wastewater
Antonio Bevilacqua, Nilde Di Benedetto, Milena Sinigaglia, Maria Rosaria Corbo (University
of Foggia, Italy)
13.15 - An approach to split 2-phase olive pomace into a polyphenol liquid and a valuable
solid phase
Steffen Hruschka (GEA Westfalia Separator Group GmbH, Germany)
13.30 - Need to have a uniform law in the Mediterranean basin for the development of ecofriendly solutions for waste, wastewater and renewable energy
Mariagrazia Chianura (Studio Legale Chianura, Grottaglie, TA, Italy)
13.45 - Concluding remarks
Poster Session
E4-1 Degradation of 2-chlorophenol by laccase-zeolite biocatalyst / Emanuela Galli1, C.M.
Polcaro2, P. Ciccioli2, E. Donati2; 1IBAF-CNR and 2IMC-CNR, Monterotondo, Rome, Italy
E4-2 IGAN ECO-POT project / Renzo Spagnesi1, Martina Lotti1, Maurizia Seggiani2, Monica
Puccini2, Sandra Vitolo2, Roberto Altieri3, Alessandro Esposito3, Francesco
Castellani3,Vitale Stanzione3, Ermanno Federici4, Laura Fidati4, Elena Montalbani4, Roberta
Pret4, Masetti Gianna5, Vincenzo Tropiano6, Michele Bellandi6, Mario Romiti7; 1Vivaio
Sandro Bruschi, Pistoia, 2University of Pisa, 3ISAFOM-CNR, Perugia, 4University of Perugia,
5Vivai Piante Masetti Sabino s.s.a, Pistoia, 6Impresa Verde Pistoia, Pistoia, 7Romiti F.lli
Mario & Marco soc.agr., Pistoia, Italy
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E4-3 The PUMAN Model. Methodology for the quantification of the wastewater volume in
dry weather. A concrete solution to elaborate a standardized definition of priorities for
actions and investments into the wastewater collection and treatment fields / Mario
Chiarugi, Oberdan Cei, Roberto Salvadori, Gianluca Baronti, Simone Lippi; Acque Spa,
Pisa, Italy
E4-4 Orange peel wastes pretreatment by acid-catalyzed steam explosion for enhancing
bioethanol production / Guglielmo Santi, Julia Jasiulewicz, Alessandro D’Annibale, Silvia
Crognale, Maurizio Petruccioli, Mauro Moresi; DIBAF, University of Tuscia, Viterbo, Italy
E4-5 Bioremediation as alternative solution for wastes management: study on potential
phenol-degrading microorganisms / Daniela Campaniello, Antonio Bevilacqua, Milena
Sinigaglia, Maria Rosaria Corbo; University of Foggia, Italy
E4-6 Fungi isolated from olive mill wastewater: Identification, phylogenetic aspects and
preliminary assessment of their effluent-degrading abilities / Vassiliki Fryssouli1, Evangelos
Dagres1, Io Kefalogianni1, Milton A. Typas2, Georgios I. Zervakis1, 1Agricultural University of
Athens (Lab of General and Agricultural Microbiology) and 2National and Kapodistrian
University of Athens, (Dept. of Genetics and Biotechnology), Athens, Greece
E4-7 Biohydrogen production from glucose, molasses and cheese whey by suspended and
attached cells of hyperthermophilic bacteria belonging to Thermotoga genus / Martina
Cappelletti1, Davide Pinelli2, Stefano Fedi1, Davide Zannoni1, Dario Frascari2; 1Dept. of
Pharmacy and BioTechnology and 2DICAM, University of Bologna, Italy
E4-8 Production of ethanol from xerofile and halo-tolerant plant biomass. The case of
Tamarix jordanis grown in the desert and irrigated with wastewater or brackish waters /
Guglielmo Santi1, Alessandro D’Annibale1, Silvia Crognale1, Amram Eshel2, Aviah
Zilberstein2, Maurizio Ruzzi1, Riccardo Valentini1, Mauro Moresi1, Maurizio Petruccioli1;
1DIBAF, University of Tuscia, Viterbo, Italy and 2Tel Aviv University, Israel
E4-9 Next generation sequencing of bacteria involved in the static composting of olive mill
waste / Laura Fidati1, Giovanni Cenci1, Alessandro Esposito2, Roberto Altieri2, Ermanno
Federici1; 1University of Perugia, and 2ISAFOM-CNR, Perugia, Italy
E4-10 Biogas production by mesophilic phase-separated anaerobic digestion of corn-DDGS
/ Botond Ráduly1, Silvia Crognale2, László Gyenge1, István Máthé1, Beáta Ábrahám1;
1Sapientia Hungarian University of Transylvania, Miercurea Ciuc, Romania and 2DIBAF,
University of Tuscia, Italy
E4-11 Mediterranean fruits wastes as biorefinery feedstocks / Ana Dionísio, Rute Neves,
Pedro Branco, Ivone Torrado, Patricia Moniz, Talita Fernandes, Florbela Carvalheiro, Luís
C. Duarte; LNEG-Unidade de Bioenergia, Lisbon, Portugal
E4-12 Potential of Opuntia ficus-indica residues for bioethanol production / T. Mesquita1,2,
M. S. Romanovich1,2, H. T. Chaves2, F. Carvalho2, M. C.Fernandes1; 1Centro de
Biotecnologia Agricola y Agro-Alimentar do Alentejo (CEBAL), Beja, Portugal,
2Departamento de Tecnologias y Ciencias Aplicadas, ESAB-IP, Beja, Portugal
E4-13 Eco-innovatve studies in Turkey for olive waste valorisation and OMWW treatment /
Erdinc Ikizoglu, Ege University, Bornova-Izmir, Turkey
An English-Italian simultaneous translation service is available
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Why a Mediterranean agro-industrial wastes cluster?
Thoughts and Insights
José Cardoso Duarte
IAMAW
“Mediterranean” it is a relatively difficult concept to use due to its different
implications from the point of view of geography, climate, the territory,
sociological or political among others. Besides the geographical concept-the
Mediterranean sea-and its evolutionary role, either historical either civilisational
we have nowadays quasi-universal concepts using the characteristic adjective
“mediterranean” from physical phenomena like “climate” to more prosaic and
familiar things such as “kitchen” all important for defining a desired and optimal
way of living. So, how this translates into action it is a recurrent problem to avoid
misusing and misunderstanding of the Mediterranean adjective. In fact the word
“Mediterranean” is used nowadays within many pratical and important initiatives
calling for a rigorous definition of its field of application. As an example we
notice that for the first time, the word Mediterranean appears on the subtitle of
ECOMONDO even if it was more or less explicitly assumed in previous
organizations of the Fair. The problem becomes even more complex when we
try to apply it to Wastes and in particular to Agro-industrial wastes of
Mediterranean origin or character. In particular we try to elaborate in the concept
for deducing its importance and for a better definition and arriving to a
reasonable consensus on the need of keeping an active cluster within the topic
of Agro-industrial wastes for the Mediterranean type agricultural productions.
This cluster should work together to promote the science, technology and
sustainability behind the exploitation of these productions. We will conclude with
recommendations for signalling its importance to the European Commission and
other important scientific and development forums.
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EVALUATION OF MEDITERRANEAN WASTES: A CASE STUDY FOR
AEGEAN REGION AGROINDUSTRIAL WASTES
Sayit Sargın, Fazilet Vardar SUKAN
Ege University Department of Bioengineering , Bornova-İzmir, TURKEY
The economies of many Mediterranean countries are agricultural and
agro-industry based and because of rapid increase in urban populations as well
as industrialization in these countries, vast amounts of residues and wastes are
generated leading to a high turnover rate of organic material with an
undisputable economic potential which is greatly undervalued. Therefore, it is
very important to identify feasible recycling and reutilization strategies, taking
into consideration the availability, convertibility, cost, existing and potential
usages of these waste materials.
Being a part of the Mediterranean basin the Aegean Region of Turkey
contributes immensely to the Turkish economy with its agro-industrial
infrastructure and consequently enormous amounts of agricultural wastes and
surpluses are produced annually. Presently, these materials are very rarely
reutilized. The fact that agro-industrial wastes contain economically valuable or
microbially metabolizable components enable them to be converted to high
value added products through biotechnological methods.
The objective of this study was to develop a data base and to determine
the sources, quantities and qualities, the physical, chemical and biological
characteristics of the Region’s agro-industrial wastes. The recycling and
reutilization potentials were investigated with respect to the information obtained
through questionnaires and the comprehensive literature survey conducted.
Wastes were grouped on the basis of their main components, industrial sector
and possible products to be produced. Existing conventional methods of
reutilization were compared with novel methods used in other countries.
Waste/Product ratios and estimated amounts of waste produced annually were
presented for all the industrial sectors included in the study.
The study clearly shows that the Aegean Region agro-industrial wastes
and surpluses have a very promising potential for reutilization. However, the
success of reutilization depends on the regional distribution, seasonality,
quantity, economic value, pretreatment requirements of the wastes as well as
the scientific and technological know-how within the country, public awareness,
governmental regulations and incentives.
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Renewable energy from thermophilic anaerobic digestion of winery
residues: batch and CSTR lab-scale study.
Bolzonella D. 1,2, Da Ros C.3,Cavinato C.3, Cecchi F. 1,2, Pavan P. 3
University of Verona, Department of Biotechnology, Strada le Grazie 1537134, Verona, Italy.
1
Interuniversity National Consortium "Chemistry for the Environment", Via delle
Industrie, 21/8-30175 Venice, Italy.
2
University Ca’ Foscari of Venice, Department of Environmental Sciences,
Informatics and Statistics, Calle Larga Santa Marta, Dorsoduro 2137-30123,
Venice, Italy. ([email protected])
3
The production of one hectolitre of wine generates about 18 kg of marcs, 4 kg
of stalks, 6 kg of lees, and 1 kg of sewage sludge from winery wastewater
treatment (ANPA, 2001). These kind of bio-wastes are characterized by high
biodegradability, but at the same time a low nutrient content. Anaerobic
digestion (AD) represents the most suitable technology for these substrates in
terms of energy saving and CO2 emission. In this study, potential methane
production of the different bio-waste was evaluated by standard biological
methanisation tests. The study shows that the most interesting substrate is wine
lees, with specific a methane production that reached 0.333 m3/kgCOD, while
the methane production from grape marc can be increased from 0.150 to 0.301
m3/kgVS after a fermentation step. Considering the amount of grape marc
available this substrate seemed to be the most interesting winery by-product.
Thus, lab-scale semi-continuous reactor was used to evaluate feasibility of AD
of grape marc alone and with sewage sludge. The process fed with grape marc
alone showed a not suitable solution: it wasn't stable at 20 days of HRT, mainly
due to the presence of slowly biodegradable compounds with low hydrolytic
rates and to the low nitrogen content. The hydrolysis can be improved by using
a pre-fermentation step, or higher HRT can be applied to stabilise the process.
Thus, a second test at HRT=40 days, feeding the reactor with fermented grape
marc showed higher biogas production (SGP), 0.3 m3/kgVS. Also in this case,
ammonium release was not enough to maintain an adequate buffer condition.
Adoption of a co-substrate as sewage sludge seems to be the most promising
approach to follow to obtain a stable process thanks to the supply of missing
nutrients, even if lower overall SGP can be obtained due to low biogas
production of sludge.
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Experiences of birth business from the recovery of waste
in the urban periphery
Graziano Bertogli
(MAN.SE.F. Onlus, Milan)
The study RECYCLING – FROM E-WASTE TO RESOURCES prepared by
UNEP in 2009 underlines that an appropriate handling of e-waste can both
prevent serious environmental damage and also recover valuable materials,
especially ferrous and non-ferrous metals and precious metals.
In fact, S.Chatterjee and Krishna Kumar, estimated in 230 kg the volume of
saleable material (steel, glass, plastic, copper, aluminum, etc) recovered from
1,000 kg of Personal Computers (PCs) and Printed Circuit Boards (PCBs) with
an estimated value of around 12.000 $ only from PCBs contained in 1,000 kg of
PCs.
In addition to usual components, in the last 5/6 years another important group of
elements became economically suitable to be recovery from e-waste: the Rare
Earth (REM) that are used in a wide range of applications, such as automotive
catalytic converters, permanent magnets, and rechargeable batteries, petroleum
cracking catalysts, flints for lighters, pigments for glass and ceramics etc.
The incresing utilization of REM coupled with the increased reduction of export
from Cina, the main producer, resulted in a growing price. For intance, the price
FOB China of cerium increase from 3.04 $/kg in 2007 till 140,6 $/kg in May
2011, opening an important opportunity for the recycling plants.
The recycling chain for e-waste is classified into three main subsequent steps:
(i) collection, (ii) sorting/dismantling and pre- processing (including sorting,
dismantling and mechanical treatment) and (iii) end- processing. Typically, the
end-processing takes place at three main destinations. Ferrous fractions are
directed to steel plants for recovery of iron, aluminum fractions are going to
aluminum smelters, while copper/lead fractions, circuit boards and other
precious metals containing fractions are going to e.g. integrated metal smelters,
which recover precious metals, copper and other non-ferrous metals, while
isolating the hazardous substances
Developing and emerging economies can play an important role in this field so
several pilote projects, easlely scalable, can be implemented mainly in urban
peripheries taking into account the present level of technology, performance of
the recycling chains of both informal and formal recyclers and other economic
and social aspects and parameters.
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Misura del potenziale metanigeno (test BMP) di sottoprodotti, scarti e
colture di integrazione tipiche dall’area del Mediterraneo.
Measure of Biochemical Methane Potential (BMP) of by-products, residues
and alternative crops typical of the Mediterranean area.
Mariangela Soldano, Lorella Rossi, Nicola Labartino, Claudio Fabbri, Sergio
Piccinini
Centro Ricerche Produzioni Animali – CRPA Lab, Sezione Ambiente ed
Energia -Reggio Emilia
Il CRPA Lab, sezione Ambiente ed Energia, ha sviluppato un sistema per
determinare il potenziale metanigeno, definito con l’acronimo inglese BMP
(Biochemical Methane Potential), di diverse matrici utilizzate negli impianti di
biogas. Il test misura la produzione massima di metano ottenibile per
degradazione anaerobica della sostanza organica contenuta nelle biomasse,
secondo la norma UNI EN ISO 11734:2004, ed espressa in Nm3 per kg solidi
volatili. Dal 2011 il laboratorio CRPA ha effettuato oltre 1000 analisi BMP di
scarti, sottoprodotti dall’agroindustria e colture energetiche.
Nei paesi dell’aerea Mediterranea, dove è in crescita il numero di impianti di
biogas e di conseguenza l’interesse verso questo settore, l’attività
agroindustriale produce quantità significative di sottoprodotti e scarti, molti dei
quali inutilizzati e che spesso rappresentano un notevole problema ambientale.
Inoltre colture “alternative” alle coltivazioni energetiche, come l’Arundo Donax,
Nicotina tabacum, Miscanthus giganteus, Jatropha curcas, prodotte in
quest’area, riscuotono sempre più interesse in digestione anaerobica.
L’articolo riporta i risultati di diversi test BMP di sottoprodotti e scarti organici
provenienti dall’area del Mediterraneo, quali sanse di olive, pastazzo di agrumi,
scarti della lavorazione del pomodoro, colture energetiche “alternative” ecc…
Dai risultati ottenuti si conferma che l’uso di queste matrici è molto promettente
in quanto di buona qualità in termini di sostanza organica e resa in metano e
quindi idonei alla digestione anaerobica. La loro trasformazione in biogas
permette di incrementare il potenziale metanigeno di piccoli impianti di
digestione anaerobica che solitamente utilizzano solo effluenti zootecnici e
diminuire l’uso delle colture energetiche dedicate nei grandi impianti.
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EFFECT OF ULTRASOUND AND MICROWAVE PRETREATMENTS ON
TWO-PHASE OLIVE MILL SOLID WASTE ANAEROBIC DIGESTIBILITY
B. Rincón*, M. González de Canales, L. Bujalance and R. Borja
Instituto de la Grasa (CSIC). Avenida Padre García Tejero, 4, 41012, Sevilla,
Spain.
This study investigated the effect of ultrasound (US) and microwave (MW)
pretreatments on two-phase olive mill solid waste (OMSW) with a view to
enhancing its anaerobic digestibility. The effect of US and MW pretreatment on
OMSW composition and subsequent anaerobic biodegradation was evaluated
by chemical oxygen demand (COD) solubilization and biochemical methane
potential (BMP) tests. Two-phase OMSW was ultrasonically pretreated at a
power of 200W and a frequency of 24 kHz for different time periods. These time
periods correspond to specific energies of 11367 (US1), 21121 (US2), 34072
(US3), 51284 (US4), 68557 (US5) and 106003(US6) kJ/kg TS, respectively. The
MW pretreatment was carried out at a power of 800 W and a targeted
temperature of 50ºC using different heating rates, ramp and holding times. The
following specific energies were applied: 4377 (MW1), 4830 (MW2), 7170 and
7660 (MW3) kJ/kg TS.
The US pretreatments that reached the highest COD solubilizations were US1,
US4 and US6, with a COD solubilization between 48-57%, these pre-treatments
were selected for BMP tests, obtaining methane yields of: 311±15, 393±14 and
370±20 mL CH4/g VSadded respectively, while the untreated OMSW gave a
methane yield of 367±4 mL CH4/g VSadded. For the MW pretreatment the COD
solubilizations achieved were very similar in all the cases and they were in the
range of 41.1%-43.1%. BMP tests revealed that US2 (pretreated at 21121kJ/kg
TS) and MW3 (pretreated at 7660kJ/kg TS) gave maximum methane yields very
similar (393-395 ± 1 mL CH4/g VSadded), which were around 7% higher than that
obtained for untreated OMSW. Methane yield for the US pretreatment increased
when the specific energy increased from 11367 to 51284 kJ/kg TS.
Nevertheless, for a specific energy of 106003 kJ/kg TS a decrease in the
methane yield was observed. For MW the maximum value was obtained for the
maximum energy tested (MW3).
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The new role of the Sewage Treatment Works:
the LIFE WW-SIP project experience
Francesca Santori
[email protected]
IAMAW, Perugia - Italy
Rethink the common notion of the sewage treatment plant, from a site devoted
to the urban waste water collection, treatment and discharge to an integrated
platform for the sustainable and profitable refining of the organic liquid waste,
should be considered a priority to face the current emergency in terms of water
scarcity, energy recovery, economic and managerial sustainability of the whole
water and waste water treatment sector. Notably, the sewage works of the
future should integrate different approaches in order to reach environmental and
economic sustainability; in actual fact, whilst in the past the works was put in
operation merely to treat urban wastewater, losing energy and valuable
components in its waste materials, at present, due to the introduction of the
anaerobic digestion process inside the works, energy is recovered from the
process residues, reducing cost and the carbon footprint of the whole process,
going towards energy neutral targets.
Based on these premises, a wastewater refinement platform, co-financed by the
LIFE+ programme, is going to be built by integrating infrastructures, processes
and workforce of the ordinary UWWTP. It will be fed with organic liquid waste
available in the surrounding area, carried via truck and pre-treated into the
Works by using the electro-coagulation process. The sludge obtained after
primary and secondary treatment, will be digested into the already constructed
anaerobic reactor for biogas production. The biogas will be conveyed to the
CHP station for energy production. The carbon dioxide produced by the CHP
station will then be directed to the PBR, fed with N, P and K present in the
sewage works effluents. The algal biomass from the PBR will be redirected to
the anaerobic reactor or used to generate algal fuel. The final products of the
whole process will be reusable water and energy; any process residue and
effluent will be re-circulated within the treatment chain.
14
CarbGrowth: Maximisation of greenhouse horticulture production with low
quality irrigation waters
Francisco M. del Amor Saavedra - Instituto Murciano de Investigación y
Desarrollo Agrario y Alimentario (IMIDA)
Agriculture in the European Union faces some serious challenges in the coming
decades: competition for water resources, rising costs, competition for
international markets, changes in climate and uncertainties in the effectiveness
of current European policies as adaptation strategies. Greenhouse production
appears as an alternative to face some of the upcoming challenges. In 2011, the
surface dedicated to greenhouse production at worldwide level was up to
3000kha, from which 5% belongs to Europe. This production is characterised by
a climate conditions associated to fourth climate regions, which allows for
diversity in technologies and practices used for greenhouse vegetable
production. On the other hand, The EU is the main export destination with
almost one-half of the world’s imports. In the period 2002-2008, fruit and
vegetables imports have experimented a constantly growth (up to 39% for this
period).Therefore, in order to ensure their competitiveness against producers
from other Non-European countries with lower labour costs as Morocco, or
Israel, European growers need to adopt new agricultural technologies to
improve net production, ensure quality and reduce production costs. On the
other hand, European aquaculture represents 20% of the total fish production.
Major environmental impacts of aquaculture have been associated mainly with
high-output of wastewater of intensive systems. In 2009, the Commission
proposed a strategy for the future of the European aquaculture which includes
new wastewater management strategies. In order to satisfy the needs of this two
different sectors, Carbgrowth aims to: (1)increase net production and tolerance
to salinity through CO2 injection (2)recover CO2 for injection by photocatalysis
(3)reusing industrial wastewater for irrigation purposes and reducing the cost
associated to water supply, (4)reduce charge loses in irrigation
systems.(5)Develop process control to optimise irrigation, CO2 injection and
greenhouse climate control.
15
Fungal laccases production using agro-food wastes: a factorial design
approach
Federica Spina1, Marcello Fidaleo2, Annalisa Nanni1, Alice Romagnolo1,
Giovanna Cristina Varese1
1Department of Life Sciences and Systems Biology, University of Torino, Viale
Mattioli 25, 10125 Torino, Italy
2Department of Innovation of Biological Systems, Food and Forestry, University
of Tuscia, Via San Camillo de Lellis, 01100 Viterbo, Italy
Fungal laccases are excellent green biocatalysts but their actual exploitation at
industrial scale is still limited by the commercial availability of large amount of
cheap enzymes of interest. Basidiomycetes are considered great enzymes
sources and, during this study, the attention was focused on a Trametes
pubescens strain: its laccases were stable, versatile and, when a direct
comparison was possible, more active than commercial formulates. The
objective was then to stimulate laccases production by controlling medium
composition and operative conditions.
Traditional C/N sources (glucose, peptone) and model inducers resulted
functional for this purpose, but they are not cost effective. For industrial
applications, the economic balance should be indeed taken into consideration.
Wastes of agrochemical and food processes may be used as alternative
nourishment sources for fungi; their lignino-cellulosic matrix mimics the natural
growth substrate of ligninolytic fungi, being able to sustain and trigger specific
metabolisms.
Tomato sauce and orange peel waste were here considered because of the
large production within Italian and, more in general, Mediterrean boundary. At
first, the concentration of both sources (C) and peptone (N) was evaluated in
presence of Cu as inorganic inducer. A 22 plus centre point replicated factorial
design was used to investigate the role of these factors and their synergic effect
on laccase production.
Both C and N concentration gave significant effects, even though N was more
important for tomato sauce. An interaction between the two factors was always
observed. The highest laccase productivity was achieved in presence of orange
peel waste (46 U/ml).
These data should be verified, and whether possible, acquiring chemical
analysis of the wastes acquired. Further experiments are then necessary to
define the optimal medium composition for laccase production. Moreover, the
effect of other factors (Cu, pH, inoculum, etc.) has to be evaluated.
16
An agro-industrial waste valorization: biopolymer production from
dephenolized and fermented grape pomace
Gonzalo A. Martinez1*, Joana B. Domingos1, Stefano Rebecchi1, Lorenzo
Bertin1,2, Fabio Fava1
1 DICAM, University of Bologna, via Terracini 28, I-40131, Bologna, Italy
2 IAMAW - International Association of Mediterranean Agro-industrial Wastes
c/o ISAFOM-CNR, Via della Madonna Alta, 128, 06128 – Perugia – Italy
The present work was dedicated to verify the possibility of extensively replacing
the utilization of simple sugars for the biotechnological production of
polyhydroxyalkanoates by pure cultures. To this aim, the utilization of a
dephenolized and fermented grape pomace (GP), i.e., a solid organic waste of
the winery industry, was proposed. The dephenolization, which represented the
first valorization step, consisted on high valuable organic molecules recovery by
super-critic CO2 extraction. Thereafter, the pomace was fermented under
anaerobically acidogenic conditions for obtaining a volatile fatty acids (VFAs)rich effluent (GPAcid), mainly containing (g/L): acetic (14.69±0.57), propionic
(0.77±0.04), iso-butyric (0.83±0.03), butyric (4.67±0.21) and caproic
(0.55±0.02) acids. Finally, a grown culture of Cupriavidus necator was fed with
the GPAcid.
Experiments were carried out in 500-mL Erlenmeyer flasks (working volume:
150-mL) at 30ºC and 180 rpm. The whole PHA production process was
separated into two stages, namely: (a) a balanced cell growth phase, using
DSMZ-81 mineral medium amended with 5 g/L of fructose, and (b) an
accumulation phase, where harvested cells were suspended in a NH4 freeDSMZ-81 medium containing 20 or 40% of GPAcid (v/v). Higher GPAcid were not
tested since VFAs concentrations would inhibit PHA accumulation. A control
experiment aimed at indirectly evaluate water matrix inhibition effect was
performed by substituting GPAcid with a VFAs water solution, wherein VFAs
concentrations were the same of GPAcid. First results showed a higher polymer
production when employing 40% of GPAcid, with a final polyhydroxybutyrate
content of 60% (cell dry weight bases). Thermo-gravimetric analyses confirmed
gas-chromatography ones and identical results were obtained in the control
experiment.
To the very best of our knowledge, this work represents the first attempt to
produce polyhydroxyalkanoates with a pure culture of Cupriavidus necator and a
dephenolized and fermented GP as an alternative carbon source.
17
Thermal application for agro industrial wastes utilization
Rodríguez-Gutiérrez, Guillermo; Fernández-Bolaños, Juan; Jimenez-Araujo,
Ana; Rodríguez-Arcos, Rocio; García-Borrego, Aranzazu; Rubio-Senent,
Fátima, Lama-Muñoz, Antonio; Mrabet, Abdessalem; and Guillén-Bejarano,
Rafael.
Instituto de la Grasa: Spanish National Research Council (CSIC). Avda.
Padre García Tejero nº 4, Seville-41012, Spain.
The agro industrial wastes generated in the Mediterranean basis countries are
based in lignocellulosic matrix that makes difficult their utilization. A novel
system based in thermal pretreatment by steam has been successfully tested to
get the olive oil waste solid utilization. Nowadays, the technology is being
studied for the utilization of other agro industrial wastes like asparagus, grapes,
tomato or dates, besides others. Although these studies are still ongoing, some
of results are showing the formation of antioxidant extracts from the liquid phase
and antioxidant fibres from the final solid. The main advantages found using the
thermal technology are based in the solubilization of interesting compounds, like
phenols and sugars to the liquid phase, the formation of antioxidant fibres in the
solid which is concentrated in other components like proteins and oil, the easier
solid-liquid separation and the possibility of further application of bioprocess to
both liquid and solid phases. In the case of asparagus and date wastes, the use
of the thermal system allows to obtain a liquid extract with antioxidant properties
and a final solid rich in fibre, part of them with also antioxidant activities. The
olive oil wastes from the two phase extraction system have been widely studied,
and the use of the thermal treatment is being transferred to industrial scale for
the production of phenols extract and a final solid rich in oil and cellulose. The
application of a physical pre-treatment, like the thermal ones, gives us the
opportunity to get a real total utilization of agro industrial wastes that are actually
causing environmental problems.
18
Valorization of corn-silage anaerobic digestate through the cultivation of
the white-rot fungus Pleurotus ostreatus
Guglielmo Santi1, Valerio G. Muzzini2, Emanuela Galli2, Simona Proietti1,
Stefano Moscatello1, Alberto Battistelli1
1Istituto di Biologia Agro-ambientale e Forestale, via Marconi 2, 05010 Porano
(TR) Italy
2Istituto di Biologia Agro-ambientale e Forestale, UOS di Montelibretti, Via
Salaria Km 29.300; 00015 Monterotondo (Roma) Italy
The white-rot fungus Pleurotus ostreatus SMR 684 was cultivated on the solid
digestate obtained from biogas production at commercial scale from corn silage
(namely corn silage digestate, CSD), with the aim of i) producing edible
mushrooms and ii) degrading lignin. The pure fungal species was grown in
sterilized Petri dishes containing 3% malt extract and 1.5% agar. When the
mycelium formed an abundant biomass, discs of 9 mm were cut and used for
inoculation of the digestate. Each circle was placed in the center of a Petri dish
(11 cm diameter) containing 15 g of wheat straw (WS, control) or CSD. Moisture
was fixed at 70% in each substrate. The fungus was incubated for 42 days and
samples were taken approximately twice per week. The mycelium in CSD grew
as fast as in the control. Each sample was extracted with 0.1 M potassium
phosphate buffer, filtered and centrifuged. The superrnatant was analyzed for
the determination of proteins, ligninolytic activities (laccase, peroxidases) and
cellulolytic and hemicellulolytic activities (endoglucanase, cellobiohydrolase,
xylanase), while the solid residue was used for the determination of lignin. It is
worth noting that the highest activities (total peroxidases: 4.06 IU/mg protein
after 24 days; xylanase: 3.03 IU/mg protein after 17 days) were found in the
mycelium grown on CSD. Lignin was reduced by 12% in CSD and 11% in WS
after 42 days. This study indicated that CSD might represent a suitable
feedstock for the production of fungal fruiting bodies. Moreover, the reduction of
lignin percentage suggests that spent CSD after mushroom cultivation, enriched
with fungal biomass, might be re-introduced into the anaerobic digester with the
consequent increase of biogas yield, since fungal biomass is reported to
represent a viable feedstock for biogas production.
19
:
Effects of dilution ratio, inoculum of yeasts and concentration of
ammonium sulphate on the bioremediation of Olive Mill Wastewater
Antonio Bevilacqua, Nilde Di Benedetto, Milena Sinigaglia, Maria Rosaria Corbo
Department of the Science of Agriculture, Food and Environment, University of
Foggia
Yeasts can play a significant role in the bioremediation of Olive Mill Wastewater
(OMW), as some strains are able to remove phenols, thus they decrease the
impact of the wastes on the soil.
This research focused on a cocktail of 5 yeast strains (Candida boidinii and
Pichia membranifaciens) inoculated into a mixture of water and OMW to reduce
the content of phenol. The ratio water/OMW, the amount of the inoculum and
the concentration of ammonium sulphate (a coadjuvant previously shown to
increase the yield in phenol removal) were modulated and combined by a
centroid. Namely, the amount of ammonium sulphate varied from 0 to 18 g/l, the
volume of inoculum from 1 to 10% and the ratio water/OMW from 0 (no dilution)
to 3 (OMW: water, 1:3).
The samples were stored at 25°C and the viable cell count and the
concentration of phenols were evaluated periodically; the results were used as
input values for multivariate analysis through the DoE approach (Design of
Experiments). The statistical analysis highlighted that the three factors of the
design affected the yield of phenol removal as single but not as interactive
terms. Namely, the effect of dilution was linear and positive, i.e. the yield of
phenol removal increased with increasing the dilution of the samples, whilst the
effect of inoculum was quadratic and the targets experienced a higher yield
when the volume of the inoculum increased up to a threshold level (2.5-4%),
after which a further increase of the inoculum caused a reduction of the yield.
In conclusion, the results of this research suggest that a suitable combination of
the targets, by modulating their level in the system , the addition of ammonium
sulphate and the dilution of OMW with water, could increase the yield and the
effectiveness of bioremediation.
20
An approach to split 2-phase olive pomace into a polyphenol liquid and a
valuable solid phase
:
Steffen Hruschka,
Product Technology Manager, Olive Oil Recovery, GEA Westfalia Separator
Group GmbH
Nowadays, the olive oil extraction process is becoming increasingly based on
centrifugal technology. Today, the majority of the oil is produced in the so
called “environmentally friendly system”, the two phase technology. Even if the
amount of mills in three phases is higher, more and more tones of olives are
being processedby separation into oil and wet pomace instead of oil water and
semi dry pomace.
It is well known that the oil quality is better due to the higher polyphenol content
inside. But it is well known, too, that the wet pomace is difficult to handle
besides the industrial processing of cogeneration for thermal and electrical
energy production after drying the sludge.
But until today after 20 years of the application of this 2-phase-process the wet
pomace remains a problem for small and medium sized olive mills. A new
patented pending process from GEA Westfalia Separator can be used for the
de-pitted pomace in order to split this pomace into dry cake with about 60-65%
dry matter (20% of the pomace), into decanter solids with 40-45% dry matter
(also 20% of the pomace) and into a liquid without solids, only with 5% soluble
materials (60% of the pomace). But the most interesting issue is that: about
80% of the polyphenols are found in the liquid. Because this liquid does not
contain any diluting water, the polyphenol concentration in this fruit water is
higher than in any previous process; for immediate and easy use.
21
Necessità di una normativa uniforme nel bacino del Mediterraneo per lo
sviluppo di soluzioni eco innovative per rifiuti, reflui e energie rinnovabili
Need to have a uniform law in the Mediterranean basin for the development
of eco-friendly solutions for waste, wastewater and renewable energy
Avv. Mariagrazia Chianura, Studio Legale Chianura via Giordano 23
Grottaglie (TA)
Tel e Fax: 099.561.02.35
Cell: 349.71.60.314; e –mail: [email protected]
Il bacino del Mediterraneo rappresenta una delle zone potenzialmente di
maggior sviluppo dal punto di vista della green economy. La vicinanza tra
l’europa e la sponda sud del Mediterraneo, unita a condizioni climatiche che ne
favoriscono attività antropiche similari, come alcuni tipi di coltivazioni – vite,
ulivo, ortaggi – favoriscono la produzione di categorie di rifiuti che nella parte
sud non trovano reimpiego, ma che invece possono essere molto appetibili per
l’europa. Le condizioni climatiche di tutto il Bacino, favorirebbero poi lo sviluppo
del settore delle rinnovabili. Ma spesso ciò che è consentito in un Paese non lo
è in un altro, col rischio che lo scambio di rifiuti - ma anche di tecnologie per il
recupero o l’utilizzo delle fonti rinnovabili - da potenziale strumento di crescita
diventi un problema. Il mercato dei rifiuti da recupero può costituire un settore di
crescita sia per l’europa che per i Paesi dell’altra sponda, sia in termini di
risparmio di risorse vergini con lo scambio di rifiuto differenziato di buona
qualità, sia come allargamento del mercato delle tecnologie per il trattamento
dei rifiuti. Stesso discorso per le rinnovabili. Ma l’ostacolo maggiore è costituito
dalla normativa frammentaria, complessa, che spesso si modifica nel breve
periodo non consentendo di attrarre investimenti stranieri di medio lungo
termine. Occorre sviluppare un sistema di norme che abbia un nucleo comune,
con lo scopo di tutelare il bacino come ecosistema, ma anche di essere
sostegno e occasione di uno sviluppo sostenibile in settori, quali quello dei rifiuti
e delle rinnovabili, che presentano ancora molte opportunità di crescita. Una
specie di carta delle buone regole ambientali: una serie di norme con principi
comuni e condivisi sulle quali sviluppare accordi bi o pluri laterali. L’europa
avrebbe il vantaggio di allargare le possibilità di sviluppo per settori che
conosciamo già e che abbiamo imparato a sviluppare (recupero rifiuti, tutela
degli ecosistemi, produzione di energia da fonti rinnovabili) e i Paesi che si
affacciano sul bacino potrebbero trovare in questo un’opportunità di sviluppo
economico e democratico, dando impulso a temi molto sentiti come quello della
tutela dell’ambiente al fine di creare e mantenere un contatto proficuo con
l’Europa.
22
E4-1
DEGRADATION OF 2-CHLOROPHENOL BY LACCASE-ZEOLITE
BIOCATALYST
E. GALLI1, C.M. POLCARO2, P. CICCIOLI2, E. DONATI2
1 Institute of Agro-Environmental and Forest Biology (IBAF), 2Institute of
Chemical Metodologies (IMC); National Research Council (CNR), Research
Area of Rome 1,
via Salaria Km 29,300; 00015 Monterotondo, Rome, Italy
Laccases (E.C.1.10.3.2.) have great biotechnological potential due to their
ability to oxidize a broad range of substrates employed in several industrial
sectors. These enzymes show broad specificity, high oxidation capacity, and
reduced formation of by-products. Several authors immobilized laccases on
different supports, synthetic and natural ones, obtaining improvements for
enzyme applications due to the enhancement of storage and operational
stabilities.
For industrial application, carrier must be chip and highly disposable. With this
aim, we have studied the possibility of using natural zeolites as a carrier for
laccase immobilization.
Zeolites are crystalline porous solids with a tridimensional structure consisting of
SiO4 tetraedra, with some tetravalent Si atoms substituted by a trivalent Al, so
producing a deficit of positive charge in the reticulum, which is compensated by
cations. The hydrophobicity is higher with a high Si/Al ratio and corresponds to
an improved proteins binding capacity at or around their isoelectric point (pI).
Clinoptilolite is a natural zeolite with a very high Si/Al2 ratio and therefore we
used it for laccase immobilization, studying both adsorption and covalent
binding.
Results obtained showed that covalent binding was the best method for laccase
immobilization, showing pH and temperature stability greater than free laccase,
an activity of about 50 U laccase g-1 of clinoptilolite (100-200 µm). When stored
in water at 4°C, the biocatalyst retained 70% of activity after three months and
30% of activity after six months.
For a practical application, the biocatalyst was used for degradation of 2chlorophenol (0.1 mM) in batch at 25°C with 40 mg biocatalyst. Samples were
periodically collected up to 48 h and analyzed by HPLC. After this time, only a
25% residual 2-chlorophenol remained in the solution.
23
E4-2
IGAN ECO-POT project
Renzo Spagnesi, Martina Lotti, Vivaio Sandro Bruschi – Pistoia
Maurizia Seggiani, Monica Puccini, Sandra Vitolo, Dipartimento di Ingegneria
Civile e Industriale (DICI) - Università di Pisa
Roberto Altieri, Alessandro Esposito, Francesco Castellani,Vitale Stanzione,
Istituto per i Sistemi Agricoli e Forestali del Mediterraneo (ISAFOM) Consiglio
Nazionale delle Ricerche (CNR), Perugia
Ermanno Federici, Laura Fidati, Elena Montalbani, Roberta Pret, Dipartimento di
Chimica, Biologia e Biotecnologie (DCBB) Università degli Studi di Perugia
Masetti Gianna, Vivai Piante Masetti Sabino s.s.a, Pistoia
Vincenzo Tropiano, Michele Bellandi, Impresa Verde Pistoia, Pistoia
Mario Romiti, Romiti F.lli Mario & Marco soc.agr., Pistoia
Plant nursery industry makes extensive use of non-biodegradable plastic pots
(polypropylene, PP); in Italy, it is estimated an annual consumption of about 440
million PP pots which, at the end of their use, represent an expensive waste to
be disposed of; therefore, the replacement of PP pot with similar products based
on biodegradable materials, may reduce significantly the environmental impact
of this sector.
ECO-POT project aims at a cost-effectively production and successive field
validation of biodegradable pots realized in different sizes; the mechanical
properties of pots, i.e. resistance to lateral deformation and crushing, and rate of
biodegradation should match needs of both operators and consumers. In
particular, the biodegradation will be tested on plantable pot which are left intact
on the root ball and planted into the field. In this case pots are designed to allow
root to grow through the container walls and to decompose rapidly after being
planted. In addition, the rate of biodegradation will be also tested on pots which
are designed to be removed before final planting, thus speculating on the
compostability of the used containers (test ISO UNI 14855-1)
Different commercial biodegradable products, alone or mixed, will be tested.
ECOPOT project involves different plant nursery farms located in Pistoia; up to
10,000 ornamental plants, placed in pots of different sizes, in different
environments and belonging to 10 different species / varieties, will be tested.
The most important expected results are: reduction in the consumption of nonrenewable resources; economic benefits for nurseries gained in the international
markets thanks to the adoption of recognizable low environmental impact
trademarks (ECOLAB).
24
E4-3
The PUMAN Model. Methodology for the quantification of the wastewater volume in
dry weather. A concrete solution to elaborate a standardized definition of priorities
for actions and investments into the wastewater collection and treatment fields.
Mario Chiarugi, Oberdan Cei, Roberto Salvadori, Gianluca Baronti, Simone Lippi.
Acque Spa, Pisa, Italy
La Direttiva 91/271/CEE indicava gli obiettivi di qualità degli scarichi da raggiungere sul
trattamento delle acque reflue urbane (assimilata dal D.Lgs 152/2006, Parte Terza del
T.U.A.). L’Italia, inadempiente, è stata oggetto di più procedure di infrazione (2004/2034,
2009/2034 e 2014/2059) inerenti la depurazione de gli scarichi civili ed industriali in
agglomerati con oltre 2000 Abitanti Equivalenti (A.E.) al fine di tutelare la qualità dei corpi
idrici ricettori, soprattutto nelle aree c.d. “sensibili”. Una delle problematiche tecniche
legate all’adempimento della direttiva è la quantificazione del carico inquinante afferente
nei vari punti di scarico che spesso viene stimato con criteri non omogenei.
Attraverso il modello matematico proposto, denominato “PUMAN” (Portate Utenze Media
Annua Nera) è possibile ottenere una stima affidabile della ripartizione idraulica delle
acque reflue tra zone fognate e non fognate ed una quantificazione in Abitanti
Equivalenti di ogni punto di scarico del sistema, sia esso un impianto di depurazione o
uno scarico non depurato.
Per stimare la portata nera in tempo asciutto, ovvero il carico idraulico prodotto nei
singoli agglomerati o in ogni unità geografica, vengono considerati i dati relativi al
fatturato di ogni singola utenza georeferenziata presente all’interno dei confini dell’unità
geografica applicando poi un coefficiente di afflusso in fognatura variabile in base alla
tipologia di utenza.
Sovrapponendo le varie utenze con il grafo del reticolo fognario è possibile associare ad
ogni ramo fognario , con criteri logici di prossimità, il carico ivi afferente e di
conseguenza, il carico afferente in ogni punto di scarico del reticolo fognario.
Il modello “PUMAN” è flessibile ed esportabile in altri contesti dove non è possibile
procedere a una misura diretta delle portate nere. Il sistema può fornire una mappatura
puntuale anche su scala nazionale, potrebbe costituire una base-dati comune per
definire i contesti critici e quantificare gli impatti in termini sociali, ambientali ed
economici. Gli amministratori delle Aziende del Servizio e il Legislatore stesso
potrebbero dotarsi di questo strumento per individuare classificare gli interventi in base a
una scala di priorità e quantificarne gli aspetti economici per gli investimenti.
Il modello, implementato su scala reale di ATO da un gestore del S.I.I., potrebbe essere
prossimamente esteso su tutto il territorio della Regione Toscana come metodo per
censire gli scarichi diretti; se esteso su scala nazionale potrebbe contribuire ad
uniformare il settore del Servizio Idrico e consentire un’oggettiva valutazione delle
necessità, delle emergenze e delle relative priorità d’intervento.
25
E4-4
:
Orange peel wastes pretreatment by acid-catalyzed steam explosion for
enhancing bioethanol production
Guglielmo Santi, Julia Jasiulewicz, Alessandro D’Annibale, Silvia Crognale,
Maurizio Petruccioli, Mauro Moresi
Department for Innovation in Biological, Agro-Food and Forest Systems (DIBAF),
University of Tuscia, Via S. Camillo De Lellis, 01100 Viterbo, Italy
Although bioethanol production from cereal grains and molasses is a mature
technology, its application raises growing socio-economic concerns due to fuel/food
competition. The second generation approach, relying on the use of lignocellulosic
wastes, might mitigate the aforementioned competition.
In this study, orange peel waste was converted into bioethanol via a process
scheme involving acid-catalyzed steam-explosion (ACSE), enzymatic
saccharification and fermentation with the industrial strain Saccharomyces
cerevisiae F15. The first step was carried in a lab-scale Direct Steam-Injection
Apparatus under constant reaction temperature (180 °C), time (150 s) and sulphuric
acid content (0.5 % v/v) and using two consistency levels, namely a solid loading of
either 160 or 480 g/L. At the latter solid loading, the solubilization of complex sugars
and the concentrations of degradation products were lower and higher, respectively,
than those found with the former consistency level. The enzymatic hydrolysis (EH)
of residual solids from ACSE pretreatment at 480 g/L with a cocktail containing
cellulase (12 FPU/ g cellulose) and pectinase (25 U/g DM) enabled the attainment
of a hydrolysate with a fermentable sugars concentration of 34.0 g/L. The sugar
concentration of these syrups were 3-fold higher than those obtained after
enzymatic hydrolysis of samples derived from ACSE at the lower solid loading.
Fermentation processes were then conducted on a 1-L Stirred Tank Reactor (STR)
operated under repeated batch mode to compare the conversion efficiency of sugar
syrups derived from ACSE and subsequent EH at the two consistency levels under
study. STR experiments showed that the use of the higher solid loading in the
ACSE step enabled to significantly increase the ethanol content in fermentation
broths with respect to that obtained from the hydrolyzate from ACSE at the lower
consistency (15.38 vs. 7.41 g/L) with a limited loss in the gravimetric ethanol yield
(0.48±0.01 vs. 0.49±0.04 g/ g) and productivity (4.07 ± 0.04 vs. 4.23 ± 0.39 g/ L
h).
26
E4-5
Bioremediation come soluzione alternativa per la gestione dei rifiuti: studio su
potenziali microrganismi “phenol-degrading”
Bioremediation as alternative solution for wastes management: study on
potential phenol-degrading microorganisms
Campaniello Daniela, Bevilacqua Antonio, Sinigaglia Milena, Corbo Maria Rosaria
Dipartimento di Scienze Agrarie, degli Alimenti e dell'Ambiente, Università di Foggia
Nel presente lavoro è stato eseguito uno screening di 119 ceppi isolati da suolo (46
identificati più generalmente come mesofili, 44 afferenti al genere Bacillus e 25 al
genere Pseudomonas spp.) al fine di individuare batteri “phenol degrading” per la
detossificazione delle acque di vegetazione dell'industria olearia. A tale scopo sono
state allestite 2 prove sperimentali. Una prima prova avente come obiettivo quello di
verificare la capacità dei batteri di degradare i composti fenolici ed una seconda
prova avente lo scopo di individuare ceppi i cui enzimi, capaci di rimuovere i fenoli,
venissero indotti da un contatto prolungato con il substrato.
In entrambi i casi è stato utilizzato un substrato colturale sintetico minimale (Mineral
Salt Medium, MSM) addizionato di acido vanillico e acido cinnamico (a 500 e
1000mg/l) come unica fonte di carbonio.
In presenza dell’acido cinnamico nessuno dei 119 ceppi testati era capace di
svilupparsi. Al contrario, in presenza dell’acido vanillico 13 ceppi rispondevano
positivamente.
Relativamente alla prova di induzione è stato osservato che in presenza dell’acido
vanillico 48 ceppi erano in grado di svilupparsi già alla concentrazione più bassa (500
mg/l); diversamente, in presenza dell’acido cinnamico solo 3 ceppi rispondevano
positivamente.
Ulteriori indagini verranno effettuate al fine di individuare ceppi batterici da impiegare
nel trattamento delle acque di vegetazione a livello industriale.
27
E4-6
Fungi isolated from olive mill wastewater: Identification, phylogenetic
aspects and preliminary assessment of their effluent-degrading abilities
Vassiliki Fryssouli1, Evangelos Dagres1, Io Kefalogianni1, Milton A. Typas2 and
Georgios I. Zervakis1
1Agricultural University of Athens, Laboratory of General and Agricultural
Microbiology, Iera Odos 75, 11855 Athens, Greece (e-mail: [email protected])
2National and Kapodistrian University of Athens, Faculty of Biology, Department
of Genetics and Biotechnology, Panepistemiopolis, 15701 Athens, Greece
Disposal of olive mill wastewater (OMW) constitutes an important environmental
issue in olive-oil producing countries since it could significantly deteriorate
quality of water and soil receptors. OMW is generated in huge quantities within a
short period of time, and presents high contents in polyphenolics and total
organic matter, which complicates treatment by either conventional
physicochemical or biological approaches.
This study focused at the diversity assessment of OMW’s cultivated mycobiota
(adapted to effectively colonize it), and at the establishment of its potential in
degrading and detoxifying the effluent. Fungi were isolated by serial dilutions
under different temperatures and in various selective media; the morphological
characteristics of their colonies were further examined for identifying them (at
least) to genus level. Then, representative isolates from each taxon and/or
morphotype were studied on the basis of their ITS1-5.8S-ITS2 rRNA sequences.
Phylogenetic analysis of the resulting data revealed the existence of four major
complexes, including Aspergillus fumigatus, A. niger, A. tubingensis, A. flavus,
Geotricum reessii, G. candidum, Penicillium crustosum, P. commune, P.
brevicompactum, Mucor racemosus, M. circinelloides and M. fragilis.
Selected strains from each species were further evaluated by examining COD,
total phenolics reduction, decolorization and phytotoxicity values of the
biotreated OMW. Preliminary results indicated that several isolates achieved
medium to high COD decrease within a two weeks cultivation period, while for a
few strains a satisfactory phenolics reduction was combined with enhancement
of plant-seed germinability. Further studies are oriented towards optimization of
individual conditions related to the effluent’s treatment process by indigenous
fungi.
Acknowledgements: This research has been co-financed by European Union
(ESF) and Greek national funds (NSRF) through the project titled
“Metagenomics of ligninolytic microorganisms – Bioconversion of plant byproducts into high-added value products” (THALIS–UOA–MIS377062).
28
E4-7
Biohydrogen production from glucose, molasses and cheese whey by
suspended and attached cells of hyperthermophilic bacteria belonging to
Thermotoga genus
Martina Cappelletti1, Davide Pinelli2, Stefano Fedi1, Davide Zannoni1, Dario
Frascari2
Dept. of Pharmacy and BioTechnology, University of Bologna, Via Irnerio 42,
40126 Bologna, Italy
2 Dept. of Chemical, Mining and Environmental Engineering, University of
Bologna, Via Terracini 28, Bologna, Italy
Abstract:
Hydrogen (H2) gas is a clean energy source with no CO2 emissions and high
energy content. As a viable alternative fuel, H2 is considered the “energy carrier”
of the future with a wider range of combustible properties than other fuels. The
major problem in the utilization of H2 is its unavailability in nature and the need
for non-expensive production methods. Fermentative production of H2 from
carbohydrate-rich organic wastes has recently received considerable attention
thanks to its possible implementation in cost-effective processes.
Hyperthermophilic anaerobic bacteria belonging to Thermotoga genus are
considered ideal organisms for H2 production because of their ability to utilize a
wide range of carbohydrates, their tolerance to moderate oxygen (O2) amount
(up to 1%) and a better resistance to high H2 partial pressure. In this work, the
feasibility of a biological H2 production process from food industry wastes was
evaluated under thermophilic conditions. The H2-producing performances of 4
Thermotoga strains (T. neapolitana, T. petrophila, T. naphtophila, T. maritima)
were compared at 77 ºC using glucose, molasses or cheese whey as carbon
source in 120 mL batch bioreactors. Optimization of growth medium
composition, biofilm support type and substrate concentration was performed
along with the analysis of inhibitory effects exerted by H2 gas accumulation and
O2 presence in fermentative cultures. Experiments testing suspended- and
attached-cells growth conditions showed a significant advantage of biofilms over
planktonic cells with glucose and molasses. Kinetic model of H2 production was
developed for the three substrates under analysis and the suspended-cell
processes were successfully scaled-up to a 19-L bioreactor. The development
of efficient H2 production processes from industrial residues will combine
economic and sustainable energy generation with waste management.
1
29
E4-8
Production of ethanol from xerofile and halo-tolerant plant biomass. The
case of Tamarix jordanis grown in the desert and irrigated with wastewater
or brackish waters
G. Santi1, A. D’Annibale1, A. Eshel2, A. Zilberstein2, S. Crognale1, M. Ruzzi1, R.
Valentini1, M. Moresi1, M. Petruccioli1
1Department for Innovation in Biological, Agro-Food and Forest systems
(DIBAF), University of Tuscia, Via S. Camillo De Lellis, 01100 Viterbo, Italy
2Department of Molecular Biology and Ecology of Plants, Tel Aviv University,
Tel-Aviv 69978, Israel
Although the production of bioethanol from cereal kernels and molasses is now
a mature technology, the consequent and possible subtraction of resources for
human consumption continues to raise large socio-economical concerns. The
approach of the second generation, based on the use of lignocellulosic
biomasses as sugar source, appears to be the way out to mitigate the problems
mentioned above. This is even more interesting if the production of plant
biomass takes place on lands that are marginal, due to soil and climate.
In the present work, 3 different Tamarix species, namely T. aphylla, T. aphylla
“Erect”-type and T. jordanis, were grown in an experimental field under extreme
desert conditions and irrigated with either reclaimed sewage or brackish water.
Depending on both species and source of irrigation, the above-ground biomass
production ranged from 18 to 36 Mg ha-1 in the first year.
Among the three chemically characterized Tamarix species, T. jordanis was
selected due to its higher cellulose content, and lower hemicellulose and phenol
contents so as to outline a preliminary process flow sheet for ethanol production.
This included steam-injection heating under acidic conditions (200 °C; 90 s;
0.5% H2SO4; 160 g L-1 solid loading) using a novel lab-scale Direct Steam
Injection Apparatus, enzymatic saccharification (50 °C; pH 5.0; 200 g L-1 solid
loading; 20 FPU g-1 cellulose) and subsequent ethanolic fermentation (30 °C;
Saccharomyces cerevisiae F-15 as the inoculum). Lab-scale fermentation runs
were carried out in a 3-L stirred bioreactor in repeated-batch mode and showed
an almost quantitative conversion of glucose into ethanol (0.507±0.006 g g-1),
thus leading to a satisfactory overall process yield of about 145 L ethanol Mg-1
Tamarix biomass.
30
E4-9
NEXT GENERATION SEQUENCING OF BACTERIA INVOLVED IN THE
STATIC COMPOSTING OF OLIVE MILL WASTE
Fidati L.1, Cenci G.1, Esposito A.2, Altieri R.2, Federici E.1
1 University Of Perugia - Chimica, Biologia e Biotecnologie - Perugia, Italy
2 Consiglio Nazionale Delle Ricerche - Istituto Per I Sistemi Agricoli e
Forestali Del Mediterraneo - Perugia, Italy
Static composting of olive mill waste (OMW) can be regarded as an easy and
convenient way for the disposal and valorisation of such agroindustrial byproduct. We investigated the bacterial community dynamics during OMW
composting by applying a culture-independent molecular approach based on
denaturing gradient gel electrophoresis (DGGE) profiling and Illumina next
generation sequencing (NGS) of amplified 16S rRNA genes. DGGE analysis
indicated the presence of a very high bacterial diversity in both the OMW and
the composting mixture and the occurrence of a rapid succession of different
bacterial populations throughout the process. Indeed, only few populations were
shared among all the samples, while most seemed peculiar of the different
phases of biotransformation of OMW into mature compost. NGS analysis
allowed the identification of these populations. Actinobacteria were abundant in
the OMW, declined at the beginning of the active/thermophilic phase and then
represented the predominant populations during the curing/mesophilic phase.
Interestingly, while in the OMW most Actinobacteria belonged to the family
Microbacteriaceae, the compost in the curing phase featured a high abundance
of Pseudonocardiaceae, mostly represented by the moderately halophilic
Prauserella genus. As expected, the most abundant populations found in the
active phase, when the temperature reached 60°C, were the Bacilli, particularly
those belonging to the thermophilic, spore-forming Geobacillus genus. Finally,
the compost in the maturing phase was mainly characterized by an even
distribution of bacteria belonging to the orders Actinomycetales,
Sphingobacteriales, Burkholderiales, Xanthomonadales and Rhizobiales.
31
E4-10
Biogas production by mesophilic phase-separated anaerobic digestion of
corn-DDGS
Botond Ráduly1, Silvia Crognale2, László Gyenge1, István Máthé1, Beáta
Ábrahám1
(1) Dept. of Bioengineering, Sapientia Hungarian University of Transylvania,
Miercurea Ciuc, Romania
(2) DIBAF, Università degli Studi della Tuscia, Viterbo, Italia
Anaerobic digestion (AD) is a promising valorization method of bioethanol
processing byproducts. By converting these byproducts into biogas, which can
be used on-site in the ethanol production line, AD is able to better the net
energy balance of the bioethanol fuel. Key parameters to the success of the
adoption of AD in the bioethanol industry are the obtainable specific methane
yield, and the concentration of methane (CH4) in the produced biogas. On these
parameters will depend the quantity of the CH4 that can be produced by the
digestion of the byproducts, as well as the required technology level of the
biogas upgrading equipment. This study addresses both aspects by proposing
phase-separated fermentation of the byproducts of bioethanol processing. By
creating optimal conditions for the different phases of the AD process in two
distinct reactors, phase separation allows for shorter retention times and better
conversion rates of the organic matter, as well as for higher CH4 content of the
produced biogas. In our experiments laboratory-scale phase-separated
digestion of corn- DDGS (distillers dried grains with solubles) has been
accomplished under mesophilic conditions. A specific methane yield of 330 mL
CH4/g VS has been obtained, at an organic loading rate of 5.21 g VS/L/d and a
total hydraulic retention time of 21 days (5.7 days in the acid, and 15.3 days in
the methane reactor). The average CH4 concentration during the experiments
was 76.4 %, with a peak value of 81.4 % CH4. Such high methane content may
enable the use of the produced biogas in cogeneration units as is, eliminating
the expensive biogas upgrading operations. The relatively short retention time
also makes the technology appealing for industrial applications.
32
E4-11
Mediterranean fruits wastes as biorefinery feedstocks
Ana Dionísio, Rute Neves, Pedro C. Branco, Ivone Torrado, Patrícia Moniz,
Talita Fernandes, Luísa Roseiro, Florbela Carvalheiro, Luís C. Duarte
LNEG – Unidade de Bioenergia, Estrada do Paço do Lumiar, 22, Edifício K2,
1649-038 Lisboa, Portugal
[email protected]
Many Mediterranean fruits are currently used in several agro-food industries,
yielding significant amounts of wastes. The majority of these waste materials do
not have any significant utilization, being mainly used as low value energy
sources, as is the case of e.g. Extracted Olive Pomace (EOP). Due to their
putative large amounts and geographical concentration at industrial sites, the
use of these residues as feedstock within the biorefinery framework can be a
potentially advantageous alternative for their valorisation, which must be
explored and validated.
As such, the available quantities of fruit wastes in Portugal and other
Mediterranean countries were estimated and are discussed. Furthermore, five
different typical wastes, namely pine nuts shells (Pinus pinea), EOP (Olea
europaea), peach pits (Prunus persica), cherimoya seeds (Annona cherimola)
and watermelon rinds (Citrullus lanatus) are used as model materials. The
materials were subjected to partition analysis and chemically characterized,
taking special interest on their extractives and hemicelluloses content and
composition.
The comparative advantages of these materials as biorefineries feedstocks are
presented and discussed using the Biotechnological Valorization Potential
Indicator (1) and guidelines are set for their management.
Acknowledgments
The financial support of Fundação para a Ciência e a Tecnologia (FCT,
Portugal) within research contract PTDC/AGR-ALI/122261/2010 is gratefully
acknowledge.
References
1. Duarte LC, Esteves MP, Carvalheiro F, Gírio FM. 2007. Biotechnological
Valorization Potential Indicator for lignocellulosic materials. Biotechnol. J.
2:1556–1563.
33
E4-12
Potential of Opuntia ficus-indica [L.] Miller residues for bioethanol
production
T. Mesquita1,2, M. S. Romanovich1,2, H. T. Chaves2, F. Carvalho2, M.
C.Fernandes1 ([email protected])
1Centro de Biotecnologia Agricola y Agro-Alimentar do Alentejo (CEBAL), Beja,
Portugal, 2Departamento de Tecnologias y Ciencias Aplicadas, ESAB-IP, Beja,
Portugal
Opuntia fícus-indica [L.] Miller (OFI) is a culture of domesticated cactus
from very ancient times, which is important for the agricultural economies
throughout arid and semiarid parts of the world. OFI plantation has been
growing in the lasts decade in Europe, especially in the exploitation of marginal
areas or lands with very dry climates.
The fruit of this plant, known as Indian-fig or prickly-pear fruit is used in
the food industry for the production of jams and juices. From this activity two
residues are produced, the peels and the seeds. Seeds constitute about 10–
15% of the edible pulp and are usually discarded as waste after extraction of the
pulp. From the seed can be extracted oil with high value. In this work the OFI
fruit peel and the oil extracted seeds were characterized according to NREL
Protocols. Also, we assessed bioethanol production, with 5% solid loading, by
direct fermentation and by Sequential Hydrolysis and Fermentation (SHF) of
both residues.
Extracted seeds presented higher glucan and Klason lignin content than
the peels. Conversely peels presented higher water extractable content that was
very rich in sugars. For the extracted seeds, the enzymatic hydrolysis, allowed a
yield of 40% of glucan conversion to glucose. On the contrary, for peels, there
was no appreciable increase in the production of glucose in the presence of
enzymes. The bioethanol production with extracted seeds was lower than with
peels. The highest bioethanol concentration achieved was 17.1 ± 0.1 g/L, for
peels in SHF mode.
34
E4-13
Eco-innovative Studies in Turkey for Olive Waste Valorisation and OMWW
Treatment
Erdinc Ikizoglu
Ege University, Bornova-Izmir, Turkey
At the last 20 years, 165 studies at the Universities of Turkey regarding to “Olive
Waste” have been published in several International Journals. However, there
are only 2 patents belonging to Universities and 3 patents belonging to the
companies.
Beside the laboratory studies at Universities, 4 pilot and industrial plants were
constructed for olive waste valorization and OMWW treatment. All these plant
studies will be reviewed at this presentation.
35
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