A #LIFEMySOIL Open Day was celebrated in Milan (Italy) with the objective to illustrate the scalability of #mycoremediation solutions for the clean-up of aged #TPH contaminated industrial soil.
The event consisted of a set of conferences on the use of the mycoremediation technology for the removal of hydrocarbons from contaminated soils and a visit to the pilot-scale mycopile set up at the Trimper S.R.L. waste treatment plant.
This video contains the first part of the agenda: the introduction and the session on mycoremediation, chaired by Silvia Crognale and Andrea Franzetti.
00:48 – INTRODUCTION TO ENI REWIND
Guido Bonfedi (Eni Rewind, Italy)
02:00 – INTRODUCTION TO UNIVERSITA’ DELLA TUSCIA
Maurizio Petruccioli (UniTus, Italy)
06:50 – INTRODUCTION TO THE LIFE MYSOIL PROJECT
Jofre Herrero (Eurecat, Spain)
18:07 – EMERGING TECHNOLOGIES DIFFUSION AND ACCEPTABILITY
Federico Silverstri (ISPRA, Italy)
37:01 – PROS & CONS OF CONVENTIONAL TREATMENTS FOR TPH REMOVAL
Laurent Thannberger (Valgo, France)
53:18 – SUSTAINABILITY OF FUNGI IN BIOREMEDIATION
Alessandro D’Annibale (UniTus, Italy)
1:14:22 – UPSCALING AND DEGRADATION EFFICIENCY OF TPH AT A FULL SCALE
Tatiana Stella (M3R, Italy)
This project has received funding from the LIFE Programme of the European Commission under contract number LIFE20 ENV/ES/000416.
Thank you so on behalf of all of the eight partners of uh the my soil project I wish to welcome you all I’m Valentina verit to this open day and be a moment to share in the advancements of this project and I hope it’ll be an interesting moment for everyone here uh
We have a very rich program so I will ask speakers to stick to the program and their lotted time in the afternoon there will be a visit who has adhered to it later on we will provide a few indications of how the afternoon will be organized and how we will get onto the
Bus and everything so again I wish you all a very fruitful day and I will hand over the floor now to Guido boned head of environmental engineering and Market development of any rewind thank you Valentina good morning everyone it’s a big pleasure to see all of you here and all the colleagues that
Are linked via streaming I hope this could be a very fruitful morning and a very fruitful day and uh uh as in your mind I’m very glad to be here with you today uh it’s uh I think it’s a a very uh important opportunity to share the
Advancement of our project and uh so I don’t want to uh steal time to the other presentation and so uh only to thank uh our Italian partner the uh University of t with professor professor Dan and our team coordinator ER that is with us uh
Today so uh just to remain in the time I only want to leave the stage to uh Professor pet choli just to introduce it to the other argument so have a fruitful morning and the fruit b day good good work thank you if I can have the presentation
Just few minutes for for presenting the University of toua and uh the department and the research group anyway I have I am starting now okay so um I shortly present the University of toua uh in vbo and the research group involved in my soil project University T of toua is in
Vbo a middle siiz City in Lao region uh in North for Rome uh vbo although is a middle siiz city is considered the largest medieval Center in Europe anyway is famous for this aspect is approximately 80 kilomet in North for Rome and uh from the 1979 uh vbo has a public medium siiz
University university of toua here you can see some uh data related to the university um not so high number of students but very motivated uh good staff six department and very good research area in terms of research activity several National and international European project uh spin-offs uh and so
On as I uh said uh the university is divided in six departments three of which are the scientific department and uh I represent here the the department of innovation in biological food AGR food and Forest systems I am the director unfortunately of the department uh as you can imagine from
The name this dep this department is a uh multidisciplinary one in which uh are merged biological biotechnological chemical uh food technology forestry uh expertise uh as for the research group involved directly in my soil project uh it is uh constituted by microbiologist by biochemists that have accumulated expertise in applied microbiology and
Particular in applied micology uh this expertise was focused directly on the my bior remediation processes uh specifically uh addressed to uh soil contaminated by persistent organic comp pollutants uh here uh you can see several expertise and methodological approaches that we can uh provided uh of course characterization of contaminated soil uh
Under different point of view ability to isolate and characterize microorganism able to uh actively uh decontaminant actively able to decontaminate uh organic pollutants ability to use a lock on that indigenous microorganism in biog gation processes and so on uh here I wanted to list some case studies and so um contaminated sites in
Italy and Europe uh on which we uh was involved uh in uh the study of uh technical um possibility to um apply bior remediation processes and finally uh here you can see the list of the participant to the project myself Professor syia cron Alexandro danal by agricultural biochemistry that
Is the respon scientific responsible of the project and the PHD student David lelli thank you for the [Applause] attention Okay so okay I can introduce myself but thank you hello everyone my name is Jero I’m the coordinator of my soul project if the presentation just appears it will be great uh I’m
Representing EA techn technological center from Catalonia okay perfect so I have the the job to introduce you to the myel project without telling you a lot about it because later on this morning you will know more about it and then at the end of the presentation I will also
Introduce aat technological Center so my soil a step further in remediation M remediation for soul recovery so we are working mainly I know in Europe in Europe we have around 2.5 million polluted sites from them we have around 14% that need remediation and from this uh 340 sites we have around
35% that are polluted by petroleum hydrocarbons uh taking into account also the polyaromatic hroc Carbones and we also know that depending of the structure of the phes the petroleum hroc Carbones one aliphatics are more easy to biodegrad aromatics a little bit harder but then asphal and resines are the uh
More unlikely to be biodegraded going further uh currently we have H several remediation strategies the first one per se it’s not a remediation strategy it’s just land filling Excavating the soil and putting somewhere else but a lot of countries are doing that in Europe a second remediation strategy is
The physical chemical remediation which is very efficient to all type of hydrocarbons but have a hike H it’s expensive plus they needs a lot of energy and then we have the conventional B remediation what we call it the bacterial biopile which is H cheaper and consumes less energy but doesn’t arrive
To the same efficiency or removal efficiency with a long chain tphs and what we’re trying to uh demonstrate in the myel project is that micr mediation the use of fungi NOA Plus organic substrate could reach similar or yeah similar uh removal efficiencies while keeping the low energy consumption and the lower price
As the conventional B remediation so after this small introduction uh we are going already to the main objective of the project which is demonstr mro remediation which is already demonstrated a lb scale but we want to arrive to the pilot so we and we want to demonstrate
It not only that works but that it’s suitable cost effective and sustainable so we are working in this uh yeah this scheme where we start with the site characterization moving to the bability test small pilots and finally to the pilot micropiles that we are moving from one liter experiment to 1
Cubic meter till 50 cubic meter this last step is the one that we are going to visit this afternoon and also uh you will see in this map where we have our pilot sites that are mainly in Italy H we have one site in Borgo fatti
And the one that we are going to buit is s i will PR pronounce badly I’m sorry H sis Lano but of course it’s not well pronounced then we have another one in France in Ro in Rua also so bad pronounced and in Spain we have two more
That these ones I will pronounce correctly that are wela and Burgos uh going further yes with the uh I won get inside any results because my colleagues are are going to present it but what we want to uh assess it’s in the pilot scale these are some pictures of RA uh Pilots
How they build their micropiles we want to assess not only the tph or petroleum hydrocarbon uh removal tax but also we want to assess it or we want to use some Advanced monitoring tools uh mot traps that are in microcos that our colleagues from is detect are going to talk about it and
Also the phospholipid fatty acids analysis and some uh isotop compounds analysis but uh we are also want to H say something about the soil health indicators that as some of you already know the European Union are working in the soil monitoring law and uh topics like Soul functionality reduction of ecotoxicity
Microbial abundance and structor are really interesting so we don’t want only the TP removal we also want that the soil that we are going to remediate is in a better conditions than the before and then other parameters like the human health risk assessment um life cycle assessment life cycle cost and socioon economic
Impact going a little bit uh through the project we have other outputs one of the main outputs will be the creation of a guidelines guidelines for implementation of mro remediation but as you are you already see we only have five pilot sites so uh we are
Working uh with uh Noom andat to doing more replicability experiments in the uh small experiments microcosmos in order to assess the different conditions of different soils also we will work in the transferability transferability mean working nior Remediation in other uh pollutants and we are going to work with the heat thermal fluids
This part of the ACT task will be held by University autonoma in Madrid and Kepler uh the Spanish colleagues then we are going to move to the H contribution to the EU standards and regulation namely white paper where we’re going to to assess the uh legal and administrative barriers that we can
Found in order to implement micr Remediation in a efficient way and all also uh by the end of the project we want to to generate a a template for describing the best uh uh available technique or emerging technique for the M Remediation in order to increase this
Uh that the mation is more used in other contexts and finally you can check uh that we are doing some dissemination activities uh not only the online presence that we have in LinkedIn in and our web page but also we are participating in conference congresses workshops this open day that is the
Second that we are helding by the Consortium plus we are also producing some articles that recently we just got almost approved a second manuscript that we were working on it so H this is uh the Consortium uhat as already introduced de the Italian colleagues that already introduced
Themselves but then we have two more uh pairs of Partners the ones in Spain University autonoma Madrid and Kepler the colleagues that are working in France that it’s the balgo plus Noom no is from Belgium but they are working in the same pilot site and is aect from
Germany you will hear more about later so I finally have I would like to introduce some few slides about erat erat is a technology Center of Catalonia uh where uh we are working in a lot of different topics but to say some ones are circular economy decarbonization Wastewater management
Air pollution reduction am among others but basically what we do is try to fill the bridge between the basic science from the universities and others research centers to the uh companies and business so that’s why uh and here and eat is in this my uh project going further uh aat have
Different areas industry digital biotechnology and sustainability mainly I come from the soil H line and in this line we are mainly focused not only in the soil sediment remediation strategies but we are also working in groundwater H in quality monitor and risk assessment and we are also working in governance and
Social engagement plus the modelization of remediation and uh the Water within the sediment finally here is the team I’m just me today here in this open uh day but you can see that we have an interdisciplinary uh team in the that it’s backing me uh thank you all [Applause]
And I think we are moving to the second block if I’m not mistaken we are doing a small change of names thank you very much so thank you very much everybody uh so I just introduce myself I’m and Franti from the University of Milo boka and I have the
Pleasure to to to have silia cron from the University of toua co-chairing with me with um with this uh this interesting session uh so in this session we have four speaker and uh the first one is federo Sylvester from uh from ispra and and the title of uh his pres
Presentation is emerging technology and diffusion and acceptability and so I give you the floor please hello everyone uh I’m fedrigo Sy from ispra I’m a biologist and I work in two departments biology and geology Department dealing with uh remediation techniques for uh for contaminated States and agroecology and Forestry and so
More things so uh I will present you uh some something about emerging Technologies diffusion and acceptability um then this is ispra is was uh built in uh 15 June of 2016 has is was called apat and ampa before is the Environmental uh research and protection uh Institute Superior Environmental Research and protection
Institute dealing with uh environmental control and monitoring and uh technical scientific support to uh to uh uh to the ministry of the environment and in in our Institute we do also uh research control uh assessment of everything so uh uh we deal with everything uh about uh environment uh snpa is the system
National of protection agency and uh dealing with 41 National interest sites uh for for contaminated size remediation risk assessment characterization risk assessment and Remediation uh uh we also write Publications such as guidelines and reports participates in meetings and technal tables everything uh about uh environment uh issues and problems so uh
We also participate to in in the international context in life projects the impel network uh I’m dealing with in network uh in particular in U fight in uh uh uh F remediation uh and in soy Hub something about uh rizon Europe projects the unip uh that is the United Nation environmental as
In in which I deal with uh nitrogen uh questions cyle nitrogen cycle questions issues so what that is prodad out in in the context of contaminated sites sites we support the environmental Ministry of national in of national interest sites and also uh environmental Regional agencies and regions to design and testing character characterization
Idation techniques and procedures proced procedures we manage the MOs moso database that is an Innovative database dealing with everything about uh the state of uh contaminated seed sites in Italy and we prepare protocols reps and guidelines this is the states of contaminated soil remediation techniques
In Italy we have uh uh um the 32% is still uh used uh to excavate and dispose in Landfield and this is not a good thing we have uh uh 22% of containment um um procedures so only to limit the the extent of contamination and uh about the biologic Tech biological techniques
Used we have only 14% And principles uh principal used for Bio remediation land fmic and phyto Remediation so so uh uh in this case we this days is dedicated to micro micro remediation that is a very Innovative tool in fact we uh I appreciate this because there is uh uh
Finally an infill application of micro remediation because as I uh research on on on net uh on the network I only find found uh some laboratorial mesom scales examples so uh the the use of funji in in fields uh uh treatment is very Innovative and we hope it will we will
Be uh uh good and uh there will be a good efficiency because uh we have as I will show later uh a lot of examples of use of plants and a few example of use of funi but funi and plants can work also together as we can see so the my
Soil project that is has been uh before uh introduced is is in a good occasion for this in this in this sense so uh I want to uh show you something about the uh normative Innovation for soil soil is is an important uh environmental Matrix
That has not been yet very uh uh um uh study yes but we we have not uh uh direct of for soil but we uh we have we will arrive on this because the European green deal in of 2019 uh is a package of policy in atives
With aims to set uh the United the the the Europe with uh to a green for a green transition in uh in the with the ultimate goal of reaching neutrality climate by 2015 so we have a zero pollution ambition for a toxic free environment and uh the the U soil
Strategy combines with u u biodiversity strategy in preserving and Resto restoring ex ecosystems and biodiversities so this package includes initiatives including also Environmental Protection this is the the good part to to have soil in the center of uh interest for environment because we have interest principally in air and water
Before so uh for hey soils in the to to reach healthy soils uh we have uh an introduced to a new soil monitor in low that is been that has been proposed to increase the value of soil and its resources we have uh something in on on on July of this year
An introduction a new uh soil monitoring l low that points the in which soil is in the center of uh interest so uh we have uh the objectives is very ambitious to reach hey soils by 2050 and we have is the soil monitoring law uh provides monitoring assessment uh
Sustainable soil management so uh to uh deal with contaminate size and then a reporting to inform public and uh and evaluation this is the problem is that in Europe in in Europe we have 60 to to 70% of soils that are currently unhealthy and we have to uh
Solve this problem so uh as nature based techniques uh as I was telling before FY remediation is one of the most uh known and used is U is pro uh is providing in Grill de for you s strategy for 2013 is the most is is one of the most
Sustainable options to to to decontaminate soils is the more sustainable and one of the most widely used most W widely used you know what is phyto remediation but he has some paths and uh one thing to say that plants that not do not work alone to to remedies so
But need funi and bacteria but in particular funi and this in this in this context is important to to link funi to plants favorite areas for phyo remediation are inherited environmental liabilities like off sites we use phyto remediation where there is liter interest in the area sites with the disproportion with with
Intervention cost and the commercial value uh wi spread contamination in rural areas and we have when we have naturalising and Landscape values to promote uh naturalis landscape wues I we have some two example of national interest sites with phyto remediation activities in by since some uh years is banol Coro
In Campania and I will show that if uh we use plants in com in combination M with m a funji to solve Organics and heavy metal contamination or also here in Lombardy bfar to we use is use contamination uh phyto remediation to for uh contaminants as PCB pcdd arenic and and
Mercury in particular the rise of degradation for PCB and pcdd and this is very interesting because uh uh roots of plants act together with funji in mikiz with mise and this is the case of Rod degradation for Organics like the my soil your my soil projects so um going
Uh going all this I start the last uh we point attention of new emerging remediation technology as phyto mining phyto mining is uh Mining and recovery of uh critical raw materials one of the most uh important issue of this years and phyto Mining is an environmental friendly techniques for
Extracting metals from ORS and contaminated sides uh as uh this these techniques that use plants to absorb in in the aerial Parts uh these critical R materials that I will show you just now as a as an ecological techniques to extract where traditional mining is not competitive the if you don’t know what
They are critical raw materials are raw materials of high economic importance for in the you in the in Europe with high risk of Supply due to their concentration source and lack of good uh and we have uh affordable substitutes so uh there is a European project in which
I am involved that is the San uh it is a plan for sustainable resource extraction to all towards the responsible sourcing of critical raw materials in protected zones this is is important and this project promotes the use of nature based solution to recover the this critical raw Mater raw material inside uh natur
2000 SE parks and and something like this because we have a need of these materials and we have the problem to uh extract this also in natural areas uh prefer and we prefer to use natural uh ways to extract so how we extract uh these crotical raw materials by uh acum
Accumulators plants that are commonly used uh and they take metals from soils and translocate to the roots and aial paths uh then uh after the upper accumulation the the aial parts are Harvest and this osed to recovery and there there are some extraction techniques that recover these materials
From uh from uh shoots Roots leaves and and so on uh these are examples of uh uh some vegetable vegetable used commonly to recover critical room materials and there is the efficiency of recover indicated the iury there ugand Dasia like no and some Fern deop linearis that recover a lot of critical
Room materials for contaminate M ARA this is a good uh example of fola micana and phip PR prium Simplex that that have also good results why phom mining is a good emerging Technologies or remediation uh because have uh an economic viability and potential for diffusion their stength is the biomass
Production uh the recovery of syml in soil and plant matter and the effectiveness of enrichment extraction Pro process the United States Department of energy suggested that the extraction of material containing uh at least 300 millgram on kilogram of critical R materan may be profitable for recovery
So the research uh on This Ss is going on uh another emerging emerging remediation technology is the one to rec to disrupt the natural origin ancestors this is a very Innovative uh technology because we have a problem as you know of obsess of fiber fibers widespread everywhere and some Studies
Have enlighted that some bacteria liens and also funi as I show you in the next slide and the prod and the production of natural acid from plants are able to remove large quantities large quantities of heavy elements reducing the danger of these fibers how do they uh act the they
Act uh REM removing iron and disrupting as obsessors obsessor fiber and uh so uh uh we arrive to have some obsessors Fiber not toxics due to the activity metabolic activities of bacteria funi and Le leans so uh this is the Cy the V virtu cycle the iron supports plants growth Plants
Provide carbon source for microbial community and microbial interaction reduce remove irons and reduce tox toxicity of course these are the studies are at laboratory scales we have not some infield application but as your uh study of for micro remediation also this uh aspect of removal of natural obsess
Abestos fiber should be uh considered in in the future so uh I show you that funi are po potential agents of bior remediation and uh how they they work fungal I can form a web te a web of te strands that bind abess those fibers making them less liable to escape in the
Air so we use the natural capacity of uh funi of making this the this these strs and uh to capture obesso fiber so uh uh for for the professor of Tusa I show you some examples of funi that are used fusarium oxis Morella helina and endrum
Myus as f as funi use in for this uh for this uh objective so I conclud you T question is emerging technologies have a societ societal acceptability and I think yes because nature-based Technologies are considered environmental beneficial cost effective with I societal acceptable and in alternative to Conventional pH physical and chemical remediation
Processes and this was tell in 20 25 and five by pylon and Smiths the trends for acceptabilities are the importance this case of use of plants but also use of funi is is similar of uh cost effectiveness no risk for environmental and health uh increas of in green areas and reforestation if we
Use plants in combination with funji and maintaining of soil structure increase in increase on biodiversity because this studies as my soil the the uh advant advantage of increased biodiversity and Improvement of landscape per perception by by by people and uh finally but uh but is not the least the contrast of
Climate change so uh this is what uh uh these are the conclusion of of of my uh of my intervention thank you for the [Applause] attention thank you so thank you very much fedo for your very complete overview of natur based Sol solution applied to to soil health I asked the
Organizer but I guess that the discussion will be at the end of the morning session so keep your question for for that time and so it’s a pleasure to introduce the second speaker of this uh of this session so the talk is entitled Pro and cons of conventional treatment for tph
Removal and will be given by Lauren tberg from uh from valgo group in France yes the floor is yours thank you so um I’m representing uh remediation Works company I’m a research director in this company and my job is to transform the academic knowledge into
A appliable Kno how we we we need to ask question to the scientist how could we manage with this and that’s why I’m participating to the to this transfer uh project uh it could be useful for the future uh for us and I will show you how
We are going for this uh I will repat a bit what the um JRA has spoken about if we check this diagram of the frequency of type of pollutant uh if you add the classical pH or classical petroleum hydrocarbons pH betex uh petroleum compounds represent
More than 50% of uh the number of poon we are encountering on on the site so because everybody use uh hydrocarbons even if you have not an industry for hydrocarbons everybody every type of Industry also private persons after for the heating for the car so there is production plant storage distribution
Until the tank you have in your home or in your car so this everywhere isus and we are facing very often to this uh issues with this pollutant um because it’s very common pollution uh we developed a lot of techniques depending uh I will classify them by the technique physical chemical and
Biological and also depending on the Matrix where you will find those pollutant in gases in liquid pure faes dissolved in in the gr water or maybe absorb to the solid Matrix uh for example as soil gases uh you may physically remove them by soy Vapor extractions is one of the name of the
Venting technique chemically you may have uh catalytic oxidation you pump and then you transform then on a platin form for example there different type of uh techniques uh if you have liquid pure phase the best way first to remove it is to pump and separate the face and to
Pump only the oil or the gasoline you have in the in the soil um if the components for example betex or low uh short chain tph you may uh sparge because they’re dissolved in in the face you’re putting BS in the water to to remove it remove them from from the
Water and also diffuse some strong oxidant to make a chemical o oxidation Inu or you may buy the same type of bubbling you may create a bio paring that stimulate uh the endogenic uh bacteria or mic microorganism and so stimulate natural biodegradation some sometimes it’s very
Suitable um we will then Focus about the absorb phase because fungal remediation will Target this uh this part of the tph pollutant I will put away the around 50% of D and Dam technology that’s just make a hole and put this to the Landfield that’s not the scope of the presentation
Today um a physical technique uh could be applied intitute or onsite we we may uh have termal dis options is one of modern recent technique achieved on the field in the last 10 years uh not before it’s a very strong possibility other physical possibility is to make a soil
Washing you just flush the soil and separate uh the coarse grain sun and grvs from the fine and Clay fraction containing the hydrocarbons and also sometimes the metals uh so it’s a very efficient way to to do this uh chemically if you want to add some
Oxidant you have to to prod produce a soil mixing is much harder than just injection you have you need special device to to mix with the soil um um biot treatment could be bioventing vation into the soil to stimulate the the microorganism in place or just build a biopile a type of uh
Solid Biore reactor and I will focus a bit of this this line to just to show you what what it is how it’s going uh if you want to apply a venting bio venting it’s simple to install a pump and then a Wells or screen Wells to
To create a Air Stream fresh air is coming with oxygen oxygen is stimulating the the biological life and you uh you you could uh um uh enhance the biod regulation in the same time if you have volatile comp comps they will go through this flow of air and could be extracted
So you need filters or sorry or vapor treatment unit after your extraction so on the field you will see some manifold like this because you need several Wells depending of the physical properties of the soil you have to to create a network of several uh Wells venting Wells it’s
Very simple technique you may run it for for for months um it’s used for maybe 20 or 30 years so it’s considered to be a very mature technique uh whatever you do if you apply biology or just physical properties you remove the existing Vapors some toxic Vapor that could
Accumulate before you start the work uh but it’s only appliable for enough volatile components uh depending of the pressure PR uh partial pressure uh vapor pressure you some compounds are not uh um feasible by by this technique it could last very long you sometimes we put a machine
During month during the building of the next uh of the next Factory or next houses uh could be longer than the time of the real work of building and physical properties of the soil strongly influence your results if you have a too clay soil you will never
Uh S have a suction effect uh big enough to produce a fast removal of the pollutant and also could have interaction for example for a very organic soil interaction between the organic Matrix and the pollutant and whatever the type of blowing you may produce you will have a low removal of the
Pollutant uh so we know that uh uh heat is influencing the volatilization and that’s uh the development of thermal dsorption uh when chemicals are not volti volatile enough we are able to remove pollutant more than C40 even if they not in in the in the legal Target
But we we know that it’s appliable at the extreme to heav or heavy pH um so the the uh panel of the target is broader uh it’s faster we are going on field for several months uh we may apply it uh in C2 we direct the burner
Inside the soil or in pile this is a big thop pile we made in the sou of France and the efficiency you just stop the heat when you overwhelm 95 or 99% depending of your of your objectives uh I told about the diversity of ation you may apply gas electricity
Fuel depending of what you have on site but the uh POS is um disadvantages expensive because of the cost of energy and in the last two years is worse than ever um and energy consumption as a cost and also environmental cost because of gas House of green house gas
Emission and at the end you have a completely dead soil cooked during month at more than 200 degrees for example so is no life in this no carbon more and it’s only a mineral part of the soil and it’s long to uh re Implement some life or something that allows just seeds to
Germinate I made some test and it’s it’s it’s quite long I told you about soil washing this is a a picture of from a lab test in one step you have this sun very clay with EVS from a lagoon and you turn it to a clean sand and you have the
Oily part is floating uh on the beaker this this is the type of machine we are using this the first one we had uh before um the first point is when you create such sand it’s valuable material you may reuse it in the next building or sell it sometimes it’s possible
Ible uh there are different effect and by chance all the effect are concentrating the pollutant in the same clay fine Po and also solubilization if you have some metals for example you may precipitate them at the end so the sludge contains the different type of pollution so it’s a convergence between
Oily and metallic pollution um but it’s limited by the clay contain if you have more than 20 or 30% of clay you don’t want to apply this technique uh because you make you don’t make profits it could work but uh the quantity of waste is too big regarding the amount of
Of the starting polluted soil it’s relatively slow this this unit is nominative 60 ton per hours but better to run 20 to 30 tons per hour so so if you have to several thousand ton of tons you will stay for very long time on this
Side to to to treat and because of the long time energy the need of people uh working on this uh plant it could be expensive and you produce also some concentrated po extract that you you have to destroy or put to a Landfield and it could participate to the
Price um I told you about the possibility of chemical action direct Inu by mixing some reagant into the soil so there are different type of machine uh those machine are not coming from the remediation work they are coming from the geot techniques so they are uh suitable for different type of soil we
Are just derivating their use normally made for uh construction uh so there have different vertical orer horizontal mixer or the lot of types depending of the soil Matrix and just we we modify with with a to to to the uh subcontractor company don’t inject uh cement or lime just take
An oxidizer or a strong reductor depending of the pollutant you want to Target and that’s the way we we are just playing this technique um but when you have several components in your soil you can’t mix several reagant you can’t mix for example an oxidant and a reductant
In the same time so it could be hard um the uh long-term efficiency is hard to demonstrate because you have not direct access you have only indirect survey of what happens so the chemical re reactions that happen in the soil uh you have to to to check them from far and
It’s not always uh easy uh depending of the weather too dry is not good too wet is not good so and also the type of soil if you have big boulders in the soil or building you can’t access under the building with this technique so the the number of case
Where it’s uh suitable is not so uh uh often uh finally uh bio biod deoration you may Implement that in biopiles some are very rough some are very technical uh depending of the country where you apply it uh relativ cheap you just put them in a pile create this reactor and
Let the microorganism work that’s not your engineers that are destroying the pollutant um it’s reputed to have a low environmental footprint and it respect biodiversity sometimes it enhance biodiversity uh because in renewing ring the soil but there are some uh uh cons also that could spend for very long time there are
Some works we are just starting for six months and every six months we are receiving or ending one of the pile and the total of the works is two years so in some case you don’t have the time to to spend two years on site uh some
People need to work on this place or to leave on this place so you have to liberate the Zone uh it’s also sensitive to weather we know that when we when we are starting in a summer sometimes the degradation is lowering during the winter and it’s no today it’s not
Efficient or not cost effective to heat biopile and um the bioavailability of the uh pollutant is seems to be the strongest limit you have the same molecule so appears to be both the same but one is removable and not the other one because of the interaction of this
Pollutant with some part of the organic Matrix of the soil and it’s very hard to anticipate it before you go on the field to to apply this so we have different advantages or disadvantages if I con compare the biodegradation and the thermal disruption that is reputed the strong
Way to remove tph from the soil and what we want is that uh M remediation hope to be all the good tick or whatever the lines and some of the points are already already demonstrated uh and some of them have to be demonstrated during the life
Myol project and that’s why I leave the floor to the next [Applause] presenters thank you luren of this uh for your for this overview of practical on practical approach on remediation and now for me it’s a pleasure to invite Professor Alexandro danal my colleagues and coordinator of Unitus steam in my
Soul project thank you okay good morning to everybody uh this short speech is expected to provide valuable argument related to the suitability of sa trophic filamentous fungi in bior remediation a brief foreground before going straight to the point uh the susceptibility of persisting organic pollutants to the biodegradation is obviously
Dependent is OBS dependent on their degree of exposure to appropriate microbial bio catalysts however in erogeneous system such as soil this is hindered by the propensity of these compounds to escape the aquous microhabitats of degrading organisms and this occurs through a variety of mechanism which include chemical precipitation sorption onto
Abiotic surfaces partitioning onto organic matter and finally interment within micro and miso soil port this obviously leads to a decline in their bi availability and to the occurrence of local accumulation phenomena and such accumulation frequently occurs in adverse environments characterized by the lack of appropriate nutrients electron acceptors and moisture conditions able
Supporting uh the contaminant degrading ability of this organisms thus an ideal biodegradation system should be able of performing a thorough exploration of the contaminated environment reaching even soed and poor entrapped contaminants moreover its catabolic capacity would not depend on the availability of contaminants as gross substrates but would be
Maintained by other compounds or by trophy Supply uh by plant rages it would also to be active under a wide range of conditions being able to perform a long range transport of water nutrients electron acceptors and even catabolic microorganism to contaminate the St spots sorry well sopy filament fun
Satisfy most of these requirements and thus in the last decades um the potential of this organism for bior remediation purposes uh have been increasingly recognized and the reasons why fungi can be excellent candidates in bi remediation is due to several peculiarities that these organisms exhibit first of all their exploratory
Growth mode which has a variety of repercussions which will be analyzed shortly then their independence from using contaminants as gross substrates so uh although several fi are able to exploit contaminants as energy sources in the majority of cases uh contaminant degradation has a CO metabolic character another Point calls into question the
Low specificity of some of their contaminant degrading enzymes and this is very functional to the removal of mixtures of um structurally related contaminants although this is not an unique prerogative of fi they are able to produce contaminant mobilizing agent and they are also able to perform the accumulation or hyper accumulation of
Metals and metalloids and this is useful in mixed contaminated scenarios such as military Refinery and Landfield waste sites last but not least several contaminant degrading species are edible species and thus the they satisfy to General regard is safe requirement and furthermore their production compost represent an affordable and ready to use in
Opulent filament fi typically exhibit a multicellular growth mode characterized by the presence of tubular elongated and Branch structures called I this I tend to form a network of interconnected and intertwined filaments that collectively make up the melium and the melum network organization enables these organism to perform an effective colonization of
Soil and to penetrate inside soil Aggregates the hore growth mode and the turo pressure developed at the Eiffel tip enable these organism to reach regions which are inaccessible to bacteria and to penetrate inside Aggregates fungal life have an extensive surface area and their biomas in soils generally range from 50 micrograms to 1
Mgram per gram of dry soil and in addition several fungal species are able to develop threadlike structures made up of parallel hi able to grow through vast distances across adverse environments allowing for the colonization of areas which are located very far from the original my sours not to be forgotten is the fact
That unlike bacteria the dispersal of The Melia does not require the presence of continuous water phases in addition to their heal growth mode filamentos fi acquire an additional chance to come into intimate contact with contaminants from the secretion of low molecular system reach amphilic proteins called hydrophobins these proteins tend to form
Amphipatic layers at the interfaces thus affect the wetability of the surfaces and mediating both addition and surface modification the ab the fungal ability of modulating surface hydrophobicity exerts a positive impact on their capacity to adhere onto hydrophobic surfaces Ting hydrophobic contaminants the melan network represents also an excellent dispersion tool for
Bacteria in particular this network can act as a highway through which motile bacteria rapidly translocate or as a Subway in which Thea just adhere to the EI being passively displaced according to the highway mechanism the water film on the Eiffel surface offers the chance to contaminate degrading bacteria
To rapidly move whereas the movement of bacteria in unsaturated environments such as the Vos is limited by the availability of continuous water Pathways a f mechanism involves the ability of the material networks to act as pipelines through which contaminants are upt uptaken and then transferred bya liit Rich vesicles with
The coplas cytoplasmatic streaming whatever the mechanism whatever the mechanism the masan networks give the chance to increase the frequency of contact between contaminant degrading bacteria and lipophilic contaminants D mitigating problems related to the low bioavailability of these contaminants and to the heterogeneous distribution of contaminants in soil Fungo life moreover and their
Expression products such as extracellular polymeric substances along with plant roots can promote the formation of stable soil Aggregates enhancing soil soil a structure and porosity this can lead to improved Iration water infiltration which can in turn support the activities of other microorganisms involved in the bio remediation process
In addition fungi produce a set of biotic compounds among which and relating to biodegradation to bior remediation surfactant and fungal enzymes are particularly relevant surfactant are reportedly anilic compounds able to perform the mobilization and P solubilization or liopic contaminants this table so this bosfa and producing cap capacity is
Rather widespread among funi and in particular among hydrocarbonoclastic species and the the this table shows how the structure variability characterizes the production of these kind of compounds and we can find mono glyer oligopeptide Theos and Soros lipids polymeric complexes so and um their surface tension reducing ability and critical missile concentration values
Valuably compete with surfactant of bacteria origin such as surfacing from basilis or rids from poninos okay and now to the enzymes several as myties and above all myties belonging to the ecological group of white Ro fungi possess an efficient Li degrading Arsenal which is in part responsible for their contaminant degrading capacity
Um this fungi do not recover energy from the degradation or green but through its degradation they acquire a better accessibility to cellulous and hemicellulose microfibrils the link between ligning Decay and bior Remediation potential of this organism is represented by the high degree of of structural similarity which between liing sub
Components and several aromatic contaminants this Li integrating Machinery exhibits several peculiar properties first of all a low substrate specificity this means that they’re able to perform the oxidation of wide array of contaminants secondly there are products and this means that their action is performed in the extracellular environment and this means that
Contaminant uptake is not a rate limiting factor in biodegradation the reaction mechanism either rely on radical mechanisms or on the production of Highly diffusible uh oxidants which can reach nonbi available Target contaminants and some enatic components of this Arsenal is such as for example laas cobbi dehyrogenase and Quon reductase are able
To produce Phantom ingredients that’s producing hydroxy radicles this table shows a list of the main components of the LI degrading system of fi which mostly include multico peroxidases and H peroxidases these enzymes share the action mechanisms based generally on the monoelectronic oxidation of aromatic contaminants and also on their ability
To extend their substrate range by using electronic mediators just such as for example aroxy radical or dixy benzil cion radicles in the case of laas and lium peroxides respectively also manganes peroxides which brings about deoxidation of balent manganes producing manganic elates which act as mono electronic oxidant is able
To extend its substrate range bya the use of a coox such as for instance Tate an iance and hop peroxy radical more interestingly there is a group of hlate peroxidases called unspecific peroxygenases which are able to perform the hydrogen dependent terminal or subterminal Hydrox silation of alkanes
So there is also a component of this ligning degrading system which is able to act on alifa compounds okay thank you thank you Sherman in addition to extracellular enzyme systems fungi also exploit cell bound and intracellular enzyme catabolic enzymes among which cytochrome p so Mis function cyto
P45 monoxygenase is one of the main components in the initial phases of attack uh this enzyme is able to perform the epoxidation and Hydrox silation of numerous contaminants with relevant examples being polycyclic aromatic hydrocarbons and dioxins in addition to so um there are several h satr p for under 50
Monoxygenase encoding genes in fungal species and these warrants are huge variability with regard their catabolic capacity okay I have to of course although some fungi are able to exploit uh organic contaminants as energy sources they their attack on this contaminants can occur extracellularly or intracellularly um metabolites or intermediates generated during extracellular oxidation
Can be further converted intracellularly or otherwise can interact with Organic Soil constituents through oxidative coupling reactions giving rise to the so-called bound resides um metabolize derived from intracellular initial attack can be excreted as such and that’s processed according to the extracellular pathway or otherwise can they they can also
Interact with the organic constituents giving rise to the formation of bound reses but uh some products can be also conjugated giving rise to meal Cil sulfate and gluton derivatives and being is created and being generally environmentally persistent or otherwise they can be further catabolized as it occurs for example for
Aliphatic compounds but also for polychoral bifs thus leading to their mineralization going to rapidly to conclusions because my time is over H although fi can be excellent degraders of persistent organic contaminants there are still few experiences at the FI scale and most of these experiences have been focused on
Wood preservation sites and Military sites thus targeted to the degradation of creot which is a mixture of polycyclic aromatic hydrocarbons and chlorophenols and to Nitro substituted explosive such as TR Nitro toin or TR Nitro triin and some of these Studies have highlighted the failure of this treatment to reach cleanup levels within
A reasonable treatment time or the lack of significant differences with the noninoculated controls and in this study however the fate of the inoculum and its interaction with the indigenous microbiota have been often neglected or even ignored um however the outcome of a micro remediation intervention strongly depends on the interaction of the
Applied inoculated species with the autous microbiota thus being able to for example to adverting the formation of dead end metabolites um but we can be optimistic since the advance and ensuing diffusion of I throughput approaches mainly relying on omic techniques are expected to provide an increased critical body of
Knowledge able to support technical choices related to the up on effective micro remediation treatment thank you for your [Applause] attention thank you Alexandro for your detailed presentation and now is uh uh the moment of last presentation for this session uh it’s a pleasure for me to introduce taana Stella uh from uh uh
Three m3r company the floor is yours so so thank you syvia for introduction yes um I’m Tana working can you hear me okay I’m Tana Stella from MTR uh I’m the executive director and MTR is a company based here in Milano and uh that is working with any rewind
In many uh projects dealing with bio remediation fyto remediation and of course micro remediation but I want to say that I’m coming from the group of morito beta Alexandra sorry it’s emotional for me and Sylvia so it’s really nice to meet you here again and I’m going to present
Something about the upscaling and theg gradation efficiency of total petrum hydrocarbon uh contaminated soil treatment at full scale so we are speaking about another scale not anymore uh laboratory or Pilot um as you know uh tph are maybe or maybe yes the most spread uh contaminants in the environment and
Especially in contaminated soil and of course how we can treat them uh we can apply inito technology if it’s possible it’s it’s of course uh the best choice because it means that we don’t need to excavate the contaminating Matrix however most of the time is not possible
So we are applying X to technology so we need to excavate the soil and when we speak about bio remediation ex2 Technologies of course we have to mention biopile which which is a technology that exploits the ability of microorganism fungi uh bacteria to degrade organic pollutant under aerobic
Condition and uh we can choose sorry this okay we can choose between two different technology we have the dynamic technology and the static one in the first case the oxygenation of the contaminating Matrix is due to a periodic mechanical turning of the contaminating Matrix on the other side
When we apply static biopile the oxygenation of the pile uh is due to a fixed aration system so these are the two technologies that we can use for the treatment of tph contaminated soil but when we have to face with this technology at industrial scale of course
We have to think about so many parameters okay that I want just to summarize in these three categories the first one the chemical and physical properties of hydrocarbons because of course we have different classes of hydrocarbons with different volatility toxicity solubility and uh you will see later
There is a point the weathering of the contamination and then of course we are speaking about biobase Technologies so we have to assess the microbial community properties and metabolic capabilities and of course we have to assess all the environmental condition that can impact on the growth and the
Activity of the microbial Community for example temperature pH but also the content of nutrients electron donors and acceptors so these are all the parameters that we have to assess before designing an efficient treatment at full scale going to chemical and physical properties as I mentioned before the weathering what is the weathering
Um n naturally several physical chemical biological processes can occur okay without any kind of uh intervention and what is the results the results is a modified hydrocarbon mixtures and as you can see here I just prepared two uh profile the first one contamination because it’s rich in Alcan
That is um the the the category I would say of hydrocarbons that are easily biodegradable so this is a fresh contamination that we can somehow easily degrade by uh biological techniques but in this case you see there are not alkanes anymore it means that there is a hold contamination where the light
Fraction the more volatile are are already removed from the mixture so it’s more recalcitrant to any kind of biological treatment so the assessment of the weathering that is possible by fingerprint analysis is a priority before designing a bi remediation approach and then microbial Community as I said we are speaking about biobased
Techniques and uh we can we use biomolecular analysis DNA based techniques because this Tech allowed us to see how many and which kind of microorganism we have already in the contaminating Matrix so we applied both qualitative analysis Next Generation sequencing so we can see which kind of microorganisms there but we can also
Perform quantitative analysis uh of Target genes it means genes that are directly involved in the biodegradation process so we can combine these uh technique and to gather a lot of information about the microbiological um status of the contaminated Matrix and as I said before we have to assess all the environmental
Conditions that can impact on the microbial growth and activity and how we can optimize we can use for example b surfactants or chemical surfactants because we can increase the B availability of contaminants and so helping let’s say microorganism to degrade them we can use nutrients because of course microorganism needs
Nutrients but sometimes we don’t have proper microorganism in the contaminating Matrix so we have to have some specific micrell Ula but of course we can use amendments when I’m speaking about amendments I’m speaking about green compost you will see spent mushroom compost they are I will say waste or
Secondary products but it’s not correct because in terms of circular economy is an added value substrate that we can use in bior remediation and it’s really cheap and this is important when we are working of course at industrial uh scale because the cost of the treatment is reduced but to practice uh this
Amendments are rich in nutrients in microbial biomass and they can act also as buing a and here the last part of my presentation I selected some pictures uh about uh some project that we were working on during the last four years uh related to biopile treatment of tph uh
Contaminated soil in different treatment plant here in Italy um in both static and dynamic configuration okay and I just selecting some picture you see in the first one uh there is a biopile where we use green compost for example as amendment in the second one the span machon compost that
Is the topic of uh this day and another case we use also an oxidant but we use the oxidant as pre-treatment because in that case we had a really H weather contamination so it was not easy to degrade only with bacteria or fungi we need to modify some somehow the
Hydrocarbon mixture to keep to make the mixture more biodegradable and then of course we apply nutrients or some microbal inoculum and here just a picture from last month uh in a treatment plant where we set up several biopiles and of course in all this application case we control all the
Conditions so the oxygenation as I said humidity and temperature these are really important however we will say that once we designed the bio remediation of a tph contaminated soil in a biopile plant we have to Define also a monitoring plan because the monitoring plan is necessary to modify several parameters when the
Process is ongoing okay so to make the process more effective more efficient so we don’t need we need of course to check the concentration of tph chemical analysis but it’s not enough we have to check several parameters such as temperature and humidity because we are speaking about uh biobased uh technology
And microorganism are related to that we can perform respirometric test so we can uh assess the amount of oxygen because of course we are speaking about aerobic processes so if we see that there is a consumption of oxygen it means that the process is going on and consequently
Also CO2 because it’s formed if the process is going and of course also M or sulfuric acids that are produced under an oxic condition so if we find these compounds something is not going well because it means there is not oxygen for microorganism and of course we also um
Apply a biomolecular monitoring to see which kind of microorganism is there the structure and the Dynamics over time so how it’s changing okay during the process and of course we perform also ecotoxicological analysis these are important because it’s to prove that any kind of toxin metabolites is produced during the biodegradation process and
This is something that authorities often requires and just to conclude I select one of this uh project uh it’s a project in collaboration with the University of Milano bck any company any wind and akap Italia that is a consulting company here in Milano uh is a project uh dealing with
The treatment of tph contaminated soil in these agricultural areas that is close to Milano close to chiso and and um the the project was dealing not only with biopile but also phyto remediation there is a full scale phyto remediation field there and but of course course we are speaking about
Biopile we selected a dynamic uh um biopile as technology we set up two biopiles the first one we didn’t had any nutrients any microbial inoc was just oxygenated in the second one we had some green compost as Amendment we monitored uh as I said before several parameters temperature
Humidity about temperature we saw for example that where we had the green compost the temperature increased it means that the process the biodegradation process start immediately and was going on here the ecotoxicological analysis we use the germination test it means that if we have an increase in the
Germination index there is a reduction in the um toxicity of the Matrix and what we observed it was an increase of the germination index it means not toxicity or anyway not toxic metabolites were formed but going to the chemical analysis I want to underline if you see
Here the red one is the uh biopile where green compost was headed the process was much much faster than in the other biop pile especially at the beginning but after two months more or less the process start to slow down but this is normal this is typical because it means
That the light fraction the more biodegradable was removed and only the more caldon still remained in The Matrix so what we did we say okay let’s try to overcome the limit of bacteria and why not let’s use funi and we use the span Mion compost of
White r fungus plotus or status that is a waste of in um food industry really rich in nutrients in enzyme cheap as substrate and so also environmentally uh sustained why we use white root fi Alexandro said everything before I want just to underline really this kind of fungi are
Characterized by excellent I would say metabolic traits uh with regard to the biodegradation of not only tph but the most persistent organic pollutants so complex mixture of hydrocarbons because of these extracellular oxidative enzymes because the highall growth so they can penetrate easily into the contaminated Matrix and of course they can tolerate
High concentration of organic but also metals and they also are involved in so humidification processes so many reason why we can use this substrate for Bio remediation but as you can see even if we had this uh substrate the rate of degradation didn’t significantly increase and this is
Probably due to the fact that we were using a dynamic biopile and probably later the colleagues from my soil project they will explain that but since we are speaking about fungi with this highfall growth the mechanical turning of the Matrix can somehow impact okay so it’s not probably the best configuration the
Dynamic biopile for the treatment for sorry for the application of this kind of substrate anyway you see uh at the end of the test we uh obtained the 90% of removal of the initial concentration um and even if okay we had a really low level our goal was 50 milligrams
Kilogram so because we were in an agricultural areas but then we uh we were going on with this uh test and of course I don’t have time to show you but using some surfactant we reach uh 70 uh milligram per kilogram during uh the process of course
We uh also uh perform some B biomolecular analysis and here I just reported the sequencing of uh two target genes so we uh studied both bacterial and fungal Community okay I want just to underline uh concerning the fungal Community you see when we had the substrate the span musroom compost
Plotus was more than 90% of the total fungal uh Community however if we look at this bar the last one at the end of the process blus was only 2% and this is the main problem as Alexandra said before the Persistence of the inoculum because of the competition
With the autocon f I and other microorganism and of course we also Quantified some Target genes we Quantified the total number of bacteria and we Quantified a biomarker that is used for the aerobic biodegradation of aliphatic hydrocarbons and also in this case we observe an increase in the copy
Of number of these genes just at the beginning of the process and then it start to to reduce as you can see in the last part of the the treatment when the biodegradation process start to slow down so to conclude I’m on time this is
My message I will say I want just to underline this is a message that I reported here for biopile treatment but is general for Bio remediation these are three points really important if you want to apply any kind of Bio remediation strategy that is effective efficient and that you
Can reach the remediation goal within a time frame that is reasonable the first one is the characterization of the contaminating Matrix we don’t we need more than the chemical analysis we don’t need just the concentration of tph we have to know which kind of contamination
We have there is old is fresh which kind of hydrocarbons are there and of course we are applying bio remediation Technologies so we have to know which kind of microorganism are already there they are prob microorganism they they are able to degrade hydrocarbons we need to head hydrocarbon so a comprehensive
Characterization is pivotal I will say for any kind of Bio remediation strategy and then of course based on that you can optimize all the environmental condition you can start your treatment and the monitoring as I said before it’s really important because you can modify some of these parameters also when the process
Is ongoing okay and this way as I said before you can achieve your bio remediation goal within a time frame that is reasonable and also reducing the cost of the treatment thank you just few words to to conclude I want to thank you all the four speaker they
Touch very very different topics but uh in a a very nice overview so now I I guess that we learn a lot and we can uh we can um understand perfectly what will be said in the next session about the the my soil project do we have to say
Something or I can just announce the the buffet the coffee break so now this is the time of the coffee break