 I'd like to welcome you all to Wednesday's parallel session number four of the 2012 global symposium on soil by diversity. And here we're going to address theme two of the meeting which is soil by diversity in action. And this has a specific focus on the links with land restoration. So it's a great honor to be here with you all today. My name is Arwin Jones, and I'm from the European Commission's joint research center. I'm a colleague of Albertor Jati who gave a presentation in the panel session just now, and I'll be moderating the next couple of hours with you. We have arrangements for this afternoon. We will have four presentations in each hour, 10 minutes each. And I'd really like to remind the presenters to keep to this 10 minutes interval. So we have plenty of time for some discussion and questions and answers afterwards. I will remind you at nine minutes that you have one minute left and please try to respect the time limitations. If you have any other participants, please have a look at the Q&A instructions in the chat. Please post your question. And please indicate the name of the presenter to whom your question is addressed and we'll try and select a few of these for discussion afterwards. Any participants, if they can afterwards, maybe to re-engage with the other questions that we won't be able to address in this session and to engage with the people afterwards. So the host of the meeting is Julia. She's here to help you, Julia Stanko. And if you have any technical problems, please send them privately. So without further delay then, I'd like to give the floor to Ms Giada Miliori from ENAIA. That's the Italian National Agency for New Technologies, Energy and Sustainable Economic Development. And she's going to tell us about her work on restoring the soil while preserving functions, a winning approach by exploiting microbial biodiversity. So the floor is yours. Good evening. Good afternoon. Better. I share my, can you see? Maybe in presentation mode. Great. Okay. So good afternoon. Let me present a winning approach to restore soil by exploiting native microbial biodiversity and preserving functions and natural equilibrium between in the soil. As well known soil is a non renewable research and degraded the soil recovery and remediation of soil contaminated by mining activities are a strategic goal for European policies. The most contaminated environments show a low microbial activity and the reduced the soil functions, the discovery of new microorganisms, heavy metal resistant and able to promote plant grow may improve fight remediation technologies for contaminated site. I want to present to show the bacteria assisted fight remediation performed in the abundant mine of ingotos in Sardinia, and the monitoring activity carried out for seven years. The Gortoso mine site was one of the largest and most productive minds of sparrita and galina in Sardinia. The plants closed in 1969. Today in Gortoso is a part of geological and mining park and in 1997 it became one of the UNESCO Networks of Geoparks. It was inserted in a highly natural environmental and landscape context. In 2009 started the umbrella project that aimed to experiment the combination of native microorganisms and plants for a fight remediation assisted by microorganisms in five European mining sites. The area of ingotos was formed by mine waste. According to the chemical analysis, it was poor in total and organic carbon and nitrogen, and the heavy metal concentration was very high, and thereby availability was from 10 to 50 times higher than other mining soils. The plants were typical of Mediterranean vegetation, and among these, aphorbia pitusa was chosen for further experiments as well adapted to the local conditions and able to accumulate heavy metals in the leaves, so useful as a phytostabilizer. The cultivable fraction of soil bacteria was characterized. Among the 41 morphotypes strains have been identified and tested for plant growth promoting functions. 90% were nitrogen fixators, nitrogen fixers, 63% were seeder offer producers, 44% phosphate mobilizer, and 32% phytoheromones producers. The nine most performing strains were selected, cultivated in the laboratory, and then used for biogmentation in a consortium. All the bacteria isolated were collected in an ea, myriad strain collection, participated by European myriad networks. The toolbox consisting of endemic plants and bacteria was firstly tested in a greenhouse pot experiment. The results supported the decision to proceed with the field trial. A 200 square meters area was divided into 27 subplots, and the experimental design included different treatments like biogmentation, mycorrhizae, and viromine in different combinations. Viromine is an alkaline byproduct of bauxite industry, obtained from the reaction between red mud and sea water. It was widely used for environmental remediation process due to its metal trapping capacity. The viromine subplots were treated mixing 10% of amendment in the first 20 centimeters of soil. The field experiment started in October 2011. After transplanting the field was regularly watered. At the start of the experimental field, the microbial activity was evaluated through biolog system. Microbial activity is an indicator of soil quality, useful for evaluate the effect of bioremediation strategy and the soil evolution by the community physiological profile. From the graph, you can see that biogmentation had an immediate positive effect on the metabolic activity in the soil. Six months after the beginning of the trial, an early assessment of the toolbox performances was carried out. As shown, despite the winter season, the plant survival was satisfactory. Soil quality was increased and biogmentation improved microbial activity towards plant interaction by root exudates. Even if phobia alone positively influenced the microbial community in the resource sphere, bacterial biogmentation further improved the microbial metabolic potential and its resilience, allowing the plant soil system to better overcome stress and critical conditions. Metal content decreased. Due to the leaching and the uptake by the plants, vioramine contributed to the reduction of metal concentration, but at the same time inhibited the plant growth. The metal content in the plant shoots confirmed that the euphorbia was able to absorb metals from the soil. Summarizing at the end of the umbrella project, it was found that euphorbia pithusa proved to be a well-adapted and a well-performing metalified species. Euphorbia and the bacterial inoculum establish a depositive association. The bacteria inoculum significantly increased soil quality. Vioramine had a positive effect in combination with bacteria and mycorrhizae. In the soil, it increased pH, total and organic carbon, reduced the mobility of heavy metals and improved microbial activity. At the same time, it limited the plant growth and hence their efficacy in phytostabilization. In 2013, started the Italian project SMERI, starting from the experience gained in the European project AMBRELLA, the project wanted to transfer their acquired knowledge to a cluster of companies. Within the project, the status of the experimental field was analyzed after about two years, and the positive trend already in Triggered has been forced to introduce the endemic plant Yunkus maritimus, able to accumulate lead and cadmium in the root tips. By augmentation was updated, adding to the previous consortium, eight strains ex-novo isolated and selected as plant growth promoters, so the experimental field was partly reorganized. The study of microbial community showed that the enhancement of plant growth promoting functions introduced by bio-relementation with native consortia triggered a positive and long-lasting process in the soil. The effect remains in the long term, only where the plants have been associated with the selected bacteria. Although the species composition changed, the introduced functions were maintained and the population was further specialized. Euphorbia better survived in the subplots treated with bio-relementation, confirming that vegetation and bacteria are closely linked and interrelated. Bacteria create favorable conditions for plants on a substrate that has none of characteristic of the real soil. In parallel, plants in the rhizosphere favor the development, the evolution and the differentiation of microbial community. At the end of the project, the vegetation showed the symptoms of stress, both in the presence and in the absence of bio-relementation. The survival of plants was greatly reduced due to the adverse environmental conditions related to climate change. Extreme drought followed by water bonds had disruptive effects in arid and poor in organic matter soils. Following this project, the experimental field was left in place but without any kind of management and control and without any kind of additional bio-relementation treatment. Between 2015 and 2018 sporadic inspection were made to follow its natural and spontaneous evolution. In 2016, the study of bacteria-culturable traction confirmed that plant growth-promoting functions were mainly concentrated in subplots treated with bacteria in ocula. The effect per system, almost about five years, and bacteria play an important role on plant survival, reducing metal toxicity and increasing the efficiency of plant stabilization. Among functions, nitrogen fixation and cedarophore production were prevalent. In the plots inoculated by bacteria, the spectrum of metabolized carbonaceous states was broader. Biogmentation enriched the metabolic capacity and introduced new metabolic functions, such as the digestion of sugar phosphate. In 2018, further analysis confirmed that although the spontaneous renaturation process was in place as detectable by bacteria metabolic functions in the control, the subplots treated with biogmentation showed a better metabolic activity and a higher level of soil recovery and maturation, as confirmed by the presence of spontaneous vegetation that here found more favorable conditions to take root and develop them in the control. So, in conclusion, it can be said that selected endemic bacteria enhance functions in the soil and allow their maintenance over time. The inoculum directs and shapes community development. The positive effect of biogmentation on plant survival gradually decreases over five years without field management. So it should be necessary to define guidelines and protocols that include repeated intervention over time in order to maintain high levels of microbial diversity and activity. The combination of biogmentation with an amendment, such as biome, gave the best results in terms of soil metabolic activity, long-term plants survival, and the stabilization of effects over time. A rational selection of the microbial ignoble that considers the ecological context can help to capture and exploit the intrinsic bioremediation potential of contaminated environments. Such processes take time and energy to achieve equilibrium and show real benefits. Anyway, they represent a sustainable low-impact and low-cost solution. Concluding the monitoring of bacterial ecosystem and plant health is essential to follow and understand the evolution of the induced plant-bacterial association. But it's also necessary to integrate these analyses with the chemical, physical and geological data in order to obtain, to complete the picture and be able to evaluate the outcome of the remediation process applied. Thank you for your attention. Thank you very much, Giada, for this presentation. If people have any questions, please can you enter them in the chat and then we can take them at the end of the session. Thank you again. So now we'll move to Nicole Barga from the University of Colorado in the USA, and she will outline some recent successes and challenges in the restoration of degraded dryland soils. So, Nicole, over to you, please. Thank you. And I'm going to make sure that we are properly set up here. Can you see my single screen now? We see two, but yeah, maybe. How about that? Okay, perfect. Great, thank you. So good morning, good afternoon, good evening, everyone. We're talking about recent successes and persistent challenges and restoration of degraded dryland soils. So what I'd like to do is just set the context first in terms of global land degradation and how it negatively impacts the well-being of 3.2 billion people globally. In drylands, which is home to 38% of the global population, I just want to stress the importance in this talk today of restoring degraded land is especially vital to human well-being in these systems. And when we look at the land base in many of these regions, it is constrained by poor soil quality and loss of soil fertility. So we're really thinking in my lab group and more broadly, how do we start restoring some of these microbial communities? And we really heard some of that in the previous talk. So the need for soil restoration is that soil surface disturbance may increase soil erosion and ultra nutrient water cycles and really be a destabilized, a lot, many of the human activities in these ecosystems may be a destabilizing factor in terms of soils and soil mobilization. And in the systems that I work in and systems globally, an important component of soil recovery is contingent on the recovery of biological soil crust. And for the remaining part of the talk, I'll be talking about biocrust communities. So what are biocrusts? Biocrusts, biological soil crusts or biocrusts are very common in dryland ecosystems. They are estimated to cover about 12% of global soil surfaces. And they are communities, diverse communities of microorganisms that are consists of wikens, mosses, santa bacteria, fungal communities and other bacterial organisms that colonize this top veneer of soil, which is approximately two millimeters of the soil surface, and are critically important for the functioning of drylands so they can alter the hydrologic cycle in terms of retaining water and they also are incredibly effective at soil stabilization. In addition to, and we also heard this in the previous talk, there are a ray of nitrogen fixers in these soils that contribute to soil fertility, in addition to playing a pretty significant role in the photosynthetic capacity of these systems. So the need for an area of soil in these ecosystems can have the same or even higher photosynthetic rates than the surrounding vegetation. So the need for restoration of biological soil crusts is that they're often very slow to recovery due to resource constraints and so there's a limited capacity for natural recovery. The recovery of a biological soil crust can be decades to centuries. And so we just really identified that, you know, after some of the soil surface disturbing activities such as oil and gas development, agricultural recreation in our region there was a real need to develop effective strategies for biocrust restoration. And we really built on this emerging field of the use of microbial inoculants in soil restoration and rehabilitation. And so we took a lot of our ideas of restoration from this field. So what I'll be presenting is some of our work in the Western United States and in particular I'll be highlighting some of our work in the cold desert region of the Great Basin of the Western US. So our approach to biocrust restoration was the staged approach which we first had to develop how develop biocrust inoculum to add to degraded soils. But we also had the question that even if we developed inoculum what are the best strategies for restoring the soils. And then we just develop inoculum and throw it out into a degraded soil. And we, we had an idea that we wouldn't be able to do that. And so we, in parallel we developed biocrust inoculum. We developed restoration strategies that then fed into these multifactorial experiments that I'll describe. I'll briefly describe how we developed biocrust nurseries or inoculum. The idea is that we wanted to take a limited amount of field collected biocrusts, because we didn't want to collect a lot of field collected biocrusts for our experiments because we would just be disturbing soil to restore degraded soil. So one pathway we took is that we collected limited amounts in the field. And then in trays in the greenhouse we just develop enhanced the biomass in the greenhouse. So going nearly tenfold over a three to four month period that then went into our field of experiments. And the second pathway as you can see going to the lower part of the screen here is that with the mixed we developed cultures from the field and and and we specifically developed cultures of the cyanobacteria, which are some of the early successional my micro organisms that that are good at stabilizing the soils. So we also have a lab grown culture. Then with our second approach to identifying the best biocrust restoration strategies. We did over a two year period, a huge array of site preparation of adding water, adding nutrients to the soil, roughening the surface, and also trying to stabilize the surface. So from those huge number of experiments, what emerged as the best candidate restoration strategy was to stabilization strategies. So what we, we learned is that with a highly eroding soil, if you add a knock inoculum that will just erode away if that soil surface is not stabilized. So one stabilization strategy we used was a straw checkerboard. So this is just when straw is inserted vertically into the soil as small silk fences to capture soil finds and also seeds. And this has been used extensively as you can see in the lower picture and parts of China and do doon stabilization. A second soil stabilization strategy was the use of a commercial product. And we, this was a polyacrylamide so you can almost think of this as a dirt glue, and that was actually the product name was dirt glue. And what that does is that we, it degrades over time but initially it will stabilize that soil surface. So those were our two stabilization strategies that then fed into our broader experiment. So what you'll be seeing in the next couple graphs are an intact control. This is our target restoration of an intact biological soil crust. Then we had a disturbed degraded soil surface where we added no inoculum. In this experiment we added field collected inoculum inoculum, because we had, we wanted to know that if that local adapted inoculum would actually be effective relative to what we had grown up in the house, which is the local biomass and what we had grown in the lab with the mixed isolates. Each one of those were inoculated onto a surface of the polyacrylamide or the straw. So some of our results. This is this first graph and well I'll be showing core fill a, which over a three year period after inoculation and this is just an indicator of the photosynthetic capacity and a loose indicator of the sign of bacterial biomass. So again we wanted to really track the sign of bacteria in these soils because those are the early colonizers that are really effective and stabilizing the soil surfaces. So what you'll see here is that we have the targets that have fairly high core fill a rates, and then all of our experiments to the right and if you can just take a look at this you can say pretty easily wow it doesn't seem like that those inoculation and soil stabilization strategies were that effective. However, if we do focus in a little bit more and we change the scale on this graph. So again we have a target about of about 20 micrograms per gram soil of core fill that what we can see is the field collected. So the the inoculate inoculum that we directly took from that field site that was locally adapted shows some evidence that that there has been more enhanced recovery relative to what we grew in the lab or grew in the greenhouse which is the yellow bars or grew in the lab which is the blue blue bars. So coming off of this work, we're, we're faced with the question what are the likely constraints to biopress recovery under these field settings. So what we did find out is that we can grow it really well in the lab, and we can also grow it really well in the greenhouse. But once we go to a field setting we're not having this great success. And so what, yes, five minutes, nine, nine, nine minutes. Okay, so I was like, Oh, that's amazing. So what we do know is that when we shade the soil surface, it does promote, in addition to inoculating it, we are able to reach our targets. So this is just illustrates when you look at that far right intact column. And then you compare it to shading plus inoculating after three years we're able to to really regrow a biocrust in the field. However, you can imagine so finally just wrapping up with some key messages, barriers and challenges still exist in biopress recovery with inoculation. And we do know that irrigation and shading likely alleviate resource constraints and UV stress, resulting enhanced biopress recovery and we can do this over a short period of time. So the challenge is scaling some of these approaches. So you can imagine, we can't shade and water large expanses of land so we're really working on scaling, thinking about how we scale some of these strategies. And I would like to acknowledge the incredible team and large team that contributed to this work, and our funding was supported by the University of Colorado and the strategic environmental research and development program in the US. And thank you. So thank you very much. They call for that presentation very interesting. Can I just remind again people to put their questions in the chat, I can see that a few of you are already doing this. So we'll move on. The next presentation is Mr Kumar is from the University of Agricultural Sciences in Bengaluru in India. And he's going to talk about the mass multiplication of native soil mezzo fauna for the reintroduction in the degraded agro ecosystems. Mr Kumar, the floor is yours. Yeah, hello. Mr Kumar, perfect. You just need to put in presentation mode. Yeah, yeah, just estimate. Yeah, the icon. Yeah, exactly. Okay. Yes, perfect. Thanks. Yeah. Mr Kumar from University of Agriculture Sciences India. Yeah, yeah, you know that biological fertility of the soil is correlated to the action between soil animals, microorganism and plants together with the abiotic factors of the soil environment to regulate the soil fertility. So you know that in nature, the detritus is converted to humans in presence of soil biota which have the different consumers levels like primary consumers, secondary consumers, tertiary consumers, microfiber feeders and ultimate decomposer. If any human activity affect the any one consumer. So that is going to affect the humans formation in the nature. So here in the natural forest, the soil biota involved in the formation of the humus in naturally. And here we know that in the during the green revolution period we introduced several agrochemicals heavy machines and cultivation practices to get a higher yield. We got self-sufficiency in the food production. However, after some time, there was a stagnation of the lower crop yields due to loss of soil biological or mineralogical fertility and sustainability. To enhance the soil biological fertility, the scientists suggested to go for organic farming, but there are some limitations because the less availability of the format menu to all agro ecosystem. And this well decomposed format menu will not support all soil invertebrates and there may be slight increase in the existing fauna of the degraded land. So here, even in Mueller in 1957 itself, so he noted the succession of animal right from fresh heaps of organic matter till the humus formation. Apart from this kissing and ratten also highlighted the importance of indigenous invertebrate component in ecological restoration in agricultural landscape for better sustainable function. So, then comes the question why mass multiplication of miso fauna is required. You know that miso fauna, along with the microflora, these are very, very fragile organisms and also involved in a lot of humus formation activity. They are present in the post space of the top layer of the soil, but these are very susceptible for abided factors like rainfall, water stagnation, soil erosion, prolonged drought, or reduced rainy days, or even termite foraging on plant residue also affect the food of miso fauna in the dry land agriculture. So here before the initiation of our experiment, we studied the factors involved in the miso fauna development in the dry land agriculture where the food is very much limited to the soil miso fauna. The onset of rainfall, it triggers the multiplication of miso fauna and reached the peak activity at the end of the rainy season and then onwards the decline in the miso fauna population was observed. So, even in the way the food is abundant in a natural grassland forest ecosystem and the Likani and Lukasafala plantation, here also the rainfall play a very important role where we observed only one single peak at the end of the rainy season. Here what we observed the food is very much important for soil organism and moisture in the soil and moisture in the food is also important for soil fauna multiplication. So, based on this experience, we formulated 13 treatments with a major medium, media like soil alone, natural one, chiropeat alone, FIM alone, and combination of soil, chiropeat, chiropeat and FIM and soil and FIM and equal proportion of soil, chiropeat and FIM. And these are the quantity of media used in different treatments per part and these are the household vegetable waste naturally occur in all households along with the tea and coffee waste and even small paper waste and bills were used as a food material for miso fauna multiplication. This is the experiment view under the greenhouse condition and this is the backyard in the open air system and the methodology the pot size was 30 centimeter width at the top and 20 centimeter width at the bottom and 30 centimeter height of the pot. The miso fauna rich soil was collected from the natural grassland forest plantation and you look in here you go, cephala plantation and mixed well about 400 grams of mixed soil per pot was placed on the surface of the medium in the pot and 250 grams of household vegetable waste were placed over the soil surface of the pots at weekly interval and these pots were watered daily approximately one liter per pot and 200 grams of soil samples were drawn per pot at monthly interval up to one year and once in two months after one year for the extraction of miso fauna. These are these are the miso fauna extracted during the experimentation period. So regarding the results. So here we got at the end of the, at the end of the 18 months. So the mean population was significantly high in the farm yard manure treatment and which was followed by equal proportion of soil, chiropeat and FOM and which was on par with the chiropeat alone. So here the soil media recorded significantly less miso fauna population compared to other media. However, all media recorded significantly higher miso fauna abundance compared to natural ecosystem. So even the best media FOM alone and the least one soil fauna, soil alone. So here also we can see the activity of miso fauna throughout the experimentation period with varied population. So regarding the diversity. So here the treatment number 1150 is to 50 soil and FOM recorded significantly higher Simpson diversity index, which was on par with chiropeat and FOM 75 is to 25 proportion and these were almost on par with the natural ecosystem. So the rest of the media recorded 0.18 to 0.27 Simpson diversity in races. So the abundance of miso fauna in the backyard. So here you can see the gradual increase in population of miso fauna in the backyard and recorded higher population at the end of the 18 months after interval and which was very close to natural ecosystem. And even in the backyard also the miso fauna multiple activity was seen throughout the experimentation period, varied from 56 to 148 animals for 400 grams of soil. So even diversity of soil animal was maximum at 18 months after interval, which was very close to natural ecosystem. So the regarding the composition. In the natural activity where it only single peak activity was observed in the nature. So here purpose in matter dominated over other. And then in the best media FOM. So here other a carry dominated over other things, and what do you read a dominated over other columbolans in the soil, the least preferred one. So where the cryptos in my time research to my tower in equal proportion and one of your day dominated over other columbolans along with. Here we also observed students scorpion isopods and isopods rest or micro funnel images in the backyard. So, again, this project matter dominated and then one of your day dominated over other column. So here also observed in phyla isopods and associates and the rest for macro fauna images. So here the conclusion, all media harbour higher miso fauna abundance throughout the year compared to miso fauna natural ecosystem at the end of rainy season only. This is suitable for urban dwelling and the soil miso fauna diversity can be maintained in an institute of different region to protect the indigenous soil miso fauna for future use. Farmers can use the backyard or format menu heaps to multiply soil miso fauna, native soil miso fauna and miso fauna rich chikai repeat or FOM or combination of these two can be easily applied in the degraded land. So the diversity of miso fauna and micro flora can be replenished introducing a small quantity of top soil and litter of the undisturbed ecosystem. Thank you for your attention. Thank you very much indeed. Mr Kumar for that very interesting presentation and for keeping exactly to time so very much appreciated to that. So again, people if you have any questions or comments, please revert to the chat. Okay, the final presentation then for this part of the session is Claudia Roas from the Institute of Agri-Food, Animal and Environmental Sciences from the University of O'Higgins in Chile. Doesn't look like Mr. Claudia. Here we go. Can you see my presentation? Yes, perfect. Yeah. There's a lot in this in the changing of the screen. So please go ahead. All right. Thank you so much. First of all, to the organizers for putting this impossible together in spite of the pandemic. And thanks to all the audience for allowing me today to share with you the experience that we have in our lab today. I'm going to be talking to you about the experience that we have when we use organic amendments, different organic amendments in soil that have been affected by fires, especially in a Mediterranean forest, which is typical of Central Chile. So here I wanted to first start by acknowledging the importance of fires in shaping the structure of forest ecosystems. We do know that they are important to shape the structure and to help to the resilience of those ecosystems. The problem is when it happened, when the fires increase in number, in frequency, intensity, and of course the amount of area that is affected. And that is what I wanted to share with you with this graph. If you take a look here, what I'm showing you in this red bar is the area affected by fires since the late 70s in Chile. And here in these lines, you can see the numbers of fires occurring from the late 70s as well. I wanted to pay attention to the last decade starting from about 2010, and we do see that the number of fires are slightly increasing in Chile. And that coincides with a decade of severe mega drought that we are experiencing in Chile. So when you take a look at the red bars, we do see that on average since the late 70s, we have had about 50,000 hectares affected by fires. And that was not the case of 2017 when we did have about almost 600,000 hectares affected by fires. So that event in 2017, it led us to conduct the research that I'm going to be sharing with you today. This mega fire, as we name it that, occurred in, particularly in central Chile, which is an area with a Mediterranean climate, and this biome is one of the five biomes on Earth. And this type is really important in terms of biodiversity and the endemisms that we do see in this area. So the research that we conducted was in one of the three main administrative region that was affected by the fires occurring in January of 2017. And actually it coincides with the summer here in Chile in the southern hemisphere. So what we did was in 2018, in June we went to a field which was affected by fires and as you can see, the vegetation is still affected. And changing the ways growing is not, they used to be trees and now they are growing as a shrub. And we do see an ecosystem that was not affected by fires. So we use this, this is in total about an hectare inside, and we did perform our experiment here. And what we did was to try different organic amendments. Yeah, we did try different organic amendments, and we set up those in small plots, small experimental plots in the affected area, and we did compare those organic amendments with the soil that were under this unaffected ecosystem. So the organic amendments that we did use were particularly animal manures that were available for the community. And here I want to emphasize the importance to work with the community and to propose restoration strategies that are effective and can be used by the community which was affected. By in this case by this land burning. So we decided to use manure that they have available at their fields. And we also use compost, this was commercial compost that has come, especially from residues from our culture. So we in these three type of amendments, we did use the same rate in terms of volume 200 cubic meter per hectare. And of course, by the bulk density and moisture content of each of these materials, the final way dry weight that we put in the field is different. But I want to also emphasize that all the treatment that we use had that covered in much because that helped us to kind of keep moisture and protect the seeds and plants that we also established here. In addition, we did have treatment with just much, nothing more than much. And of course the control plots, which were burned, and we did not add anything there and of course the ecosystem of reference that I mentioned before. So for the experiment what I'm going to be showing you today here we sample in January of 2019. So that means two years after the chorus of fires, and after eight months after the application of the organic amendments that we try. So what we can see first is that I want to point out that we did use basal respiration using the methods of a substrate induced respiration with glucose. And what we observe here is that after two years, remember the fire occurring in 2017 and we sample in 2019, and after two years we still see fire disturbance in soil respiration. So the soils that were affected by fire still respire significantly more than the ecosystem of reference. Now, when we take a look at other soil conditions that is really important for us in terms of biological conditions, we do see that I want you to pay attention to these two treatments and we do see that after two years of fire occurrence, we still see effects of fires in terms of organic carbon content, we have less organic carbon in after two years of fires occurrence in the soils that were affected. And also we still see lower amount of nitrogen, total nitrogen in soils that were affected by land burning. Now, when we take a look at the treatments, we do see that as we the soils that we see fresh organic amendments, that means manure, especially swine manure and then followed by poultry manure are the soils that are still respiring more. They are liberating more CO2 than for example compost, which is a material more stabilized, the organic manure there is more stabilized. And we do see also that the treatment with just much, nothing more than much, also has more basal respiration than the control. So, from this graph, we can see that soil after two years still show the effect of fires and when we add organic matter, fresh materials resulting more soil respiration. So now when we take a look at the microbial biomass, we don't see that clear pattern that we do see here with the addition of organic amendment or just the control with a fire affected soil. We do see that almost all the treatment are similar in terms of microbial biomass, but again, the two treatments that are resulting in more microbial biomass in the soils are the ones that are aware treated with with fresh organic amendment, especially against sline manure. And another point that I wanted to highlight here is that when we try to restore soil, especially biological related conditions, using organic amendments, we do need to be aware of the immediate and long term responses of the soil to these different amendments. Why I am saying that is because we do see that fresh materials, immediately increased, when I say immediately eight months, increased basal respiration and microbial biomass. But in the long term, the carbon that is getting into the soil is less in fresh materials. When I compare that with a more stabilized material as compost. So we do need to take into account what are the goals, the overall goals of the restoration approach that we are we are using. So in this case, the use of compost is allowing us to predict that the restoration effort will last longer in the soil as more organic carbon is getting into the soil. And this result is eight months and we do see we are seeing the same results 20 months later too. So this is a really good notice. So here in just in terms of carbon mineralization and metabolic quotients, we do see again the same thing, the fresh organic material are activating more the soil than the compost material. So this is something that we need to take into account. And just to this is the last for the results and we do see that the organic amendment so is separate apart from those that did not receive amendment this is the barn soil and this is the control and all the parameters that we measure were affected, especially by treatments and all the biological condition were affected by treatment in among the physical condition, chemical condition we measure electrical conductivity was the one that most affected biological condition is in spite of the low level that we do see. And out of the treatment all affected the biological condition and interestingly enough we we confirm with the statistical analysis that the burn soil still affecting biological condition. So with that, it is important to highlight that is the type of organic matter matters, because the response is different in soil. And then, from the organic amendment that we use fresh material, inspire immediate response in microbial activity, but carbon sources more stabile carbon sources, like compost, my reflect reflect longer a positive effect in restoration of these soils. So that's why it's important it's important to understand, which is the organic matter that I'm adding. And for the future work we are combining these ecological functions. I do want to understand how the community structure of these ecosystem are shaped by the organic amendments and the fire gardens. So with that, I would like to send thanks to the group in Spain, which is part of this work and my group here in Chile, the fun finding funding agency and with that I will leave it here and thanks to all for their attendance. Okay, thank you very much Claudia for that presentation. And again, please, if you have any questions or comments to put them in the chat and maybe we don't have so much time but at least we can now try to have a number of points raised for for discussion. So for Giada and Nicole, maybe there's a common question on the possibility to scale up your treatments or your experiments would like to comment on that maybe Giada first and then Nicole. For me, actually the scale up is not so difficult to do. And there are more and more cases in which the biogmentation and especially phytoremediation applied together with the biogmentation has been successfully applied. So the critical points are to the time of recovery and the second the choice of plants and microorganisms. So it's important to choose plants and microorganisms while integrated in the environment and that not have any negative impact. It's important to do good characterization in the site and not only at the site but even in the landscape context in order to make a right choice. Thank you. Nicole. Yeah, thank you. I, as I said, it is a real challenge to think about doing microbial inoculation at the scale of, you know, what would be needed in some of these degraded lands. But I think one, one answer is to think about the development of inoculum so we already have with bio biofuels and cyanobacteria, we have facilities in several countries that we are able to scale up and be able to grow the cyanobacterial population so the challenge still is is that once it goes to the field, making that inoculum successful so we had some really great suggestions here in the chat of how do you mix with other materials such as biochar or something that will alter that that the conditions at the surface when inoculating. And so I think that that still is a real challenge and I don't have a great answer but for that at this point but it is something that we are thinking about is that that scaling at this point. Thank you very much. There's a question here for Mr Kumar. What would you say is the most important role of soil mesophony on soil health and functioning. Okay, thank you sir. I know that I show that the interlink of different soil animals in nature for the formation of humans. So here they have their feeding behavior different primary secondary tertiary consumer primary the decomposer and secondary and tertiary are the predators. Here we have microfiber feeders which regulate the soil microbes so which the disease outbreak will be maintained and apart from that the dead bodies of these mesophony and fecal material acts as a medium for soil microflora. So where whatever we introduce the microbes into the soil so their natural survival on this dead body and fecal material will be there apart from that the fecal material also helps in stable soil aggregate formation which helps in soil structure formation which is very important for natural condition of the soil apart from that here it helps in nutrient retention and sustainability in the nature. So these soil mesophony are working at the ground level at soil level when compared to the macrophony they come and go where they have higher mobility but these are a very fragile organism along with the mesophony they play very very very important. That's why we have to multiply and introduce especially the native one which are related to the native region so that we have to multiply that one to get sustainability unless this one so we don't get much things. Thank you. Thank you very much. And maybe a final question for for Claudia. Is it practical to use manure. Or other amendments for soil restoration at a larger scale so for example, more than 100 hectares. Thanks for the question because it's allowing me to highlight something which is very important in a large scale. If you're thinking to cover all the area or the land of course is not approachable from the economic point of view. But we what we are proposing is to use small slots. Many small slot rather than covering all the surface because we have seen edge positive effect that when you amend the soil not only you get positive effect in the soil that you use amendment but also at the ages. So you have like an island of fertility that is expanding so that is an interesting question because I have had it many times and when you think in the large scale covering everything is not economically feasible, but when you try small spots that can be a more reasonable approach. Okay. So, unfortunately, I think we've run out of time for the first first hour. I would still encourage you to keep chatting and exchanging via the via the chat. I see that there is a proposal for people to join an internet cafe for people to network and continue discussions after the session. So, I think that was a really interesting session on the group of papers on showing the value of sole biodiversity and land restoration in a range of different climate conditions and issues. So thank you very much for your willingness and to participate in the effort you've made to present your findings clearly and interestingly as well. So, now we'll move on to the to the second part of the session. Well, we will have said welcome back but I think you all stay here anyway so nobody goes. So we have four more presentations. Again, I like to just remind people to keep to the allotted 10 minutes I will remind you when you have one minute to go. Thanks in FNO have said actually that there is no formal deadline to the to the end of the session so if people are interested in discussion is still ongoing we can maybe bring back some of the questions from the first session as well for discussion at the end of the end of this one hour. So we'll see how we how we get on. So then the first presentation this afternoon of the second session rather is given by Sarah Pellaz from the University of Limerick in Ireland, and she's also going to talk about soil macrofauna and mesophora diversity in rehabilitated mind tailings so Sarah, the floor is yours. Thanks Arwin. Can you see my screen. Yes. So thanks to the organizers as well. And good afternoon morning and evening to everyone. I'm Sarah Pellaz. I'm PhD student on soil biodiversity and ecosystem restoration from the University of Limerick and today. I'm going to guide you through some of the results from my sampling campaign of last year. My PhD focuses on understanding ecosystem development in with a special emphasis on soil macrofauna and mesophora diversity in rehabilitated mind tailings. As you all might know, mining is a necessary good but very controversial activity. Most of the commodities we enjoy on our daily life highly depends on metal mining. Let me just give you an example. Do you really know where the materials of your mobile come from? I can tell you that more than one half of all the components are common for minerals and metals coming from a mine. However, metal mining is one of the most ecologically damaging activities worldwide. So that is why industry needs to come up with sustainable, long term and effective rehabilitation strategies. Traditionally, rehabilitation assessment had focused on monitoring of all ground vegetation and soils with less attention to soil fauna diversity. Now we know that soil fauna are critical for rehabilitation strategies to become sustainable. And that is why in our study, we did a general evaluation of soil invertebrates diversity, focusing on three of the most important ecosystem engineers, earthworms, spritz tails and ants. We wanted to answer the following research question. Are soil invertebrates diversity and abundance a good indicator of rehabilitation success? To answer this, we identified key soil fauna group in a chrono sequence of 5, 15 and 30 years of rehabilitated mind tailings. With this data, we calculate several diversity indexes to evaluate the rehabilitation success. Let me just introduce you the study side. The study sites are rehabilitated metalliferous tailings located at an industrially-sensitive site. So these three locations are made of tailings from lead and zinc extraction. It is important to notice that the substrate is a techno soil whose physical chemical properties and pedogenesis are not compared to that of actual soils. You can see the structure of these techno soils in the diagram below. During the sampling of a foreground invertebrates, we did de-back sampling with a de-back which is a vacuum cleaner device. We also installed 10 pitfalls per location and to sampling the earthworms, we did hand sorting. All the samples were identified up to morpho-species with species level when possible. Let me show you some of the results. Here we compare the results from the diversity indexes in these three locations. We can see that there is not huge homogeneity in the results on the indexes. For example, Simpson points toward a decrease in diversity from years of rehabilitation, richness just toward the opposite trend, and Sanon and evening's results here look to be more aligned. Tradition on Sanon has been the most used in diversity assessment because it accounts for entropy rather than species number. So this might represent reality better. So according to these results from Sanon, we have that biodiversity invertebrates biodiversity is at its highest at the beginning, then decreases and goes up again. But what is going on with ecosystem engineers in these three sites? We have at the beginning in the early sites that columbola communities are characterized by opportunistic species. Ants were absent and earthworms are species that are disturbing tolerance. However, in the older sites we have more complex columbola communities. Ants appear in higher number and we have earthworms characterized from well-developed soils. Just to walk out the results, I can tell you that with years from rehabilitation, the diversity is at its highest at the beginning, then decreases and goes up again. The data on ecosystem engineers shows that there is more abundance and complex community in older sites. So how do we explain these results? Let me show you two pictures from the field. On the left you have location A and on the right you have location B. And if we only look at the information from diversity indexes to evaluate rehabilitation, we could misinterpret these results. Saying that rehabilitation at location A was more success because it presented a higher diversity than in location B. And we know that this is not what is actually happening here because data on abundance and diversity of ecosystem engineers points toward the opposite. The results from biodiversity indexes are very explained and respond better to the classical model of succession in which when a new habitat became available, many species can potentially colonize it and in diversity rises. So if we come back to the question at the beginning of the slide, are soil invertebrates diversity and abundance a good indicator? The data from our sites points toward the direction that these indexes alone are not a good indicator of rehabilitation success in these sites. We also like to highlight that this kind of large general invertebrates diversity studies are not transferred to industry since they require large number of taxonomy specialists and sampling effort. We would like to point our research for the future and our next steps will be that rehabilitation assessment will focus more on grouping soil fauna into functional rather than taxonomic because it's more ecologically meaningful. And we would like to evaluate the soil food web in this chrono sequence. We would like to look at the species specific feeding with isotope analysis. And we also think that more research is needed on the role of ecosystem engineers as good long term indicators of rehabilitation success. So thanks very much for your attention and my contact details are in this last slide. Thank you very much Sarah for both an interesting presentation and also for keeping so succinct into time so thank you very much. And so people if you have any questions or comments, please post in the chat and we'll come back to Sarah at the end of the session. So moving on. And the next presentation is by Mr Raul Ortega from the University of Almeria in Spain. And he will present his work on the recovery of microbial status with organic amendments on soils affected by mining activity, the Decadal Temporal Scale. Thank you, Edwin. Good afternoon to everyone. Let me share my screen. Can you see the presentation? Yes, it's clear. Okay, thank you. As you say, the title of this work is recovery of microbiological status with organic amendments on soils affected by mining activity in a Decadal Temporal Scale. This research is hosted by the Soy Microbiological Lab of the University of Almeria in Spain, of which Dr. Isabel Miralles and I are the leaders. Our study area is located in the southeast of Spain in the province of Almeria. And we have work in a limestone extraction quarry, where the ecosystem is totally degraded by mining activities. The climatic conditions are extreme, only 230 millimeters of annual precipitation, high solar radiation, and completely degraded soils without organic matter structure, very fragile. Where erosion processes and the certification can occur, as we can see in the picture. The use of organic amendments in soil restoration is of great interest, because it has been shown that they improve physical, chemical, and biological soil properties. Besides, the soil bacteria communities are favored by these inputs of organic matter together with the addition of other taxa included in the amendments. So this bacteria can contribute to the plant establishment and favor the mineralization and unification of organic matter through the improvement of biochemical cycles. In Barbi, we have established two experimental parser, one in 2008, here in the left picture, and another one in 2018, right, with two different research projects here in the right. Now I'm going to focus on the data from the long term experiment. In both experimental parser, we are studying the evolution of the microbiological communities after the addition of organic amendments. On the workshop here, we have studied if the addition of compost derived from urban waste permitted the development of soil chemical properties and bacteria communities similar to those of the natural soils of the surroundings of in a temporal scale of a decade. So we have, we established several clubs of 15 meter long and five meter width, which accounts for a total surface of 75 square meters. Three replicates of these parsers were used for each of the following conditions. One, soils with organic amendments. Two, soils without amendment or control soils. And three, non-degrade natural soils. The input of organic amendment, the soils were applied, was estimated to increase at 2% of total organic carbon in the first 20 centimeter of the soil. We also restored soil with three different plant species, adapted to the Mediterranean semi-ariconditions of the area, steep at the Nazisman, until the Fittisoides and until the Thermiflora. Ten years later, we sampled soils up to a depth of 10 centimeters. We studied some chemical properties and bacterial communities, and we analyzed differences among treatment for these variables and relationships between chemical properties and bacterial tassonomy. The chemical properties analyzed in this work were total organic carbon, pH, and total nitrogen, and the bacterial tassonomy were determined by metagenomic analysis. We extracted the DNA of the bacteria using a key agent commercial key, and the amplicons were later sequenced using the Lumina sequencing platform. Bioinformatic analysis were carried on with the software China tool, and statistical analysis include Permanoba, general linear models and Pearson correlation coefficients. About our results, we found 162 soil bacteria taxa up to a level of genus with an abundance higher than 0.1% in all the samples. General linear models show it that compost from urban waste significantly influenced the 59% of all bacteria taxa, meanwhile natural soils only influenced the 14%. We found some common bacterial taxa in amended and natural soils, but not in controlled soils. And here we have an example of this tassa where we can see that they both appear in amended soils and natural soils, but not in controlled soils without inputs of organic matter. About the chemical parameters, we can say that all pH of the soil were basic because of the carbonate contents of the barren material, the line stone quarry, but not amended soils, control control soils show it higher values, and statistically were different to the soil with compost and natural soils, where the decrease in pH level can be explained by the higher and similar values of total organic matter, higher than 3%. Meanwhile, in controlled soils, the values were near two zero, despite of the meaning activity is 10 years ago. So with compost show it the highest nitrogen contents compared with other treatments and again control soil non amended show it nearly nothing nitrogen content. On this table, we show bacterial taxa that on one hand they were common in natural and amended soils, but they were not present in controlled soils. I mean soils without input of organic matter. And on another hand, they show it statistical significance with chemical properties. We can see that all these taxa are positively correlated with total organic carbon and total nitrogen, which clearly shows the establishment of bacterial taxa in composted soils with similar behavior to other more mature soils, in this case, natural soils. From our source, we can say that composting in totally degraded soil promoted in a decade scale, chemical properties similar or even better to the natural soils. Luna at all show it in this same soils that the improvement of the chemical properties was already significant after five years of restoration. We have confirmed that after 10 years this effect is maintained about the microbial populations, all the authors note the addition of organic amendments in restore soils, increase and improve the soil microbial communities, and the origin of the compost influence the component, the composition of the communities. We also confirm that compost have an important influence on the bacterial taxa, 59% of the common taxa between natural soils and amended soils show a similar behavior with chemical parameters as described before positive correlation with total organic carbon and nitrogen content and negative correlation with pH. Some bacterial taxa that we found, like pedomicrobium was already observed in soils rich in organic matter, and terremonase was found in developed soils near our study area, corroborating the compost treat soils, permit the establishment of taxa present in more mature soils. In conclusion, we can say that our resource suggests that soil restored with urban waste compost has established chemical properties and microbial, microbial communities similar to the surrounding natural soils in a decade scale. The soils without organic amendments used as control did not improve their chemical and biological properties. This data and other have been published in one in this paper, I think I can later copy and paste it in the chat, in case someone is interested. And then I have one more minute. I'd like to say that at present we are working on plants and on the blocks I showed at the beginning, starting in 2018, the same line stone quarry and we are studying the fact of several organic amendments in physical, chemical, biochemical and biological property, biological properties where microbial community and CO2 loses these there are three, and now we are using three inputs of organic matter and vegetable compost from garden waste, vegetable compost from article to greenhouse studios and establish seed waste sludge. Here are some pictures of the works. Here, here, a picture of the evolution of the soil restoration from 2019 to nowadays. And with that I finish, I hope I'm on time. Thank you very much, Raul, for that, for a very striking demonstration of the power of soil biodiversity. And also a very strong links to the circular economy and the bioeconomy sector and the reuse there of sewage sludge, which is actually quite a topical issue. So indeed, yes, thank you for keeping the time. As with the previous speakers, please put your questions to Raul in the chat and we'll come back to those in 20 minutes or so. So thank you again. So moving on, the next presentation that is by Mr. Daniel Castro from the Instituto Amazonica de Investigaciones científicas from the National University of Columbia. And he will outline how much so the biodiversity is restored after a cattle ranching pasture is abandoned. So, Daniel, the floor is yours. Thank you. Can you see my screen? Yeah, it's very clear. Okay. Good afternoon everyone and good morning from Columbia. My name is Daniel Castro, a researcher at Institute in Amazon Institute, since you from Columbia, I will present this is to the title, how much a soil diversity is restored after a cattle ranching pasture is abandoned for its natural regeneration in the Amazon region of Columbia. Soils like diversity, biological communities are considered healthy soils in tropical soils, there are two groups of great importance. This group is the soil micro fauna that contribute to the organic matter cycling and the efficient return to the nutrient of plants. And the second group is the arouscule micro risal fungi that contribute significantly in the efficient post-produce acquisition of the plants to micro risal associations. The forestation of natural forest to transform in pastures is commonly in the great Amazonia and also in the Colombian Amazon region. Soil reporting out is biological community restored themselves after a natural after pastures abandoned for their natural regeneration. Meanwhile, soil micro fauna and arouscule fungal communities are high sensitive to the channel life use. Soil in a bunch of the potential that soil micro fauna has an indicator of soil quality and the permanently present of the arouscule micro risal fungi association to the plant communities. We have a right how soil micro fauna and arouscule micro risal fungi diversity is affected by the forestation and how much of is restored through a natural regeneration process. This is to be was conduit in the Kakaeta state in the northwestern of Colombian Amazon region. We evaluate five different restoration different restoration stage. We evaluate pastures that have recently livestock activity. We also evaluate young secondary vegetation vegetation from natural regeneration between zero to 10 years. Also secondary vegetation between 10 to 20 years. Also secondary old forest between 20 to 40 years and finally my two forests more than a for a 40 years old six plot a of each regeneration age were sample for a total of 30 plots. Soil micro fauna something was done following the methodology of tropical soil biology and fertility program ESB a on each restoration age upload of 60 per 60 mirrors well landed. There are five a T SBA monoliths of a 25 per 25 centimeters what made in each monolith samples were collected for different beans. This is toy unfortunately a wasn't possible the collection. Another hand for Arbusco micro risal fungal sampling a top soil sample of approximately of 100 range was collected at the same at the site of the of the each T SBI monoliths. And therefore classification of a soil micro fauna morphological identification has has we carry out for micro risal fungal a we identify a specific autos using you know you do in a sequencing methods and specific primers. You know what this was performant for soil micro fauna the US city and species richness and Chris Caldwell it is what you said for a micro risal fungal abundance sports and species richness. The city analysis was done through a verification and extrapolation curves. So our results in soil micro fauna community a 2528 taxonomic groups, a them, the most sound and groups where and their might and larbis, especially larbis for colioptera and lepioptera. All the specific specimens a we identify at least two to family, and we found 791 morphotypes in 124 families in 30, 37 orders in nine class and two films. The termites and ants were the most dense groups, a termites was the highest, the highest diversity values in soil in restoration at age, but less representation was show it in pastures, and was more abundant and to the micro fauna abundance didn't change of the years. The density show a big difference between termites and ants, big, the two, the two most dominant groups in density, but in abundance other groups are important such as colioptera, clopoda, diplopoda are the spiders. The soil micro fauna species richness increase with the age of restoration regeneration present a significant difference in sucessional forest of the 10 years old with the free sucessional age is micro fauna density and species richness were not correlation in the natural regeneration consequence, where in all regeneration stage we present high values of species richness, significantly different for pastures. Pioner species are abundant in the second stage of regeneration and therefore high values of species richness are present. But over the time, these species are less abundant or disappear, another species big to appear that not adapt well to degraded soils. For example, in June, in young stage we found many occurrence of other species as Guzmania or Puntaata, Paratochina longicornis and Solonopsis derminata that species are considered as urban pests in Colombia. In the summer mites, we found a riskidermis boteroi and a paternal medianusa and a teratermis tiniis, and these species adapt better to low fertility soils, and these species disappear or are greater species in mature forests. As was expected, the lowest diversity occur in pastures. After 40 years of natural generation, soil micro fauna diversity is not restored completely. However, some authors indicate that soil micro fauna communities could be restored in a short time when activities process and performance are agroforestic system, forest plantation, and other crop systems. So, for our words, mycorrhizal fungi, we have obtained a 44-bisculptaxa. Mycorrhizal genus was the most abundant genus followed by the alcohol spore genus. Mycorrhizal fungi communities were different at the 5th forest stage of the current sequence in the diversity of mycorrhizal fungi is projected to be much greater than obtained. In the soil, the abundance of spores and the virtual taxa richness of arbustular mycorrhizal fungi was higher in young regeneration states than in mature forests. In mature forests, mycorrhizal is mainly in ruts and not in the soil metrics. We also found that some virtual taxa were present in all chrono sequence, while other virtual taxa, as you can see in green in the table, only appear in the latest stage of regeneration. Finally, we conclude that the graduate pastures that Araban donated for the natural generation could restore their soil biological communities. The natural truth, the time. A restoration of soil biological communities take more than 20 years to reach a similar species region that not used to raid a mature forest, but species composition would not be the same. A change in the soil and the muscular mycorrhizal fungi composition and the biomass could have consequences that how don't be estimated. Thank you to our founding institute and thank you for your attention. Thank you very much for that presentation. And again also for for for keeping to time. And please, people, if anybody has any questions, please put them in the chat and we'll come back to them in about 10 minutes or so. So the final presentation for this session is from Mr. Warshidan Dania. Apologies for the pronunciation if it's incorrect from the factory of agriculture from the University of Peradena in Sri Lanka. And Warshid will give a presentation on the effect of shifting cultivations on bacterial communities in the forests of Sri Lanka. Warshid, the floor is yours. Thank you for the opportunity given to present in this forum. Good morning, good afternoon and good evening to everyone. I'm going to present our work on bacterial community diversity as affected by the shifting cultivation on Kuru Lu International Biosphere Reserve, which we did using a metagenomic approach. The dry forested ecosystems in Sri Lanka are constantly under pressure. In 1920, we had about 50% of our land area in this country under forest. However, it has reduced to 26% by now. And from this land cover, if you look at the dry forested ecosystems, it covers 22% of the land area in the country. We have very less information about the soil biodiversity in our forested ecosystems, especially in the dry forested ecosystems. In this study, we targeted the Huru Lu International Biosphere Reserve, which is located in the northeastern part of the country, and also in relatively dry environment. These foresties are the high pressure of socioeconomic stressors and experiencing a range of forest disturbances. So there are studies that indicate that forest disturbances affect soil microbial diversity in evergreen forests. And also it is important to understand the factors governing soil biodiversity in these forest ecosystems when we are formulating conservation measures. Therefore, the objective of a study was to assess the effect of forest disturbances, mainly because of the shifting cultivation practices on diversity of soil bacterial communities in this international biosphere reserve in Sri Lanka. So we have selected three types of land covers in this area. One is relatively undisturbed forest and then regenerating forest after clearing the land for shifting cultivation and then sites under active shifting cultivation. So these three types of land covers were selected for the study and we collected soil samples from 0 to 5 and 5 to 10 centimeter depth classes. You can see the location of samples in this map and when we were collecting soil samples we used 10 meter by 10 meter quadrants and five samples were collected per site to make a representative sample of the site. And the information on vegetation cover was also recorded at the same time. The soil was used for characterization first by analyzing chemical, physical and microbiological properties. And then by looking at the data we decided to use only 0 to 5 centimeter depth class for molecule analysis because of the financial limitations we could not process a large number of samples as we have used the metagenomic approach. So we collected, we extracted DNA in replicate from each soil and then pull the replicated DNA extracts to represent one side and then perform metagenomic analysis using next generation sequencing technique targeting V3, V4 region of the 16S-RRNA gene. And applicant size we used after trimming was 250 base sphere. So if we look at the soil characteristics, it is clear that the 0 to 5 centimeter depth has been, has shown more changes in the properties that we have measured with respect to the land cover. So for example the active carbon content, the bio density and soil electrical conductivity, these properties were affected by the land cover type at the 0 to 5 centimeter depth, more than 5 to 10 centimeter depth. And potential nitrogen mineralization, which is an indicator of the microbial, one side of microbial functionality in the soil, also shown changes with respect to land cover. So with this information we have hypothesized that the change, the microbial community diversity would change with the land cover type at 0 to 5 centimeter depth. And the active carbon, which comprises a major component of the soil carbon, which is a major soil carbon fraction influenced and interacting with the soil microbial community is, has shown significant correlations with electrical conductivity, bio density and potential nitrogen mineralization rate. So let's move into the bacterial diversity information. So from the sequences we have generated in this study, after quality control, we have recovered more than 75% of the sequences, which is a good amount in next generation sequencing technique. And the refraction curves indicated that the species coverage obtained from each sequence run was optimum and species richness for each site was exhaustingly sampled. For taxonomic assignment, we have considered 97% similarity at 95% confidence interval and on average, only about 16% of the sequences were assigned up to general level. So there was high rate of mismatches, which is common in next generation sequencing technique. However, this may lead to some important biological signals of novel species as well. So this 16% of the sequences identified were represented by 84 different genera. Let's look at the alpha diversity measurements. So we have calculated the Shannon and Simpson index, both Shannon and Simpson indexes indices indicated higher values for the soil from bacterial diversity in soil from relatively undisturbed soil compared to regenerating forest and the shifting cultivation. So the high bacterial diversity and less species dominance was there in the undisturbed forest soils compared to other two land covers. So if we look into detail about the species composition, actinomycetale, basilarle, clostridialae and bercolderialae were the commonly found bacterial orders in all soils. However, the abundance of members of actinomycetale were higher in disturbed soils. And if we look at the actinomycetale order and look at the different genera present, actino plants were more dominant in soils under shifting cultivation than in soil under forest cover. These are just a few examples to show that how the species composition may change with the land cover in this forested system. So if we take a look at the order basilarle, the genus basillus was the most dominant genera in this order. And the dominant species in genus basillus are indicated in this graph. And if we look at basillus megaterium and basillus primilus, they were found only under shifting cultivation in this particular study. The beta diversity was also calculated. So in this study we have looked at both forward and reverse primus, forward and reverse sequences as a quality control measure. And the results from forward and reverse sequence analysis for regenerating forest and for shift, sorry, from shifting cultivation were more similar. However, in undisturbed forest systems, the results from forward and reverse sequence analysis were slightly different. There was a slight difference in that analysis, which indicate some interferences in the data analytical steps in this platform. So, however, when we look as a whole, we can see that samples came from sites in active shifting cultivation was significantly different from the samples that came from relatively undisturbed forest and the regenerating forest soils. So what our study indicated is, in conclusion, the forest soil with minimum anthropological soil disturbances harbored the highest bacterial diversity at genus level and less domination of single genus over others in soil bacterial communities. High mismatches in sequence alignment may be partly due to the fact that a large majority has not been previously reported and may have novel species as well. So it may not on not necessarily totally because of the data analysis errors in next generation sequence. And also disturbing the forest ecosystems in Hulu dry mixed evergreen forest in Sri Lanka for cultivation has reduced soil bacterial diversity. With this, I would like to acknowledge my collaborators, Professor Pushpa Kumara, Dr. Rainu Karthanaika and Mr. Bhagya, and the assistance provided by a number of people in this study and the financial assistance from Biodiversity Secretariat of Ministry of Environment Sri Lanka. Thank you for your attention. Thank you very, very much, Washi, for that really clear and interesting presentation. Just highlight the impacts of land cover change on soil microbial communities. So, again, if anybody has any questions to the last presentation, please now add them to the chat. So we have about 10, 15 minutes maybe of time for discussion and questions. I see already that we have some some exchanges in the chat but maybe I'll just pick up on one for Sarah based on the very first presentation. We were talking about the difficulties of assessing the success of restoration programs. So how would you actually go about quantifying the success rate of the restoration project. Well, I would say that, apart from these biological indexes, we obviously we need to include physical measurements of these sites. In the case of post mining sites, soil organic matter and decomposition and water holding are very important, and they could give you good indications of the status of the restoration in the sites. Okay. Thank you very much. Thank you. Thank you for the second. So, there's a question to round wall as well about the impressive increase in total organic matter content after the restoration process was very impressive. But can you say anything about other soil properties like like the stability of aggregates, and so on. In the paper, I said that the resources were included. We also studied the electrical conductivity and the resources again were similar of compost, amended soils with natural soils. But now I pasted several papers. Now with the new project that is already going on where we are performing many physical and chemical analysis. So there is more information from the new study area of physical and chemical parameters about the stability of aggregate. We haven't already done it, but it's true that we would like to buy the wood sieve machine to perform this kind of analysis too. Okay, thank you very much. There's also a question to Daniel in that the degraded pastures do not allow natural secondary forest regrowth in short period. And if you also need additional interventions such as cover crops. Do you have any research information about this information, about these interventions and how to accelerate the recovery of degraded lands? Yeah, the Amazon have a different coverage and it depends of the coverage. You have more difficult to threat that the pastures can restore it through the time in short of in long time. In this case, our study are in the north part of the Amazonian is they have a very, very high influence to the Andean Amazon Piedmont and this influence in a short time, we have a high restored in all communities in this site. But it's important to know that the connection of biological between natural fragments and these pastures and the gradate coverage have a good connectivity for all the species can mix it in these matrix soils for a better recovery of the biological communities. I, from one, I showed a two publication of our Institute that showed not only in the soils, but in the so many biological components. All we can restore it these communities in a short time and what is the world practice for you cans made in the in the courage with the communities and all these biological connectivity in the courage for have a better results in the in a short time to restore all communities. Okay, thank you very much. I mean, just a general question, maybe to all four of the presenters then I mean, I think you've all demonstrated that your approaches work and are successful. Have you had any positive feedback, say from policymakers or planners that they see then the greater potential of your approaches? Again, it relates a little bit to the upscaling question from the first session. Do you have any positive feedback, let's say, from beyond the scientific community of the work that you've been doing? Maybe we start with Sarah. Yeah, thanks, Arwin. I would say that for the mining industry it's very important to get to prove the long-term sustainability of these rehabilitation strategies. So I think there is a lot of hope in the role of these ecosystem engineers, soil fusions to be able to demonstrate that long-term. Yeah, okay. Daniel, okay, you're working with mining as well, more in a rock quarrying than on the metal deposits, maybe of Sarah, do you have a similar sort of experience? Yeah, I think that I agree with Sarah and all soil microfauna, not only the ecosystem engineers, but the guilds of all soil microfauna that can improve the restoration in very courage degraded and all these mix it between the soil microfauna with engineers, the creditors, and the microbiological components and also a micro and miso fauna can show it so many things about we can restore it or we can, all the tools we can make, I include politician tools, technical tools and social tools for made more efficient to recover the soils that is degraded and all these components are very useful with all the studies that my partners made today show that the results that the soil biodiversity is very important for the seed, the state of the soil in their data states and natural state and all these a consequence restoration issues. Thank you. Raul? Yes, here in Spain the regulation says that mining companies must restore degraded soils after their activity and that's why we are working with the Femex company in this experiment I show here and our demonstration there, we have two more projects, we are restoring soils that were agricultural soils that were abandoned some decades ago and now we are restoring them and administration is really, really interesting because it's in public forest and also now we are studying how the soils evolve after important perturbations like fires. There has been an important fire here in the province and we are working together with the nation of the administration to follow the evolution of the soils. Okay. And finally, Washi from your perspective what's the involvement of the policymakers and the broader community to your work in Sri Lanka? Yes, this study that I presented it was funded by the minister of environment which indicate their keen interest on soil biodiversity as well. Until very recently I would say about five years back all the biodiversity communication even the policies regulations were only about about ground visible diversity that means like plants, animals and insects. However, soil biodiversity was recognized as an important component in biodiversity conservation measures with these studies. Actually, we were able to show the immense diversity in our soils in terms of bacteria and also we did study the fungal diversity in these dry sun forested soils which I did not present here. This indicated we have very much rich diversity in Sri Lanka which is unexplored. There's so much potential and the ministry is really interested. And now we are formulating even the biodiversity policies amending it with the information that we generated through these studies on soil biodiversity. So this was kind of an eye opener even for the ministry to acknowledge the soil biodiversity and what these organisms might be doing in our ecosystems, yeah. Okay, thank you very much. Actually, I've just been scanning through the chat and I see that actually a lot of the questions have been answered in real time during the presentation. So thank you very much for that engagement by all of the speakers. So I think we've come really to the end of the session. I think it's been a fascinating two hours. I think it's been a very positive two hours. I think it's really enlightening to see let's say the positive demonstrations of the work and really to show what we all really know in our hearts that by considering the biodiversity and the life in the soil, we actually do end up with a better, healthier soil. And I think all your presentations and including also the four in the previous session clearly show that and the benefits that they bring to society as a whole. So I'd just like to express my appreciation for your work in the preparation in your presentations and in keeping to time and in engaging with the more than 120 people who have been following this session. So it's really good to have that number of people much more probably than if we were in a physical meeting, although it's always much nicer to be face-to-face with your wall. Just the colleagues from FAO have asked me just to remind you of a few things before we close that the presentations will be available on the symposium website after the meeting ends and the meeting this afternoon was recorded so you can always listen back to that. Certificates of attendance will be granted for people who have followed the four days of the symposium and if you need one, you can just write to the Global Soil Biodiversity Symposium in the mailbox and it's in the addresses in the chat there so that you can request that. And then also just to remind you that the poster session is still ongoing. So please take a moment to browse through some of those posters. There's a lot of interesting material there and give your same votes of appreciation to the posters. So again, I'll just remind you again in the chat there is from Erika the offer to maintain the conversation and the discussions going through the web-based cafe. So please feel free to join that. I encourage you very much to do so. So again, I'd just like to thank you very, very much and we'll see you back in the symposium at one o'clock tomorrow. So have a good morning, afternoon, evening, wherever you are. Thank you very much indeed. Bye for now. Thank you very much to everyone. Thank you to all.