Does poly-3-hydroxybutyrate biodegradation affect the quality of soil organic matter?

The search for eco-friendly substitutes for traditional plastics has led to the production of biodegradable bioplastics. However, concerns have been raised about the impact of bioplastic biodegradation on soil health. Despite these concerns, the potential negative consequences of bioplastics during various stages of biodegradation remain underexplored. Therefore, this study aims to investigate the impact of micro-bioplastics made of poly-3-hydroxybutyrate (P3HB) on the properties of three different soils. In our ten-month experiment, we investigated the impact of poly-3-hydroxybutyrate (P3HB) on Chernozem, Cambisol, and Phaeozem soils. Our study focused on changes in soil organic matter (SOM), microbial activity, and the level of soil carbon and nitrogen. The observed changes indicated an excessive level of biodegradation of SOM after the soils were enriched with micro-particles of P3HB, with concentrations ranging from 0.1% to 3%. The thermogravimetric analysis confirmed the presence of residual P3HB (particularly in the 3% treatment) and underscored the heightened biodegradation of SOM, especially in the more stable SOM fractions. This was notably evident in Phaeozem soils, where even the stable SOM pool was affected. Elemental analysis revealed changes in soil organic carbon content following P3HB degradation, although nitrogen levels remained constant. Enzymatic activity was found to vary with soil type and responded differently across P3HB concentration levels. Our findings confirmed that P3HB acts as a bioavailable carbon source. Its biodegradation stimulates the production of enzymes, which in turn affects various soil elements, indicating complex interactions within the soil ecosystem.

Biodegradation of poly-3-hydroxybutyrate after soil inoculation with microbial consortium: Soil microbiome and plant responses to the changed environment.

Biodegradable plastics play a vital role in addressing global plastics disposal challenges. Poly-3-hydroxybutyrate (P3HB) is a biodegradable bacterial intracellular storage polymer with substantial usage potential in agriculture. Poly-3-hydroxybutyrate and its degradation products are non-toxic; however, previous studies suggest that P3HB biodegradation negatively affects plant growth because the microorganisms compete with plants for nutrients. One possible solution to this issue could be inoculating soil with a consortium of plant growth-promoting and N-fixing microorganisms. To test this hypothesis, we conducted a pot experiment using lettuce (Lactuca sativa L. var. capitata L.) grown in soil amended with two doses (1 % and 5 % w/w) of P3HB and microbial inoculant (MI). We tested five experimental variations: P3HB 1 %, P3HB 1 % + MI, P3HB 5 %, P3HB 5 % + MI, and MI, to assess the impact of added microorganisms on plant growth and P3HB biodegradation. The efficient P3HB degradation, which was directly dependent on the amount of bioplastics added, was coupled with the preferential utilization of P3HB as a carbon (C) source. Due to the increased demand for nutrients in P3HB-amended soil by microbial degraders, respiration and enzyme activities were enhanced. This indicated an increased mineralisation of C as well as nitrogen (N), sulphur (S), and phosphorus (P). Microbial inoculation introduced specific bacterial taxa that further improved degradation efficiency and nutrient turnover (N, S, and P) in P3HB-amended soil. Notably, soil acidification related to P3HB was not the primary factor affecting plant growth inhibition. However, despite plant growth-promoting rhizobacteria and N2-fixing microorganisms originating from MI, plant biomass yield remained limited, suggesting that these microorganisms were not entirely successful in mitigating the growth inhibition caused by P3HB.

Possibility of using tannins to control greenhouse gas production during digestate storage.

The presented paper deals with the testing of a possibility to reduce emissions of undesirable greenhouse gases (CH4, CO2; NOx) and their mixture (biogas) during the storage of digestate using applications of secondary plant metabolites (tannins). The experiment was conducted in laboratory conditions in which the digestate was placed in fermentation chambers. Prior to the fermentation process, preparations were applied to the digestate, which contained tannins: Tanenol Antibotrytis (TA), Tanenol Clar (TC) and Tanenol Rouge (TR) in three concentrations (0.5, 1.0 and 2.0% w/w). The application of these preparations demonstrably affected the production of biogas and the contents of CH4, CO2 and N therein. The application of TR preparation in the concentration of 1.0% and 2.0% significantly reduced the production of biogas as compared with all variants. The preparation further inhibited the process of CH4 development. In contrast, the other preparations with the content of different kinds of TA and TC increased the production of biogas (on average by 15%), CH4 (on average by 7%) and CO2 (on average by 12%) as compared with the control variant and TR variant. These two variants reduced the concentration of N in biogas on average by 38%. Thus, the tested Tanenol tannin preparations can be used in different concentrations either to control emissions of greenhouse gases during the storage of digestate or, in case of increased production of CO2 for its reuse in order to increase methane yields in the process of anaerobic fermentation.

Effect of Digestate Modified with Amendments on Soil Health and Plant Biomass under Varying Experimental Durations.

A digestate with amendments provides plants with available nutrients and improves the microbiological properties of treated soil. Modification of a digestate through the addition of a biochar and sulphur source is less well-known. This pot experiment aimed at comparing the short- and long-time fertilization effects of a digestate enriched with biochar, with elemental sulphur, or with a combination of both on soil health and plant biomass. The experiment was carried out with maize, cultivated twice (1st-12th week = pre-cultivation; re-sowing after shoot harvest, 13th-24th = main cultivation) in soil amended with prepared digestate. The digestate used in pre-cultivation was incubated untreated (D) and was then treated with biochar (D + B), with elemental sulphur at a low (LS) and high (HS) dose, or with a combination of both (D + B + LS and D + B + HS). An additional unamended digestate (D) was added to each soil variant before the main cultivation. The application of digestate with a high dose of elemental sulphur and biochar mediated the most significant differences in the soil. The increase (compared to the unamended soil) was of short-term type (+11% and +6% increased total nitrogen and carbon after 12 weeks), then of long-term type (+54% and +30% increased sulphur and arylsulfatase activity after 24 weeks), and later emerged in the 13th to the 24th week of the experiment (+57% and +32% non-inhibited urease, increased N-acetyl-ß-D-glucosaminidase and phosphatase). No significant differences in the effect of the applied amendments on dry aboveground plant biomass were observed.

The combined effect of graphene oxide and elemental nano-sulfur on soil biological properties and lettuce plant biomass.

The impact of graphene oxide (GO) nanocarbon on soil properties is mixed, with both negative and positive effects. Although it decreases the viability of some microbes, there are few studies on how its single amendment to soil or in combination with nanosized sulfur benefits soil microorganisms and nutrient transformation. Therefore, an eight-week pot experiment was carried out under controlled conditions (growth chamber with artificial light) in soil seeded with lettuce (Lactuca sativa) and amended with GO or nano-sulfur on their own or their several combinations. The following variants were tested: (I) Control, (II) GO, (III) Low nano-S + GO, (IV) High nano-S + GO, (V) Low nano-S, (VI) High nano-S. Results revealed no significant differences in soil pH, dry plant aboveground, and root biomass among all five amended variants and the control group. The greatest positive effect on soil respiration was observed when GO was used alone, and this effect remained significant even when it was combined with high nano-S. Low nano-S plus a GO dose negatively affected some of the soil respiration types: NAG_SIR, Tre_SIR, Ala_SIR, and Arg_SIR. Single GO application was found to enhance arylsulfatase activity, while the combination of high nano-S and GO not only enhanced arylsulfatase but also urease and phosphatase activity in the soil. The elemental nano-S probably counteracted the GO-mediated effect on organic carbon oxidation. We partially proved the hypothesis that GO-enhanced nano-S oxidation increases phosphatase activity.

Time-dependent impact of co-matured manure with elemental sulfur and biochar on the soil agro-ecological properties and plant biomass.

Farmyard manure is the most common type of organic fertilizer, and its properties depend mainly on the type of livestock, bedding material and the conditions of fermentation. Co-maturing of manure with other amendments to modify its final properties has been seen as a win-win strategy recently. This study aimed to evaluate the differences in the effect of unenriched manure and manures co-matured with biochar, elemental sulfur or both amendments on the soil physico-chemical and biological properties, and plant (barley, maize) biomass production. For this purpose a pot experiment was carried out in a time-dependent way. Samples were taken from 12 week-lasting (test crop barley) and 24 week-lasting (test crop maize) pot cultivation carried out in a growth chamber. Co-matured manure with biochar showed the highest rate of maturation expressed as humic to fulvic acid ratio, its amendment to soil significantly increased the dry aboveground biomass weight in the half-time (12 weeks) of experiment. However, the effect vanished after 24 weeks. We received for this variant highest long-term (24 weeks) contents of total carbon and nitrogen in soil. Contrarily, co-matured manure with biochar and elemental sulfur led to short-term carbon sequestration (the highest total carbon in 12 weeks) due to presumed retardation of microbial-mediated transformation of nutrients. We conclude that the prolonged pot experiment with biochar or elemental sulfur enriched manure led to the increased recalcitrancy of soil organic matter and retardation of soil nutrient transformation to the plant-available form.

Effect of Mycotoxins in Silage on Biogas Production.

Mycotoxins can pose a threat to biogas production as they can contaminate the feedstock used in biogas production, such as agricultural crops and other organic materials. This research study evaluated the contents of deoxynivalenol (DON), zearalenone (ZEA), fumonisin (FUM), and aflatoxin (AFL) mycotoxins in maize silage prior to it being processed in a biogas plant and in digestate produced at the end of the anaerobic digestion (AD) process. In the experiment, three samples of silage were collected from one silage warehouse: Variant 1 = low contamination, Variant 2 = medium contamination, and Variant 3 = heavy contamination, which were subjected to investigation. A significantly reduced biogas production was recorded that was proportional to the increasing contamination with molds, which was primarily due to the AD of silage caused by technologically erroneous silage treatment. The AD was connected with changes in silage composition expressed by the values of VS content, sugar content, lactic acid content, acetic acid content, and the ratio of lactic acid content to acetic acid content. The production of biogas and methane decreased with the increasing contents of NDF, ADF, CF, and lignin. The only exception was Variant 2, in which the content of ADF, CF, and lignin was lower (by 8-11%) than that in Variant 1, and only the content of NDF was higher (by 9%) than that in Variant 1. A secondary factor that also correlated with changes in the composition of the substrate was the development of undesirable organisms, which further contributed to its degradation and to the production of mycotoxins. It was also demonstrated in this study that during the AD process, the tested mycotoxins were degraded, and their content was reduced by 27-100%. Only the variant with low mold contamination showed a DON concentration increase of 27.8%.

Bentonite as a Functional Material Enhancing Phytostabilization of Post-Industrial Contaminated Soils with Heavy Metals.

Growing awareness of the risks posed by pollution of the soil environment is leading to the development of new remediation strategies. The technique of aided phytostabilization, which involves the evaluation of new heavy-metal (HM)-immobilizing amendments, together with appropriately selected plant species, is a challenge for environmental protection and remediation of the soil environment, and seems to be promising. In this study, the suitability of bentonite for the technique of aided phytostabilization of soils contaminated with high HM concentrations was determined, using a mixture of two grass species. The HM contents in the tested plants and in the soil were determined by flame atomic absorption spectrometry. The application of bentonite had a positive effect on the biomass of the tested plants, and resulted in an increase in soil pH. The concentrations of copper, nickel, cadmium, lead and chromium were higher in the roots than in the above-ground parts of the plants, especially when bentonite was applied to the soil. The addition of the analyzed soil additive contributed significantly to a decrease in the levels of zinc, copper, cadmium and nickel in the soil at the end of the experiment. In view of the above, it can be concluded that the use of bentonite in the aided phytostabilization of soils polluted with HMs, is appropriate.

Food and agricultural wastes-derived biochars in combination with mineral fertilizer as sustainable soil amendments to enhance soil microbiological activity, nutrient cycling and crop production.

The ever-increasing human population associated with high rate of waste generation may pose serious threats to soil ecosystem. Nevertheless, conversion of agricultural and food wastes to biochar has been shown as a beneficial approach in sustainable soil management. However, our understanding on how integration of biochar obtained from different wastes and mineral fertilizers impact soil microbiological indicators is limited. Therefore, in the present study the effects of agricultural (AB) and food waste derived (FWB) biochars with and without mineral fertilizer (MF) on crop growth and soil health indicators were compared in a pot experiment. In particular, the impacts of applied amendments on soil microbiological health indicators those related to microbial extracellular (C, N and P acquiring) enzymes, soil basal as well as different substrate induced respirations along with crop's agronomic performance were explored. The results showed that compared to the control, the amendment with AB combined with MF enhanced the crop growth as revealed by higher above and below ground biomass accumulation. Moreover, both the biochars (FWB and AB) modified soil chemical properties (pH and electric conductivity) in the presence or absence of MF as compared to control. However, with the sole application of MF was most influential strategy to improve soil basal and arginin-induced respiration as well as most of the soil extracellular enzymes, those related to C, N and P cycling. Use of FWB resulted in enhanced urease activity. This suggested the role of MF and FWB in nutrient cycling and plant nutrition. Thus, integration of biochar and mineral fertilizers is recommended as an efficient and climate smart package for sustainable soil management and crop production.

Plant-endophyte mediated improvement in physiological and bio-protective abilities of marigold (Tagetes patula).

Endophytic bacteria improve the growth, physiology, and metabolite profile of plants. They are known as potential biocontrol agents of soil-borne diseases. This study evaluated the effects of endophytic bacterial strains on growth, vase life, biochemical attributes, and antioxidant and nematicidal activities of French marigold (Tagetes patula). French marigold seeds were sole and consortium inoculated with three promising endophytic bacterial strains, Burkholderia phytofirmans (PsJN), Enterobacter sp. (MN17), and Bacillus sp. (MN54). The vase life of French marigold was promoted by 66.6% in the individual application of PsJN and 100% in plants treated with consortium compared to the uninoculated control. The shoot and root fresh weights were also increased by 65.9 and 68.7%, with the combined application of all three strains. The total phenolics, flavonoid, and protein contents were higher in consortium treatment with an increase of up to 38.0, 55.9, and 65.9%, respectively, compared to the uninoculated control. Furthermore, combined application of endophytic bacterial strains promoted DPPH radical scavenging, mortality of plant-parasitic nematodes, and ferric reducing antioxidant power activities with increase of up to 278.0, 103.8, and 178.0%, respectively, compared to uninoculated control. An increase in antioxidant activities of ascorbate peroxidase (APX), catalase (CAT), glutathione peroxidase (GPX), and superoxide dismutase (SOD) were observed up to 77.3, 86.0, 91.6, and 102.9%, respectively by combined application of endophytic bacterial strains. So, given the economic importance of floriculture crops, endophytic bacterial isolates studied here have shown a great potential for improving the productivity of cultivated ornamental French marigold.

Co-composting of cattle manure with biochar and elemental sulphur and its effects on manure quality, plant biomass and microbiological characteristics of post-harvest soil.

Improvement of manure by co-composting with other materials is beneficial to the quality of the amended soil. Therefore, the manure was supplied with either biochar, elemental sulphur or both prior to fermentation in 50 L barrels for a period of eight weeks. The manure products were subsequently analyzed and used as fertilizers in a short-term pot experiment with barley fodder (Hordeum vulgare L.). The experiment was carried out under controlled conditions in a growth chamber for 12 weeks. The sulphur-enriched manure showed the lowest manure pH and highest ammonium content. The co-fermentation of biochar and sulphur led to the highest sulphur content and an abundance of ammonium-oxidizing bacteria in manure. The biochar+sulphur-enriched manure led to the highest dry aboveground plant biomass in the amended soil, whose value was 98% higher compared to the unamended control, 38% higher compared to the variant with biochar-enriched manure and 23% higher compared to the manure-amended variant. Amendment of the sulphur-enriched manure types led to the highest enzyme activities and soil respirations (basal, substrate-induced). This innovative approach to improve the quality of organic fertilizers utilizes treated agricultural waste (biochar) and a biotechnological residual product (elementary sulphur from biogas desulphurization) and hence contributes to the circular economy.

Soil Properties and Maize Yield Improvement with Biochar-Enriched Poultry Litter-Based Fertilizer.

Conversion of poultry litter into fertilizer presents an environmentally friendly way for its disposal. The amendment of stabilizing sorption materials (e.g., biochar) to broiler chicken rearing seems promising, as it protects produced litter from nutrient losses and improves fertilizing efficacy. Thus, a pot experiment was carried out with maize and organic fertilizers produced from biochar-amended chicken bedding. The properties of three types of poultry-matured litter, amended with biochar at 0%, 10% and 20% dose, were analyzed. These matured litters were added to soil and physicochemical, biological properties and dry aboveground crop biomass yield were determined. Both biochar doses improved matured litter dry matter (+29%, +68% compared to unamended litter) and organic carbon (+5%, +9%). All three fertilizers significantly increased dry plant aboveground biomass yield (+3% and +42% compared to control litter-treated variant) and N-acetyl-ß-D-glucosaminidase activity (+51%, +57%) compared to unamended control soil. The 20% biochar poultry-matured litter derived the highest dry plant aboveground biomass, highest respiration induced by D-glucose (+53%) and D-mannose (+35%, compared to control litter-treated variant), and decreased pH (-6% compared to unamended control). Biochar-derived modification of poultry litter maturation process led to organic fertilizer which enhanced degradation of soil organic matter in the subsequently amended soil. Furthermore, this type of fertilizer, compared to conventional unamended litter-based type, increased microbial activity, nutrient availability, and biomass yield of maize in selected biochar doses, even under conditions of significant soil acidification.

Assessment of digestates prepared from maize, legumes, and their mixed culture as soil amendments: Effects on plant biomass and soil properties.

Digestate prepared from anaerobic digestion can be used as a fertilizer, as it contains ample amounts of plant nutrients, mainly nitrogen, phosphorous, and potassium. In this regard, digestates produced from mixed intercropped cereal and legume biomass have the potential to enrich soil and plants with nutrients more efficiently than monoculture-based digestates. The objective of this study was to determine the impact of different types of digestates applied at a rate of 40 t·ha-1 of fresh matter on soil properties and crop yield in a pot experiment with lettuce (Lactuca sativa) as a test crop. Anaerobic digestion of silages was prepared from the following monocultures and mixed cultures: broad bean, maize, maize and broad bean, maize and white sweet clover, and white sweet clover. Anaerobic digestion was performed in an automatic custom-made system and applied to the soil. Results revealed that fresh and dry aboveground biomass as well as the amount of nitrogen in plants significantly increased in all digestate-amended variants in comparison to control. The highest content of soil total nitrogen (+11% compared to the control) and urease (+3% compared to control) were observed for maize digestate amendment. Broad bean digestate mediated the highest oxidizable carbon (+48%), basal respiration (+46%), and N-acetyl-ß-D-glucosamine-, L-alanine-, and L-lysine-induced respiration (+22%, +35%, +22%) compared to control. Moreover, maize and broad bean digestate resulted in the highest values of N-acetyl-ß-D-glucosaminidase and ß -glucosidase (+35% and +39%), and maize and white sweet clover digestate revealed the highest value of arylsulfatase (+32%). The observed differences in results suggest different effects of applied digestates. We thus concluded that legume-containing digestates possibly stimulate microbial activity (as found in increased respiration rates), and might lead to increased nitrogen losses if the more quickly mineralized nitrogen is not taken up by the plants.

Using Waste Sulfur from Biogas Production in Combination with Nitrogen Fertilization of Maize (Zea mays L.) by Foliar Application.

In Europe, mainly due to industrial desulfurization, the supply of soil sulfur (S), an essential nutrient for crops, has been declining. One of the currently promoted sources of renewable energy is biogas production, which produces S as a waste product. In order to confirm the effect of the foliar application of waste elemental S in combination with liquid urea ammonium nitrate (UAN) fertilizer, a vegetation experiment was conducted with maize as the main crop grown for biogas production. The following treatments were included in the experiment: 1. Control (no fertilization), 2. UAN, 3. UANS1 (N:S ratio, 2:1), 4. UANS2 (1:1), 5. UANS3 (1:2). The application of UAN increased the N content in the plant and significantly affected the chlorophyll content (N-tester value). Despite the lower increase in nitrogen (N) content and uptake by the plant due to the application of UANS, these combinations had a significant effect on the quantum yield of PSII. The application of UANS significantly increased the S content of the plant. The increase in the weight of plants found on the treatment fertilized with UANS can be explained by the synergistic relationship between N and S, which contributed to the increase in crop nitrogen use efficiency. This study suggests that the foliar application of waste elemental S in combination with UAN at a 1:1 ratio could be an effective way to optimize the nutritional status of maize while reducing mineral fertilizer consumption.

Effect of Seed Coating and PEG-Induced Drought on the Germination Capacity of Five Clover Crops.

The effect of coating the seed of clover crops by water absorbing seed process (WASP) technology pelletization on its germination capacity was studied in conditions of diverse drought intensities simulated by different concentrations of polyethylenglycol (PEG) 8000 solution. Drought resistance was monitored in the seed of five fodder clover species: Anthyllis vulneraria L., Medicago lupulina L., Trifolium repens L., Melilotus albus Medik. and Onobrychis viciifolia Scop. In the seed of given plant species, germination capacity was determined along with the share of dead and hard seeds. Although the coating significantly (p < 0.05) affected the drought resistance of seeds, the germination capacity increased only in conditions of milder drought (simulation with PEG: 0.1-0.3 mol). With the increasing intensity of drought induced by higher PEG concentrations (0.4-0.7 mol) the number of germinable seeds demonstrably decreased and the number of dead seeds increased in the coated seed as compared with the uncoated seed. The coated seed can be appropriate for use in M. lupulina, M. albus and T. repens, while the uncoated seed can be used in A. vulneraria and O. viciifolia.

A critical review of the possible adverse effects of biochar in the soil environment.

Biochar has received extensive attention because of its multi-functionality for agricultural and environmental applications. Despite its many benefits, there are concerns related to the long-term safety and implications of its application, mainly because the mechanisms affecting soil and organism health are poorly quantified and understood. This work reviews 259 sources and summarises existing knowledge on biochar's adverse effects on soil from a multiangle perspective, including the physicochemical changes in soil, reduced efficiency of agrochemicals, potentially toxic substances in biochar, and effects on soil biota. Suggestions are made for mitigation measures. Mixed findings are often reported; however, the results suggest that high doses of biochar in clay soils are likely to decrease available water content, and surface application of biochar to sandy soils likely increases erosion and particulate matter emissions. Furthermore, biochar may increase the likelihood of excessive soil salinity and decreased soil fertility because of an increase in the pH of alkaline soils causing nutrient precipitation. Regarding the impact of biochar on (agro)chemicals and the role of biochar-borne toxic substances, these factors cannot be neglected because of their apparent undesirable effects on target and non-target organisms, respectively. Concerning non-target biota, adverse effects on reproduction, growth, and DNA integrity of earthworms have been reported along with effects on soil microbiome such as a shift in the fungi-to-bacteria ratio. Given the diversity of effects that biochar may induce in soil, guidelines for future biochar use should adopt a structured and holistic approach that considers all positive and negative effects of biochar.

Effect of carbon-enriched digestate on the microbial soil activity.

OBJECTIVES: As a liquid organic fertilizer used in agriculture, digestate is rich in many nutrients (i.e. nitrogen, phosphorus, sulfur, calcium, potassium); their utilization may be however less efficient in soils poor in organic carbon (due to low carbon:nitrogen ratio). In order to solve the disadvantages, digestate enrichment with carbon-rich amendments biochar or humic acids (Humac) was tested. METHODS: Soil variants amended with enriched digestate: digestate + biochar, digestate + Humac, and digestate + combined biochar and humic acids-were compared to control with untreated digestate in their effect on total soil carbon and nitrogen, microbial biomass carbon, soil respiration and soil enzymatic activities in a pot experiment. Yield of the test crop lettuce was also determined for all variants. RESULTS: Soil respiration was the most significantly increased property, positively affected by digestate + Humac. Both digestate + biochar and digestate + Humac significantly increased microbial biomass carbon. Significant negative effect of digestate + biochar (compared to the control digestate) on particular enzyme activities was alleviated by the addition of humic acids. No significant differences among the tested variants were found in the above-ground and root plant biomass. CONCLUSIONS: The tested organic supplements improved the digestate effect on some determined soil properties. We deduced from the results (carbon:nitrogen ratio, microbial biomass and activity) that the assimilation of nutrients by plants increased; however, the most desired positive effect on the yield of crop biomass was not demonstrated. We assume that the digestate enrichment with organic amendments may be more beneficial in a long time-scaled trial.

Nano Zero Valent Iron (nZVI) as an Amendment for Phytostabilization of Highly Multi-PTE Contaminated Soil.

In recent years, a lot of attention has been given to searching for new additives which will effectively facilitate the process of immobilizing contaminants in the soil. This work considers the role of the enhanced nano zero valent iron (nZVI) strategy in the phytostabilization of soil contaminated with potentially toxic elements (PTEs). The experiment was carried out on soil that was highly contaminated with PTEs derived from areas in which metal waste had been stored for many years. The plants used comprised a mixture of grasses-Lolium perenne L. and Festuca rubra L. To determine the effect of the nZVI on the content of PTEs in soil and plants, the samples were analyzed using flame atomic absorption spectrometry (FAAS). The addition of nZVI significantly increased average plant biomass (38%), the contents of Cu (above 2-fold), Ni (44%), Cd (29%), Pb (68%), Zn (44%), and Cr (above 2-fold) in the roots as well as the soil pH. The addition of nZVI, on the other hand, was most effective in reducing the Zn content of soil when compared to the control series. Based on the investigations conducted, the application of nZVI to soil highly contaminated with PTEs is potentially beneficial for the restoration of polluted lands.

Fourier Transform Infrared Spectroscopy vibrational bands study of Spinacia oleracea and Trigonella corniculata under biochar amendment in naturally contaminated soil.

Fourier transform infrared spectroscopy (FTIR) spectroscopy detects functional groups such as vibrational bands like N-H, O-H, C-H, C = O (ester, amine, ketone, aldehyde), C = C, C = N (vibrational modes of a tetrapyrrole ring) and simply C = N. The FTIR of these bands is fundamental to the investigation of the effect of biochar (BC) treatment on structural changes in the chlorophyll molecules of both plants that were tested. For this, dried leaf of Spinacia oleracia (spinach) and Trigonella corniculata (fenugreek) were selected for FTIR spectral study of chlorophyll associated functional groups. The study's primary goal was to investigate the silent features of infrared (IR) spectra of dried leave samples. The data obtained from the current study also shows that leaf chlorophyll can mask or suppress other molecules' FITR bands, including proteins. In addition, the C = O bands with Mg and the C9 ketonic group of chlorophyll are observed as peaks at1600 (0%BC), 1650 (3%BC) and 1640, or near to1700 (5%BC) in spinach samples. In fenugreek, additional effects are observed in the FTIR spectra of chlorophyll at the major groups of C = C, C = O and C9 of the ketonic groups, and the vibrational bands are more evident at C-H and N-H of the tetrapyrrole ring. It is concluded that C-N bands are more visible in 5% BC treated spinach and fenugreek than in all other treatments. These types of spectra are useful in detecting changes or visibility of functional groups, which are very helpful in supporting biochemical data such as an increase in protein can be detected by more visibility of C-N bands in FTIR spectra.