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Article

Potential Effect of Biochar on Soil Properties, Microbial Activity and Vicia faba Properties Affected by Microplastics Contamination

by
Heba Elbasiouny
1,*,
Azza A. Mostafa
1,
Amina Zedan
1,
Hala M. Elbltagy
1,
Sherifa F. M. Dawoud
1,
Bodor A. Elbanna
1,
Safaa A. El-Shazly
1,
Amany A. El-Sadawy
1,
Aisha M. Sharaf-Eldin
1,
Marwa Darweesh
1,
Al-Zahraa E. E. Ebrahim
1,
Sara Mohamed Amer
1,
Nour Osman Albeialy
1,
Doaa Shokry Alkharsawey
1,
Nora Rabiea Aeash
1,
Amira Osama Abuomar
1,
Rehab Elsayed Hamd
1 and
Fathy Elbehiry
2,*
1
Environmental and Biological Science, Home Economics Faculty, Al-Azhar University, Tanta 31732, Egypt
2
Basic and Applied Sciences Department, Higher Institute for Agricultural Co-Operation, Shubra El-Kheima 13766, Egypt
*
Authors to whom correspondence should be addressed.
Agronomy 2023, 13(1), 149; https://doi.org/10.3390/agronomy13010149
Submission received: 1 October 2022 / Revised: 15 December 2022 / Accepted: 24 December 2022 / Published: 2 January 2023
Browse Figures
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Abstract

:
Microplastics (MPs) contamination is an emerging issue globally; however, adverse impacts of MPs on soil, plants and microbial activity have not been intensively studied. In this study, the potential effect of different levels of MPs (1.5, 7.5, 15%) has been investigated on soil properties, plant properties (Vicia Faba) and microbial activities through a pot experiment. The effect of biochar (BC: 2%) to mitigate the adverse effects of MP has also been examined. Soil properties (pH, EC, OM, CaCO3 and some elements) have significantly differed due to contamination of soil by MPs as well as by adding BC to the soil. The pH and CaCO3 were significantly increased more than in the control, while EC, TDS, available P, Mn and Fe were significantly decreased lower than the control, which implies adsorption on microplastic. Plant properties, such as enzymes, chlorophyll and fresh and dry weight in roots, were adversely affected by MPs contamination; however, BC mitigated this effect, especially with low contamination levels of MPs. The fresh and dry weight of the shoot was not significantly affected by MPs. The cytogenetic analysis showed that the mitotic index was significantly reduced compared to the control (9.39%), while BC increased the mitotic index at 1.5% MPs (7.11%) although it was less than the control. The percentage of abnormalities of V. faba root tip cells under different levels of MPs was significantly increased more than the control; however, BC mitigated this effect, especially at 7.5% MPs. The total count of bacteria and fungi even in soil or in the rhizosphere area did not follow a clear trend; however, the effect of BC was clear in increasing their activities. Microbial biomass carbon and nitrogen were also significantly affected by MPs and BC. In this study, the BC level was low, however, it mitigated some adverse effects of MPs, especially at 1.5 and 7.5% of MPs. Thus, the BC could be promising in mitigating the negative impacts of MPs when applied with suitable levels that need more future studies.

1. Introduction

Plastics have become an indispensable aspect of modern life. Plastic is frequently utilized in modern civilization. Current production and consumption, however, are unsustainable. The global usage of plastics in 2019 was estimated to be 368 million tons [1,2] but is expected to be doubled within 20 years [2]. Plastic waste is recognized as toxic, and the risk grows when polymers degrade in nature to secondary microplastics (MPs) or even nanoplastics [3]. Microplastics are often described as plastic trash with dimensions less than 5 mm. They either are the product of larger plastic fragmentation (secondary MPs) or the direct release of small plastic fragments from anthropogenic activities, which are considered the major contributor to global contamination [1,4]. The primary causes of MP accumulation in diverse ecosystems of the environment include urban sprawl, industrialization, uncontrolled usage and inadequate waste management of plastic items. Microplastics in the soil are seen as an emerging danger to agro-ecosystems. The majority of MPs research has been conducted in aquatic environments, while the eco-toxicological impacts of these pollutants in terrestrial ecosystems, particularly agro-ecosystems, are still little understood. The harmful impacts of MPs on soil’s physical, chemical and biological characteristics are gradually becoming clear. [5]. Thus, MP and residual plastic in soil can effect soil fertility, plant health and plant production [6,7,8]. Soil contamination with MPs implies suffering plant communities in this soil from various degrees of MP contamination. Because of the importance of plants in terrestrial ecosystems and the continued release of MPs, the potential effects of MPs on terrestrial plants are causing significant worry. The MP exposure can have a number of effects on terrestrial plants’ physiology, morphology and community structure. Mulching agricultural soil with plastic films, for example, might enhance water evaporation from this soil, resulting in more severe droughts. They can also be absorbed by the roots, fruits and vegetables or adsorbed onto the surface of the roots [9,10,11].
Microplastics pose significant threats to terrestrial ecosystems due to their abundance and persistence [12]. The broad presence of MPs in many ecosystems makes them accessible to a diverse spectrum of species living in those ecosystems, including plants, animals, and then, even humans. As a result, their existence and possible detrimental impacts on all living species should be carefully examined [13]. Higher plants, which constitute an essential component of the terrestrial ecosystem, are invariably subjected to MPs [12]. Given that the great majority of higher plants are terrestrial plants and that soil is a significant sink for MPs, soil cultivation experiments are required to expose the impacts of MPs on plants. In most soil cultivation experiments, the investigated MPs were typically applied to soils [12]. Knowledge of the ecotoxicology of MP is advancing as well; however, information regarding its impacts on plants is still lacking [3].
In addition, contamination of soil by MP has many different impacts on the microbial community structure and activity by changing soil parameters [14]. Changes of microbial communities because of MP may alter biogeochemical cycling, which possibly results in the influence on the whole functions and services of soil ecosystem [15]. As well, altering the microbial soil ecosystems due to MP may also increase or decrease enzymatic processes based on MP type [16].
MPs can be degraded naturally by some processes, such as photodegradation and thermo-oxidative degradation. During these processes, the MPs’ polymer can be degraded into smaller pieces with a molecular weight that is favorable to microbial decomposition. However, the C in the polymer may be converted by microbes into CO2. In addition, the long period (more than 50 years) for totally decomposing the MPs is another disadvantage of these processes [17]. Furthermore, some physical techniques and synthesis of some chemicals are used in this purpose; however, complexity, non-greener character and polymer and environmental variability are among other disadvantages for MPs’ remediation [18]. Thus, it is critical to develop MPs’ removal technologies; for instance, including MPs’ removal procedures in wastewater treatment will decrease the quantity of MPs that reach soil ecosystems as a result of sewage irrigation [19].
Biochar is one of the most effective and widely available biomass adsorbents. It has a stable and unique structure and many properties, such as abundance, carbon-rich and high porosity. In addition, BC is less expensive, more eco-friendly and needs less energy-intensive manufacturing technology. All of these features make it effective in the removal of numerous types of pollutants, including MPs. As a result, investigating approaches for removal of MPs utilizing BC can assist researchers in developing and improving novel mitigation technologies [20]. Some researchers addressed the using of BC or modified BC for MP removal with high efficiency; however, most of these trials were conducted on water such as [21,22,23]. Wang et al. [22] recommended further research for more confidence on the effectiveness of BC to remove MP. Little is known about using BC in this purpose in soil because of some experiments performed as incubation experiments with the acceleration of MP removal of BC such as [24,25,26] or microcosm experiment [15] and little others on cultivated plants. Palansooriya et al. [16] and Dissanayake et al. [26] reported that the potential use of BC as a soil amendment can generally improve the soil quality of contaminated soil with MP particles; however, this impact differs depending on the temperature and feedstock type.
Although the quantity of research on the toxicological effects of MPs on organisms has increased dramatically, there is a need to better understand the potential mitigating effect of BC of date palm nuclei in this approach. This will achieve two benefits: utilizing the date palm nuclei and avoiding their accumulation in the arid environments especially and ecofriendly remediation of MP-contaminated soil. Thus, this study aims to: (1) study the effects of MPs and the toxicity, enzyme activity and properties of Vicia faba planted in pots; (2) examine the effects of MPs on the soil properties; (3) investigate the effects of MPs on microbial count and microbial biomass C and N; and (4) study the remediating effect of BC on the harmful effects on soil, plant and microbial activity. Thus, the novelty of this study is to better understand the effect of BC on mitigating the impacts of MPs on soil, plant and microbes, focusing on some aspects that have not been studied well before, such as cytogenetic effect of MPs in Vicia faba and investigating the effect of MP on soil C and N biomass.

2. Materials and Methods

The experiment was carried out at the Environmental and Biological Sciences Dept., Fac. of Home Economics, Al-Azhar Univ., and Tanta, Egypt.

2.1. Microplastic and Biochar Preparation

The residues of plastic banners (acrylic plastic) were obtained locally, manually cut with scissors into small pieces and ground by special grind into pieces less than 5 mm. The Biochar used in this study was made from date nuclei which were washed after collection, oven dried at 105°C and then pyrolyzed at 500°C in a muffle furnace for 4 h.

2.2. Soil and Seeds Preparation

Vicia Faba bean (sakha 1) were obtained from the agricultural research center, Sakha, Kafr El-Sheikh, Egypt, sorted in equal size and used in the experiment. The seeds were soaked for 24 h in water and dried by filter paper. Bulk soil (0–30 cm) was collected from agricultural soil from El-Gharbia governorate, Egypt, air-dried, ground, sieved (2 mm mesh size) and kept in plastic bags until analysis and use in the experiment.

2.3. Experimental Design

The current experiment was designed using a completely randomized design. First, microplastics were mixed with soil before being placed into each individual pot to help provide an equal distribution of MPs particles in the soil. Seven treatments with three replicates for each concentration were established: control (soil without MPs), 1.5% MPs, 7.5% MPs, 15% MPs, 1.5% MPs + 2% BC, 7.5% MPs + 2% BC, 15% MPs + 2% BC. Each pot contained 300 g of soil mixed with a treatment and four seeds of Vicia Faba beans.
In December 2021, the seeds were planted in pots and watered with 100 mL twice a week (approximately 70% of soil field capacity) during the first 3 weeks of growth and then every week. Pots were watered gently by hand by spraying distilled water on the soil surface. After 45 days, plants were carefully removed from the soil, washed by tap water and rinsed many times with distilled water.

2.4. Soil, Biochar and Microplastic Analysis

The physiochemical soil properties were performed. Soil pH, EC, TDS and organic matter were measured according to standard method. Soil pH was determined by pH-meter (JENWAY 3510, UK) in 1: 2.5 (soil: water) (w/v) suspensions [27]. Electrical conductivity (EC) of soil and TDS were measured in 1: 5 (soil: water extract) (w/v) using EC meter (Mi170, Milwaukee, Italy) [27]. Organic matter (OM) in soil (OM) was determined using a muffle furnace at 400 °C for 4 h by the loss on ignition method [28,29]. Total CaCO3 was measured by a Collins calcimeter. Soil available phosphorus (P) was extracted using ammonium bicarbonate–diethylene triaminepentaacetic (AB–DTPA) and calorimetrically determined by the ascorbic acid method. Available Fe, Mn and Zn were extracted using AB–DTPA [30]. The total metals were measured after extraction by HNO3 and HCl acids and 30% hydrogen peroxide [31]. The metals concentration was measured by atomic absorption spectrometry AAS (GBC Avanta E, Victoria, Australia).
The functional groups of BC and MPs and treated soil samples were analyzed by fourier transform infrared (FTIR) spectra of specimens, Jasco FT/IR 4100 spectrometer in the wavelength range of 4000–400 cm−0 with equipping all the samples on KBr tablets.

2.5. Plant Analysis

2.5.1. Germination Percentage

The germination percentage was calculated as in the following equation:
Germination rate% = (Number of germinated seeds/total number of seeds) * 100

2.5.2. Phytotoxicity

The phytotoxicity percentage is described as a percentage of phytotoxicity compared to phytotoxicity of untreated controls and is defined as follows [32,33,34]:
Phytotoxicity% = (Length of control root − Length of treated root)/(Length of control root) × 100

2.5.3. Morphological Traits

Morphological traits were directly determined (after plant removing from soil and washing) for fine roots (i.e., <2 mm in diameter), root length, shoot length, surface area and number of root nodules on a fresh sample of the plants. Fresh weight was recorded and plants were dried at 60°C for 72 h for recording dry weight. Plant leaves were randomly sampled from different parts of the plants. Maximum leaflet width (W) and length (L) were measured, and leaf area was calculated as described by [35].

2.5.4. Chlorophyll Content Measurement

Equal circle pieces of leaves were treated by 1: 1 v/v of 80% acetone and absolute ethyl alcohol to extract chlorophyll. Chlorophyll content was spectrophotometrically determined using a chlorophyll meter (Konica-Minolta, Osaka, Japan) [36].

2.5.5. Enzymes Activity

After the experiment, samples from fresh leaves were collected to measure the total soluble enzymes activity of Catalase (CAT) activity [37], peroxidases (POD) [38] and polyphenol oxidase [39].

2.5.6. Metals in Plants

Metals in plants were determined by the ash drying of 1 g of plant in a muffle furnace at 450 °C for 4 h, extracting with 20% HCl [40] and then the metal measured by the same previous ASS.
Bioaccumulation factor (BAF) was determined as an indicator to plat efficiency to accumulate metals from the soil and was calculated as follow [30]:
Bioaccumulation factor = metal in plant/metal in soil * 100

2.5.7. Cytological Analysis

Root tips of germinated seeds (1.5–2 cm) were cut and fixed in Carnoy’s fixative solution (1:3 glacial acetic acid: ethyl alcohol absolute) for 24 h. Carnoy’s fixed root tips were kept in ethyl alcohol (70%) at 4°C till using for cytological analysis.
Aceto–carmine stain (2%) was used for cytological preparation [41]. Mitotic index, numbers and types of abnormalities were scored in at least 3000 examined cells/treatment (1000 cell/replicate) using light microscope.
Percentage of abnormal cells and mitotic index (MI) were estimated by the following equation:
Mitotic index (MI) = (Total dividing cell/total dividing and non-dividing cell) * 100
Percentage of abnormal cell = (Total abnormal cell/total dividing cell) * 100

2.6. Microbial Activity

2.6.1. Total Count of Bacteria and Fungi

Standard dilution technique was used in which 10 g of soil were suspended in 90 mL of sterile distilled water and shaken for 20 min. Tenfold serial dilution was used for bacterial and fungal counts, as follows:
  • Total count of bacteria was estimated through plate count on media of nutrient agar [42]. This media were made at pH 7.4 ± 0.2 and incubated at 30 °C for 2 days.
  • Total count of fungi was estimated by plate count on media of Potato-dextrose agar after incubation of 5 days of incubation at 28°C [42] and adjusting pH at 5.2.

2.6.2. Soil Microbial Biomass Carbon and Nitrogen

Exactly 5 g of microwaved and field-moist soils were placed into 50 mL centrifuge tubes with 20 mL 0.5 M K2SO4 (adjusted at pH 7.0) and extracted by horizontal shaking at 250× g rpm for 60 min. The soil suspension was filtered to obtain soil-free filtrate. A few drops of concentrated sulfuric acid were added to the filtrates to inhibit microbial breakdown of organic C; then the samples were frozen until analyzed. Total organic C by TOC analyzer, Sievers 5310 C, while total organic N in these extracts were analyzed by Kjeldahl digestion and both calculated as in [43].

2.6.3. Statistical Analysis

The obtained data were statistically analyzed for One-Way Analysis of Variance and Duncan test. The results significance was considered at p < 0.05 by statistical package for social sciences (SPSS) software (for windows version 18). The results are presented as a mean ± standard deviation (SD).

3. Results

3.1. Soil Properties

As soil properties influence plant growth, MPs may indirectly affect plant growth by affecting soil characteristics. In this study, MP in soil resulted in significant increase in soil pH of all treatments more than the control (Table 1 and Table 2); however, there was insignificant difference between MP treatments and MP+BC. The increase of pH in some BC treatments was higher soil treated with MPs; this may be attributed to the alkaline nature of BC. Dissanayake et al. [26] reported that shape and type of MP affects the soil pH; however, the mechanism is still unclear. Although the EC values in soil were low, all treatment decreased significantly more than the control in all MP treatment (1.5, 7.5, and 15%) and 1.5%MP+BC. TDS values followed the same trend of EC. Soil organic matter decreased significantly more than the control, especially in 1.5%MP, 7.5%MP and 1.5%MP+BC. Zhang et al. [8] reported a decline in soil OM due to using plastic film in agricultural soils and plastic residue, and they found an inverse relationship between soil properties, crop production and plastic residue. It is clearly observed that BC application increased significantly the OM in soil, especially in 7.5%MP+BC. This may be explained by that MP at this level may because of MP is a source of organic C (which is highly correlated with organic matter) thus releasing dissolving OM as mentioned in the following sections regarding enzyme activities and total count of bacteria and fungi which increased by addition of BC also (where is has high organic matter). This may be supported by the data of microbial biomass C (which is a part of OC that correlated positively with OM) that followed the same trend. On the other hand, the higher level of MP may lead to more inverse impacts on soil organic matter which impeded the microbial and enzyme activity and declined the release of organic C and dissolving OM. As well, the nature of the soil (alkaline soil) play an important role in this matter, where Elasiouny et al. [44] reported that, in alkaline soils, widespread organic biomass is susceptible to speedy mineralization thus declining OM in soil. The data in the following table revealed that increased OM significantly in all treatments more than in the control; however, BC application decreased the treatment of 15%MP+BC significantly than its corresponding of 15%MP. The rate of degradation of MP in soil is currently unknown. However, it is assumed that the physical, chemical, biological and enzymatic characteristics of soils may provide a degradative environment for MPs degradation in topsoil. These mechanisms break down big plastic particles into smaller pieces, which accumulate in the soil and eventually reach plants, invertebrates, vertebrates and freshwater species [5].
The results also showed that available P decreased significantly in all treatments more than in the control; however, the BC application increased P concentration to some extent (with insignificant difference), especially in 1.5%MP+BC and 7.5%MP+BC over 1.5%MP and 7.5%MP. This indicates that the pollution with MP in soil may decrease P concentration in soil by 70%. Available Mn concentration in soil decreased significantly in soil after adding MP and MP+BC. The same trend is observed in available Fe concentration except 7.5% MP. Available Zn followed a different trend than Fe and Mn, where its concentration fluctuated between the treatment and it increased significantly more than in the control in all treatments except 15%MP and 15%MP+BC. The BC application increased Zn concentration in 1.5% MP+BC and 7.5%MP+BC than 1.5%MP and 7.5%MP, respectively. Zhu et al. [45] stated that MPs could disturb the quality of soil and nutrient cycles through changing the soil properties, hence reducing its fertility and disturbing the local microbial community. However, Osman et al. [46] reported that BC can modify MPs in the medium through sorption and/or implicit microbial biodegradation.
The results may be interpreted by the FTIR images, which include the function groups of BC, MP and treated samples as shown in Figure 1. The FTIR spectra in MPs showed the presence of C=O and O–H functional groups especially, which referees to the presence of esters and phenols compounds. Whereas, the C=C and C–H functional groups was dominated in BC referring to the presence of alkene and aromatic compounds. In all treated soil, the dominated functional groups were C=C and O–H. Usman et al. [47] confirmed the presence of C=C group at 400–600°C as well as the presence of O–H, which disappeared at the high temperature of 800°C in data palm BC.
It is reported that MP adds surface with unique properties to the soil, which enable it to transfer and adsorb substances from soil, especially heavy metals [48]. Thus, the decreasing of studied elements, especially P, Mn and Fe, may be accounted for enhancing the adsorption capacity of MPs due to the presence of O-containing groups that were confirmed also by He et al. [49] in their research on Mn adsorption by aged polystyrene MP.
Since both BC and MPs particles’ surfaces have negative charge, the force between both would be Van der Waals force [22]. It was concluded that electrostatic interaction and chemical bonding between BC and MP particles controlled their retention mechanisms [20]. Wang et al. [22] investigated the filtration of MPs on BC and found that the MPs’ retention mechanisms onto BC were described as “stuck: where the large particles of MPs were stuck in the gaps between the filtration particles”, “trapped: where the particles were trapped inside the BC pores” and “entangled: where the BC formed flaky small particles that displayed colloidal properties entangling the MPs and increased their size, hence, triggering their immobility”.

3.2. Plant Properties

3.2.1. Germination Percentage of Seeds

Microplastic did not adversely affect the germination percentage of seeds where all seeds were grown. The germination rate is a critical indicator used for measuring seed vitality and germination ability under different environmental stressors [50]. It is clear that the examined doses of MP did not have any effects on the plant in the initial stage. On the contrary of this study, Huang et al. [51] found that seeds exposed to MPs for 8 h suffered late germination. This may be because the MPs physically blocked the pores in the seed capsule, inhibiting the seeds to absorb water.

3.2.2. Phytotoxicity Percentage of Root

The percentage of phytotoxicity (Table 3) of treated root demonstrated higher phytotoxicity with lower MPs level. The BC application decreased the phytotoxic effect of MPs to roots especially at the 7.5% MP+BC. These results indicate that the 7.5% level of MPs was the highest influence on the root of Vicia faba. Although 15% MP is a higher level of MP, its effect was lower than 1.5% and 7.5% MP. This may have implied that the plant in the low level can adsorb higher levels of MPs through its roots to some extent, and after this extent, it can adsorb other MPs particles. This was clear to some extent because, in the first weeks of the experiment, the MPs particles can be seen, while when harvesting most of the MPs were broken and could not be seen by eyes on the soil particles. This indicates the breakdown of MPs particles and adsorption by plants. Li et al. [13] stated that current findings have demonstrated that submicron (i.e., 0.2 μm) MPs can enter the stele of plants via the crack-entry mechanism at locations of lateral root emergence, providing MPs with a novel route into the food web and/or the human body. These submicron MPs in plants may break down and thus emit hazardous substances (e.g., benzene), interfering with plant metabolism. Although MPs with relatively large particle sizes cannot be ingested by plant roots, they may have an impact on the survival and growth of the targeted plants through a variety of mechanisms, such as contouring sunlight, causing damage to root tissue, blocking cell wall pores and changing soil physicochemical properties.

3.2.3. Morphological Traits

Vicia faba plant in this study responded to the MPs addition to soil in different ways as seen in the following results. It is also reported that different plant species respond differently to the same MPs exposure [13]. The data in Table 4 and Table 5 show that MP affected positively on the shoot length and all treatment of MP was longer than the control. In addition, BC application increased the shoot length significantly more than the control; however, there was insignificant difference between MP treatment and MP+BC. Although most of the treatments were longer than control treatment in root length, the difference was insignificant. Fresh and dry weight was also not significantly affected by addition of MP although the low level of MP was lower than control even after BC application. On the other hand, dry and fresh weight of root was significantly increased more than the control by MP addition. The physical soil alterations help to interpret the decline in root weight, hence affecting the adsorption of crop nutrient and, ultimately, crop yield [7]. This may be also interpreted by the strongest stimulation [52]; however, BC application significantly decreased fresh and dry weight of root more than MP treatments. The number of root nodules significantly increased more than the control after addition of MP and MP+BC, and the highest increase was noticed at the level 7.5%MP+BC. This indicates that the BC application affected the root nodules number positively although it is not clearly affected on the other morphological traits. Yadav et al. [5] stated that the effects of MP on plants are often unknown; hence, more studies are needed in this area as the presence of MPs in agroecosystems can be a major impediment to sustainable agriculture output. The influence of MP on plants and soil productivity is poorly understood. To comprehend and forecast probable long-term repercussions with specific reference to the agroecosystem, pioneering work in the field is required [5]. Therefore, this study aims to contribute in achieving this objective.

3.2.4. Chlorophyll Content

It is indicated from Figure 2 that chlorophyll content (a, b and total) flocculated between the treatment and decreased more than the control in all treatments even with BC application. The highest decrease of chlorophyll was recorded with 1.5%MP and 1.5% MP+BC. This is not in accordance with the statement of Li et al. [13] in which they stated that, in general, a low concentration of pollutants has little or insignificant negative effects on plants. This also is implied on MPs. It is reported by [53] the contradictory results of chlorophyll content after exposure to MPs, and the decrease of chlorophyll content was markedly observed. This was similar to our results.

3.2.5. Enzyme Activity

Plant enzymes (catalase, peroxidase and polyphenol oxidase) were evaluated as bioindicators for survival of Vicia faba plants under MPs contamination stress. The addition of MPs to soil in this experiment changes the activity of the previous mentioned enzymes as in Figure 3. Catalase activity was increased more than the control with increasing MPs from 1.5% to 7.5%, while it decreased at 15% of MPs. On the other hand, the BC application in soil decreased the catalase activity with the level of MP (1.5%) lower than the control, while it increased more than the control with the higher level of MPs, especially 7.5%. Peroxidase activity followed the opposite trend than catalase, and the level 7.5% was the most special. The observed results of increasing catalase and decreasing peroxidase was also recorded by Lian et al. [52] in the leaves of Glycine max affected by MPs. Polyphenol oxidase followed a different trend than both previous enzymes; it increased clearly more than the control with 1.5% MP, while it did not change more than the control with other levels of MPs. On the other hand, after application of BC and MP+BC, the polyphenol oxidase decreased more than the control with 1.5%MP+BC; increased clearly more than the control with 7.5%MP+BC; and did not change more than the control with 15%MP+BC as in 15%MP only. Soil enzyme activity, which is mediated by soil microflora, is critical for sustaining nutrient cycling in the terrestrial environment, such as the carbon, phosphorus and nitrogen cycles [5]. Liang et al. [54] reported that MPs, as an organic carbon source, may add an artificial carbon source, potentially altering soil enzyme function as a possible substrate [54], which subsequently affects the microbial activity and the plant. They found in their experiment on the effect of MPs fiber on soil enzyme activities that adding organic materials (leaves and straws of various plants) to soil increased enzyme activity, but MPs fibers reduced enzyme activity in some circumstances. The impacts of MPs fibers on enzyme activities were neutral in soil without OM addition, whereas negative effects of MPs fibers were seen with OM addition [54]. In brief, Pérez-Reverón et al. [53] reported that the presence of MPs results in greater levels of oxidative stress markers. As a result, antioxidant enzymes, such as catalase and peroxidase, generally exhibit higher concentrations. This is similar to some treatments in our study. Li et al. [13] stated that large MPs cannot be adsorbed by roots; they may accumulate on the roots. Thus, MPs, mainly those that have a coarse surface and sharp angles, may physically harm a plant root which limits root activity and impedes root growth. It is supposed that MPs induce physical damage to roots which causes ROS generation. The high ROS concentration exceeds cell tolerance, and antioxidant enzyme synthesis is reduced. However, when soil amendments such as BC are applied, the inhibitory effects of MPs on plant roots may be reduced. This is how it is in some of the cases in our study. Wang et al. [22] conducted a leaching column experiment and found that BC filters had a capacity higher than 95% to remove and immobilize MPs with 10 μm diameters.

3.2.6. Bioaccumulation Factor (BAF) of Some Metals in Plants

The BAF is used to evaluate the efficiency of a plant to accumulate and translocate metals [55]. According to [56,57], BAF is used for sorting plants as excluders, accumulators and hyperaccumulators of metals based on their accumulated concentration as follows: <1, <5 and >1, >5, respectively. Thus, in this study, Vicia Faba plant is considered an accumulator to Mn in all treatments, especially MPs only (Figure 4). However, the BC decreased the BAF values of Mn in plant that those of MPs only which means that BC can mitigate the effect of MP. The BAF of Fe as higher than 5 indicates that Vicia Faba plant is considered a hyperaccumulator to Fe in these conditions. As well, all treatments were higher than in the control except in 1.5%MP+BC and 7.5%MP+BC. The data also showed that the Vicia Faba plant is considered a hyperaccumulator Zn, with values higher than control except in 15%MP and 7.5%MP+BC. As well, it is shown that BC decreased the BAF in some cases than MP only. It can be concluded that MP increased the accumulation of these metals in plants, and the idea is supported that MP can facilitate the entrance of metals and pollutants to plant tissues. This was clear in this study, where during the experiment, no deficiency symptoms on grown plants were noticed.
Although BC decreased the accumulation of metals in some cases in this study, it is clear that its concentration was not enough to avoid all negative effects of MP. The toxicity of MPs to living organisms is most likely caused by the adsorption of various chemicals to their surface, followed by the processes of their adverse effects on biota survival and reproduction. Furthermore, toxicity mechanisms are related to oxidative stress and aberrant immune systems to describe the negative consequences of microplastics. Physical features of plastics within the body, in addition, might be another reason for observing unfavorable consequences. When MPs are consumed, they can cause harm such as obstruction in the organism’s body due to their size and structure [13]. Elbasiouny et al. [58] also stated that MPs tend to absorb and accumulate metals and contaminants from surrounded environments, causing not only physical harm to species but also the possible transfer of pollutants to organisms and the food chain.

3.2.7. Cytological Analysis

In the present study, phase indices were calculated, where the highest percentage of prophase was recorded in control, whereas the highest percentage of metaphase was recorded in 1.5%MP+BC and the highest percentage of anaphase and telophase was recorded in 15%MP+BC (Table 6). The Increase in the cytogenotoxicity of toxicants can be indicated by the decrease in the cells mitotic index (MI) [59]; it is considered a good indicator of cytotoxicity [60]. The results in Table 6, Table 7 and Table 8 showed the effect of different concentrations of MPs and MPs+BC on mitotic index (%) and abnormalities (%) in V. faba root tips. Cytological analysis showed the highest value of mitotic index in control, followed by 1.5%MP+BC with significant differences between control and other treatments. The results illustrated insignificant (p > 5%) differences between treatments before and after BC application except in the treatment of 7.5%MP and 7.5%MP+BC. It is clear that MPS induces an increase in the mitotic index than MPs+BC, as well, MPs increased the chromosomal aberration%, as well BC application significantly reduced the toxicity of MPs. It is noticed that BC with its constant percentage (2%) did not affect the higher amount of MPs, indicating that studying the effect of the application of a higher amount of BC on mitigating the negative impacts of MP is critical particularly with the positive effect of BC at this studied level. The most frequent types of mitotic abnormalities induced by MPs were stickiness, chromosomal disturbances, pole-to-pole metaphase, fragments, bridges and c-metaphase (Figure 5). These results are agreed with Kaur et al. [61] who reported that MPs decreased the mitotic index. In addition, most of the MP concentration caused cytotoxic and nuclear damage by adversely impacting the spindle formation and induction of micro-nucleated cells [60]. Gopinath et al. [62] attributed the decrease in the MI of root cells to the potential of MPs in DNA synthesis inhibition, abduction of mitotic phases and slow cell progression. Although the studies on cytotoxicity are rare, some works indicated significant genotoxic effects of MP on wheat [63], spring onion [64], and ryegrass [65], A. cepa [66] and V. faba [67]. The primary mechanism following the MP genotoxic effects is mostly unknown; however, it might be due to the MP accumulation of MPs in the root tissues, hence blocking cell wall pores and disrupting the transport of essential nutrients [67].

3.3. Microbial Activity in Rhizosphere Zone and Soil

3.3.1. Total Count of Bacteria and Fungi in Rhizosphere Zone and Soil

Plant growth and development are heavily reliant on soil structure and biota, and any changes in the soil microbiota may have a direct impact on plant growth, productivity, population and diversity. Thus, microbes are key drivers in soil ecosystem activities; however, they are extremely vulnerable to environmental pollutants and stress [5]. The influence of the BC in soil treated with MPs on the quantification of bacteria and fungi populations in rhizosphere zone and soil was evaluated. Table 9 and Table 10 show that the treatment of soil with BC significantly increased the total count of bacteria and fungi in rhizosphere zone compared to the control and MP treatments only; however, their values with MPs are higher than the control. Yadav et al. [3] reported that MPs have an impact on the soil microbial community and organisms by altering soil microbial diversity, inducing oxidative stress, causing death, decreasing soil fauna activity and growth and so on. It is noticed in our study also that the highest bacterial and fungal count in rhizosphere zone was recorded with 7.5%MP+BC. An insignificant difference was noticed in total count of bacteria in soil treated with 7.5%MP and 15%MP. The effect of BC was markedly noticed also on the total count of fungi in rhizosphere and soil. In the present study, a difference in the soil’s microbial load was observed with the addition of MPs, with increasing MP a microbial count increased initially followed by a decline. This result might be because of the increase in content of dissolved soil organic N, P and C after addition of MP in soil, as reported by Liu et al. [68]. The MP particles, because of their ubiquity and relatively large surface area, provide a new ecological habitat for bacterial populations in soil conditions. Changes in soil porosity caused by MP addition may affect O fluxes and, as a result, the distribution of aerobic and anaerobic bacteria. Furthermore, it has been shown that the quantity of MPs in soils has a growing influence on the nutritional contents of dissolved OM; hence, they promote soil microbial activity [53]. Several studies showed that application of OM in agriculture soil leads to increased release of growth regulators in root zone, owing to the increase of microbial load in rhizospher soil [69]. This may be one of the reasons that soil treated with BC was higher in its microbial count. Ren et al. [25] also found that the BC application in soil increased the OM of rhizosphere′s soil, P and N, in addition to increasing the microorganisms’ count that has the capability of degradation and utilization of cellulose and OM. It is noticed from the data in the following table that microbial count in rhizosphere region is higher than soil. Yadav et al. [3] reported that the plant rhizosphere is a zone of complex biological and ecological activities that covers the soil around the roots. Plants absorb minerals/nutrients and water from the rhizosphere to grow and develop. Microplastics and nanoplastics may impede plant development by modifying soil microbiota. Osman et al. [46] stated that BC can be utilized to boost the microbial niche during anaerobic digestion as well as the adsorption mechanism, thus helping the decomposition of MPs. Particularly, BC can enhance direct interspecies electron transfer in anaerobic digestion processes by boosting the growth pseudomonas stutzeri and pseudomonas putid as an effective plastic biodegrading bacterium. Also, one of the main reasons that may influence the microbial activity is pH (which increased in this study). It is reported by Meng et al. [70] that soil pH is an important component associated with soil nutrient dynamics, microbial activity and decomposition of SOC. As a result, MP pollution can influence microbial populations both directly and indirectly by influencing physicochemical processes in the soil.

3.3.2. Soil Microbial Biomass C and N

It is clear from Figure 6 that microbial biomass C (MBC) increased markedly in all treatments more than in the control because of the addition of MPs and BC also. The highest MBC was observed with 7.5%MP+BC followed by 7.5%MP. It is shown also that MBC increased gradually when MPs level increases. Although the known relationship between soil C and N, microbial biomass N (MBN) did not follow the same trend of MBC; MBN has a flocculation trend. However, all treatment increased more than the control except 1.5%MP+BC. The highest level of MBN was recorded with 15%MP followed by 1.5%MP = 15%MP+BC. The studies that interesting in MBC and MBN in soil treated with MPs are very rare and may this point be not considered yet. Thus, this study is contributing to fill the knowledge gap in this regard. However, Ren et al. [71] reported that BC was shown to accelerate the breakdown rate of organic pollutants via increasing microbial activity or microbial utilization of the organic pollutants on the BC, therefore impacting microorganisms either directly or indirectly. By modifying soil conditions, BC may either directly provide nutrients and protection to soil microorganisms or provide a better living environment for them. This may be one of the reasons that aid to enhance MBC or N in soil treated with MP and BC.

4. Conclusions

This study confirmed some negative impacts of MP on soil properties, especially soil pH, OM and CaCO3 as well as the association of MP in transferring Fe, Mn and Zn based on the resulting high bioaccumulation factors of Fe and Zn. Physiological and cytological properties of Vicia faba are affected negatively by MP, especially at high level of MPs. The study markedly supported some previous statement regarding enhancing microbial count and biomass due to the presence of MP in soil. Despite these negative impacts of MPs, biochar revealed some positive effects in mitigating the impacts of MPs on soil properties, Vicia faba properties and microorganisms’ activity. Biochar in this study was used with a concentration of 2%; this may be little to mitigate the negative impacts of MPs, so it is recommended to use higher concentration of biochar rather alone or mixed with other eco-friendly materials to maximize the positive effects of soil and plants. Although, it is clear from the results that biochar could be promising in MPs remediation from soil, more studies are also recommended to better understand the BC mechanisms in this process, especially with different types of biochar and in a nano-form of biochar considering the effect of different soil properties and surrounding environmental conditions, especially higher temperature.

Author Contributions

H.E.: Conceptualization, methodology, validation, formal analysis, investigation, resources, data curation, writing—original draft preparation, writing—review and editing, supervision, project administration; A.A.M.: Conceptualization, methodology, validation, formal analysis, investigation, resources, data curation supervision, project administration; A.Z.: Conceptualization, methodology, validation, formal analysis, resources, data curation, writing—original draft preparation, writing—review and editing, supervision, project administration; H.M.E.: methodology, formal analysis, investigation, resources, data curation, writing—original draft; S.F.M.D.: methodology, formal analysis, investigation, resources, data curation, writing—original draft preparation; B.A.E.: methodology, formal analysis, investigation, resources, writing—original draft; S.A.E.-S.: methodology, formal analysis, investigation, resources, writing—original draft; A.A.E.-S.: methodology, formal analysis, investigation, resources, writing—original draft; A.M.S.-E.: methodology, formal analysis, investigation, resources, writing—original draft; M.D.: methodology, formal analysis, data curation; A.-Z.E.E.E.: methodology, formal analysis, data curation, writing—original draft; S.M.A.: methodology, resources, formal analysis, writing—original draft preparation; N.O.A.: methodology, formal analysis, resources writing—original draft preparation; D.S.A.: methodology, resources, formal analysis, writing—original draft preparation; N.R.A.: methodology resources, formal analysis, writing—original draft preparation; A.O.A.: methodology, formal analysis, resources, writing—original draft preparation; R.E.H.: methodology, formal analysis, resources, writing—original draft preparation; F.E.: Conceptualization, methodology, formal analysis, validation, investigation, data curation, writing—review and editing. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by The Academy of Scientific Research & Technology—ASRT-Graduation Project (Bedaity project), Egypt for financiering and supporting this work.

Data Availability Statement

The data are available when required under the responsibility of the corresponding author.

Acknowledgments

The authors acknowledge ASRT and Environmental and Biological Science Department, Home Economics faculty, Al-Azhar University, Egypt for supporting and facilitating this work.

Conflicts of Interest

The authors declare no conflict of interest.

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Figure 1. FTIR spectra of biochar, microplastic and all treated soils. (a) Control; (b) 1.5% MP; (c) 7.5% MP; (d) 15% MP; (e) 1.5% MP +BC; (f) 7.5% MP + BC; (g) 15% MP + BC; (h) microplastic; (i) biochar.
Figure 1. FTIR spectra of biochar, microplastic and all treated soils. (a) Control; (b) 1.5% MP; (c) 7.5% MP; (d) 15% MP; (e) 1.5% MP +BC; (f) 7.5% MP + BC; (g) 15% MP + BC; (h) microplastic; (i) biochar.
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Figure 2. Biochar effect on the chlorophyll content in Vicia faba plants affected by microplastic in soil. Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
Figure 2. Biochar effect on the chlorophyll content in Vicia faba plants affected by microplastic in soil. Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
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Figure 3. Biochar effects on the enzymes’ activity in Vicia faba plants affected by microplastic in soil. CAT: Catalase, POX: Peroxidase, PPO: Polyphenol Oxidase.
Figure 3. Biochar effects on the enzymes’ activity in Vicia faba plants affected by microplastic in soil. CAT: Catalase, POX: Peroxidase, PPO: Polyphenol Oxidase.
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Figure 4. Biochar effects on bioaccumulation factor (BAF) of studied metals in Vicia faba plants affected by microplastic in soil.
Figure 4. Biochar effects on bioaccumulation factor (BAF) of studied metals in Vicia faba plants affected by microplastic in soil.
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Figure 5. Types of abnormalities observed in V. faba root tips cells due to microplastic effect in soil and mitigation by biochar. (a) Stickiness prophase and anaphase with bridge and vagrant chromosome, (b) C–metaphase, (c) Disrupted metaphase, anaphase with fragment and C-metaphase, (d) Anaphase with laggard chromosome, (e) Telophase with laggard chromosome, (f) Scattered ball metaphase, (g) Pole-to-pole metaphase, (h) Star anaphase (i) Multipolar anaphase.
Figure 5. Types of abnormalities observed in V. faba root tips cells due to microplastic effect in soil and mitigation by biochar. (a) Stickiness prophase and anaphase with bridge and vagrant chromosome, (b) C–metaphase, (c) Disrupted metaphase, anaphase with fragment and C-metaphase, (d) Anaphase with laggard chromosome, (e) Telophase with laggard chromosome, (f) Scattered ball metaphase, (g) Pole-to-pole metaphase, (h) Star anaphase (i) Multipolar anaphase.
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Figure 6. Biochar effects on microbial biomass C (MBC) and microbial biomass nitrogen (MBN) in soil affected by microplastic.
Figure 6. Biochar effects on microbial biomass C (MBC) and microbial biomass nitrogen (MBN) in soil affected by microplastic.
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Table
Table 1. Biochar effects on soil properties treated with different levels of microplastic.
Table 1. Biochar effects on soil properties treated with different levels of microplastic.
TreatmentpHEC d Sm−1TDS (mg L−1)OM%CaCO3%Avail. P mg kg−1Avail. Mn mg kg−1Avail. Fe mg kg−1Avail. Zn mg kg−1
Control7.49 d* ± 0.040.46 a ± 0.008231.50 a ± 4.503.36 b ± 0.148.71 c ± 0.411.20 a ± 1.0018.32 a ± 4.2719.03 a ± 7.530.91 e±0.07
1.5%MP7.53 cd ± 0.040.44 b ± 0.004221.00b ± 2.002.20 c ± 0.4010.99 a ± 0.620.20 b ± 0.007.85 b ± 0.068.02 c ± 0.572.34 b ± 0.21
7.5%MP7.63 ab ± 0.060.41 d ± 0.004206.00 d ± 2.002.85 bc ± 0.6510.37 ab ± 0.410.25 b ± 0.057.95 b ± 0.0616.12 ab ± 6.291.38 d ± 0.05
15%MP7.61 ab ± 0.020.44 b ± 0.001217.50 b ± 0.503.10 bc ± 0.8010.99 a ± 0.200.20 b ± 0.007.18 b ± 0.219.95 bc ± 0.740.82 e ± 0.02
1.5%MP+BC7.61 ab ± 0.060.42 c ± 0.003211.50 c ± 1.502.57 bc ± 0.6210.99 a ± 0.200.35 b ± 0.156.67 b ± 0.449.34 bc ± 1.342.60 a ± 0.28
7.5%MP+BC7.57 bc ± 0.020.47 a ± 0.003233.50 a ± 1.505.10 a ± 0.4010.37 ab ± 0.410.30 b ± 0.009.76 b ± 0.239.34 bc ± 0.211.79 c ± 0.028
15%MP+BC7.65 a ± 0.020.47 a ± 0.006234.50 a ± 3.503.15 bc ± 0.1510.16 b ± 0.200.20 b ± 0.097.03 b ± 0.4210.05 bc ± 0.140.84 e ± 0.015
EC: Electrical conductivity; TDS: Total dissolved salts; OM: Organic matter; Avail: available. * Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
Table
Table 2. One way analysis of variance of soil properties affected by microplastics and biochar.
Table 2. One way analysis of variance of soil properties affected by microplastics and biochar.
Soil PropertyFSig.
pH7.3830.001
EC63.2210.000
TDS59.2430.000
OM9.9500.000
CaCO32.6800.000
Avail. P13.4200.060
Avail. Mn19.1080.000
Avail. Fe3.6700.021
Avail. Zn0.2850.000
Table
Table 3. Biochar effect on the phytotoxicity of soil microplastic on Vicia faba roots.
Table 3. Biochar effect on the phytotoxicity of soil microplastic on Vicia faba roots.
TreatmentControl1.5% MP7.5% MP15% MP1.5%MP+BC7.5%MP+BC15% MP+BC
Phytotoxicity%020.9223.4812.1317.4313.2910.82
Table
Table 4. Biochar effect on the morphological traits of Vicia faba that affected by microplastic addition to soil.
Table 4. Biochar effect on the morphological traits of Vicia faba that affected by microplastic addition to soil.
TreatmentsShoot Length (cm)Root Length (cm)Fresh Weight of Shoot (g)Dry Weight of Shoot (g)Fresh Weight of Root (g)Dry Weight of Root (g)Number of Root Nodules
Control19.6 b* ± 1.8917.18 a ± 1.227.11 a ± 1.690.68 a ± 0.274.21 d ± 0.550.26 d ± 0.047.0 b ± 5.91
1.5%MP23.3 ab ± 2.2815.76 a ± 3.046.90 a ± 1.570.67 a ± 0.275.71 bc ± 1.130.39 bcd ± 0.0812.8 ab ± 4.20
7.5%MP27.0 a ± 2.6220.34 a ± 7.457.99 a ± 1.490.65 a ± 0.246.20 ab ± 1.150.40 ab ± 0.099.4 b ± 7.23
15% MP22.9 ab ± 3.4118.32 a ± 2.107.01 a ± 0.670.74 a ± 0.077.51 a ± 0.650.47 a ± 0.0513.6 ab ± 2.40
1.5%MP+BC25.9 a ± 2.4016.90 a ± 3.976.71 a ± 0.880.60 a ± 0.214.67 cd ± 0.790.29 cd ± 0.0610.6 b ± 6.54
7.5%MP+BC26.2 a ± 4.8017.20 a ± 2.387.82 a ± 2.320.80 a ± 0.375.65 bc ± 1.700.36 bc ± 0.1118.8 a ± 6.76
15%MP+BC25.6 a ± 1.1921.00 a ± 4.757.54 a ± 1.680.65 a ± 0.296.63 ab ± 0.810.43 ab ± 0.0712.4 ab ± 4.39
* Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
Table
Table 5. One way analysis of variance of plant properties affected by microplastic and biochar.
Table 5. One way analysis of variance of plant properties affected by microplastic and biochar.
Plant PropertyFSig.
Shoot length4.0560.005
Root length1.1200.376
Fresh weight of shoot0.4940.807
Dry weight of shoot0.3260.918
Fresh weight of root5.8770.000
Dry weight of root5.0630.001
Number of root nodules2.2220.070
Table
Table 6. Cytological indicators of examined root tips of Vicia faba affected by biochar after microplastic addition to soil.
Table 6. Cytological indicators of examined root tips of Vicia faba affected by biochar after microplastic addition to soil.
TreatmentsNumber of Examined CellsNumber of Dividing CellsNumber of Abnormal CellsMitotic Phase Index
PMAT
Control3278309-62.4510.0312.9414.56
1.5%MP30211778244.6316.3823.1615.81
7.5%MP302620010054.5017.5020.507.50
15% MP32341596352.2017.6116.3513.83
1.5%MP+BC31881687934.5226.7822.6116.07
7.5%MP+BC3117126464.4419.015.0721.42
15%MP+BC31161466441.0913.0123.9721.91
P: Prophase; M: Metaphase; A: Anaphase; T: Telophase.
Table
Table 7. Biochar effect on mitotic index, types and percentage of abnormalities of V. faba root tip cells under different level of microplastic.
Table 7. Biochar effect on mitotic index, types and percentage of abnormalities of V. faba root tip cells under different level of microplastic.
TreatmentsStickinessDisruptedMitotic Aberration (%)Mitotic Index
(%)		Abnormalities
(%)
Multipolar AnaphaseFragmentBridgeC-MetaphasePole-to-Pole MetaphaseStar
Anaphase		Scattered Ball MetaphaseLaggard
Control------- -9.39 ± 2.42 a*0.00 ± 0.00 e
1.5%MP42.2624.39-13.414.874.871.212.431.214.875.85 ± 0.15 bc69.91 ± 5.31 a
7.5%MP3425-9712-1666.60 ± 1.08 b57.38 ± 2.03 b
15% MP25.3923.804.766.3411.1115.879.521. 58-1.584.89 ± 0.56 bc46.77 ± 3.23 c
1.5%MP+BC14.4514.454.8115.6614.4515.668.437.221.203.617.11 ± 1.02 b49.30 ± 5.57 c
7.5%MP+BC26.0810.86-13.0415.2117.394.342.174.346.524.06 ± 1.23 c29.45 ± 2.98 d
15%MP+BC32.8126.56-12.518.756.253.125---4.69 ± 1.24 bc47.77 ± 4.33 c
* Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
Table
Table 8. One-way analysis of variance of cytological indicators of examined root tips of Vicia faba affected by biochar after microplastic addition to soil.
Table 8. One-way analysis of variance of cytological indicators of examined root tips of Vicia faba affected by biochar after microplastic addition to soil.
Cytological IndicatorsFSig.
Mitotic index5.9890.003
Abnormalities104.7850.000
Table
Table 9. Effect of microplastic and biochar treatment on total count of bacteria and fungi.
Table 9. Effect of microplastic and biochar treatment on total count of bacteria and fungi.
TreatmentsBacteria C.F.U × 107Fungi C.F.U × 103
Rhizosphere ZoneSoilRhizosphere ZoneSoil
Control10.3 ± 2.51 b*5.3 ± 3.0 c9.6 ± 3.21 d3.0 ± 1.00 c
1.5%MP18.6 ± 3.51 b10.3 ± 1.52 b23.6 ± 4.50 bc8.3 ± 2.51 bc
7.5%MP17.3 ± 9.29 b5.0 ± 1.00 c13.6 ± 3.51 d10.3± 4.04 b
15%MP14.3 ± 6.65 b2.6 ± 1.52 c17.6 ± 6.65 cd8.0 ± 3.00 bc
1.5%MP+BC56.6 ± 16.74 a4.0 ± 1.73 c27.6 ± 2.51 b19.0 ± 3.00 a
7.5%MP+BC73.3 ± 23.86 a11.6 ± 5.13 b33.0 ± 5.56 ab23.3 ± 4.50 a
15%MP+BC51.0 ± 7.93 a16.3 ± 2.08 a38.6 ± 4.72 a23.0 ± 4.58 a
Values with the same letter within column are insignificantly different according to Duncan multiple range test (p ≤ 005). C.F.U: Colony Forming Unit. * Different letters in each column indicate significant difference at p < 0.05 (based on Duncan test) between treatments.
Table
Table 10. One way analysis of variance of total count of bacteria and fungi in rhizosphere and soil affected by biochar after microplastic addition to soil.
Table 10. One way analysis of variance of total count of bacteria and fungi in rhizosphere and soil affected by biochar after microplastic addition to soil.
PropertyFSig.
Total bacteria in rhizosphere12.5490.000
Total bacteria in soil10.7030.000
Total fungi in rhizosphere15.7070.000
Total fungi in soil16.6310.000
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Elbasiouny, H.; Mostafa, A.A.; Zedan, A.; Elbltagy, H.M.; Dawoud, S.F.M.; Elbanna, B.A.; El-Shazly, S.A.; El-Sadawy, A.A.; Sharaf-Eldin, A.M.; Darweesh, M.; et al. Potential Effect of Biochar on Soil Properties, Microbial Activity and Vicia faba Properties Affected by Microplastics Contamination. Agronomy 2023, 13, 149. https://doi.org/10.3390/agronomy13010149

AMA Style

Elbasiouny H, Mostafa AA, Zedan A, Elbltagy HM, Dawoud SFM, Elbanna BA, El-Shazly SA, El-Sadawy AA, Sharaf-Eldin AM, Darweesh M, et al. Potential Effect of Biochar on Soil Properties, Microbial Activity and Vicia faba Properties Affected by Microplastics Contamination. Agronomy. 2023; 13(1):149. https://doi.org/10.3390/agronomy13010149

Chicago/Turabian Style

Elbasiouny, Heba, Azza A. Mostafa, Amina Zedan, Hala M. Elbltagy, Sherifa F. M. Dawoud, Bodor A. Elbanna, Safaa A. El-Shazly, Amany A. El-Sadawy, Aisha M. Sharaf-Eldin, Marwa Darweesh, and et al. 2023. "Potential Effect of Biochar on Soil Properties, Microbial Activity and Vicia faba Properties Affected by Microplastics Contamination" Agronomy 13, no. 1: 149. https://doi.org/10.3390/agronomy13010149

APA Style

Elbasiouny, H., Mostafa, A. A., Zedan, A., Elbltagy, H. M., Dawoud, S. F. M., Elbanna, B. A., El-Shazly, S. A., El-Sadawy, A. A., Sharaf-Eldin, A. M., Darweesh, M., Ebrahim, A. -Z. E. E., Amer, S. M., Albeialy, N. O., Alkharsawey, D. S., Aeash, N. R., Abuomar, A. O., Hamd, R. E., & Elbehiry, F. (2023). Potential Effect of Biochar on Soil Properties, Microbial Activity and Vicia faba Properties Affected by Microplastics Contamination. Agronomy, 13(1), 149. https://doi.org/10.3390/agronomy13010149

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