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Abstract

The aim of the study was to determine the time-delayed (after three years from the moment of soil pollution) effect of petroleum-derived products (PDPs) (petrol, diesel fuel and used engine oil) on the interaction between selected host plant (broad bean) and a herbivorous insect closely related to it (Sitona spp.). We assessed the condition of the plant exposed to pollutants (i.e. its growth and chemical composition), then we evaluated the attractiveness of the plant for both larvae and adults of the insect. The evaluation covered also the effect of bioremediation by using ZB-01 biopreparation. The results showed that after 3 years from soil contamination, engine oil and diesel fuel limited the feeding of adult sitona weevils while petrol caused increase in the attractiveness of plants for these insects. The PDPs negatively affected the growth of plants. The changes in element content depended on the type of pollutant. The biopreparation ZB-01 eliminated or reduced the differences caused by the presence of PDPs in the soil regarding the chemical composition of the host plant, and limited feeding by both the larvae and adult individuals of sitona weevils. The negative relationships between the contents of both some macroelements (Mg, S) and heavy metals (Zn, Ni), and feeding of imago of Sitona were observed. The obtained results indicate that PDPs remain for a long time in the environment and adversely affect not only the organisms directly exposed to the pollution – plants growing on polluted soil but also further links of the trophic chain, i.e. herbivores
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Authors and Affiliations

Milena Rusin
1
Janina Gospodarek
1
Aleksandra Nadgórska-Socha
2

  1. Department of Microbiology and Biomonitoring, University of Agriculture, Kraków, Poland
  2. Department of Ecology, University of Silesia in Katowice, Poland
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Abstract

Using microorganisms in eliminating or reducing the impacts of harmful remnants is very ancient manner. The current study was conducted to explore the potential of utilizing some fungi species that isolated from the main sewage treatment plant in Al-Muamirah area, Babylon-Iraq, in reducing some pollutants. Six fungi taxa Aspergillus flavus, A. niger, A. terrues, Candida albicans, C. krusei, and Penicillium digitatum were identified before any treatment process, whereas only four fungi species A. flavus 20%, A. niger 20%, A. terrues 10%, and P. digitatum 18% were recognized after completing the physical and chemical treatment stages. Only three taxa A. niger, A. terrues, and P. digitatum were employed to reveal their capability in treating the sewage water, which represent the biological treatment stage as the final step of the treatment processes. The results showed a considerable capability of these fungi species in decreasing many variables values such as pH, total soluble solids (TSS), electrical conductivity (EC), salinity, total alkalinity, chlorides, nitrite, and phosphate. Where, slightly low reduction was detected in TSS value in all experiments (1.1–5.9%), similarly, both EC and salinity which were decreased with low ratios (6.6%, 3.9%, respectively). Taxon A. terrues exhibited high ability in reducing the total alkalinity and chloride ions in the treated water (30.9%, 43%, respectively) in comparison with the other two fungi species. Furthermore, all three fungi species were posed high capability in decreasing nutrients, where both nitrite and phosphate ions were highly reduced (87–97% and 22.8–32.1%, respectively). Based on these findings, we suggest using other microorganisms and exploring their capacity in removing the pollutants, and revealing the ability of the above fungi taxa in removing other pollutants.
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Authors and Affiliations

Nuha F. Kadhim
1
Wathiq J. Mohammed
1
ORCID: ORCID
Ibtihal M. Al Hussaini
1
Hala M.N. Al-Saily
1
Rasha N. Ali
1

  1. The University of Babylon, College of Science, Department of Biology, PO Box: 4 Iraq – Babylon – Hillah, Babilon, Iraq
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Abstract

Heavy metal contamination often accompanies pollution with petroleum oil derivatives. Metal ions may considerably affect the process of hydrocarbon biodegradation. The results obtained for bacterial strains Gordon ia a/kanivorans S7 and Pseudom on as fluorescens SL3 indicated the limitation of cell growth and reduced ability of degradation of petroleum oil hydrocarbons in the presence ferric ions in the range of 15-100 mg Fe/dm3. However, the addition of these ions in the range of 1-7 mg Fe/dm3 might appear to be advantageous for biodegradation process. Supplementation with ferric ions in the range of 50-100 mg Fe/dm' decreases emulsifying activity of tested strains. The excess of these ions is accumulated in the bacterial cells.
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Authors and Affiliations

Ewa Kwapisz
Aneta Piątkowska
Małgorzata Piotrowicz-Wasiak
Jacek Polak
Stanisław Bielecki
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Abstract

The paper presents results or laboratory tests or solid-phase anaerobic biorerncdiation or soil contaminated with chlorinated pesticides. It was shown that using methanogcnic granular sludge as inoculum and lactate as electron donor, it is possible to remove 80% or y-HCH, 94% 01· methoxychlor and 93% or DDT against control sample, with DDD accumulation much less than stoichiometric. Pesticides removal was practically completed alter 4-6 weeks of incubation at 22°C. Additional application of nonionic surfactant Tween 80 resulted in about one and a half-fold decrease or residual concentrations or some corn pounds. It also enhanced DDT conversion to some extent, decreasing DDD accumulation and intensifying production of D8P, the terminal metabolite or DDT anaerobic degradation pathway. Use of methanol as electron donor produced effects quite similar to these obtained with lactate, however with reduced results scatter.
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Authors and Affiliations

Tomasz Baczyński
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Abstract

The selection of bioremediation techniques is important for purification of contaminated soil for agricultural use. Studies on soil contaminated with petroleum substances have indicated that the applied method of remediation has a bigger impact on the development of oat seedlings than the level of contamination. A yeast inoculum appeared to be a technique which was the friendliest to vegetation of oat

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Authors and Affiliations

Małgorzata Pawełczak
Barbara Dawidowska-Marynowicz
Bartosz Oszywa
Magdalena Koszałkowska
Łukasz Kręcidło
Teresa Krzyśko-Łupicka
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Abstract

Bioremediation is based on microorganisms able to use pollutants either as a source of carbon or in co-metabolism, and is a promising strategy in cleaning the environment. Using soil contaminated with petroleum products from an industrial area in Saudi Arabia (Jubail), and after enrichment with the polycyclic aromatic hydrocarbon (PAH) naphthalene, a Methylobacterium radiotolerans strain (N7A0) was isolated that can grow in the presence of naphthalene as the sole source of carbon. M. radiotolerans is known to be resistant to gamma radiation, and this is the first documented report of a strain of this bacterium using a PAH as the sole source of carbon. The commonly reported Pseudomonas aeruginosa (strain N7B1) that biodegrades naphthalene was also identified, and gas chromatography analyses have shown that the biodegradation of naphthalene by M. radiotolerans and P. aeruginosa did follow both the salicylate and phthalate pathways.

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Authors and Affiliations

Alexis Nzila
Assad Thukair
Saravanan Sankara
Basheer Chanbasha
Musa M. Musa
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Abstract

Changes in activity of amylase, cellulase, protease and dehydrogenase during biodegradation of petroleum contaminants were investigated in laboratory studies. Refinery soil polluted for a long time was modified by commercial biological preparation and natural biological preparation (prepared from this soil). These modifications did not influence cellulase and protease activity. Activity of amylase and dehydrogenase changed in these samples. In the sixth week the highest activity of dehydrogenase was noticed, especially in soil with commercial biological preparation. Modifications had a great influence on contaminants degradation. The highest elimination of TPH (20,4%) and heavy fractions (10,5%) took place in soil modified by commercial biological preparation. The highest PAH elimination was in soil with natural bacterial preparation
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Authors and Affiliations

Wioletta Przystaś
Korneliusz Miksch
Anna Małachowska-Jutsz
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Abstract

The effects of using three different biopreparations (one natural N2 and two commercial Kl and K2) in petroleum hydrocarbons age-polluted clay soil were studied. The samples of soil were taken from refinery in Czechowice-Dziedzice and classified as heavily degraded in proportion C:N = 100:0,7. Changes after introducing biopreparations into soil (bioaugmentation), their influence on microflora development and effects of removing polluting substances were studied. Bioaugmentation did not result in intensive growth of bacterial number, which was on the control sample's level during experiment. Soil's biodegradation activity also increased in level smaller than expected.
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Authors and Affiliations

Ewa Zabłocka-Godlewska
Korneliusz Miksch
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Abstract

In the present work, bacteria of the Pseudomonas genus native to the Ecuadorian Amazon with the capacity to treat contaminated water and soils were selected. For this purpose, 20 soil samples from Amazon region with evidence of contamination were analysed. For identification, each sample was assigned a code according to the sampling area: Joya de los Sachas (S), Minga (M) and Siete de Julio-Shushufindi (SH). The cultures were performed in the combination of Bushnell Hass (BH) + Luria Bertani (LB) and Müeller-Hinton (MH) + Brucella agar (BA) media, all with the addition of diesel to verify their efficacy in the growth of bacteria capable of surviving in contaminated media. The combination with ideal results was that of BH + LB, by means of Gram-staining it was determined that 19 of the samples had interest microorganisms. To characterize the isolates at the species level, biochemical tests of: catalase, citrate, glucose, hemolytic activity and urease were applied, which allowed to confirm the existence of the Pseudomonas of interest. The results indicated that P. stutzeri (in samples S1 and M1), P. aeruginosa (in SH2 and SH5) and P. putida (in S7, S8, S10 and SH4) obtaining a total of 8 isolates (40%) of interest from the initial 19. With the results obtained from this work, an optimal culture method was standardized for the selection of bacteria with potential for treating contaminated soils and water.
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Authors and Affiliations

Henry Joel Escudero-López
1
ORCID: ORCID
Karla Anabel Serrano-Carrillo
2
ORCID: ORCID
Carlos Rodrigo Jácome-Pilco
1
ORCID: ORCID
Herminia del Rosario Sanaguano-Salguero
1
ORCID: ORCID
Isidro Favian Bayas-Morejón
1
ORCID: ORCID

  1. State University of Bolívar, Faculty of Agricultural Sciences, Agroindustry Career, Biotechnological Research and Development Center, Km 3 1/2 sector Alpachaca, CP: 020150, Guaranda, Ecuador
  2. San Pedro Educational Unit, Department of Education, Guaranda, Ecuador
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Abstract

The Kalina pond has been well known as a severely degraded area in the Silesia region, Poland. The environmental deterioration results from high contamination of water and bottom sediments with recalcitrant and toxic organic compounds, mainly phenol. The study was aimed at developing a bioremediation-based approach suitable for this type of polluted areas, involving microbiological treatment of water as a key and integral part of other necessary actions: mechanical interventions and the use of physical methods. During the initial biological treatment stage, autochthonous microorganisms were isolated from contaminated samples of water, soil and sediment, then subjected to strong selective pressure by incubation with the pollutants, and finally, cultivated to form a specialised microbial consortium consisting of five extremophilic bacterial strains. Consortium propagation and its biodegradation activity were optimised under variant conditions enabling bacteria to proliferate and to obtain high biomass density at large volumes allowing for the in situ application. After installing aeration systems in the pond, the consortium was surface-sprinkled to launch bioremediation and then both bacterial frequency and the contaminant level was systematically monitored. The complex remediation strategy proved efficient and was implemented on an industrial scale enabling successful remedial of the affected site. Treatment with the specifically targeted and adapted microbial consortium allowed for removal of most organic pollutants within a four-month season of 2022: the chemical oxygen demand (COD) value decreased by 72%, polyaromatic hydrocarbon (PAH) level by 97%, while the content of total phenols and other monoaromatic hydrocarbons (BTEX) dropped below the detection thresholds.
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Authors and Affiliations

Katarzyna Starzec
1
ORCID: ORCID
Emilia Stańkowska
2
Paulina Supel
1
ORCID: ORCID
Robert Mazur
3
ORCID: ORCID
Piotr Surma
2
Paweł Kaszycki
1
ORCID: ORCID

  1. University of Agriculture in Kraków, Faculty of Biotechnology and Horticulture, Department of Plant Biology and Biotechnology, al. Mickiewicza 21, 31-120 Kraków, Poland
  2. Remea Sp. z o. o., ul. Bonifraterska 17, 00-203 Warszawa, Poland
  3. AGH University of Science and Technology, Faculty of Mining Geodesy and Environmental Engineering, Department of Environmental Protection and Landscaping, al. Mickiewicza 30, 30-059 Kraków, Poland
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Abstract

Three chromium resistant bacterial strains, Pseudomonas fluorescens PF28, Enterobacter amnigenus EA31 and Enterococcus gallinarum S34 isolated from tannery waste contaminated soil were used in this study. All strains could resist a high concentration of K2Cr2O7 that is up to 300 mg/L. The effect of these strains on clover plants (Trifolium campestre) in the presence of two chromium salts CrCl3 and K2Cr2O7 was studied in soil microcosm. Application of chromium salts adversely affected seed germination, root and shoot length. Bacterial inoculation improved the growth parameters under chromate stress when compared with non inoculated respective controls. There was observed more than 50% reduction of Cr(VI) in inoculated soil microcosms, as compared to the uninoculated soil under the same conditions. The results obtained in this study are significant for the bioremediation of chromate pollution.

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Authors and Affiliations

Hanane Sayel
Nezha Tahri Joutey
Wifak Bahafid
Naima El Ghachtouli
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Abstract

The aim of this work was to determine the influence of native and alien keratinolytic fungi inocula on the petroleum hydrocarbon removal rate in soil covered and not covered with hair. The hair was the major nutrient for keratinolytic fungi and other soil microorganisms. The fungal inocula accelerated the petroleum hydrocarbon biodegradation process during the first month of the experiment. During the second month, TPH removal rates were similar for soil inoculated and not inoculated with fungi. The highest petroleum hydrocarbon removal rate was observed in soil inoculated with native fungal strains. The TPI-I removal rates were about 64% in soil covered with hair and 77% in soil not covered with hair. The lowest removal rate was observed in soil not inoculated with fungi (60%). The hair applied as additional nitrogen, sulfur and carbon source did not impact, or slightly inhibited, the petroleum hydrocarbon biodegradation process. The fungal inocula caused dramatic changes in soil fungal qualitative composition.
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Authors and Affiliations

Wioletta Przystaś
Krzysztof Ulfig
Korneliusz Miksch
Aleksandra Witała
Jerzy Szdzuj
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Abstract

This study is aimed at measuring the effect of pig, cow, horse and poultry manures on the degradation of

selected Polycyclic Aromatics Hydrocarbons present in oil sludge. Four kilograms of soil amended with 1.2 kg of oil

sludge was mixed with wood chips in a ratio of 1:2 (w:v) soil mixture: wood chips. The mixture was divided into fi ve parts

and four parts were separately mixed with pig, cow, horse or poultry manures in a ratio of 2:1 (w:w) and the fi fth portion

was used as the control with no manure added. All experiments were incubated for 10 months at room temperature.

Compost piles were turned weekly for aeration and moisture level was maintained by adding deionised water enough

to prevent the compost from getting dry. Moisture level, pH, temperature, CO2

evolution and oxygen consumption

were measured monthly and the ash content of the compost at the end of experimentation. Highest temperature reached

was 27.5°C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2

evolution was highest in poultry manure at

18.78 μg/dwt/day. Microbial growth and activities were enhanced as indicated by increase in temperature, moisture

level, pH value and respiration rate in all the compost piles. Bacteria capable of utilizing PAHs were isolated, purifi ed

and characterized by molecular techniques using polymerase chain reaction with specifi c universal primers and the

amplicons were sequenced. Bacteria identifi ed were Bacillus, Arthrobacter and Staphylococcus species. Percentage

reduction in PAHs was measured using automated soxhlet extractor with Dichloromethane and gas chromatography/mass

spectrometry. Results from PAH concentration measurements showed reduction of between 77% and 99%. Co-

-composting of contaminated soil with animal manures enhanced the reduction in PAHs.

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Authors and Affiliations

Onyedikachi Ubani
Harrison Ifeanyichukwu Atagana
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Abstract

Compounds present in oil sludge such as polycyclic aromatic hydrocarbons (PAHs) are known to be cytotoxic, mutagenic and potentially carcinogenic. Microorganisms including bacteria and fungi have been reported to degrade oil sludge components to innocuous compounds such as carbon dioxide, water and salts. In the present study, we isolated different bacteria with PAH-degrading capabilities from compost prepared from oil sludge and animal manures. These bacteria were isolated on a mineral base medium and mineral salt agar plates. A total of 31 morphologically distinct isolates were carefully selected from 5 different compost treatments for identification using polymerase chain reaction (PCR) of the 16S rRNA gene with specific primers (universal forward 16S-P1 PCR and reverse 16S-P2 PCR). The amplicons were sequenced and sequences were compared with the known nucleotides from the GenBank. The phylogenetic analyses of the isolates showed that they belong to 3 different clades; Firmicutes, Proteobacteria and Actinobacteria. These bacteria identified were closely related to the genera Bacillus, Arthrobacter, Staphylococcus, Brevibacterium, Variovorax, Paenibacillus, Ralstonia and Geobacillus. The results showed that Bacillus species were predominant in all composts. Based on the results of the degradation of the PAHs in the composts and results of previous studies on bacterial degradation of hydrocarbons in oil, the characteristics of these bacterial isolates suggests that they may be responsible for the breakdown of PAHs of different molecular weights in the composts. Thus, they may be potentially useful for bioremediation of oil sludge during compost bioremediation.

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Authors and Affiliations

Onyedikachi Ubani
Harrison Ifeanyichukwu Atagana
Mapitsi Silvester Thantsha
Adeleke Rasheed
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Abstract

The aim of this work is purification of contaminated soil from manufactured gas plants MGP, which arc not used. Prepared chromatographic methodologies, which allow qualitative identification and quantitative determination of individual aliphatic and aromatic (TAH, PAH) hydrocarbons in the soil from the selected MGP, arc presented. The results of the research on remediation of the soil polluted with petroleum hydrocarbons (TAH, PAHs) in semi-field conditions arc discussed. Application of basic biorcmcdiation and bioaugmcntation with indigenous microorganisms, enriched with PAH biodegradable fungi, resulted in reduction in amount of 5- and 6-polycyclic aromatic hydrocarbons. The research enables control of remediation progress, selection of optimal doses of biogenie compounds and determines the time of the process. The entire cycle of soil remediation was monitored with the use of gas chromatography (GC). Estimation ofbiodcgradation degree of individual aliphatic hydrocarbons (alkancs) was based on changes in their concentration in the soil and determined by biodegradation indicators: C,/pristanc and C,/phytanc ratios. Attention was paid particularly to biodcgradation of polycyclic aromatic hydrocarbons (PAH), which were observed in substantial concentrations in the polluted soil. The elaborated chromatographic methodology of PAH determination in the soil enabled estimation of a biodcgradation rate referring to individual compounds. Moreover, attempts to create a TAH biodcgradation model with the use of C30-l 7a(H),2 l l3(H)-hopanc were undertaken.
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Authors and Affiliations

Teresa Steliga
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Abstract

Petroleum products are complex mixture of compounds of varied biological properties. They can

cause harmful changes in contaminated ecosystems and threaten humans and living organisms as well. Bioremediation (including bioremediation stimulated by biogenic substances and inoculation with biopreparations

from autochthonous bacteria and fungi) can result in creation of metabolites of a varied structure and biological

activeness, which has been partly recognised. Some of them are more toxic than an initial substrate. Besides,

they might have mutagenic features and be responsible for cancer. Estimation of bioremediation effectiveness in

waste pits was completed with toxicological monitoring. It was led with the use of living organisms as biomarkers representing all trophic levels of a chosen ecosystem: producers, consumers and reducers. This process enables total estimation of natural environment conditions. The aim of the research was to determine the influence

of petroleum contaminants and indirect metabolites (produced during bioremediation) on soil biocenose. The

results of biotests (toxicity, phytotoxicity and genotoxity) have been taken into account. The following biotests,

prepared and produced by Microbiotest, have been applied: PhytotoxkitTM, Ostracodtoxkit FTM, acute toxicity

tests Microtox® Solid Phase and Ames mutagenicity tests. The obtained results enabled observation of changes

in toxic properties during purification of the soil taken from waste pits. In addition, it can be claimed whether

the areas are suitable for forest usage.

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Authors and Affiliations

T. Steliga
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Abstract

Groundwater contamination due to leakage of gasoline is one of the several causes which affect the groundwater environment by polluting it. In the past few years, In-situ bioremediation has attracted researchers because of its ability to remediate the contaminant at its site with low cost of remediation. This paper proposed the use of a new hybrid algorithm to optimize a multi-objective function which includes the cost of remediation as the first objective and residual contaminant at the end of the remediation period as the second objective. The hybrid algorithm was formed by combining the methods of Differential Evolution, Genetic Algorithms and Simulated Annealing. Support Vector Machines (SVM) was used as a virtual simulator for biodegradation of contaminants in the groundwater flow. The results obtained from the hybrid algorithm were compared with Differential Evolution (DE), Non Dominated Sorting Genetic Algorithm (NSGA II) and Simulated Annealing (SA). It was found that the proposed hybrid algorithm was capable of providing the best solution. Fuzzy logic was used to find the best compromising solution and finally a pumping rate strategy for groundwater remediation was presented for the best compromising solution. The results show that the cost incurred for the best compromising solution is intermediate between the highest and lowest cost incurred for other non-dominated solutions.

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Authors and Affiliations

Deepak Kumar
Sudheer Ch
Shashi Mathur
Jan Adamowski

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